WorldWideScience

Sample records for acellular bone explants

  1. Adaptive bone formation in acellular vertebrae of sea bass (Dicentrarchus labrax L.)

    NARCIS (Netherlands)

    Kranenbarg, S.; Cleynenbreugel, van T.; Schipper, H.; Leeuwen, van J.L.

    2005-01-01

    Mammalian bone is an active tissue in which osteoblasts and osteoclasts balance bone mass. This process of adaptive modelling and remodelling is probably regulated by strain-sensing osteocytes. Bone of advanced teleosts is acellular yet, despite the lack of osteocytes, it is capable of an adaptive

  2. Comparison of structural, architectural and mechanical aspects of cellular and acellular bone in two teleost fish.

    Science.gov (United States)

    Cohen, Liat; Dean, Mason; Shipov, Anna; Atkins, Ayelet; Monsonego-Ornan, Efrat; Shahar, Ron

    2012-06-01

    The histological diversity of the skeletal tissues of fishes is impressive compared with that of other vertebrate groups, yet our understanding of the functional consequences of this diversity is limited. In particular, although it has been known since the mid-1800s that a large number of fish species possess acellular bones, the mechanical advantages and consequences of this structural characteristic - and therefore the nature of the evolution of this feature - remain unclear. Although several studies have examined the material properties of fish bone, these have used a variety of techniques and there have been no direct contrasts of acellular and cellular bone. We report on a comparison of the structural and mechanical properties of the ribs and opercula between two freshwater fish - the common carp Cyprinus carpio (a fish with cellular bone) and the tilapia Oreochromis aureus (a fish with acellular bone). We used light microscopy to show that the bones in both fish species exhibit poor blood supply and possess discrete tissue zones, with visible layering suggesting differences in the underlying collagen architecture. We performed identical micromechanical testing protocols on samples of the two bone types to determine the mechanical properties of the bone material of opercula and ribs. Our data support the consensus of literature values, indicating that Young's moduli of cellular and acellular bones are in the same range, and lower than Young's moduli of the bones of mammals and birds. Despite these similarities in mechanical properties between the bone tissues of the fish species tested here, cellular bone had significantly lower mineral content than acellular bone; furthermore, the percentage ash content and bone mineral density values (derived from micro-CT scans) show that the bone of these fishes is less mineralized than amniote bone. Although we cannot generalize from our data to the numerous remaining teleost species, the results presented here suggest

  3. Histologic and morphologic evaluation of explanted bone anchors from bone-anchored hearing aids.

    Science.gov (United States)

    Mlynski, Robert; Goldberg, Eva; Ebmeyer, Joerg; Scheich, Matthias; Gattenlöhner, Stefan; Schwager, Konrad; Hagen, Rudolf; Shehata-Dieler, Wafaa

    2009-05-01

    Bone-anchored hearing aids are a standard option in rehabilitation of patients with conductive or mixed hearing loss, and also CROS fitting. However, the skin-penetrating bone anchor repeatedly gives reason for discussion about the risk of infection of surrounding tissues as a major cause of malfunction. In the present study, explanted bone anchors with surrounding bone and soft tissue were examined and compared with the morphology of lost implants. The anchors originated from five patients. Two needed explantation due to deafness with the need of cochlea implantation. A third patient underwent explantation due to meningeal irritation by the bone anchor. Another patient lost the implant due to mechanical stress shortly after implantation. The last implant was lost in a child without apparent reason. All implants were clinically free of infection and had been stable for a median implantation period of 12 months. During the explantation procedure, the fixtures were recovered together with the attached soft tissue and bone. The specimens were examined by light microscopy or scanning electron microscopy (SEM). Sectioning for light microscopy was performed with a diamond-coated saw microtome. Histopathologic examination of the surrounding skin and subcutaneous soft tissue showed slight inflammation in one case only. The bone was regularly vital, presenting no signs of inflammation. The threads of the fixtures were filled with bone, with particularly strong attachment to the flank of traction. The SEM investigation exposed the ultrastructural interaction of bone with the implant surface. Filiform- and podocyte-like processes of osteocytes attach to the implant; lost implants did not reflect these features. Implant integration involves both osseointegration as well as soft tissue integration. Titanium oxide as the active implant surface promotes this integration even in unstable implants. The morphologic analysis exposed structural areas of the implant with weak bone

  4. Ectopic bone formation in rapidly fabricated acellular injectable dense collagen-Bioglass hybrid scaffolds via gel aspiration-ejection.

    Science.gov (United States)

    Miri, Amir K; Muja, Naser; Kamranpour, Neysan O; Lepry, William C; Boccaccini, Aldo R; Clarke, Susan A; Nazhat, Showan N

    2016-04-01

    Gel aspiration-ejection (GAE) has recently been introduced as an effective technique for the rapid production of injectable dense collagen (IDC) gel scaffolds with tunable collagen fibrillar densities (CFDs) and microstructures. Herein, a GAE system was applied for the advanced production and delivery of IDC and IDC-Bioglass(®) (IDC-BG) hybrid gel scaffolds for potential bone tissue engineering applications. The efficacy of GAE in generating mineralizable IDC-BG gels (from an initial 75-25 collagen-BG ratio) produced through needle gauge numbers 8G (3.4 mm diameter and 6 wt% CFD) and 14G (1.6 mm diameter and 14 wt% CFD) was investigated. Second harmonic generation (SHG) imaging of as-made gels revealed an increase in collagen fibril alignment with needle gauge number. In vitro mineralization of IDC-BG gels was confirmed where carbonated hydroxyapatite was detected as early as day 1 in simulated body fluid, which progressively increased up to day 14. In vivo mineralization of, and host response to, acellular IDC and IDC-BG gel scaffolds were further investigated following subcutaneous injection in adult rats. Mineralization, neovascularization and cell infiltration into the scaffolds was enhanced by the addition of BG and at day 21 post injection, there was evidence of remodelling of granulation tissue into woven bone-like tissue in IDC-BG. SHG imaging of explanted scaffolds indicated collagen fibril remodelling through cell infiltration and mineralization over time. In sum, the results suggest that IDC-BG hybrid gels have osteoinductive properties and potentially offer a novel therapeutic approach for procedures requiring the injectable delivery of a malleable and dynamic bone graft that mineralizes under physiological conditions.

  5. Guided bone regeneration using acellular bovine pericardium in a rabbit mandibular model: in-vitro and in-vivo studies.

    Science.gov (United States)

    Bai, M; Zhang, T; Ling, T; Zhou, Z; Xie, H; Zhang, W; Hu, G; Jiang, C; Li, M; Feng, B; Wu, H

    2014-08-01

    To investigate the feasibility of acellular bovine pericardium (BP) for guided bone regeneration (GBR) in vitro and in vivo. The success of GBR relies on the fact that various cellular components possess different migration rates into the defect site and that a barrier membrane plays a significant role in these processes. BP membrane was isolated and decellularized using an enzymatic method. The microarchitecture, mechanical properties, cytotoxicity and cell chemotaxis properties of the acellular BP were evaluated in vitro, and the in-vivo efficacy of the acellular BP was also investigated in a rabbit mandibular model. The acellular BP membrane possessed an interconnected fibrous structure. Glutaraldehyde (GA) treatment was efficient for enhancement of the mechanical properties of the acellular BP bur and resulted in negligible cytotoxicity. After 16 wk, standardized osseous defects created in the rabbit mandible, and covered with acellular BP, were associated with an enhanced deposition of mineralized tissue when compared with defects left to spontaneous healing. GA-treated acellular BP is promising as a barrier membrane for GBR for further in-vivo and clinical studies. © 2013 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  6. Acellular allogeneic nerve grafting combined with bone marrow mesenchymal stem cell transplantation for the repair of long-segment sciatic nerve defects: biomechanics and validation of mathematical models

    Directory of Open Access Journals (Sweden)

    Ya-jun Li

    2016-01-01

    Full Text Available We hypothesized that a chemically extracted acellular allogeneic nerve graft used in combination with bone marrow mesenchymal stem cell transplantation would be an effective treatment for long-segment sciatic nerve defects. To test this, we established rabbit models of 30 mm sciatic nerve defects, and treated them using either an autograft or a chemically decellularized allogeneic nerve graft with or without simultaneous transplantation of bone marrow mesenchymal stem cells. We compared the tensile properties, electrophysiological function and morphology of the damaged nerve in each group. Sciatic nerves repaired by the allogeneic nerve graft combined with stem cell transplantation showed better recovery than those repaired by the acellular allogeneic nerve graft alone, and produced similar results to those observed with the autograft. These findings confirm that a chemically extracted acellular allogeneic nerve graft combined with transplantation of bone marrow mesenchymal stem cells is an effective method of repairing long-segment sciatic nerve defects.

  7. Synergistic effects of bone mesenchymal stem cells and chondroitinase ABC on nerve regeneration after acellular nerve allograft in rats.

    Science.gov (United States)

    Wang, Ying; Jia, Hua; Li, Wen-Yuan; Tong, Xiao-Jie; Liu, Gui-Bo; Kang, Si-Wen

    2012-04-01

    This study aimed to evaluate whether combination therapy of bone marrow stromal cells (BMSCs) transplantation and chondroitinase ABC (ChABC) treatment further enhances axonal regeneration and functional recovery after acellular nerve allograft repair of the sciatic nerve gap in rats. Eight Sprague-Dawley rats were used as nerve donors, and 32 Wistar rats were randomly divided into four groups: Group I: acellular rat sciatic nerve (ARSN) group; Group II: ChABC treatment; Group III: BMSCs transplantation; and Group IV: ChABC treatment and BMSCs transplantation. The results showed that compared with ARSN control group, BMSC transplantation promoted axonal regeneration, the secretion of neural trophic factors NGF, BDNF and axon angiogenesis in nerve graft. ChABC treatment degraded chondroitin sulfate proteoglycans in ARSN in vitro and in vivo and improved BMSCs survival in ARSN. The combination therapy caused much better beneficial effects evidenced by increasing sciatic function index, nerve conduction velocity, restoration rate of tibialis anterior wet muscle weight, and myelinated nerve number, but did not further boost the therapeutic effects on neurotrophic factor production, axon angiogenesis, and sensory functional recovery by BMSC transplantation. Taken together, for the first time, we demonstrate the synergistic effects of BMSC transplantation and BMSCs treatment on peripheral nerve regeneration, and our findings may help establish novel strategies for cell transplantation therapy for peripheral nerve injury.

  8. Repair of peripheral nerve defects with chemically extracted acellular nerve allografts loaded with neurotrophic factors-transfected bone marrow mesenchymal stem cells

    Institute of Scientific and Technical Information of China (English)

    Yan-ru Zhang; Ka Ka; Ge-chen Zhang; Hui Zhang; Yan Shang; Guo-qiang Zhao; Wen-hua Huang

    2015-01-01

    Chemically extracted acellular nerve allografts loaded with brain-derived neurotrophic fac-tor-transfected or ciliary neurotrophic factor-transfected bone marrow mesenchymal stem cells have been shown to repair sciatic nerve injury better than chemically extracted acellular nerve allografts alone, or chemically extracted acellular nerve allografts loaded with bone marrow mesenchymal stem cells. We hypothesized that these allografts compounded with both brain-derived neurotrophic factor- and ciliary neurotrophic factor-transfected bone marrow mesenchymal stem cells may demonstrate even better effects in the repair of peripheral nerve injury. We cultured bone marrow mesenchymal stem cells expressing brain-derived neuro-trophic factor and/or ciliary neurotrophic factor and used them to treat sciatic nerve injury in rats. We observed an increase in sciatic functional index, triceps wet weight recovery rate, myelin thickness, number of myelinated nerve ifbers, amplitude of motor-evoked potentials and nerve conduction velocity, and a shortened latency of motor-evoked potentials when al-lografts loaded with both neurotrophic factors were used, compared with allografts loaded with just one factor. Thus, the combination of both brain-derived neurotrophic factor and cili-ary neurotrophic factor-transfected bone marrow mesenchymal stem cells can greatly improve nerve injury.

  9. Acellular bone marrow extracts significantly enhance engraftment levels of human hematopoietic stem cells in mouse xeno-transplantation models.

    Directory of Open Access Journals (Sweden)

    Kazem Zibara

    Full Text Available Hematopoietic stem cells (HSC derived from cord blood (CB, bone marrow (BM, or mobilized peripheral blood (PBSC can differentiate into multiple lineages such as lymphoid, myeloid, erythroid cells and platelets. The local microenvironment is critical to the differentiation of HSCs and to the preservation of their phenotype in vivo. This microenvironment comprises a physical support supplied by the organ matrix as well as tissue specific cytokines, chemokines and growth factors. We investigated the effects of acellular bovine bone marrow extracts (BME on HSC in vitro and in vivo. We observed a significant increase in the number of myeloid and erythroid colonies in CB mononuclear cells (MNC or CB CD34+ cells cultured in methylcellulose media supplemented with BME. Similarly, in xeno-transplantation experiments, pretreatment with BME during ex-vivo culture of HSCs induced a significant increase in HSC engraftment in vivo. Indeed, we observed both an increase in the number of differentiated myeloid, lymphoid and erythroid cells and an acceleration of engraftment. These results were obtained using CB MNCs, BM MNCs or CD34(+ cells, transplanted in immuno-compromised mice (NOD/SCID or NSG. These findings establish the basis for exploring the use of BME in the expansion of CB HSC prior to HSC Transplantation. This study stresses the importance of the mechanical structure and soluble mediators present in the surrounding niche for the proper activity and differentiation of stem cells.

  10. Synergistic effects of ultrashort wave and bone marrow stromal cells on nerve regeneration with acellular nerve allografts.

    Science.gov (United States)

    Pang, Chao-Jian; Tong, Lei; Ji, Li-Li; Wang, Zhen-Yu; Zhang, Xu; Gao, Hai; Jia, Hua; Zhang, Li-Xin; Tong, Xiao-Jie

    2013-10-01

    Acellular nerve allografts (ANA) possess bioactivity and neurite promoting factors in nerve tissue engineering. Previously we reported that low dose ultrashort wave (USW) radiation could enhance the rate and quality of peripheral nerve regeneration with ANA repairing sciatic nerve defects. Meanwhile, ANA implanted with bone marrow stromal cells (BMSCs) exhibited a similar result. Thus, it is interesting to know whether it might yield a synergistic effect when USW radiation is combined with BMSCs-laden ANA. Here we investigated the effectiveness of ANA seeded with BMSCs, combined with USW therapy on repairing peripheral nerve injuries. Adult male Wistar rats were randomly divided into four groups: Dulbecco's modified Eagle's medium (DMEM) control group, BMSCs-laden group, ultrashort wave (USW) group and BMSC + USW group. The regenerated nerves were assayed morphologically and functionally, and growth-promoting factors in the regenerated tissues following USW administration or BMSCs integration were also detected. The results indicated that the combination therapy caused much better beneficial effects evidenced by increased myelinated nerve fiber number, myelin sheath thickness, axon diameter, sciatic function index, nerve conduction velocity, and restoration rate of tibialis anterior wet weight. Moreover, the mRNA levels of brain-derived neurotrophic factor (BDNF) and vascular endothelial growth factor (VEGF) in the spinal cord and muscles were elevated significantly. In conclusion, we found a synergistic effect of USW radiation and BMSCs treatment on peripheral nerve regeneration, which may help establish novel strategies for repairing peripheral nerve defects. Copyright © 2013 Wiley Periodicals, Inc.

  11. Acellular allogeneic nerve grafting combined with bone marrow mesenchymal stem cell transplantation for the repair of long-segment sciatic nerve defects:biomechanics and validation of mathematical models

    Institute of Scientific and Technical Information of China (English)

    Ya-jun Li; Bao-lin Zhao; Hao-ze Lv; Zhi-gang Qin; Min Luo

    2016-01-01

    We hypothesized that a chemically extracted acellular allogeneic nerve graft used in combination with bone marrow mesenchymal stem cell transplantation would be an effective treatment for long-segment sciatic nerve defects. To test this, we established rabbit models of 30 mm sciatic nerve defects, and treated them using either an autograft or a chemically decellularized allogeneic nerve graft with or without simultaneous transplantation of bone marrow mesenchymal stem cells. We compared the tensile properties, electrophysiological function and morphology of the damaged nerve in each group. Sciatic nerves repaired by the allogeneic nerve graft combined with stem cell trans-plantation showed better recovery than those repaired by the acellular allogeneic nerve graft alone, and produced similar results to those observed with the autograft. These ifndings conifrm that a chemically extracted acellular allogeneic nerve graft combined with transplanta-tion of bone marrow mesenchymal stem cells is an effective method of repairing long-segment sciatic nerve defects.

  12. Ridge preservation with acellular dermal matrix and anorganic bone matrix cell-binding peptide P-15 after tooth extraction in humans. A histologic and morphometric study

    Directory of Open Access Journals (Sweden)

    Arthur B. Novaes Jr

    2012-06-01

    Full Text Available Aim: The aim of this study was to analyze by histomorphometric parameters the use of acellular dermal matrix (ADM with or without anorganic bovine bone matrix (ABM / synthetic cell-binding peptide P-15 in the formation of bone in human alveoli. Materials and methods: Eighteen patients in need of extraction of maxillary anterior teeth were selected and randomly assigned to the test group (ADM plus ABM/P-15 or the control group (ADM only. Histomorphometric measurements and histological analysis were recorded about 6 months after ridge preservation procedures in ten patients. The amount of newly formed bone, the most recently formed bone, fibrous tissue plus marrow spaces and remaining graft particles were measured and analyzed. Results: At 6 months, the new bone area parameter and the percentage of fibrous tissue plus marrow space areas showed higher values to the control group, and statistically significant differences when compared with the test group (p=0.03. Conclusion: The ADM acted as a membrane. The association of ABM/P-15 with ADM resulted in new bone formation within the alveoli, but the results were not considered relevant when used in this indication.

  13. Improved cartilage regeneration by implantation of acellular biomaterials after bone marrow stimulation: a systematic review and meta-analysis of animal studies

    Directory of Open Access Journals (Sweden)

    Michiel W. Pot

    2016-09-01

    Full Text Available Microfracture surgery may be applied to treat cartilage defects. During the procedure the subchondral bone is penetrated, allowing bone marrow-derived mesenchymal stem cells to migrate towards the defect site and form new cartilage tissue. Microfracture surgery generally results in the formation of mechanically inferior fibrocartilage. As a result, this technique offers only temporary clinical improvement. Tissue engineering and regenerative medicine may improve the outcome of microfracture surgery. Filling the subchondral defect with a biomaterial may provide a template for the formation of new hyaline cartilage tissue. In this study, a systematic review and meta-analysis were performed to assess the current evidence for the efficacy of cartilage regeneration in preclinical models using acellular biomaterials implanted after marrow stimulating techniques (microfracturing and subchondral drilling compared to the natural healing response of defects. The review aims to provide new insights into the most effective biomaterials, to provide an overview of currently existing knowledge, and to identify potential lacunae in current studies to direct future research. A comprehensive search was systematically performed in PubMed and EMBASE (via OvidSP using search terms related to tissue engineering, cartilage and animals. Primary studies in which acellular biomaterials were implanted in osteochondral defects in the knee or ankle joint in healthy animals were included and study characteristics tabulated (283 studies out of 6,688 studies found. For studies comparing non-treated empty defects to defects containing implanted biomaterials and using semi-quantitative histology as outcome measure, the risk of bias (135 studies was assessed and outcome data were collected for meta-analysis (151 studies. Random-effects meta-analyses were performed, using cartilage regeneration as outcome measure on an absolute 0–100% scale. Implantation of acellular

  14. The application of acellular dermal matrix combined with coralline hydroxyapatite in guided bone regeneration%异种脱细胞真皮基质联合珊瑚羟基磷灰石在GBR术中的应用

    Institute of Scientific and Technical Information of China (English)

    汪竹红; 康博; 黄达鸿; 管红雨; 温玉洁; 林天赐; 林丽娥

    2014-01-01

    Objective: To evaluate the clinical effectiveness of acellular dermal matrix combined with coralline hydroxyapatite for guided bone regeneration. Methods: 17 patients with 27 lost teeth were included in this study. 10 anterior lost teeth area with the alveolar bone thickness about 4mm was placed ankylos implants using bone condensing technique followed by guided bone generation. The other lost teeth area with bone defect was placed ankylos implants using routine method followed by GBR. 6-8 months later, the second-stage operation was performed and the condition of the new bone was observed. Results: All implants showed good osseointegration and were covered by alveolar bone except one implant whose labial neck about 1.5mm height wasn't covered by bone. Conclusion:Acellular dermal matrix combined with coralline hydroxyapatite can achieve good bone formation in guided bone regeneration.%目的:评价异种脱细胞真皮基质联合珊瑚羟基磷灰石在引导骨组织再生术中的应用效果。方法:17例共27颗牙缺失患者作为研究对象,其中10颗上前牙牙槽骨宽度约4mm的延期种植先行骨挤压术植入种植体再行GBR术,其余12颗延期即刻种植上前牙及5颗环状骨缺损后牙常规植入种植体后行GBR术。6-8m后观察成骨效果。结果:除一例患者右上侧切牙植体颈部唇侧暴露约1.5mm左右,其余患者植体均被新生骨包绕,成骨效果显著。结论:异种脱细胞真皮基质联合珊瑚羟基磷灰石在牙种植术中引导骨组织再生效果良好。

  15. Guide bone regeneration with acellular dermal matrix in the maxillary anterior region%脱细胞真皮基质在上前牙GBR种植术中的临床研究

    Institute of Scientific and Technical Information of China (English)

    董强; 夏茜; 马洪; 王小玲; 杨红; 周成菊; 毛久凤

    2014-01-01

    目的:研究采用脱细胞真皮基质进行引导骨组织再生技术(GBR)并同期种植体植入的短期临床效果。方法29例上前牙脱细胞真皮基质进行 GBR并同期植入种植体,经软组织塑形后,完成最终上部结构。随访3~9个月,对种植体周围软硬组织进行评价。结果29例均获得良好骨整合,种植体无松动脱落。种植体周围软硬组织状态良好,美学效果满意。结论采用脱细胞真皮基质进行上前牙 GBR并同期植入种植体,短期内可获得较满意的临床效果。%Objective To evaluate the clinical and aesthetic results of guide bone regeneration(GBR)with acellular dermal matrix and implant placement in the maxillary anterior region.Methods 29 cases in the maxillary anterior region were selected carefully, GBR with acellular dermal matrix were processed and the implants were placed immediately.Impressions were taken after soft tis-sue development with provisional implant restorations and the definitive restorations were finished.The follow-up time was 3 to 9 months.The evaluated indexes involved marginal bone level at mesial and distal aspects of the implants,the interproximal papilla in-dex score of Jemt′s classification and the level of the labial soft-tissue margin.Results 29 cases were good bone integration,implant without mobility.Hard and soft tissue around implants in good condition,aesthetic effect was satisfied.Conclusion Using acellular dermal matrix to come forward to tooth GBR and implanted implant during this period,can obtain satisfactory clinical effect in the short term.

  16. Clinical application of heterogeneous acellular dermal matrix used in alveolar bone grafting%异种脱细胞真皮基质膜在齿槽裂植骨术中的临床应用

    Institute of Scientific and Technical Information of China (English)

    李可兴; 刘曙光

    2014-01-01

    Objective To observe the effect of bone repair and evaluate its esthetic outcome with heterogeneous acellular dermal matrix cover the alveolar cleft bone grafting area in the alveolar cleft operation.Methods In 67 cases,unilateral cleft palate,were treated by alveolar cleft conventional surgical method.Cancellous iliac bone grafting were control group,heterogeneous acellular dermal matrix cover the alveolar cleft bone grafting area were treatment group.Radiographs was taken at 1 st,3 rd,6 th,12 th,18 th,24 th month postoperatively to observe the bone regeneration alveolar cleft zone.Results The alveolar cleft graft area new bone formation with Ⅰ,Ⅱ,Ⅲ,Ⅳ grade after 6 months in control group was 15,11,9,6 cases and in treatment group was 13,9,3,1 case.The graft survival rate and success rate (97.8%,84.3%) of treatment group were higher than that of control group (84.5%,63.7%),the difference was statistically significant (P < 0.05).Conclusion The successful rate of operation could be warranted,by the joint application of autogenous iliac bone grafts and heterogeneous acellular dermal matrix in the alveolar cleft operation.%目的 在齿槽裂手术中,将异种脱细胞真皮基质膜覆盖齿槽裂植骨区,观察新骨形成状况,评价植骨修复效果.方法 选择67例单侧齿槽裂患者,按治疗方法不同分为对照组和试验组.对照组单纯应用髂骨骨松质移植41例,试验组应用异种脱细胞真皮基质膜加髂骨骨松质移植26例.术后1,3,6,12,18,24个月随访,X线片观察齿槽裂植骨区新骨生成情况.结果 对照组病例术后6个月齿槽裂植骨区新骨形成Ⅰ,Ⅱ,Ⅲ,Ⅳ级分别为15,11,9,6例,齿槽裂植骨成活率为84.5%,临床成功率为63.7%.而试验组病例植骨区新骨形成Ⅰ,Ⅱ,Ⅲ,Ⅳ级分别为13,9,3,1例.齿槽裂植骨成活率为97.8%,临床成功率为84.3%.两组植骨成活率、临床成功率比较差异有统计学意义(P<0.05).结论 自体髂骨加异

  17. Acellular dermis-assisted prosthetic breast reconstruction: a systematic and critical review of efficacy and associated morbidity.

    Science.gov (United States)

    Sbitany, Hani; Serletti, Joseph M

    2011-12-01

    The use of acellular dermal matrix to assist in two-stage expander/implant breast reconstruction has increased over recent years. However, there are questions regarding the potential for increased morbidity when using these techniques relative to standard submuscular coverage techniques. This systematic review combines published data comparing the techniques, to compare morbidity and advantages of acellular dermal matrix relative to standard submuscular coverage techniques. An English language literature search was performed to find articles reporting outcomes of two-stage expander/implant reconstruction using acellular dermal matrix. The outcome categories analyzed were patient/treatment demographics, tissue expander characteristics, and complications. Nine articles met inclusion criteria for this analysis. Six of these were matched cohort studies comparing outcomes of acellular dermal matrix techniques to standard submuscular techniques. The remaining three were case series of acellular dermal matrix techniques. The only difference found in complications was a higher rate of seroma for the acellular dermal matrix group (4.3 percent versus 8.4 percent, p = 0.03). Despite this, both groups illustrated similar rates of infection leading to explantation (3.2 percent for submuscular and 3.4 percent for acellular dermal matrix, p = 0.18). In addition, acellular dermal matrix techniques illustrated greater intraoperative fill volumes and consistently fewer fills required to reach expander capacity. The use of acellular dermal matrix in two-stage expander/implant reconstruction offers a safety profile similar to that of standard submuscular techniques. Both techniques have shown similar rates of infection ultimately requiring explantation. In addition, acellular dermal matrix offers the advantage of a more rapid reconstruction with less need for manipulation of the prosthetic through filling. Therapeutic, III.

  18. 异种脱细胞真皮基质联合Bio-oss Collagen修复牙槽骨缺损的临床研究%Clinical application of acellular dermal matrix combined with Bio-oss Collagen to repair alveolar bone defects

    Institute of Scientific and Technical Information of China (English)

    杨春羚; 林良缘; 庄亮亮; 曾金表

    2011-01-01

    Objective: To investigate the effect of guided bone regeneration of acellular dermal matrix (ADM) combined with Bio -oss Collagen in alveolar defect. Method: 18 cases patients with severe alveolar bone resorption or damage were included.Bone defect after teeth extraction were very serious and the residual height and width of the alveolar bone were very limited in all these cases. Bio-oss Collagen was delivered into the sockets immediately after tooth extraction and covered with acellular dermal matrix membrane.Suture was removed 2 weeks postop.and the alveolar bone was regularly examined at the 3rd month. Result:New bone was found to form well in both physical examination and X-ray examination in all the 18 cases in the 3rd month.The height and width of the alveolar bone were significantly increased, providing a good bone condition for later FPD prosthetic treatment. Conclusion: Acellular dermal matrix combined with Bio-oss Collagen can improve the bone condition before restoration in clinical.%目的:通过引导骨再生(GBR)技术评估异种脱细胞真皮基质(acellular dermal matrix,ADM)与Bio-oss Collagen联合应用在修复牙槽骨缺损中的作用.方法:选择拔牙术后牙槽骨缺损严重的病例18例,拔牙同期在拔牙创植入Bio-oss Collagen并覆盖异种脱细胞真皮基质(海奥生物膜),术后2周拆线,3个月复诊并拍摄X线片.结果:术后经临床检查和X线检查,18例患者植骨区新骨形成良好,牙槽骨高度与丰满度明显改善,术区骨生成良好.结论:临床上异种脱细胞真皮基质与Bio-oss Collagen联合应用能有效修复牙槽骨缺损,改善修复前的骨条件.

  19. Improved cartilage regeneration by implantation of acellular biomaterials after bone marrow stimulation: a systematic review and meta-analysis of animal studies

    NARCIS (Netherlands)

    Pot, M.W.; Gonzales, V.K.; Buma, P.; Hout, J. in't; Kuppevelt, T.H. van; Vries, R.B. de; Daamen, W.F.

    2016-01-01

    Microfracture surgery may be applied to treat cartilage defects. During the procedure the subchondral bone is penetrated, allowing bone marrow-derived mesenchymal stem cells to migrate towards the defect site and form new cartilage tissue. Microfracture surgery generally results in the formation of

  20. In vivo bone regeneration using tubular perfusion system bioreactor cultured nanofibrous scaffolds.

    Science.gov (United States)

    Yeatts, Andrew B; Both, Sanne K; Yang, Wanxun; Alghamdi, Hamdan S; Yang, Fang; Fisher, John P; Jansen, John A

    2014-01-01

    The use of bioreactors for the in vitro culture of constructs for bone tissue engineering has become prevalent as these systems may improve the growth and differentiation of a cultured cell population. Here we utilize a tubular perfusion system (TPS) bioreactor for the in vitro culture of human mesenchymal stem cells (hMSCs) and implant the cultured constructs into rat femoral condyle defects. Using nanofibrous electrospun poly(lactic-co-glycolic acid)/poly(ε-caprolactone) scaffolds, hMSCs were cultured for 10 days in vitro in the TPS bioreactor with cellular and acellular scaffolds cultured statically for 10 days as a control. After 3 and 6 weeks of in vivo culture, explants were removed and subjected to histomorphometric analysis. Results indicated more rapid bone regeneration in defects implanted with bioreactor cultured scaffolds with a new bone area of 1.23 ± 0.35 mm(2) at 21 days compared to 0.99 ± 0.43 mm(2) and 0.50 ± 0.29 mm(2) in defects implanted with statically cultured scaffolds and acellular scaffolds, respectively. At the 21 day timepoint, statistical differences (pbioreactor to improve bone tissue regeneration and highlights the benefits of utilizing perfusion bioreactor systems to culture MSCs for bone tissue engineering.

  1. In Vivo Bone Regeneration Using Tubular Perfusion System Bioreactor Cultured Nanofibrous Scaffolds

    Science.gov (United States)

    Yeatts, Andrew B.; Both, Sanne K.; Yang, Wanxun; Alghamdi, Hamdan S.; Yang, Fang; Jansen, John A.

    2014-01-01

    The use of bioreactors for the in vitro culture of constructs for bone tissue engineering has become prevalent as these systems may improve the growth and differentiation of a cultured cell population. Here we utilize a tubular perfusion system (TPS) bioreactor for the in vitro culture of human mesenchymal stem cells (hMSCs) and implant the cultured constructs into rat femoral condyle defects. Using nanofibrous electrospun poly(lactic-co-glycolic acid)/poly(ɛ-caprolactone) scaffolds, hMSCs were cultured for 10 days in vitro in the TPS bioreactor with cellular and acellular scaffolds cultured statically for 10 days as a control. After 3 and 6 weeks of in vivo culture, explants were removed and subjected to histomorphometric analysis. Results indicated more rapid bone regeneration in defects implanted with bioreactor cultured scaffolds with a new bone area of 1.23±0.35 mm2 at 21 days compared to 0.99±0.43 mm2 and 0.50±0.29 mm2 in defects implanted with statically cultured scaffolds and acellular scaffolds, respectively. At the 21 day timepoint, statistical differences (pbioreactor to improve bone tissue regeneration and highlights the benefits of utilizing perfusion bioreactor systems to culture MSCs for bone tissue engineering. PMID:23865551

  2. Explant cultures of human colon

    DEFF Research Database (Denmark)

    Autrup, Herman; Barrett, L.A.; Jackson, F.E.

    1978-01-01

    Human colonic epithelium has been cultured as explants in a chemically defined medium for periods of 1 to 20 days. The viability of the explants was shown by the preservation of the ultrastructural features of the colonic epithelial cells and by active incorporation of radioactive precursors into...

  3. 脱细胞真皮基质对骨质疏松大鼠骨缺损愈合影响%Effect of acellular dermal matrix on osteoporosis rats bone defect healing

    Institute of Scientific and Technical Information of China (English)

    王晓晗; 赵志国; 王智明; 王文茉; 张力平

    2016-01-01

    Objective To observe the characteristics and capabilities of acellular dermal matrix( ADM) in osteoporosis rats cranial parietal bone defect repair guided bone regeneration( GBR) ,to explore the biocompatibility and effects on bone re-generation. Methods A total of 26 SD female rats were randomly divided into the control group(Sham group:n=13)and the ovariectomized group(VOX group:n=13). Conventional breeding for 3 months after the surgery,after the success of the building,in skull of rats,there were 2 defective holes with 5 mm preparation on both sides of central line,one side was cov-ered with ADM,the other side was control blank( CK) . In Sham group,the CK side was Group A,the ADM cover side was Group B. In the CK side in OVX group was Group C,the ADM cover side was Group D. In 6 and 12 weeks postoperatively, the clinical features such as the bone defect healing,bone tissue HE and masson trichromatic dyeing,new bone lengths were compared,the mineralization rate,immunohistochemical method was to detect callus osteocalcin expression in different peri-ods. Results Among the gross observation experimental animals,there were 2 deaths caused by bowel bilges gas in. Other animals healed within a week without infection and wound dehiscence,visible sutures were not fallen off. 6 weeks after sur-gery,the blank defects naked eye obvious difference between the two groups,the defect area was covered with transparency, defect edge was clear. ADM cover side was with visible white ADM,defect edge was clear. After 12 weeks,there was no na-ked eye obvious difference between two groups of blank defect,the defect area was covered with transparency,defect edge was clear. ADM cover side with visible white ADM film was thinner,harder to hit,defect edge was obtuse. The tissue mor-phology observation 6 weeks when two groups of new bone gap defect was not obvious,the broken end by fibrous tissue pack-age. Sham group of bone defect end osteogenesis was dense, ADM retained membrane

  4. Guided bone regeneration with acellular dermal matrix as a barrier for bone defects%脱细胞真皮基质膜引导骨缺损成骨变化

    Institute of Scientific and Technical Information of China (English)

    贾仁杰; 任玉卿; 徐昊; 王维英; 弋中萍; 赵保东

    2016-01-01

    BACKGROUND:Acel ular dermal matrix has good biocompatibility and absorbability and exhibits superiority in the guided bone regeneration. OBJECTIVE:To compare the histological changes and osteogenic effects in bone defects after guided bone regeneration with acel ular dermal matrix and Bio-Gide membrane. METHODS:Mandibular second, third and fourth premolars and the first molars bilateral y were extracted from 12 beagle dogs. Three months later, four three-wal bone defect models in the mandible of each dog were made, and randomized into acel ular dermal matrix plus bone graft group (acel ular dermal matrix group), Bio-Gide plus bone graft group (Bio-Gide group), bone graft group, and blank control group (no treatment). In the former two groups, acel ular dermal matrix and Bio-Gide were used to cover the bone grafts, respectively. RESULTS AND CONCLUSION:After surgery, al the beagle dogs recovered wel . Al the groups except the control group showed dramatical improvement in histological changes and percentage of new bone area, and this improvement was more significant in the Bio-Gide and acel ular dermal matrix groups. Moreover, there was no significant difference between the Bio-Gide and acel ular dermal matrix groups. Therefore, the acel ular dermal matrix can be a candidate for bone repair instead of Bio-Gide membrane in the clinical practice.%背景:脱细胞真皮基质膜具有良好的生物相容性、可吸收性、引导骨再生性能。目的:比较脱细胞真皮基质膜和Bio-Gide膜引导骨缺损成骨的组织学变化及引导骨再生的效果的差异。  方法:12只比格犬拔除双侧下颌骨第二、三、四前磨牙及第一磨牙3个月后,在每只犬的下颌骨各建立4处标准的三壁骨缺损模型,随机分为脱细胞真皮基质膜联合骨修复材料组、Bio-Gide膜联合骨修复材料组、骨修复材料组、空白对照组。除空白对照组不做任何处理外,将骨修复材料充实于其余3组骨

  5. Osteogenic function of human acellular bone loaded with bone marrow stromal cells%骨髓基质细胞复合人脱细胞骨的成骨活性

    Institute of Scientific and Technical Information of China (English)

    张旗涛; 于有; 杨林; 姚猛; 陶天遵

    2006-01-01

    BACKGROUND: To search for an alloxenogeneic bone with good load bearing function and osteoblastic activity for treating bone defects is an important study issue. We have made a comparative study on its biome chanical characteristics and found that there was no significant difference in maximum load stress, maximum pressure as compared with fresh bone of the same size. Clinicians are concerned about the osteoblastic activity and whether the osteoblastic activity can be reserved after human allogenous a cellular bone (HAB) loaded with bone marrow stromal cells (BMSCs). OBJECTIVE: To investigate the experimental effect of HAB loaded with induced BMSCs, and observe the cellular adherence and growth as well as detect its osteoblastic activity. DESIGN: Single sample experiment. SETTING: Second Affiliated Hospital of Harbin Medical University. MATERIALS: This experiment was conducted at the Experimental Center of the Second Affiliated Hospital of Harbin Medical University between January 2003 and August 2004. HAB was obtained from fresh corpse iliac bones (donated voluntarily). METHODS: Connective tissues and cell compounds of the iliac bones were removed by processing with hydroperoxide andether solution and sterilized for preparing HAB. BMSCs from living femoral shaft bone marrow were cultured immediately in ordinary and mineralized medium containing DMEM, fetal bovine serum, dexomethasone, β-glycerophophate and ascor bic acid. Proliferation and differentiation of bone stromal cells were deter mined by detecting the level of alkaline phosphatase (ALP) and osteocalcin (OCN) in the culture medium. Induced bone stromal cells solution was condensed and implanted within HAB scaffold. Cellular osteoblastic activ ity was determined through morphological observation under the light mi croscope and electron microscope as well as biochemical index detection. MAIN OUTCOME MEASURES: ① Detection results of ALP and OCN of BMSCs/HAB composite. ② Histological observation results

  6. Experimental total wrapping of breast implants with acellular dermal matrix: a preventive tool against capsular contracture in breast surgery?

    Science.gov (United States)

    Schmitz, Marweh; Bertram, Martin; Kneser, Ulrich; Keller, Andrea K; Horch, Raymund E

    2013-10-01

    Capsular contracture remains a hitherto unsolved complication after implantation of silicone gel-filled breast prostheses. Based on clinical and experimental data, the use of an acellular dermal matrix as a sheath around implants may lead to lesser capsular contracture acting as a proposed biological environment mimicking wound bed tissue. The aim of our study was to analyse the tissue reaction after implantation of silicone prosthesis with and without an envelope of acellular dermal matrix. Implantation of 60 silicone prostheses in the back of Lewis rats were carried out, randomly paired taking one rat from group A and one from group B. Group A included implants completely enveloped with xenogenic acellular dermis and group B undraped silicone implants. At 3, 6 and 12 weeks postoperatively, the samples were explanted and subjected to histological and immunohistochemical evaluation. A new myofibroblast tissue layer was identified in proximity to the implant in both groups. The thickness of the layer in group A was continuously thinner than in group B regarding the different explantation time points. Implants completely wrapped with acellular dermal matrix showed significantly lesser inflammatory signs at 3 and 12 weeks after implantation compared to controls. Cell proliferation after 12 weeks was significantly decreased in group A. The slight myofibroblast layer and reduced rate of inflammation and proliferation in the treatment group show a positive effect of total acellular dermal matrix envelope and hypothesise the decrease of capsular contracture in long-term periods. Copyright © 2013 British Association of Plastic, Reconstructive and Aesthetic Surgeons. Published by Elsevier Ltd. All rights reserved.

  7. Acellular nerve allograft promotes selective regeneration

    Institute of Scientific and Technical Information of China (English)

    Haili Xin; Guanjun Wang; Xinrong He; Jiang Peng; Quanyi Guo; Wenjing Xu

    2011-01-01

    Acellular nerve allograft preserves the basilar membrane tube and extracellular matrix, which pro-motes selective regeneration of neural defects via bridging. In the present study, a Sprague Dawley rat sciatic nerve was utilized to prepare acellular nerve allografts through the use of the chemical extraction method. Subsequently, the allograft was transplanted into a 10-mm sciatic nerve defect in Wistar rats, while autologous nerve grafts from Wistar rats served as controls. Compared with autologous nerve grafts, the acellular nerve allografts induced a greater number of degenerated nerve fibers from sural nerves, as well as a reduced misconnect rate in motor fibers, fewer acetyl-choline esterase-positive sural nerves, and a greater number of carbonic anhydrase-positive senso-ry nerve fibers. Results demonstrated that the acellular nerve allograft exhibited significant neural selective regeneration in the process of bridging nerve defects.

  8. 无细胞神经移植物复合骨髓间充质干细胞修复大鼠坐骨神经缺损%Repairing rat sciatic nerve defects with the compound of acellular nerve graft and bone marrow mesenchymal stem cells

    Institute of Scientific and Technical Information of China (English)

    张彩顺; 吕刚; 张基仁; 肖强

    2009-01-01

    BACKGROUND: Previous studies have successfully prepared the natural and biologically degraded acellular nerve graft and have proved the effect of promoting neural regeneration.OBJECTIVE: To construct tissue engineered artificial nerve with acellular nerve graft and bone marrow mesenchymal stem cells, and to observe the effect of promoting motor functional recovery and repairing rat sciatic nerve defects. DESIGN, TIME AND SETTING: Randomized control animal experiment was performed in the Medical TIssue Engineering Laboratory of the First Affiliated Hospital of Liaoning Medical University between June 2008 and February 2009. MATERIALS: Wistar adult healthy male rats weighing 180-200 g were used to prepare acellular nerve graft, while Wistar adult healthy male rats weighing 100-120 g were used to prepare bone marrow mesenchymal stem cells. Tissue engineered artificial nerve was produced with acellular nerve graft co-cultured with bone marrow mesenchymal stem cells. METHODS: Sixty Wistar adult healthy male rats weighing 180-200 g were induced sciatic nerve defect models, 15 mm long. SD rats were divided into three groups at random with 20 animals in each group. ①Experiment group: Rat sciatic nerve defects were bridged with tissue engineered artificial nerve. ②Blank control group: Rat sciatic nerve defects were bridged with tissue engineered nerve scaffold. ③Autologous nerve control group: Rat sciatic nerve defects were bridged with autologous nerve graft. MAIN OUTCOME MEASURES: At 12 weeks postoperation, the recovery of motor function was evaluated with gross observation, electrophysiology, histological observation and triceps surae wet weight.RESULTS: ①At 12 weeks postoperation, the toes at the operation side could separate and supported to the ground in the experiment group; there was no significant difference in the regenerated nerve conduction velocity between experimental group and autologous nerve graft group. ②At 12 weeks postoperation, histochemical

  9. Acellular nerve scaffold loading bone marrow mesenchymal stem cells to prepare artificial nerve for sciatic nerve defects%无细胞神经支架复合骨髓间充质干细胞构建组织工程人工神经修复坐骨神经缺损

    Institute of Scientific and Technical Information of China (English)

    张彩顺; 吕刚

    2011-01-01

    背景:作者已经成功制备了无细胞神经移植物,并且复合骨髓间充质干细胞构建组织工程人工神经桥接大鼠坐骨神经缺损.目的:无细胞神经移植物复合骨髓间充质干细胞构建组织工程人工神经修复大鼠坐骨神经缺损后运动功能的恢复.方法:成年雄性SD大鼠构建大鼠坐骨神经15 mm缺损模型,分别应用组织工程人工神经、组织工程神经支架或自行神经桥接坐骨神经缺损.桥接后20周再生神经电生理学测定,手术侧胫骨前肌湿质量、腓肠肌组织学及透视电镜分析.结果与结论:桥接20周后,组织工程人工神经与自体神经移植组胫骨前肌湿质量比较,差异无显著性意义(P > 0.05),神经干传导速度为(30.56±2.15)m/s.结果提示,无细胞神经移植物复合骨髓间充质干细胞构建的组织工程人工神经桥接大鼠坐骨神经缺损后,可以促进再生神经运动功能的恢复.%BACKGROUND: The authors have successfully prepared an acellular nerve graft to be combined with bone marrow mesenchymal stem cells in artificial nerve bridge of rat sciatic nerve defects.OBJECTIVE: To investigate the motor function of rats with sciatic nerve defects following repairing using acellular nerve graft combined with bone marrow mesenchymal stem cells.METHODS: Adult SD rats were used to prepare rat models of 15-mm sciatic nerve defects. Tissue-engineered nerve, tissue-engineered nerve scaffold, and autogenous nerve were separately applied to bridge sciatic nerve defects. After 20 weeks, electrophysiological determination of regenerated nerves was conducted, wet weight of the anterior tibial muscle was determined,and histological and transmission electron microscopy analyses of the gastrocnemius was done.RESULTS AND CONCLUSION: Twenty weeks after nerve grafting, no difference in the wet weight of the anterior tibial muscle was shown to be significant between tissue-engineered nerve and autogenous nerve grafts (P > 0

  10. Acellular dermis-assisted breast reconstruction.

    Science.gov (United States)

    Spear, S L; Parikh, P M; Reisin, E; Menon, N G

    2008-05-01

    In 2004, the authors reported their findings with placement of tissue expanders for breast reconstruction in the partial submuscular position, the equivalent of the "dual-plane" technique for breast augmentation. Limitations with subpectoral expander placement include difficulty controlling the lower pole of the pocket during expansion, unprotected device coverage by a thin inferior mastectomy flap, possible effacement of the inframammary fold, and limited control over the superior migration of the pectoralis major muscle. This study aimed to examine the safety and efficacy of an acellular dermal sling in providing inferolateral support to the device during immediate breast reconstruction and expansion. This study prospectively investigated 58 breasts of 43 consecutive women who underwent immediate breast reconstruction with tissue expanders and acellular dermis. After completion of adjuvant therapy and expansion, the devices were exchanged for implants. The patients were tracked through January, 2007. The study parameters included demographic information, oncologic data, complications, and aesthetic outcomes. The mean time required to complete reconstruction was 8.6 months. The overall complication rate after expander/acellular dermis placement was 12%, whereas the complication rate after exchange to implants was 2.2%. The aesthetic outcome for reconstructed breasts did not differ significantly from that for the control subjects who had no surgery. Acellular dermis appears to be a useful adjunct in immediate prosthetic breast reconstruction. Acellular dermis-assisted breast reconstruction has a low complication rate, helps to reconstruct an aesthetically pleasing breast, and facilitates expeditious completion of the reconstruction.

  11. Alternatives to Acellular Dermal Matrix: Utilization of a Gore DualMesh Sling as a Cost-Conscious Adjunct for Breast Reconstruction.

    Science.gov (United States)

    Grow, Jacob N; Butterworth, James; Petty, Paul

    2017-01-01

    Objective: This study seeks an alternative to acellular dermal matrix in 2-staged breast reconstruction while minimizing cost. It was hypothesized that use of a Gore DualMesh would allow for similar intraoperative tissue expander fill volumes, time to second-stage reconstruction, and number of postoperative fills compared with acellular dermal matrix at only a fraction of the expense. Methods: Retrospective analysis comparing Gore DualMesh (59 breasts, 34 patients), acellular dermal matrix (13 breasts, 8 patients), and total muscle coverage (25 breasts, 14 patients) for postmastectomy breast reconstruction was performed. Time to second-stage reconstruction, number of expansions, and relative initial fill volumes were compared between the 3 groups. Secondarily, complication rates were also considered, including seroma, infection, expander/implant explantation, removal of mesh, and capsular contracture. Statistical analysis was performed utilizing the Fisher exact test and the χ(2) test for categorical variables and the Mann-Whitney U test for continuous variables. Results: Relative initial fill volumes, number of expansions, and time to second-stage reconstruction showed no statistical difference between the acellular dermal matrix and Gore DualMesh groups (P = .494, P = .146, and P = .539, respectively). Furthermore, the Gore DualMesh group underwent significantly fewer fills (P Gore DualMesh represents a safe alternative to acellular dermal matrix for breast reconstruction with similar aesthetic results in certain patients at a fraction of the cost.

  12. Efficient Isolation of Mesenchymal Stem Cells from Human Bone Marrow by Direct Plating Method Combined with Modified Primary Explant Culture%直接铺种结合改良组织块培养法可有效分离人骨髓中的间充质干细胞

    Institute of Scientific and Technical Information of China (English)

    邢文; 庞爱明; 姚剑峰; 李园; 石慧; 盛梦瑶; 周圆; 赵迎旭; 许明江

    2013-01-01

    Human bone marrow is the major source of mesenchymal stem cells (MSC). It was reported that the standard density gradient centrifugation method was not efficient in isolating MSC and it may be caused by the existing of bone marrow particles. In previous studys, a lot of MSC were obtained by culturing bone marrow particles alone combined with standard method. However, it is time- and labor-consuming to obtain bone marrow particles by filtering and to isolate MNC by density gradient centrifugation. This study was purposed to explore the more simple and efficient method to isolate MSC from bone marrow. Seven normal bone marrow aspirates were collected and centrifugated. The bone marrow particles floated on surface layers were cultured by modified primary explant culture, whereas the bone marrow aspirates deposited were cultured by direct plating method, then the immun phenotype and differentiation capability of isolated cells were analyzed. The results showed that in 3 of 7 aspirates, bone marrow particles were floated on surface layers, whereas the other bone marrow cells and some particles were deposited after centrifugation. The MSC were reliably isolated from the floating layers or deposited aspirates by modified primary explant culture and direct plating method separately. After 3 passages the isolated MSC did not express CD45 and CD34, but expressed CD105 ,CD73, CD44,CD90,CD49e and they could differentiate into chondrocytes and adipocytes. It is concluded that normal human bone marrow MSC can be isolated simply and efficiently by direct plating method in combination with modified primary explant culture.%骨髓是间充质干细胞(MSC)的重要来源.研究显示,标准密度梯度离心法分离骨髓MSC的效率不高,骨髓小粒是造成该法低效的原因.通过组织块法分离骨髓小粒,再结合标准法,可从单份骨髓标本分离获得更多MSC,然而这种方法费时费力.本研究探求分离骨髓MSC更简单、更有效的方法.收集7

  13. Acellular Dermal Matrix in Postmastectomy Breast Reconstruction

    NARCIS (Netherlands)

    A.M.S. Ibrahim (Ahmed)

    2014-01-01

    markdownabstract__Abstract__ Over the last decade the use of acellular dermal matrix (ADM) in reconstructive breast surgery has been transformative. Some authors have gone as far as to suggest that it is the single most important advancement in prosthetic breast reconstruction. ADMs are able

  14. Enzymatic Cell Isolation and Explant Cultures of Rat Calvarial Osteoblast Cells

    Institute of Scientific and Technical Information of China (English)

    2005-01-01

    Osteoblast cells were isolated from the calvarial bones of newborn Wistar rats and cultured in vitro via both collagenase digestion method and explant technique, and a comparative study was carried out on the two culture methods. The biologic characteristics of tbs osteoblast cells were studied via cell number counting,morphology observation, alkaline phosphatase staining of the cells and alizarine- red staining of the calcified nodules. The results show that osteoblast cells can be cultured in vitro via collagenase digestion method and explant technique, and the obtained cells are of good biologic characteristics. In comparison with the explant techniqne,the operative procedure of the enzymatic digestion method is more complicated. The digestion time must be carefully controlled. However, with this method, one can obtain a lager number of cells in a short time. The operative procedure of the explant technique is simpler, but it usually takes longer time to obtain cells of desirable number.

  15. Morphology of spinal cord extracellular matrixderived acellular scaffolds fabricated in rats

    Institute of Scientific and Technical Information of China (English)

    Wenhua Yin; Kaiwu Lu; Dadi Jin

    2011-01-01

    Acellular peripheral allograft scaffolds can be fabricated using chemical extraction techniques, but methods for producing acellular scaffold derived from spinal cord tissue are not currently available.The present study demonstrated that chemical extraction using Triton X-100 and sodium deoxycholate could be used to completely remove the cells, axons and neural sheaths in spinal cord extracellular matrix-derived scaffolds. The matrix fibers were longitudinally arranged in a wave-like formation, and were connected by fiber junctions. Lattice-shaped fiber cages appeared and developed into bone trabecula-like changes. The natural structure of matrix fibers in the scaffolds was maintained; this helps to guide the differentiation and migration of implanted stem cells. Decellularized spinal cord extracellular matrix-derived scaffolds can provide an ideal substance for fabricating tissue-engineered spinal cord.

  16. Cartilage oligomeric matrix protein enhances the vascularization of acellular nerves

    Directory of Open Access Journals (Sweden)

    Wei-ling Cui

    2016-01-01

    Full Text Available Vascularization of acellular nerves has been shown to contribute to nerve bridging. In this study, we used a 10-mm sciatic nerve defect model in rats to determine whether cartilage oligomeric matrix protein enhances the vascularization of injured acellular nerves. The rat nerve defects were treated with acellular nerve grafting (control group alone or acellular nerve grafting combined with intraperitoneal injection of cartilage oligomeric matrix protein (experimental group. As shown through two-dimensional imaging, the vessels began to invade into the acellular nerve graft from both anastomotic ends at day 7 post-operation, and gradually covered the entire graft at day 21. The vascular density, vascular area, and the velocity of revascularization in the experimental group were all higher than those in the control group. These results indicate that cartilage oligomeric matrix protein enhances the vascularization of acellular nerves.

  17. Cartilage oligomeric matrix protein enhances the vascularization of acellular nerves

    Institute of Scientific and Technical Information of China (English)

    Wei-ling Cui; Long-hai Qiu; Jia-yan Lian; Jia-chun Li; Jun Hu; Xiao-lin Liu

    2016-01-01

    Vascularization of acellular nerves has been shown to contribute to nerve bridging. In this study, we used a 10-mm sciatic nerve defect model in rats to determine whether cartilage oligomeric matrix protein enhances the vascularization of injured acellular nerves. The rat nerve defects were treated with acellular nerve grafting (control group) alone or acellular nerve grafting combined with intraperitoneal injection of cartilage oligomeric matrix protein (experimental group). As shown through two-dimensional imaging, the vessels began to invade into the acellular nerve graft from both anastomotic ends at day 7 post-operation, and gradually covered the entire graft at day 21. The vascular density, vascular area, and the velocity of revascularization in the experimental group were all higher than those in the control group. These results indicate that cartilage oligomeric matrix protein enhances the vascularization of acellular nerves.

  18. Central role of pyrophosphate in acellular cementum formation.

    Directory of Open Access Journals (Sweden)

    Brian L Foster

    Full Text Available BACKGROUND: Inorganic pyrophosphate (PP(i is a physiologic inhibitor of hydroxyapatite mineral precipitation involved in regulating mineralized tissue development and pathologic calcification. Local levels of PP(i are controlled by antagonistic functions of factors that decrease PP(i and promote mineralization (tissue-nonspecific alkaline phosphatase, Alpl/TNAP, and those that increase local PP(i and restrict mineralization (progressive ankylosis protein, ANK; ectonucleotide pyrophosphatase phosphodiesterase-1, NPP1. The cementum enveloping the tooth root is essential for tooth function by providing attachment to the surrounding bone via the nonmineralized periodontal ligament. At present, the developmental regulation of cementum remains poorly understood, hampering efforts for regeneration. To elucidate the role of PP(i in cementum formation, we analyzed root development in knock-out ((-/- mice featuring PP(i dysregulation. RESULTS: Excess PP(i in the Alpl(-/- mouse inhibited cementum formation, causing root detachment consistent with premature tooth loss in the human condition hypophosphatasia, though cementoblast phenotype was unperturbed. Deficient PP(i in both Ank and Enpp1(-/- mice significantly increased cementum apposition and overall thickness more than 12-fold vs. controls, while dentin and cellular cementum were unaltered. Though PP(i regulators are widely expressed, cementoblasts selectively expressed greater ANK and NPP1 along the root surface, and dramatically increased ANK or NPP1 in models of reduced PP(i output, in compensatory fashion. In vitro mechanistic studies confirmed that under low PP(i mineralizing conditions, cementoblasts increased Ank (5-fold and Enpp1 (20-fold, while increasing PP(i inhibited mineralization and associated increases in Ank and Enpp1 mRNA. CONCLUSIONS: Results from these studies demonstrate a novel developmental regulation of acellular cementum, wherein cementoblasts tune cementogenesis by modulating

  19. Data from acellular human heart matrix

    Directory of Open Access Journals (Sweden)

    Pedro L Sánchez

    2016-09-01

    Full Text Available Perfusion decellularization of cadaveric hearts removes cells and generates a cell-free extracellular matrix scaffold containing acellular vascular conduits, which are theoretically sufficient to perfuse and support tissue-engineered heart constructs. This article contains additional data of our experience decellularizing and testing structural integrity and composition of a large series of human hearts, “Acellular human heart matrix: a critical step toward whole heat grafts” (Sanchez et al., 2015 [1]. Here we provide the information about the heart decellularization technique, the valve competence evaluation of the decellularized scaffolds, the integrity evaluation of epicardial and myocardial coronary circulation, the pressure volume measurements, the primers used to assess cardiac muscle gene expression and, the characteristics of donors, donor hearts, scaffolds and perfusion decellularization process.

  20. Differentiation of mesenchymal stem cells into neuronal cells on fetal bovine acellular dermal matrix as a tissue engineered nerve scaffold

    Institute of Scientific and Technical Information of China (English)

    Yuping Feng; Jiao Wang; Shixin Ling; Zhuo Li; Mingsheng Li; Qiongyi Li; Zongren Ma; Sijiu Yu

    2014-01-01

    The purpose of this study was to assess fetal bovine acellular dermal matrix as a scaffold for supporting the differentiation of bone marrow mesenchymal stem cells into neural cells fol-lowing induction with neural differentiation medium. We performed long-term, continuous observation of cell morphology, growth, differentiation, and neuronal development using several microscopy techniques in conjunction with immunohistochemistry. We examined speciifc neu-ronal proteins and Nissl bodies involved in the differentiation process in order to determine the neuronal differentiation of bone marrow mesenchymal stem cells. The results show that bone marrow mesenchymal stem cells that differentiate on fetal bovine acellular dermal matrix display neuronal morphology with unipolar and bi/multipolar neurite elongations that express neuro-nal-speciifc proteins, includingβIII tubulin. The bone marrow mesenchymal stem cells grown on fetal bovine acellular dermal matrix and induced for long periods of time with neural differen-tiation medium differentiated into a multilayered neural network-like structure with long nerve ifbers that was composed of several parallel microifbers and neuronal cells, forming a complete neural circuit with dendrite-dendrite to axon-dendrite to dendrite-axon synapses. In addition, growth cones with filopodia were observed using scanning electron microscopy. Paraffin sec-tioning showed differentiated bone marrow mesenchymal stem cells with the typical features of neuronal phenotype, such as a large, round nucleus and a cytoplasm full of Nissl bodies. The data suggest that the biological scaffold fetal bovine acellular dermal matrix is capable of supporting human bone marrow mesenchymal stem cell differentiation into functional neurons and the subsequent formation of tissue engineered nerve.

  1. 引导种植牙区骨再生的异种脱细胞真皮基质%Acellular dermal matrix used for guiding bone regeneration in the dental implant area

    Institute of Scientific and Technical Information of China (English)

    周静; 邓蔡; 张进锋

    2013-01-01

      BACKGROUND: Acel ular dermal matrix is a kind of prosthodontics membrane material which has been widely used due to the good biocompatibility. OBJECTIVE: To evaluate the effect of acel ular dermal matrix in guiding bone regeneration in the dental implant area. METHODS: Immunohistochemical staining was performed, and the microscope was used to observe the microstructure and cel compatibility of acel ular dermal matrix, in order to evaluate the feasibility of acel ular dermal matrix in guiding bone regeneration. The dental implantation patients who received bone regeneration with acel ular dermal matrix were fol owed-up to evaluate the osteogenesis effect and the effect on soft tissue defects. Then, the effects of Bio-Gide membrane and Bot medical col agen membrane on guiding bone regeneration were compared. RESULTS AND CONCLUSION: The microstructure of acel ular dermal matrix showed there was basement membrane surface and tissue surface. The stylode-like structure and hair fol icle could be observed on the basement membrane surface and the tissue surface was scaly structure, and acel ular dermal matrix had no influence on the proliferation activity osteoblast-like cel s and the alkaline phosphatase activity, but had good cel compatibility. The clinical researches showed that acel ular dermal matrix used in dental implantation was effective for bone regeneration, and there was no significant difference in the effect on guiding bone regeneration when compared with Bio-Gide membrane and Bot medical col agen membrane. The acel ular dermal matrix had good bone regeneration effect in repairing soft tissue deficiencies after bone augmentation.%  背景:异种脱细胞真皮基质属于口腔修复膜材料,因具有良好的生物相容性而被广泛应用。目的:评价异种脱细胞真皮基质在牙区引导牙种植骨再生的效果。方法:以免疫组化染色后显微镜观察异种脱细胞真皮基质的显微结构和细胞相容性,评

  2. Exogenic Acellular Dermal Matrix in Guided Bone Regeneration of Dental Implant%异种脱细胞真皮基质在牙种植中引导骨再生的临床观察

    Institute of Scientific and Technical Information of China (English)

    韦丽萍; 左陈启; 王远勤

    2011-01-01

    Objective: To identify the clinical effect of a homemade exogenic acelluar dermal matrix as a barrier membrane in guide bone regeneration (GBR) of dental implants. Methods: Seventy eight dental implants/cases with limited bone-bed were divided into 2 groups, and treated by GBR technology with different barrier membrane. Experiment group (38 cases) used homemade exogenic acelluar dermal matrix, while in the control group (40 cases) Bio-Gide biofilm was used. Results: The difference of bone harvested in two groups was not statistically significant (P>0.05). All patients were healed with first intention and the healing rate of 2 groups were all 100%. One case complicated with facial swelling in control group, which was cured after anti-inflammatory and symptomatic treatment. There was no statistically significant between two groups in the demographic basic information, such as, adverse events, healing of incision, bone growth effect,and the like. Conclusion: This alternative homemade material should be considered in GBR by practitioners.%目的:观察国产异种脱细胞真皮基质修复膜作为屏障膜,在牙种植的引导骨再生中应用的临床效果.方法:78例需要引导骨再生的种植病例,均为前牙区或前磨牙区单牙或连续多牙位(3牙)缺失;其中前牙52颗,前磨牙26颗.缺牙区牙槽嵴主要为唇颊侧垂直性吸收,牙槽嵴顶宽度约1~3 mm,牙槽嵴水平吸收量在2 mm以内.分为2组,一组采用国产异种脱细胞真皮基质修复膜作为GBR技术的屏蔽膜(38例),另一组采用Bio-Gide生物膜作为对照(40例),比较二者的临床效果.结果:2组在骨生长效果之间的差异无统计学意义(P>0.05).2组患者切口均甲级愈合,切口愈合率均为100%.术后3 d,对照组有1例发生面部肿胀,经抗感染治疗和对症处理后缓解.本研究的其他病例均未出现感染等不良事件.结论:使用国产异种脱细胞真皮基质修复膜在牙种植术中进行骨引

  3. Acellular organ scaffolds for tumor tissue engineering

    Science.gov (United States)

    Guller, Anna; Trusova, Inna; Petersen, Elena; Shekhter, Anatoly; Kurkov, Alexander; Qian, Yi; Zvyagin, Andrei

    2015-12-01

    Rationale: Tissue engineering (TE) is an emerging alternative approach to create models of human malignant tumors for experimental oncology, personalized medicine and drug discovery studies. Being the bottom-up strategy, TE provides an opportunity to control and explore the role of every component of the model system, including cellular populations, supportive scaffolds and signalling molecules. Objectives: As an initial step to create a new ex vivo TE model of cancer, we optimized protocols to obtain organ-specific acellular matrices and evaluated their potential as TE scaffolds for culture of normal and tumor cells. Methods and results: Effective decellularization of animals' kidneys, ureter, lungs, heart, and liver has been achieved by detergent-based processing. The obtained scaffolds demonstrated biocompatibility and growthsupporting potential in combination with normal (Vero, MDCK) and tumor cell lines (C26, B16). Acellular scaffolds and TE constructs have been characterized and compared with morphological methods. Conclusions: The proposed methodology allows creation of sustainable 3D tumor TE constructs to explore the role of organ-specific cell-matrix interaction in tumorigenesis.

  4. The effects of acellular amniotic membrane matrix on osteogenic differentiation and ERK1/2 signaling in human dental apical papilla cells.

    Science.gov (United States)

    Chen, Yi-Jane; Chung, Min-Chun; Jane Yao, Chung-Chen; Huang, Chien-Hsun; Chang, Hao-Hueng; Jeng, Jiiang-Huei; Young, Tai-Horng

    2012-01-01

    The amniotic membrane (AM) has been widely used in the field of tissue engineering because of the favorable biological properties for scaffolding material. However, little is known about the effects of an acellular AM matrix on the osteogenic differentiation of mesenchymal stem cells. In this study, it was found that both basement membrane side and collagenous stroma side of the acellular AM matrix were capable of providing a preferential environment for driving the osteogenic differentiation of human dental apical papilla cells (APCs) with proven stem cell characteristics. Acellular AM matrix potentiated the induction effect of osteogenic supplements (OS) such as ascorbic acid, β-glycerophosphate, and dexamethasone and enhanced the osteogenic differentiation of APCs, as seen by increased core-binding factor alpha 1 (Cbfa-1) phosphorylation, alkaline phosphatase activity, mRNA expression of osteogenic marker genes, and mineralized matrix deposition. Even in the absence of soluble OS, acellular AM matrix also could exert the substrate-induced effect on initiating APCs' differentiation. Especially, the collagenous stroma side was more effective than the basement membrane side. Moreover, the AM-induced effect was significantly inhibited by U0126, an inhibitor of extracellular signaling-regulated kinase 1/2 (ERK1/2) signaling. Taken together, the osteogenic differentiation promoting effect on APCs is AM-specific, which provides potential applications of acellular AM matrix in bone/tooth tissue engineering.

  5. Induction of embryogenics calli in cupuassu explants

    OpenAIRE

    Maria das Graças Rodrigues Ferreira; Fernando Enrique Ninamango Cárdenas; Carlos Henrique Siqueira de Carvalho; Andréa Almeida Carneiro; Carlos Ferreira Damião Filho

    2004-01-01

    Objetivou-se a indução de calos embriogênicos em cupuaçuzeiro, em função do tipo de explante e meio de cultura. Foram testados como explantes, segmentos cotiledonares e eixos embrionários divididos em três partes: região da plúmula, radícula e hipocótilo. Os explantes foram cultivados em 2 diferentes meios de cultura: 1) MS suplementado com 2,4-D (1 mg L-1) e Cinetina (0,25 mg L-1); 2) MS acrescido de ANA (5 mg L-1) e Cinetina (0,25 mg L-1). Constatou-se que a região do hipocótilo foi a parte...

  6. Bone

    Science.gov (United States)

    Helmberger, Thomas K.; Hoffmann, Ralf-Thorsten

    The typical clinical signs in bone tumours are pain, destruction and destabilization, immobilization, neurologic deficits, and finally functional impairment. Primary malignant bone tumours are a rare entity, accounting for about 0.2% of all malignancies. Also benign primary bone tumours are in total rare and mostly asymptomatic. The most common symptomatic benign bone tumour is osteoid osteoma with an incidence of 1:2000.

  7. 化学去细胞异体神经周围复合BMSCs生物蛋白胶复合物促周围神经缺损修复%EFFECT OF CHEMICAL EXTRACTED ACELLULAR NERVE ALLOGRAFT SUPPLEMENTING WITH BONE MARROW MESENCHYMAL STEM CELLS EMBEDDED IN FIBRIN GLUE ON FUNCTIONAL RECOVERY OF TRANSECTED SCIATIC NERVES

    Institute of Scientific and Technical Information of China (English)

    赵喆; 许文静; 卢世璧; 王玉; 彭江; 赵斌; 赵庆; 刘炎; 任志午; 詹胜峰; 张莉

    2011-01-01

    目的 将BMSCs复合在化学去细胞异体神经(chemical extracted acellular nerve allograft,CEANA)周围,观察对CEANA修复周围神经缺损效果的影响.方法 成年雄性C57小鼠21只,体重25~30 g;成年雄性Balb/c小鼠15只,体重25~30 g.取Balb/c小鼠双侧坐骨神经,制备CEANA.取C57小鼠3只,分离培养BMSCs,取5 x 106个第3代BMSCs添加到500μL生物蛋白胶制备BMSCs生物蛋白胶复合物,共培养3、7、14、21 d后,分别取其上清与PC12细胞共培养,观察对PC12细胞的影响.取成年雄性C57小鼠18只,制备小鼠左侧坐骨神经10mm缺损模型,随机分成3组(n=6),分别采用自体神经移植复合生物蛋白胶(A组)、CEANA移植复合BMSCs生物蛋白胶复合物(B组)、CEANA移植复合生物蛋白胶(C组)修复坐骨神经缺损;实验动物右侧切开暴露坐骨神经,作为正常对照.术后行大体观察;术前及术后2、4、6、8周测量小鼠坐骨神经指数(static sciatic index,SSI);术后8周取材计算术侧小腿三头肌湿重恢复率并行小腿三头肌Masson染色观察,吻合口远端神经行甲苯胺蓝染色和透射电镜观察.结果 BMSCs在生物蛋白胶内均匀分布,外观呈球形,培养3d后可见BMSCs呈多个长突起.加入BMSCs生物蛋白胶复合物共培养3、7、14、21 d的上清,PC12细胞均分化为类神经元样细胞.术后各组动物切口愈合良好.各组SSI随时间延长逐渐增加,术后4、6、8周A组SSI均高于B、C组,差异有统计学意义(P0.05).术后8周,B组小腿三头肌湿重恢复率、有髓神经纤维总数均优于C组,但较A组差,差异有统计学意义(P0.05).结论 在CEANA周围添加BMSCs生物蛋白胶复合物可提高周围神经损伤修复效果.%Objective To investigate the effect of bone marrow mesenchymal stem cells (BMSCs) embedded in fibrin glue around chemical extracted acellular nerve allograft (CEANA) on the peripheral nerve regeneration. Methods Twenty-one adult male C57 mice (weighing 25-30 g

  8. CALLUS INDUCTION FROM FLORAL EXPLANTS OF CUPUASSU

    Directory of Open Access Journals (Sweden)

    MARIA DAS GRAÇAS RODRIGUES FERREIRA

    2013-01-01

    Full Text Available There are few studies related to the in vitro cultivation of plants from theTheobroma genus and no effective micropropagation protocols for T.grandiflorum. The aim of this study was to evaluate the calli formation in cupuassu floral explants, targeting their organogenic or embryogenicdevelopment. Experiments were conducted in the Plant Tissue Culture Laboratory of EMBRAPA, Porto Velho, Rondônia, Brazil. Floral parts from unopened immature flower buds taken from seedless cupuassu trees were sterilized and employed as a source of explants. These explants were cultivated in Petri dishes in an induction medium consisting of MS salts and vitamins, supplemented with glycine(3 mg.L-1, lysine (0,4 mg.L-1, leucine (0,4 mg.L-1, arginine (0,4 mg.L-1, tryptophan (0,2 mg.L-1, 2,4-D (1 mg.L-1, kinetin (0,25 mg.L-1, coconut water (50 ml.L-1, sucrose (40 g.L-1, Gelrite (2,2 g.L-1 and pH adjusted to 5,8. Cultures were maintained in the dark for 3 weeks at 27°C and then subcultured for six weeks in medium without growth regulators supplemented with glycine (1 mg.L-1, lysine (0,2 mg.L-1, leucine (0,2 mg.L-1, arginine (0,2 mg.L-1, tryptophan (0,1 mg.L-1, coconut water (100 ml.L-1, sucrose (40 g.L-1, Gelrite (2,2 g.L-1 and pH 5,8. We used a completely randomized design with 10 replications of 5 explants per plate and four different explant sources: staminode, petal, ligule and ovary. As a result, we obtained a highercalli formation in theinduction medium when ovaries were used as source of explants. However, there was no development of somatic embryosor organogenic response in medium without growth regulators and further studies are being conducted.

  9. Successful breast reconstruction using acellular dermal matrix can be recommended in healthy non-smoking patients

    DEFF Research Database (Denmark)

    Gunnarsson, Gudjon Leifur; Børsen-Koch, Mikkel; Arffmann, Susanne

    2013-01-01

    We present Scandinavia's first series of immediate alloplastic breast reconstructions with an acellular dermal matrix.......We present Scandinavia's first series of immediate alloplastic breast reconstructions with an acellular dermal matrix....

  10. Pulmonary heart valve replacement using stabilized acellular xenogeneic scaffolds; effects of seeding with autologous stem cells

    Directory of Open Access Journals (Sweden)

    Harpa Marius Mihai

    2015-12-01

    Full Text Available Background: We hypothesized that an ideal heart valve replacement would be acellular valve root scaffolds seeded with autologous stem cells. To test this hypothesis, we prepared porcine acellular pulmonary valves, seeded them with autologous adipose derived stem cells (ADSCs and implanted them in sheep and compared them to acellular valves.

  11. Hypoxia inhibits hypertrophic differentiation and endochondral ossification in explanted tibiae.

    Directory of Open Access Journals (Sweden)

    Jeroen C H Leijten

    Full Text Available PURPOSE: Hypertrophic differentiation of growth plate chondrocytes induces angiogenesis which alleviates hypoxia normally present in cartilage. In the current study, we aim to determine whether alleviation of hypoxia is merely a downstream effect of hypertrophic differentiation as previously described or whether alleviation of hypoxia and consequent changes in oxygen tension mediated signaling events also plays an active role in regulating the hypertrophic differentiation process itself. MATERIALS AND METHODS: Fetal mouse tibiae (E17.5 explants were cultured up to 21 days under normoxic or hypoxic conditions (21% and 2.5% oxygen respectively. Tibiae were analyzed on growth kinetics, histology, gene expression and protein secretion. RESULTS: The oxygen level had a strong influence on the development of explanted fetal tibiae. Compared to hypoxia, normoxia increased the length of the tibiae, length of the hypertrophic zone, calcification of the cartilage and mRNA levels of hypertrophic differentiation-related genes e.g. MMP9, MMP13, RUNX2, COL10A1 and ALPL. Compared to normoxia, hypoxia increased the size of the cartilaginous epiphysis, length of the resting zone, calcification of the bone and mRNA levels of hyaline cartilage-related genes e.g. ACAN, COL2A1 and SOX9. Additionally, hypoxia enhanced the mRNA and protein expression of the secreted articular cartilage markers GREM1, FRZB and DKK1, which are able to inhibit hypertrophic differentiation. CONCLUSIONS: Collectively our data suggests that oxygen levels play an active role in the regulation of hypertrophic differentiation of hyaline chondrocytes. Normoxia stimulates hypertrophic differentiation evidenced by the expression of hypertrophic differentiation related genes. In contrast, hypoxia suppresses hypertrophic differentiation of chondrocytes, which might be at least partially explained by the induction of GREM1, FRZB and DKK1 expression.

  12. Composition of acellular pertussis and combination vaccines: a general review.

    Science.gov (United States)

    Jadhav, S S; Gairola, S

    1999-06-01

    Since the development and introduction of the acellular pertussis vaccine in Japan in the early eighties, we have come a long way in using this component in combination with other vaccines. However, the basic problem in development of an effective and safe pertussis vaccine is that the antigens to induce complete protection against clinical pertussis and the precise mechanism by which pertussis vaccine confers immunity is yet unknown. Hence, the composition of future acellular pertussis vaccine remains an open issue. Recently, acellular pertussis vaccine has been licensed for the booster doses in the U.S.A. and for primary immunization of infants in Italy and Germany. A multicentric trial has been carried out to compare the serological response and adverse reactions of 13 acellular pertussis vaccines. These vaccines contained one or more of the four components, i.e. FHA, PT, 69 kDa OMP and fimbriae. All vaccines were associated with substantially fewer and less adverse reactions and were more immunogenic with respect to antibodies against the added antigens. DTP vaccines in the near future will have combinations of other components and the key antigen for combination will be acellular pertussis component which is going to replace whole cell pertussis component in DTP vaccines. In view of this, manufacturers like ourselves from the developing countries are still groping in the dark, uncertain whether we should have a single component acellular pertussis vaccine or multicomponent one. This will have a major impact on the cost of production, the final cost of the combination vaccines and the regulatory issues that we will have to tackle in view of the recent thinking on harmonization in the pharmaceutical industry. Copyright 1999 The International Association for Biologicals.

  13. Abdominal wall repair with human acellular dermal autograft

    Directory of Open Access Journals (Sweden)

    Roel E. Genders

    2011-12-01

    Full Text Available Repair of abdominal wall defects in the presence of contamination or infection is a significant problem. The loss of tissue warrants enforcement of the abdominal wall, preferably by autologous material. However, autologous repair often requires extensive surgery. This paper presents a review of available literature of placement of an acellular human dermis to repair an abdominal fascia defect, in contaminated as well as in non-contaminated surgical fields. It is illustrated with a case report that describes the successful reconstruction of an infected abdominal wall defect with a human acellular dermis allograft. A systematic literature review was undertaken with searches performed in the Pubmed and Cochrane databases for the period up till March 2009, using the search terms Alloderm [Substance Name], Hernia [Mesh] and the key words acellular dermis, acellular dermal matrix, human acellular dermal allograft and abdominal wall defect. To assess methodological quality, each article was subjected to a modification of the methodological index for non-randomized studies (MINORS according to Slim et al. Two items from the original index were not included because none of the studies selected had an unbiased assessment of the study end points and in none of the studies was a prospective calculation of the study size performed. Seventeen studies were included in the review. Data were extracted regarding study design, number of patients, surgical technique, followup period, contaminated or non-contaminated area of the fascia defect, mortality and morbidity (hemorrhage, seroma, wound dehiscence, infection of the operative procedure, the longterm results (removal of the graft, reherniation and bulging and level of evidencey. A total of 169 short-term complications and 151 longterm complications occurred after 643 surgical procedures reconstructing both contaminated and clean abdominal wall defects by implantation of an HADA. Human acellular dermal allograft

  14. Treatment of amalgam tattoo with a subepithelial connective tissue graft and acellular dermal matrix.

    Science.gov (United States)

    Thumbigere-Math, Vivek; Johnson, Deborah K

    2014-04-01

    A 54-year-old female was referred for management of a large amalgam tattoo involving the alveolar mucosa between teeth #6 and #9. The lesion had been present for over 20 years following endodontic treatment of teeth #7 and #8. A two-stage surgical approach was used to remove the pigmentation, beginning with removal of amalgam fragments from the underlying bone and placement of a subepithelial connective tissue graft and acellular dermal matrix to increase soft tissue thickness subadjacent to the amalgam. Following 7 weeks of healing, gingivoplasty was performed to remove the overlying pigmented tissue. At the 21-month follow-up appointment, the patient exhibited naturally appearing soft tissue with no evidence of amalgam tattoo.

  15. Making post-mortem implantable cardioverter defibrillator explantation safe

    OpenAIRE

    Räder, Sune B.E.W.; Zeijlemaker, Volkert; Pehrson, Steen; Svendsen, Jesper H

    2009-01-01

    Aims The aim of this study is to investigate whether protection with rubber or plastic gloves during post-mortem explantation of an implantable cardioverter defibrillator (ICD) offers enough protection for the explanting operator during a worst-case scenario (i.e. ICD shock). Methods and results We investigated the insulating properties of rubber and plastic gloves (double layer) within the first 60 min exposure (mimicking the maximum time of an explantation procedure) to saline (simulating t...

  16. Evaluation of lymphangiogenesis in acellular dermal matrix

    Directory of Open Access Journals (Sweden)

    Mario Cherubino

    2014-01-01

    Full Text Available Introduction: Much attention has been directed towards understanding the phenomena of angiogenesis and lymphangiogenesis in wound healing. Thanks to the manifold dermal substitute available nowadays, wound treatment has improved greatly. Many studies have been published about angiogenesis and cell invasion in INTEGRA® . On the other hand, the development of the lymphatic network in acellular dermal matrix (ADM is a more obscure matter. In this article, we aim to characterize the different phases of host cell invasion in ADM. Special attention was given to lymphangiogenic aspects. Materials and Methods: Among 57 rats selected to analyse the role of ADM in lymphangiogenesis, we created four groups. We performed an excision procedure on both thighs of these rats: On the left one we did not perform any action except repairing the borders of the wound; while on the right one we used INTEGRA® implant. The excision biopsy was performed at four different times: First group after 7 days, second after 14 days, third after 21 days and fourth after 28 days. For our microscopic evaluation, we used the classical staining technique of haematoxylin and eosin and a semi-quantitative method in order to evaluate cellularity counts. To assess angiogenesis and lymphangiogenesis development we employed PROX-1 Ab and CD31/PECAM for immunohistochemical analysis. Results: We found remarkable wound contraction in defects that healed by secondary intention while minor wound contraction was observed in defects treated with ADM. At day 7, optical microscopy revealed a more plentiful cellularity in the granulation tissue compared with the dermal regeneration matrix. The immunohistochemical process highlighted vascular and lymphatic cells in both groups. After 14 days a high grade of fibrosis was noticeable in the non-treated group. At day 21, both lymphatic and vascular endothelial cells were better developed in the group with a dermal matrix application. At day 28

  17. Acellular Nerve Allografts in Peripheral Nerve Regeneration: A Comparative Study

    Science.gov (United States)

    Moore, Amy M.; MacEwan, Matthew; Santosa, Katherine B.; Chenard, Kristofer E.; Ray, Wilson Z.; Hunter, Daniel A.; Mackinnon, Susan E.; Johnson, Philip J.

    2011-01-01

    Background Processed nerve allografts offer a promising alternative to nerve autografts in the surgical management of peripheral nerve injuries where short deficits exist. Methods Three established models of acellular nerve allograft (cold-preserved, detergent-processed, and AxoGen® -processed nerve allografts) were compared to nerve isografts and silicone nerve guidance conduits in a 14 mm rat sciatic nerve defect. Results All acellular nerve grafts were superior to silicone nerve conduits in support of nerve regeneration. Detergent-processed allografts were similar to isografts at 6 weeks post-operatively, while AxoGen®-processed and cold-preserved allografts supported significantly fewer regenerating nerve fibers. Measurement of muscle force confirmed that detergent-processed allografts promoted isograft-equivalent levels of motor recovery 16 weeks post-operatively. All acellular allografts promoted greater amounts of motor recovery compared to silicone conduits. Conclusions These findings provide evidence that differential processing for removal of cellular constituents in preparing acellular nerve allografts affects recovery in vivo. PMID:21660979

  18. Stabilization of gene expression and cell morphology after explant recycling during fin explant culture in goldfish

    Energy Technology Data Exchange (ETDEWEB)

    Chenais, Nathalie; Lareyre, Jean-Jacques; Le Bail, Pierre-Yves; Labbe, Catherine, E-mail: catherine.labbe@rennes.inra.fr

    2015-07-01

    The development of fin primary cell cultures for in vitro cellular and physiological studies is hampered by slow cell outgrowth, low proliferation rate, poor viability, and sparse cell characterization. Here, we investigated whether the recycling of fresh explants after a first conventional culture could improve physiological stability and sustainability of the culture. The recycled explants were able to give a supplementary cell culture showing faster outgrowth, cleaner cell layers and higher net cell production. The cells exhibited a highly stabilized profile for marker gene expression including a low cytokeratin 49 (epithelial marker) and a high collagen 1a1 (mesenchymal marker) expression. Added to the cell spindle-shaped morphology, motility behavior, and actin organization, this suggests that the cells bore stable mesenchymal characteristics. This contrast with the time-evolving expression pattern observed in the control fresh explants during the first 2 weeks of culture: a sharp decrease in cytokeratin 49 expression was concomitant with a gradual increase in col1a1. We surmise that such loss of epithelial features for the benefit of mesenchymal ones was triggered by an epithelial to mesenchymal transition (EMT) process or by way of a progressive population replacement process. Overall, our findings provide a comprehensive characterization of this new primary culture model bearing mesenchymal features and whose stability over culture time makes those cells good candidates for cell reprogramming prior to nuclear transfer, in a context of fish genome preservation. - Highlights: • Recycled fin explants outgrow cells bearing stable mesenchymal traits. • Cell production and quality is enhanced in the recycled explant culture system. • Fresh fin primary culture is highly variable and loose epithelial traits over time.

  19. Applications of acellular dermal matrix in revision breast reconstruction surgery.

    Science.gov (United States)

    Spear, Scott L; Sher, Sarah R; Al-Attar, Ali; Pittman, Troy

    2014-01-01

    Acellular dermal matrix has been used for over a decade in primary breast reconstruction. Few articles have specifically examined its use in revision breast reconstruction for fold malposition, capsular contracture, rippling, and symmastia. One hundred thirty-five revision breast reconstructive procedures using acellular dermal matrix (AlloDerm) in 118 patients (154 breasts) over a 5-year period were reviewed. Most procedures were revisions or part of the second stage of previous mastectomy reconstructions; three were revisions after reconstruction of congenital chest wall deformities. Fifty-seven revisions (37 percent) were for inferior fold malposition, followed by 40 (25.9 percent) for inferior pole support, 42 (27.2 percent) for capsular contracture, 10 (6.4 percent) for rippling, and five (3.2 percent) for symmastia. The overall complication rate was 5 percent. Revisions with acellular dermal matrix were successful in 147 of 154 breasts (95.5 percent). The most common complication was capsular contracture, occurring in five breasts (3.2 percent). There was one infection (0.6 percent), failure to lower the inframammary fold in one breast (0.6 percent), and one persistence of rippling (0.6 percent). The mean follow-up was 207 days. Acellular dermal matrix has proven to be a reliable tool for managing some of the most common and challenging problems in implant-based breast reconstruction. Although there are few published data on the success of more conventional solutions to fold malposition, lower pole support, and capsular contracture, the addition of acellular dermal matrix to buttress these repairs has been shown to provide a high likelihood of success with a low risk of complications.

  20. Subcutaneous Implant-based Breast Reconstruction with Acellular Dermal Matrix/Mesh: A Systematic Review.

    Science.gov (United States)

    Salibian, Ara A; Frey, Jordan D; Choi, Mihye; Karp, Nolan S

    2016-11-01

    The availability of acellular dermal matrix (ADM) and synthetic mesh products has prompted plastic surgeons to revisit subcutaneous implant-based breast reconstruction. The literature is limited, however, with regards to evidence on patient selection, techniques, and outcomes. A systematic review of the Medline and Cochrane databases was performed for original studies reporting breast reconstruction with ADM or mesh, and subcutaneous implant placement. Studies were analyzed for level of evidence, inclusion/exclusion criteria for subcutaneous reconstruction, reconstruction characteristics, and outcomes. Six studies (186 reconstructions) were identified for review. The majority of studies (66.7%) were level IV evidence case series. Eighty percent of studies had contraindications for subcutaneous reconstruction, most commonly preoperative radiation, high body mass index, and active smoking. Forty percent of studies commenting on patient selection assessed mastectomy flap perfusion for subcutaneous reconstruction. Forty-five percent of reconstructions were direct-to-implant, 33.3% 2-stage, and 21.5% single-stage adjustable implant, with ADM utilized in 60.2% of reconstructions versus mesh. Pooled complication rates included: major infection 1.2%, seroma 2.9%, hematoma 2.3%, full nipple-areola complex necrosis 1.1%, partial nipple-areola complex necrosis 4.5%, major flap necrosis 1.8%, wound healing complication 2.3%, explantation 4.1%, and grade III/IV capsular contracture 1.2%. Pooled short-term complication rates in subcutaneous alloplastic breast reconstruction with ADM or mesh are low in preliminary studies with selective patient populations, though techniques and outcomes are variable across studies. Larger comparative studies and better-defined selection criteria and outcomes reporting are needed to develop appropriate indications for performing subcutaneous implant-based reconstruction.

  1. Stabilization of gene expression and cell morphology after explant recycling during fin explant culture in goldfish.

    Science.gov (United States)

    Chenais, Nathalie; Lareyre, Jean-Jacques; Le Bail, Pierre-Yves; Labbe, Catherine

    2015-07-01

    The development of fin primary cell cultures for in vitro cellular and physiological studies is hampered by slow cell outgrowth, low proliferation rate, poor viability, and sparse cell characterization. Here, we investigated whether the recycling of fresh explants after a first conventional culture could improve physiological stability and sustainability of the culture. The recycled explants were able to give a supplementary cell culture showing faster outgrowth, cleaner cell layers and higher net cell production. The cells exhibited a highly stabilized profile for marker gene expression including a low cytokeratin 49 (epithelial marker) and a high collagen 1a1 (mesenchymal marker) expression. Added to the cell spindle-shaped morphology, motility behavior, and actin organization, this suggests that the cells bore stable mesenchymal characteristics. This contrast with the time-evolving expression pattern observed in the control fresh explants during the first 2 weeks of culture: a sharp decrease in cytokeratin 49 expression was concomitant with a gradual increase in col1a1. We surmise that such loss of epithelial features for the benefit of mesenchymal ones was triggered by an epithelial to mesenchymal transition (EMT) process or by way of a progressive population replacement process. Overall, our findings provide a comprehensive characterization of this new primary culture model bearing mesenchymal features and whose stability over culture time makes those cells good candidates for cell reprogramming prior to nuclear transfer, in a context of fish genome preservation.

  2. Explant culture: An advantageous method for isolation of mesenchymal stem cells from human tissues.

    Science.gov (United States)

    Hendijani, Fatemeh

    2017-04-01

    Mesenchymal stem cell (MSC) research progressively moves towards clinical phases. Accordingly, a wide range of different procedures were presented in the literature for MSC isolation from human tissues; however, there is not yet any close focus on the details to offer precise information for best method selection. Choosing a proper isolation method is a critical step in obtaining cells with optimal quality and yield in companion with clinical and economical considerations. In this concern, current review widely discusses advantages of omitting proteolysis step in isolation process and presence of tissue pieces in primary culture of MSCs, including removal of lytic stress on cells, reduction of in vivo to in vitro transition stress for migrated/isolated cells, reduction of price, processing time and labour, removal of viral contamination risk, and addition of supporting functions of extracellular matrix and released growth factors from tissue explant. In next sections, it provides an overall report of technical highlights and molecular events of explant culture method for isolation of MSCs from human tissues including adipose tissue, bone marrow, dental pulp, hair follicle, cornea, umbilical cord and placenta. Focusing on informative collection of molecular and methodological data about explant methods can make it easy for researchers to choose an optimal method for their experiments/clinical studies and also stimulate them to investigate and optimize more efficient procedures according to clinical and economical benefits. © 2017 John Wiley & Sons Ltd.

  3. Characterization of Myelomonocytoid Progenitor Cells with Mesenchymal Differentiation Potential Obtained by Outgrowth from Pancreas Explants

    Directory of Open Access Journals (Sweden)

    Marc-Estienne Roehrich

    2012-01-01

    Full Text Available Progenitor cells can be obtained by outgrowth from tissue explants during primary ex vivo tissue culture. We have isolated and characterized cells outgrown from neonatal mouse pancreatic explants. A relatively uniform population of cells showing a distinctive morphology emerged over time in culture. This population expressed monocyte/macrophage and hematopoietic markers (CD11b+ and CD45+, and some stromal-related markers (CD44+ and CD29+, but not mesenchymal stem cell (MSC-defining markers (CD90− and CD105− nor endothelial (CD31− or stem cell-associated markers (CD133− and stem cell antigen-1; Sca-1−. Cells could be maintained in culture as a plastic-adherent monolayer in culture medium (MesenCult MSC for more than 1 year. Cells spontaneously formed sphere clusters “pancreatospheres” which, however, were nonclonal. When cultured in appropriate media, cells differentiated into multiple mesenchymal lineages (fat, cartilage, and bone. Positive dithizone staining suggested that a subset of cells differentiated into insulin-producing cells. However, further studies are needed to characterize the endocrine potential of these cells. These findings indicate that a myelomonocytoid population from pancreatic explant outgrowths has mesenchymal differentiation potential. These results are in line with recent data onmonocyte-derivedmesenchymal progenitors (MOMPs.

  4. Tetanus–diphtheria–acellular pertussis vaccination for adults: an update

    Science.gov (United States)

    2017-01-01

    Although tetanus and diphtheria have become rare in developed countries, pertussis is still endemic in some developed countries. These are vaccine-preventable diseases and vaccination for adults is important to prevent the outbreak of disease. Strategies for tetanus, diphtheria, and pertussis vaccines vary from country to country. Each country needs to monitor consistently epidemiology of the diseases and changes vaccination policies accordingly. Recent studies showed that tetanus–diphtheria–acellular pertussis vaccine for adults is effective and safe to prevent pertussis disease in infants. However, vaccine coverage still remains low than expected and seroprevalence of protective antibodies levels for tetanus, diphtheria, and pertussis decline with aging. The importance of tetanus–diphtheria–acellular pertussis vaccine administration should be emphasized for the protection of young adult and elderly people also, not limited to children. PMID:28168170

  5. Engineering an improved acellular nerve graft via optimized chemical processing.

    Science.gov (United States)

    Hudson, Terry W; Liu, Stephen Y; Schmidt, Christine E

    2004-01-01

    The long-term goal of our research is to engineer an acellular nerve graft for clinical nerve repair and for use as a model system with which to study nerve-extracellular matrix interactions during nerve regeneration. To develop this model acellular nerve graft we (1) examined the effects of detergents on peripheral nerve tissue, and (2) used that knowledge to create a nerve graft devoid of cells with a well-preserved extracellular matrix. Using histochemistry and Western analysis, the impact of each detergent on cellular and extracellular tissue components was determined. An optimized protocol was created with the detergents Triton X-200, sulfobetaine-16, and sulfobetaine-10. This study represents the most comprehensive examination to date of the effects of detergents on peripheral nerve tissue morphology and protein composition. Also presented is an improved chemical decellularization protocol that preserves the internal structure of native nerve more than the predominant current protocol.

  6. Biomechanical properties of peripheral nerve after acellular treatment

    Institute of Scientific and Technical Information of China (English)

    MA Xin-long; SUN Xiao-lei; YANG Zhao; LI Xiu-lan; MA Jian-xiong; ZHANG Yang; YUAN Zhen-zhen

    2011-01-01

    Background Peripheral nerve injury causes a high rate of disability and a huge economic burden,and is currently one of the serious health problems in the world.The use of nerve grafts plays a vital role in repairing nerve defects.Acellular nerve grafts have been widely used in many experimental models as a peripheral nerve substitute.The purpose of this study was to test the biomechanical properties of acellular nerve grafts.Methods Thirty-four fresh sciatic nerves were obtained from 17 adult male Wistar rats (age of 3 months) and randomly assigned to 3 groups:normal control group,nerve segments underwent no treatment and were put in phosphate buffered saline (pH 7.4) and stored at 4℃ until further use; physical method group,nerve segments were frozen at -196℃ and then thawed at 37℃; and chemical method group,nerve segments were chemically extracted with the detergents Triton X-200,sulfobetaine-10 (SB-10) and sulfobetaine-16 (SB-16).After the acellularization process was completed,the structural changes of in the sciatic nerves in each group were observed by hematoxylin-eosin staining and field emission scanning electron microscopy,then biomechanical properties were tested using a mechanical apparatus (Endura TEC ELF 3200,Bose,Boston,USA).Results Hematoxylin-eosin staining and field emission scanning electron microscopy demonstrated that the effects of acellularization,demyelination,and integrity of nerve fiber tube of the chemical method were better than that of the physical method.Biomechanical testing showed that peripheral nerve grafts treated with the chemical method resulted in some decreased biomechanical properties (ultimate load,ultimate stress,ultimate strain,and mechanical work to fracture) compared with normal control nerves,but the differences were not statistically significant (P >0.05).Conclusion Nerve treated with the chemical method may be more appropriate for use in implantation than nerve treated with the physical method.

  7. Effectiveness of acellular pertussis vaccination during childhood (Spain).

    Science.gov (United States)

    Plans, P; Toledo, D; Sala, M R; Camps, N; Villanova, M; Rodríguez, R; Alvarez, J; Solano, R; García-Cenoz, M; Barrabeig, I; Godoy, P; Minguell, S

    2016-12-01

    Pertussis vaccination with 4-5 doses of acellular vaccines is recommended in Spain to all children at 2 months to 6 years of age. The effectiveness of the acellular pertussis vaccination was assessed in this study by comparing the incidence of secondary pertussis in vaccinated (4-5 doses) and unvaccinated or partially vaccinated (0-3 doses) household contacts 1-9 years old of confirmed cases of pertussis in Spain in 2012-13. Eighty-five percent of contacts had been vaccinated with 4-5 doses of acellular pertussis vaccines. During the 2-year study period, 64 cases of secondary pertussis were detected among 405 household contacts 1-9 years old: 47 among vaccinated and 17 among unvaccinated or partially vaccinated contacts. The effectiveness for preventing secondary pertussis, calculated as 1 minus the relative risk (RR) of secondary pertussis in vaccinated vs. unvaccinated/partially vaccinated contacts, was 50 % [95 % confidence interval (CI): 19-69 %, p Spain.

  8. Making post-mortem implantable cardioverter defibrillator explantation safe

    DEFF Research Database (Denmark)

    Räder, Sune B E W; Zeijlemaker, Volkert; Pehrson, Steen;

    2009-01-01

    AIMS: The aim of this study is to investigate whether protection with rubber or plastic gloves during post-mortem explantation of an implantable cardioverter defibrillator (ICD) offers enough protection for the explanting operator during a worst-case scenario (i.e. ICD shock). METHODS AND RESULTS...... that the resting voltage over the operating person would not exceed 50 V. CONCLUSION: The use of intact medical gloves made of latex, neoprene, or plastic eliminates the potential electrical risk during explantation of an ICD. Two gloves on each hand offer sufficient protection. We will recommend the use...

  9. Tissue Culture Responses from Different Explants of Rice

    Institute of Scientific and Technical Information of China (English)

    WANG Xiu-hong; SHI Xiang-yuan; WU Xian-jun

    2005-01-01

    Different culture explants, including anther, young panicle, young embryo, and mature embryo, from 19 rice varieties were used for callus induction and green plantlet differentiation. The culture efficiency differed significantly among the four types of explants, and varied from genotype to genotype. Callus induction frequency presented significantly positive correlation each between anther and young panicle, anther and mature embryo, and young panicle and young embryo. Green plantlet differentiation showed no relationship between different types of explants. In addition, no relationship was found between callus induction frequency and green plantlet differentiation frequency.

  10. Respiration rate in human pituitary tumor explants.

    Science.gov (United States)

    Anniko, M; Bagger-Sjöbäck, D; Hultborn, R

    1982-01-01

    Studies on the respiration rate of human pituitary tumor tissue have so far been lacking in the literature. This study presents the results from four adenomas causing acromegaly, all with different clinical degrees of the disease. Determination of oxygen uptake was performed in vitro with a spectrophotorespirometric system. Pieces of the tumors were explanted to an organ culture system with a high degree of stability. The secretion rate of growth hormone (GH) and prolactin (PRL) was determined. After 4-8 days in vitro, specimens were analyzed for respiration rate. This was approximately 1-1.5 microliters O2/h/micrograms dry weight. The activity of the pituitary tumor tissue was characterized by both the hormone secretion rate and the respiration rate. Particularly active foci were found to occur in the adenoma tissue. Depending on the individual tumor, the GH secretion rate was approximately 0.1-100 pmol/micrograms dry weight/h and PRL secretion rate approximately 0.4-18 micrograms/micrograms dry weight/h. The respiration rate--as is also the hormone secretion rate--is dependent on the time in vitro prior to analysis. The respiration rate in individual tumors is a parameter which does not reflect GH or PRL serum levels or clinical activity of the disease.

  11. An ovine tracheal explant culture model for allergic airway inflammation

    Directory of Open Access Journals (Sweden)

    Abeynaike Latasha

    2010-08-01

    Full Text Available Abstract Background The airway epithelium is thought to play an important role in the pathogenesis of asthmatic disease. However, much of our understanding of airway epithelial cell function in asthma has been derived from in vitro studies that may not accurately reflect the interactive cellular and molecular pathways active between different tissue constituents in vivo. Methods Using a sheep model of allergic asthma, tracheal explants from normal sheep and allergic sheep exposed to house dust mite (HDM allergen were established to investigate airway mucosal responses ex vivo. Explants were cultured for up to 48 h and tissues were stained to identify apoptotic cells, goblet cells, mast cells and eosinophils. The release of cytokines (IL-1α, IL-6 and TNF-α by cultured tracheal explants, was assessed by ELISA. Results The general morphology and epithelial structure of the tracheal explants was well maintained in culture although evidence of advanced apoptosis within the mucosal layer was noted after culture for 48 h. The number of alcian blue/PAS positive mucus-secreting cells within the epithelial layer was reduced in all cultured explants compared with pre-cultured (0 h explants, but the loss of staining was most evident in allergic tissues. Mast cell and eosinophil numbers were elevated in the allergic tracheal tissues compared to naïve controls, and in the allergic tissues there was a significant decline in mast cells after 24 h culture in the presence or absence of HDM allergen. IL-6 was released by allergic tracheal explants in culture but was undetected in cultured control explants. Conclusions Sheep tracheal explants maintain characteristics of the airway mucosa that may not be replicated when studying isolated cell populations in vitro. There were key differences identified in explants from allergic compared to control airways and in their responses in culture for 24 h. Importantly, this study establishes the potential for the

  12. Calcification preceding new bone formation induced by demineralized bone matrix gelatin.

    Science.gov (United States)

    Yamashita, K; Takagi, T

    1992-03-01

    Demineralized bone matrix gelatin (BMG) was implanted into the skeletal muscle of Sprague-Dawley (S.D.) rats, and histological changes were examined 3, 5, 7, 10 and 15 days later. Before bone formation, a specific calcification process was found in most of the BMG from day 5 and 7 after implantation. The heterotopic calcified sites were not always consistent with the sites of the alkaline phosphatase activity. It was considered that this calcification progresses without any cellular components, and we distinguished this type of calcification as "acellular mineral deposition" from the calcification which occurs in new bone formation. This "acellular mineral deposition" was first observed as small spherical calcified deposits in the BMG on day 7 after implantation; these deposits then gradually grew and fused with each other. Some multinucleated cells appeared near the site of calcification on day 7 after implantation, but osteoblasts or osteoblast-like cells were scarcely observed around the calcified deposits in BMG until day 7. Vascularization was often observed near the "acellular mineral deposition" and the new bone formation. Fourier transform infrared spectroscopy showed that the calcified deposits in BMG were composed of hydroxyapatite, carbonateapatite and other calcium phosphate components, and that the first two components became prominent with time. It is believed that the "acellular mineral deposition" is due to the deposition of calcium and phosphate into the BMG by a process of heterogenic nucleation that does not involve osteoblasts or matrix vesicles. Bone formation induced by the BMG occurred after the "acellular mineral deposition." The experimental calcification shown in this paper seems a useful model for the study of biocalcification.

  13. A Meta-analysis of Postoperative Complications of Tissue Expander/Implant Breast Reconstruction Using Acellular Dermal Matrix.

    Science.gov (United States)

    Zhao, Xiangyi; Wu, Xiaowei; Dong, Jie; Liu, Yingying; Zheng, Liang; Zhang, Liming

    2015-12-01

    Acellular dermal matrix (ADM) is commonly used for tissue expander/implant breast (TE/I-based) reconstruction. But the relation between ADM and postoperative complications remains controversial. A few meta-analyses were conducted in 2011-2012 and the result revealed that ADM can increase the risk of complications. The purpose of our study is to offer updated evidence for ADM clinical application by analyzing the effect of ADM on complications of TE/I-based breast reconstruction. The literature published from January 2010 to February 2015 was searched in EMbase, Medline, Science Direct, the Cochrane Central Register of Controlled Trials (CENTRAL), CBMdisc, CNKI, VIP, and the references of those included studies were also searched by hand. According to inclusive criteria, 11 studies were selected and the values were extracted from the included literature. Complications with four different categories assigned for overall complications, infection, hematoma/seroma, and explantation were collected. RevMan 5.1 was used for meta-analysis. The evidence level was assessed by using the GRADE system. Eleven published studies were included. The results showed that compared to the control group, the ADM group increased the rate of overall complications (OR = 1.33, 95% CI 1.03-1.70, p = 0.03), infection (OR = 1.47, 95% CI 1.04-2.06, p = 0.03), hematoma/seroma (OR = 1.66, 95% CI 1.13-2.44, p = 0.01), but there was no significant difference in explantation (OR = 1.37, 95% CI 0.89-2.11, p = 0.15). Based on the GRADE system, all the evidence was at level C and weak recommendation. In TE/I-based breast reconstruction, ADM increased the incidence of overall complications, infection, and hematoma/seroma; the incidence of explantation remains unknown. For the poor quality of the original studies, a prudent choice is suggested; and more high-quality, large-sample studies are needed. This journal requires that authors assign a level of evidence to each submission to which Evidence

  14. Intraocular lens explantation or exchange: indications, postoperative interventions, and outcomes

    Directory of Open Access Journals (Sweden)

    Refik Oltulu

    2015-06-01

    Full Text Available ABSTRACT Purpose: To analyze the indications for explantation or exchange of intraocular lenses (IOLs, which were originally implanted for the correction of aphakia during cataract extraction. Methods: All cases that involved intraocular lens explantation or exchange in one institution between January 2008 and December 2014 were analyzed retrospectively. Results: In total, 93 eyes of 93 patients were analyzed. The median time interval between implantation and explantation of the anterior chamber intraocular lenses (AC IOL and posterior chamber intraocular lenses (PC IOL was 83.40 ± 83.14 months (range: 1-276 months and 55.14 ± 39.25 months (range: 1-168 months, respectively. Pseudophakic bullous keratopathy (17 eyes, 38.6% and persistent iritis (12 eyes, 27.8% in the AC IOL group and dislocation or decentration (30 eyes, 61.2% and incorrect IOL power (nine eyes, 18.4% in the PC IOL group were the most common indications for explantation of IOLs. The mean logMAR best corrected visual acuity (BCVA improved significantly from 1.30 preoperatively to 0.62 postoperatively in the PC IOL group (p<0.001 but did not improve significantly in the AC IOL group (p=0.186. Conclusions: The primary indication for IOL explantation or exchange was pseudophakic bullous keratopathy in the AC IOL group and was dislocation or decentration in the PC IOL group. PC IOL explantation or exchange is safe and improves visual acuity.

  15. Chemically extracted acellular allogeneic nerve graft combined with ciliary neurotrophic factor promotes sciatic nerve repair

    Institute of Scientific and Technical Information of China (English)

    Yanru Zhang; Hui Zhang; Kaka Katiella; Wenhua Huang

    2014-01-01

    A chemically extracted acellular allogeneic nerve graft can reduce postoperative immune re-jection, similar to an autologous nerve graft, and can guide neural regeneration. However, it remains poorly understood whether a chemically extracted acellular allogeneic nerve graft combined with neurotrophic factors provides a good local environment for neural regenera-tion. This study investigated the repair of injured rat sciatic nerve using a chemically extracted acellular allogeneic nerve graft combined with ciliary neurotrophic factor. An autologous nerve anastomosis group and a chemical acellular allogeneic nerve bridging group were prepared as controls. At 8 weeks after repair, sciatic functional index, evoked potential amplitude of the soleus muscle, triceps wet weight recovery rate, total number of myelinated nerve fibers and myelin sheath thickness were measured. For these indices, values in the three groups showed the autologous nerve anastomosis group > chemically extracted acellular nerve graft + ciliary neurotrophic factor group > chemical acellular allogeneic nerve bridging group. These results suggest that chemically extracted acellular nerve grafts combined with ciliary neurotrophic factor can repair sciatic nerve defects, and that this repair is inferior to autologous nerve anasto-mosis, but superior to chemically extracted acellular allogeneic nerve bridging alone.

  16. Whooping cough, twenty years from acellular vaccines introduction.

    Science.gov (United States)

    Greco, D; Esposito, S; Tozzi, A; Pandolfi, E; Icardi, G; Giammanco, A

    2015-01-01

    Clinical pertussis resulting from infection with B. pertussis is a significant medical and public health problem, despite the huge success of vaccination that has greatly reduced its incidence. The whole cell vaccine had an undeniable success over the last 50 years, but its acceptance was strongly inhibited by fear, only partially justified, of severe side effects, but also, in the Western world, by the difficulty to enter in combination with other vaccines: today multi-vaccine formulations are essential to maintain a high vaccination coverage. The advent of acellular vaccines was greeted with enthusiasm by the public health world: in the Nineties, several controlled vaccine trials were carried out: they demonstrated a high safety and good efficacy of new vaccines. In fact, in the Western world, the acellular vaccines completely replaced the whole cells ones. In the last years, ample evidence on the variety of protection of these vaccines linked to the presence of different antigens of Bordetella pertussis was collected. It also became clear that the protection provided, on average around 80%, leaves every year a significant cohort of vaccinated susceptible even in countries with a vaccination coverage of 95%, such as Italy. Finally, it was shown that, as for the pertussis disease, protection decreases over time, to leave a proportion of adolescents and adults unprotected. Waiting for improved pertussis vaccines, the disease control today requires a different strategy that includes a booster at 5 years for infants, but also boosters for teenagers and young adults, re-vaccination of health care personnel, and possibly of pregnant women and of those who are in contact with infants (cocooning). Finally, the quest for better vaccines inevitably tends towards pertussis acellular vaccines with at least three components, which have demonstrated superior effectiveness and have been largely in use in Italy for fifteen years.

  17. Porosity of porcine bladder acellular matrix: impact of ACM thickness.

    Science.gov (United States)

    Farhat, Walid; Chen, Jun; Erdeljan, Petar; Shemtov, Oren; Courtman, David; Khoury, Antoine; Yeger, Herman

    2003-12-01

    The objectives of this study are to examine the porosity of bladder acellular matrix (ACM) using deionized (DI) water as the model fluid and dextran as the indicator macromolecule, and to correlate the porosity to the ACM thickness. Porcine urinary bladders from pigs weighing 20-50 kg were sequentially extracted in detergent containing solutions, and to modify the ACM thickness, stretched bladders were acellularized in the same manner. Luminal and abluminal ACM specimens were subjected to fixed static DI water pressure (10 cm); and water passing through the specimens was collected at specific time interval. While for the macromolecule porosity testing, the diffusion rate and direction of 10,000 MW fluoroescein-labeled dextrans across the ACM specimens mounted in Ussing's chambers were measured. Both experiments were repeated on the thin stretched ACM. In both ACM types, the fluid porosity in both directions did not decrease with increased test duration (3 h); in addition, the abluminal surface was more porous to fluid than the luminal surface. On the other hand, when comparing thin to thick ACM, the porosity in either direction was higher in the thick ACM. Macromolecule porosity, as measured by absorbance, was higher for the abluminal thick ACM than the luminal side, but this characteristic was reversed in the thin ACM. Comparing thin to thick ACM, the luminal side in the thin ACM was more porous to dextran than in the thick ACM, but this characteristic was reversed for the abluminal side. The porcine bladder ACM possesses directional porosity and acellularizing stretched urinary bladders may increase structural density and alter fluid and macromolecule porosity.

  18. Risk of Febrile Seizures and Epilepsy After Vaccination With Diphtheria, Tetanus, Acellular Pertussis, Inactivated Poliovirus, and Haemophilus Influenzae Type b

    DEFF Research Database (Denmark)

    Sun, Yuelian; Christensen, Jakob Christensen; Hviid, Anders

    2012-01-01

    Context Vaccination with whole-cell pertussis vaccine carries an increased risk of febrile seizures, but whether this risk applies to the acellular pertussis vaccine is not known. In Denmark, acellular pertussis vaccine has been included in the combined diphtheria-tetanus toxoids-acellular pertus......Context Vaccination with whole-cell pertussis vaccine carries an increased risk of febrile seizures, but whether this risk applies to the acellular pertussis vaccine is not known. In Denmark, acellular pertussis vaccine has been included in the combined diphtheria-tetanus toxoids...

  19. Human acellular dermal wound matrix: evidence and experience.

    Science.gov (United States)

    Kirsner, Robert S; Bohn, Greg; Driver, Vickie R; Mills, Joseph L; Nanney, Lillian B; Williams, Marie L; Wu, Stephanie C

    2015-12-01

    A chronic wound fails to complete an orderly and timely reparative process and places patients at increased risk for wound complications that negatively impact quality of life and require greater health care expenditure. The role of extracellular matrix (ECM) is critical in normal and chronic wound repair. Not only is ECM the largest component of the dermal skin layer, but also ECM proteins provide structure and cell signalling that are necessary for successful tissue repair. Chronic wounds are characterised by their inflammatory and proteolytic environment, which degrades the ECM. Human acellular dermal matrices, which provide an ECM scaffold, therefore, are being used to treat chronic wounds. The ideal human acellular dermal wound matrix (HADWM) would support regenerative healing, providing a structure that could be repopulated by the body's cells. Experienced wound care investigators and clinicians discussed the function of ECM, the evidence related to a specific HADWM (Graftjacket(®) regenerative tissue matrix, Wright Medical Technology, Inc., licensed by KCI USA, Inc., San Antonio, TX), and their clinical experience with this scaffold. This article distills these discussions into an evidence-based and practical overview for treating chronic lower extremity wounds with this HADWM. © 2013 The Authors. International Wound Journal © 2013 Medicalhelplines.com Inc and John Wiley & Sons Ltd.

  20. A novel method for coral explant culture and micropropagation.

    Science.gov (United States)

    Vizel, Maya; Loya, Yossi; Downs, Craig A; Kramarsky-Winter, Esti

    2011-06-01

    We describe here a method for the micropropagation of coral that creates progeny from tissue explants derived from a single polyp or colonial corals. Coral tissue explants of various sizes (0.5-2.5 mm in diameter) were manually microdissected from the solitary coral Fungia granulosa. Explants could be maintained in an undeveloped state or induced to develop into polyps by manipulating environmental parameters such as light and temperature regimes, as well as substrate type. Fully developed polyps were able to be maintained for a long-term in a closed sea water system. Further, we demonstrate that mature explants are also amenable to this technique with the micropropagation of second-generation explants and their development into mature polyps. We thereby experimentally have established coral clonal lines that maintain their ability to differentiate without the need for chemical induction or genetic manipulation. The versatility of this method is also demonstrated through its application to two other coral species, the colonial corals Oculina patigonica and Favia favus.

  1. Nanofibers coated on acellular tissue-engineered bovine pericardium supports differentiation of mesenchymal stem cells into endothelial cells for tissue engineering.

    Science.gov (United States)

    Mathapati, Santosh; Bishi, Dillip Kumar; Venugopal, Jayarama Reddy; Cherian, Kotturathu Mammen; Guhathakurta, Soma; Ramakrishna, Seeram; Verma, Rama Shanker

    2014-04-01

    This study aimed to develop biodegradable, polymer-based nanofibers coated on acellular tissue-engineered bovine pericardium (ATEBP) for cell interfaces, enabling more exquisite functionality, such as mesenchymal stem cell (MSC) adhesion, proliferation and differentiation into endothelial cells for tissue engineering. ATEBP coated with nanofibers of poly(L-lactic acid)-co-poly(ε-caprolactone) (PLACL) and a blend of PLACL and gelatin were analyzed for human bone marrow-derived MSC adhesion, proliferation and differentiation into endothelial cells. The cell culture-based approach showed an increase in human bone marrow-derived MSC adhesion, proliferation and differentiation into endothelial cells on ATEBP coated with PLACL/gelatin nanofibers compared with ATEBP and PLACL nanofibers coated on ATEBP. ATEBP coated with PLACL/gelatin nanofibrous scaffolds, along with human bone marrow-derived MSCs differentiated into endothelial cells, might improve the scaffolds' functionality for tissue engineering.

  2. Citrus tissue culture employing vegetative explants.

    Science.gov (United States)

    Chaturvedi, H C; Singh, S K; Sharma, A K; Agnihotri, S

    2001-11-01

    Citrus being a number one fruit of the world due to its high nutritional value, huge production of fruits and fruit products, the citrus industry may be considered a major fruit industry. Though citrus orchard area in India is comparable to USA, the produce is far less, while its export is nil. Biotechnology has played an outstanding role in boosting the citrus industry, e.g., in Spain, which is now the biggest exporter of citrus fruit with the application of micrografting. Amongst the fruit trees, perhaps the maximum tissue culture research has been done in citrus during the past four decades, however, the results of practical value are meagre. The shortfalls in citrus tissue culture research and some advancements made in this direction along with bright prospects are highlighted, restricting the review to vegetative explants only. Whilst utilization of nucellar embryogenesis is limited to rootstocks, the other aspects, like, regeneration and proliferation of shoot meristems measuring 200 microm in length--a global breakthrough--of two commercially important scion species, Citrus aurantifolia and C. sinensis and an important rootstock, C. limonia, improvement of micrografting technique, cloning of the same two scion species as well as some Indian rootstock species, employing nodal stem segments of mature trees, of immense practical value have been elaborated. A rare phenomenon of shift in the morphogenetic pattern of differentiation from shoot bud differentiation to embryoid formation occurred during the long-term culture of stem callus of C. grandis. Stem callus-regenerated plants of C. aurantifolia, C. sinensis and C. grandis showed variation in their ploidy levels and a somaclonal variant of C. sinensis, which produced seedless fruits was isolated. Tailoring of rooting in microshoots to a tap root-like system by changing the inorganic salt composition of the rooting medium, resulting in 100% transplant success, and germplasm preservation through normal growth

  3. REACTOGENICITY OF ACELLULAR PERTUSSIS VACCINE AND THE POSSIBILITY OF ITS USE IN ELDER CHILDREN

    Directory of Open Access Journals (Sweden)

    M.G. Galitskaya

    2008-01-01

    Full Text Available As is know, in the past few years, the incidence of pertussis has increased again. The article reveals the reasons of this phenomenon and the possible solutions for this problem. Besides, comparative analysis of the whole cell vaccine used in this country as within the framework of the national immunizations schedule and modern acellular vaccines is made. Results of multicenter research, convincingly proving the safety and efficiency of acellular pertussis vaccine, are presented.Key words: pertussis, prophylaxis, whole cell vaccine, acellular vaccine, efficiency, children.

  4. Correlated response of in vitro regeneration capacity from different source of explants inCucumis melo.

    Science.gov (United States)

    Molina, R V; Nuez, F

    1995-01-01

    The variation among and within different populations of the regeneration ability from leaf, cotyledon and hypocotyl explants has been studied. A control population and two lines selected by their regeneration capacity from leaf explants were used. Significant differences among the plants of the control population,for the organogenic response, were detected. The regeneration capacity varies depending on the type of explant. Selection in order to improve the regeneration frequency from leaf explants also raises the organogenic response in the other explant types. This result suggests the presence of a partial common genetic system controlling the regeneration frequency of the diverse types of explants.

  5. Schwann-like cells seeded in acellular nerve grafts improve nerve regeneration.

    Science.gov (United States)

    Fan, Lihong; Yu, Zefeng; Li, Jia; Dang, Xiaoqian; Wang, Kunzheng

    2014-05-21

    This study evaluated whether Schwann-like cells (SLCs) induced from bone marrow-derived mesenchymal stem cells (BM-MSCs) transplanted into acellular nerve grafts (ANGs) could repair nerve defects compared with nerve isografts and ANGs with BM-MSCs. BM-MSCs extracted, separated and purified from the bone marrow of rats, and some of the BM-MSCs were cultured with mixed induction agents that could induce BM-MSCs into SLCs. Either SLCs or BM-MSCs were seeded onto 10-mm ANGs, and the isografts were chosen as the control. The walking-track test, tibialis anterior muscle weight measurement, electrophysiological examination, toluidine blue staining, transmission electron micrographs and immunostaining of S-100 and VEGF in these three groups were evaluated in a 10-mm rat sciatic injury-repair model. The walking-track test, tibialis anterior muscle weight measurement and electrophysiological examination of the sciatic nerve suggested the groups of ANGs with SLCs and isografts obtained better results than the BM-MSC group (P0.05). All the histomorphometric analyses (toluidine blue staining, transmission electron micrographs and immunostaining of S-100 and VEGF) showed that there were more regenerating nerve fibers in the group of ANGs with SLCs than the BM-MSCs (P0.05). SLCs seeded in ANGs and isografts show better functional regeneration compared with BM-MSCs seeded in ANGs. Additionally, SLCs combined with ANGs present almost the same outcome as the isografts. Therefore, SLCs with ANGs can be a good choice in nerve defect repairs.

  6. Engineering 3D bio-artificial heart muscle: the acellular ventricular extracellular matrix model.

    Science.gov (United States)

    Patel, Nikita M; Tao, Ze-Wei; Mohamed, Mohamed A; Hogan, Matt K; Gutierrez, Laura; Birla, Ravi K

    2015-01-01

    Current therapies in left ventricular systolic dysfunction and end-stage heart failure include mechanical assist devices or transplant. The development of a tissue-engineered integrative platform would present a therapeutic option that overcomes the limitations associated with current treatment modalities. This study provides a foundation for the fabrication and preliminary viability of the acellular ventricular extracellular matrix (AVEM) model. Acellular ventricular extracellular matrix was fabricated by culturing 4 million rat neonatal cardiac cells around an excised acellular ventricular segment. Acellular ventricular extracellular matrix generated a maximum spontaneous contractile force of 388.3 μN and demonstrated a Frank-Starling relationship at varying pretensions. Histologic assessment displayed cell cohesion and adhesion within the AVEM as a result of passive cell seeding.

  7. Risk of Brain Damage Following Pertussis Immunization with Whole-Cell cf Acellular Vaccines

    Directory of Open Access Journals (Sweden)

    J Gordon Millichap

    2004-06-01

    Full Text Available Serious neurological disorders reported following whole-cell (WC in comparison to acellular (AC pertussis vaccines (PV were evaluated by the Genetic Centers of America, Silver Spring, MD.

  8. IN VITRO REGENERATION CASTOR (RICINUS COMMUNIS L.) USING COTYLEDON EXPLANTS

    Science.gov (United States)

    A novel plant regeneration protocol was established for castor (Ricinus communis L.), an important oilseed crop. Mature seed-derived cotyledon explants produced adventitious shoots when placed on Murashige and Skoog (MS) medium containing thidiazuron (TDZ). The rate of shoot regeneration was maximal...

  9. Simple, effective and economical explant-surface sterilization ...

    African Journals Online (AJOL)

    STORAGESEVER

    2009-10-19

    Oct 19, 2009 ... ISSN 1684–5315 © 2009 Academic Journals. Full Length Research ... highest reduction in bacterial and fungal contamination (0%) at time intervals between 20 - 45 min. The ... Key words: Explant, surface sterilization, cowpea, rice, sorghum, JIK® ... majority of commercial and scientific plant tissue culture.

  10. Bovine versus Porcine Acellular Dermal Matrix: A Comparison of Mechanical Properties

    Directory of Open Access Journals (Sweden)

    David M. Adelman, MD, PhD, FACS

    2014-05-01

    Conclusions: Before implantation, BADM is inherently stronger than PADM at equivalent thicknesses and considerably stronger at increased thicknesses. These results corroborate clinical data from a previous study in which PADM was associated with a higher intraoperative device failure rate. Although numerous properties of acellular dermal matrix contribute to clinical outcomes, surgeons should consider initial mechanical strength properties when choosing acellular dermal matrices for load-bearing applications such as hernia repair.

  11. The Removal of Hydrogel Explants: An Analysis of 467 Consecutive Cases

    NARCIS (Netherlands)

    Crama, N.; Klevering, B.J.

    2016-01-01

    PURPOSE: To describe the complications associated with hydrogel explants and to describe the indications, surgical technique, and risks involved in the removal of a hydrogel explant. DESIGN: Single-center, retrospective interventional case series. PARTICIPANTS: Patients who underwent surgical remova

  12. Interaction of cochlear nucleus explants with semiconductor materials.

    Science.gov (United States)

    Mlynski, Robert; Volkenstein, Stefan; Hansen, Stefan; Brors, Dominik; Ebmeyer, Joerg; Dazert, Stefan

    2007-07-01

    Implantable hearing devices such as cochlear implants and auditory brainstem implants deliver auditory information through electrical stimulation of auditory neurons. The combination of microelectronic electrodes with auditory nerve cells may lead to further improvement of the hearing quality with these devices. Whereas several kinds of neurons are known to grow on semiconductor substrates, interactions of cochlear nucleus (CN) neurons with such materials have yet to be described. To investigate survival and growth behavior of CN neurons on different semiconductor materials. CN explants from postnatal day 10 Sprague-Dawley rats were cultured for 96 hours in Neurobasal medium on polished and unpolished silicon wafers (p-type Si [100] and p-type Si3N4[100]) as well as plastic surface. These surfaces had been coated with poly-L-lysine and laminin. Neuronal outgrowth was examined using image analysis software after immunohistologic staining for neurofilament. Neurite length and directional changes were quantified. Additionally, neurite morphology and adhesion to the semiconductor material was evaluated by scanning electron microscopy. Although proper adhesion of CN explants was seen, no neurite growth could be detected on unpolished silicon wafers (Si and Si3N4). Compared with the other test conditions, polished, laminin-coated Si3N4 wafers showed best biocompatibility regarding neurite length and number per explant. CN explants developed a mean of eight neurons with an average length of 236 mum in 96 hours of culture on these wafers. The results of this study demonstrate the general possibility of CN neuron growth in culture on semiconductors in vitro. The differences in neuron length and number per explant indicate that the growth of CN neurons is influenced by the semiconductor substrate as well as extracellular matrix proteins, with laminin-coated p-type Si3N4[100] being a preferable material for future hybrid experiments on auditory-neuron semiconductor chips.

  13. Absorção de macronutrientes por explantes de bananeira in vitro Macronutrient absorption by banana explants in vitro

    Directory of Open Access Journals (Sweden)

    Josefa Diva Nogueira Diniz

    1999-07-01

    Full Text Available Com o objetivo de estudar a absorção de macronutrientes (N, P, K, Ca, Mg e S em explantes de bananeira cv. Prata Anã, foram utilizados explantes de plantas estabelecidas in vitro, inoculados em meio básico de Murashige & Skoog (1962 contendo sacarose (30 g/L, e BAP (3,5 mg/L com sete tratamentos, representados pelos períodos de 0, 10, 20, 30, 40, 50 e 60 dias de cultivo e três repetições. As quantidades de macronutrientes totais absorvidas pelos explantes seguiram a ordem: K > N > Ca > ou = P > Mg @ S. O P foi o nutriente absorvido mais rapidamente pelos explantes, com 75% extraído do meio de cultivo nos primeiros 30 dias, cessando sua absorção aos 50 dias, restando ainda 9% no meio de cultivo. A absorção do S cessou também aos 50 dias, quando 66% deste nutriente ainda permanecia no meio de cultivo. Este resultado sugere haver uma relação, quanto à absorção, entre esses dois nutrientes. As maiores taxas de absorção de todos os nutrientes foram verificadas nos primeiros 20 dias. O rizoma, o pseudocaule e as folhas, se diferenciaram quanto à concentração e extração ou acúmulo de nutrientes.The absorption of the nutrients (N, P, K, Ca, Mg and S by banana (Musa sp. cv. Prata Anã explants on the basic medium of Murashige & Skoog (1962 supplemented with sucrose (30 g/L and BAP (3.5 mg/L were evaluated at 0, 10, 20, 30, 40, 50 and 60 days after inoculation. The seven treatments were arranged on a completely randomized design with three replicates. The sequence of nutrient absorption by the explants was K > N > Ca > or = P > Mg @ S. The P was the nutrient with the fastest absorption rate and at the 30th day the explants had already absorbed 75% of the P from the medium. The P absorption stopped by the 50th day. The S absorption stopped at the 50th day with 66% of it remaining in the medium. The results suggested a close relationship between these two nutrients. The highest rates of nutrient absorption were observed during the

  14. Morphologic differences observed by scanning electron microscopy according to the reason for pseudophakic IOL explantation

    DEFF Research Database (Denmark)

    Fernandez-Buenaga, Roberto; Alio, Jorge L.; Ramirez, Jose M.

    2015-01-01

    Purpose To compare variations in surface morphology, as studied by scanning electron microscopy (SEM), of explanted intraocular lenses (IOLs) concerning the cause leading to the explantation surgery. Methods In this prospective multicenter study, explanted IOLs were analyzed by SEM and energy-dis...

  15. Quality of placental RNA: Effects of explant size and culture duration.

    Science.gov (United States)

    Brew, O; Nikolopoulou, E; Hughes, A; Christian, M; Lee, Y; Oduwole, O; Sullivan, M H F; Woodman, A

    2016-10-01

    We evaluated the impact of placental micro (≤50 mg) and macro (∼200 mg) explants, oxygen concentration and culture method on placental RNA quality after long-term culture. Our findings show that micro explants cultured at 8% oxygen have the best RNA quality and tissue structure. Macro explants were less viable after long-term culture. Macro explants and explants undergoing syncytial degeneration produced poor quality RNA and should be avoided. Copyright © 2016 Elsevier Ltd. All rights reserved.

  16. Chondrogenesis of human infrapatellar fat pad stem cells on acellular dermal matrix

    Directory of Open Access Journals (Sweden)

    Ken eYe

    2016-01-01

    Full Text Available Acellular dermal matrix (ADM has been in clinical use for decades in numerous surgical applications. The ability for ADM to promote cellular repopulation and revascularisation, and tissue regeneration is well documented. Adipose stem cells have the ability to differentiate into mesenchymal tissue types, including bone and cartilage. The aim of this study was to investigate the potential interaction between ADM and adipose stem cells in vitro using TGFβ3 and BMP6.Human infrapatellar fat pad derived adipose stem cells (IPFP-ASC were cultured with ADM derived from rat dermis under chondrogenic (TGFβ3 and BMP6 in vitro for 2 and 4 weeks. Histology, qPCR and immunohistochemistry were performed to assess for markers of chondrogenesis (collagen Type II, SOX9 and proteoglycans. At 4 weeks, cell-scaffold constructs displayed cellular changes consistent with chondrogenesis, with evidence of stratification of cell layers and development of a hyaline-like cartilage layer superficially which stained positively for collagen Type II and proteoglycans. Significant cell-matrix interaction was seen between the cartilage layer and the ADM itself with seamless integration between each layer. Real time qPCR showed significantly increases of COL2A1, SOX9, and ACAN gene expression over 4 weeks when compared to control. COL1A2 gene expression remained unchanged over 4 weeks.We believe the principles which make ADM versatile and successful for tissue regeneration are application to cartilage regeneration. This study demonstrates in vitro the ability for IPFP-ASCs to undergo chondrogenesis, infiltrate and interact with ADM. These outcomes serve as a platform for in vivo modelling of ADM for cartilage repair.

  17. Shoot Regeneration from Leaf Explants of Withania somnifera (L. Dunal

    Directory of Open Access Journals (Sweden)

    Aruna Girish JOSHI

    2010-03-01

    Full Text Available Regeneration from leaf explants of Withania somnifera (L. for mass propagation was studied on Murashige and Skoog�s medium supplemented with Kinetin (Kn and 6-benzylaminopurine (BAP alone or in combination. Shoot buds were induced from the midrib on the abaxial side in presence of Kn and BAP (4 �M. These shoot buds developed into shoots on the same medium. Rooting of these shoots was achieved in 0.5 �M of IBA.

  18. Foldable Lens Explantation and Exchange:The Reason and Solution

    Institute of Scientific and Technical Information of China (English)

    Danying Zheng; Zhenpin Zhang; Wenhui Yang; Weirong Chen

    2001-01-01

    Objective: To report the explantation and exchange of Hydrophilic Acrylic foldable intraocular lens (IOL) on 14 patients who had visual disturbances caused by the change of transparence on optic. Methods: Sixteen Hydrophilic Acrylic foldable intraocular lenses from 14 patients who presented with decreased visual acuity from 6 months to 1 year after normal phacoemulsification and IOL implantation associated with extensive transparent change on optic of the lens. The lenses were explanted with the bisection technique. All the eyes were reinserted with Acrysof foldable lenses. Results: Sixteen lenses were removed successfully and exchanged with the new lens in the capsule. The posterior capsular rupture and vitreous loss were found in the first two cases. One of them had the zonulysis due to the radial tear of the anterior capsule during the enlargement of the capsular opening. The anterior vitrectomy was performed with IOL fixed on the ciliary sulcus. The visual acuity of all the patients improved obviously without posterior complication. Conclusion: Foldable lens explantation with the bisection technique and exchange had a successful outcome with improvement of ocular condition. Eye science 2001; 17:54 ~56.

  19. Progress in various crosslinking modification for acellular matrix

    Institute of Scientific and Technical Information of China (English)

    Yang Haitang; Tan Qiang; Zhao Heng

    2014-01-01

    Objective To review the current crosslinking strategies for acelluar matrix scaffold,laying the foundation for subsequent experiment.Data sources Data were mainly obtained from recent papers published in PubMed or indexed by Web of Science,with keyword like crosslinking.Results Various crosslinking strategies,including chemical,physical and biological methods,have been introduced to facilitate the performance of fresh acellular matrix.Chemical crosslinking reagents,involved in synthetic and naturally derived agents,need to be eliminated before implantation in case of their potential biotoxicity,although several crosslinking agents with less toxicity and specific characteristics have been developed.Physical crosslinking methods present to be safe,additive-free and relatively controllable for rapid surface functionalization with no consideration of remaining radioactivity.Biological crosslinking strategies have attracted great interest,and have been demonstrated to enhance collagen-based crosslinking since their preparations do not need toxic or potentially biologically contaminated substances and can be carried out under physiological conditions.Conclusions Kinds of crosslinking methods with its potential advantages have been developed to modify raw acelluar matrix,of which the performance are promising after being crosslinked by several crosslinking treatments.Further preclinical and clinical evaluations should be taken to vertify their safety and efficacy for the tissues and organs substitutes in tissue and regenerative medicine.

  20. Porcine bladder acellular matrix (ACM): protein expression, mechanical properties.

    Science.gov (United States)

    Farhat, Walid A; Chen, Jun; Haig, Jennifer; Antoon, Roula; Litman, Jessica; Sherman, Christopher; Derwin, Kathleen; Yeger, Herman

    2008-06-01

    Experimentally, porcine bladder acellular matrix (ACM) that mimics extracellular matrix has excellent potential as a bladder substitute. Herein we investigated the spatial localization and expression of different key cellular and extracellular proteins in the ACM; furthermore, we evaluated the inherent mechanical properties of the resultant ACM prior to implantation. Using a proprietary decellularization method, the DNA contents in both ACM and normal bladder were measured; in addition we used immunohistochemistry and western blots to quantify and localize the different cellular and extracellular components, and finally the mechanical testing was performed using a uniaxial mechanical testing machine. The mean DNA content in the ACM was significantly lower in the ACM compared to the bladder. Furthermore, the immunohistochemical and western blot analyses showed that collagen I and IV were preserved in the ACM, but possibly denatured collagen III in the ACM. Furthermore, elastin, laminin and fibronectin were mildly reduced in the ACM. Although the ACM did not exhibit nucleated cells, residual cellular components (actin, myosin, vimentin and others) were still present. There was, on the other hand, no significant difference in the mean stiffness between the ACM and the bladder. Although our decellularization method is effective in removing nuclear material from the bladder while maintaining its inherent mechanical properties, further work is mandatory to determine whether these residual DNA and cellular remnants would lead to any immune reaction, or if the mechanical properties of the ACM are preserved upon implantation and cellularization.

  1. Acellular matrix of bovine pericardium bound with L-arginine

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Hyo Joo [Department of Molecular Science and Technology, Ajou University, Suwon 443-749 (Korea, Republic of); Bae, Jin Woo [Department of Molecular Science and Technology, Ajou University, Suwon 443-749 (Korea, Republic of); Kim, Chun Ho [Laboratory of Tissue Engineering, Korea Cancer Center Hospital, Seoul 139-240 (Korea, Republic of); Lee, Jin Woo [Department of Orthopaedic Surgery, College of Medicine, Yonsei University, Seoul 120-749 (Korea, Republic of); Shin, Jung Woog [Department of Biomedical Engineering, Inje University, Gimhae 621-749 (Korea, Republic of); Park, Ki Dong [Department of Molecular Science and Technology, Ajou University, Suwon 443-749 (Korea, Republic of)

    2007-09-15

    Surface immobilization of bioactive molecules onto natural tissues has been interestingly studied for the development of new functional matrices for the replacement of lost or malfunctioning tissues. In this study, an acellular matrix of bovine pericardium (ABP) was chemically modified by the direct coupling of L-arginine after glutaraldehyde (GA) cross-linking. The effects of L-arginine coupling on durability and calcification were investigated and the biocompatibility was evaluated in vitro and in vivo. A four-step detergent and enzymatic extraction process has been utilized to remove cellular components from fresh bovine pericardium (BP). Microscopic observation confirmed that nearly all cellular constituents are removed. Thermal and mechanical properties showed that the durability of L-arginine-treated matrices increased as compared with control ABP and GA-treated ABP. Resistance to collagenase digestion revealed that modified matrices have greater resistance to enzyme digestion than control ABP and GA-treated ABP. The in vivo calcification study demonstrated much less calcium deposition on L-arginine-treated ABP than GA-treated one. In vitro cell viability results showed that ABP modified with L-arginine leads to a significant increase in attachment of human dermal fibroblasts. The obtained results attest to the usefulness of L-arginine-treated ABP matrices for cardiovascular bioprostheses.

  2. Acellular dermal matrix based nipple reconstruction: A modified technique

    Directory of Open Access Journals (Sweden)

    Raghavan Vidya

    2017-09-01

    Full Text Available Nipple areolar reconstruction (NAR has evolved with the advancement in breast reconstruction and can improve self-esteem and, consequently, patient satisfaction. Although a variety of reconstruction techniques have been described in the literature varying from nipple sharing, local flaps to alloplastic and allograft augmentation, over time, loss of nipple projection remains a major problem. Acellular dermal matrices (ADM have revolutionised breast reconstruction more recently. We discuss the use of ADM to act as a base plate and strut to give support to the base and offer nipple bulk and projection in a primary procedure of NAR with a local clover shaped dermal flap in 5 breasts (4 patients. We used 5-point Likert scales (1 = highly unsatisfied, 5 = highly satisfied to assess patient satisfaction. Median age was 46 years (range: 38–55 years. Nipple projection of 8 mm, 7 mm, and 7 mms were achieved in the unilateral cases and 6 mm in the bilateral case over a median 18 month period. All patients reported at least a 4 on the Likert scale. We had no post-operative complications. It seems that nipple areolar reconstruction [NAR] using ADM can achieve nipple projection which is considered aesthetically pleasing for patients.

  3. Porcine bladder acellular matrix (ACM): protein expression, mechanical properties

    Energy Technology Data Exchange (ETDEWEB)

    Farhat, Walid A [Department of Surgery, Division of Urology, University of Toronto and Hospital for Sick Children, Toronto, ON M5G 1X8 (Canada); Chen Jun; Haig, Jennifer; Antoon, Roula; Litman, Jessica; Yeger, Herman [Department of Developmental and Stem Cell Biology, Research Institute, Hospital for Sick Children, Toronto, ON M5G 1X8 (Canada); Sherman, Christopher [Department of Anatomic Pathology, Sunnybrook and Women' s College Health Sciences Centre, Toronto, ON (Canada); Derwin, Kathleen [Department of Biomedical Engineering, Lerner Research Institute and Orthopaedic Research Center, Cleveland Clinic Foundation, Cleveland, OH (United States)], E-mail: walid.farhat@sickkids.ca

    2008-06-01

    Experimentally, porcine bladder acellular matrix (ACM) that mimics extracellular matrix has excellent potential as a bladder substitute. Herein we investigated the spatial localization and expression of different key cellular and extracellular proteins in the ACM; furthermore, we evaluated the inherent mechanical properties of the resultant ACM prior to implantation. Using a proprietary decellularization method, the DNA contents in both ACM and normal bladder were measured; in addition we used immunohistochemistry and western blots to quantify and localize the different cellular and extracellular components, and finally the mechanical testing was performed using a uniaxial mechanical testing machine. The mean DNA content in the ACM was significantly lower in the ACM compared to the bladder. Furthermore, the immunohistochemical and western blot analyses showed that collagen I and IV were preserved in the ACM, but possibly denatured collagen III in the ACM. Furthermore, elastin, laminin and fibronectin were mildly reduced in the ACM. Although the ACM did not exhibit nucleated cells, residual cellular components (actin, myosin, vimentin and others) were still present. There was, on the other hand, no significant difference in the mean stiffness between the ACM and the bladder. Although our decellularization method is effective in removing nuclear material from the bladder while maintaining its inherent mechanical properties, further work is mandatory to determine whether these residual DNA and cellular remnants would lead to any immune reaction, or if the mechanical properties of the ACM are preserved upon implantation and cellularization.

  4. Management of gingival recession with acellular dermal matrix graft: A clinical study

    Directory of Open Access Journals (Sweden)

    V R Balaji

    2016-01-01

    Full Text Available Aims and Objectives: Obtaining root coverage has become an important part of periodontal therapy. The aims of this studyare to evaluate the clinical efficacy of acellular dermal matrix graft in the coverage of denuded roots and also to examine the change in the width of keratinized gingiva. Materials and Methods: A total of 20 sites with more than or equal to 2 mm of recession depth were taken into the study, for treatment with acellular dermal matrix graft. The clinical parameters such as recession depth, recession width, width of keratinized gingiva, probing pocket depth (PD, and clinical attachment level (CAL were measured at the baseline, 8th week, and at the end of the study (16th week. The defects were treated with a coronally positioned pedicle graft combined with acellular dermal matrix graft. Results: Out of 20 sites treated with acellular dermal matrix graft, seven sites showed complete root coverage (100%, and the mean root coverage obtained was 73.39%. There was a statistically significant reduction in recession depth, recession width, and probing PD. There was also a statistically significant increase in width of keratinized gingiva and also gain in CAL. The postoperative results were both clinically and statistically significant (P < 0.0001. Conclusion: The results of this study were esthetically acceptable to the patients and clinically acceptable in all cases. From this study, it may be concluded that acellular dermal matrix graft is an excellent substitute for autogenous graft in coverage of denuded roots.

  5. Improvement in nerve regeneration through a decellularized nerve graft by supplementation with bone marrow stromal cells in fibrin.

    Science.gov (United States)

    Zhao, Zhe; Wang, Yu; Peng, Jiang; Ren, Zhiwu; Zhang, Li; Guo, Quanyi; Xu, Wenjing; Lu, Shibi

    2014-01-01

    Acellular nerve grafting is often inferior as well as an inadequate alternative to autografting for the repair of long gaps in peripheral nerves. Moreover, the injection method is not perfect. During the injection of cells, the syringe can destroy the acellular nerve structure and the limited accumulation of seed cells. To resolve this problem, we constructed a nerve graft by acellular nerve grafting. Bone marrow-mesenchymal stromal cells (BM-MSCs) were affixed with fibrin glue and injected inside or around the graft, which was then used to repair a 15-mm nerve defect in rats. The acellular nerve graft maintained its structure and composition, and its tensile strength was decreased, as determined by two-photon microscopy and a tensile testing device. In vitro, MSCs embedded in fibrin glue survived and secreted growth factors such as nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF). We repaired 15-mm Sprague-Dawley rat sciatic nerve defects using this nerve graft construction, and MSCs injected around the graft helped improve nerve regeneration and functional recovery of peripheral nerve lesions as determined by functional analysis and histology. Therefore, we conclude that supplying MSCs in fibrin glue around acellular nerves is successful in maintaining the nerve structure and can support nerve regeneration similar to the direct injection of MSCs into the acellular nerve for long nerve defects but may avoid destroying the nerve graft. The technique is simple and is another option for stem cell transplantation.

  6. Tissue engineering of the small intestine by acellular collagen sponge scaffold grafting.

    Science.gov (United States)

    Hori, Y; Nakamura, T; Matsumoto, K; Kurokawa, Y; Satomi, S; Shimizu, Y

    2001-01-01

    Tissue engineering of the small intestine will prove a great benefit to patients suffering from short bowel disease. However cell seeding in tissue engineering, such as fetal cell use, is accompanied by problems of ethical issues, rejection, and short supply. To overcome these problems, we carried out an experimental study on tissue engineering of the small intestine by acellular collagen sponge scaffold grafting. We resected the 5 cm long jejunum from beagle dogs and reconstructed it by acellular collagen sponge grafting with a silicon tube stent. The graft was covered with the omentum. At 1 month after operation, the silicon stent was removed endoscopically. Animals were sacrificed 1 and 4 months after operation, and were examined microscopically. Neo-intestinal regeneration was observed and the intestinal mucosa covered the luminal side of the regenerated intestine across the anastomosis. Thus, the small intestine was regenerated by tissue engineering technology using an acellular collagen sponge scaffold.

  7. Acellular comet assay: a tool for assessing variables influencing the alkaline comet assay.

    Science.gov (United States)

    Kennedy, Erin K; McNamee, James P; Prud'homme Lalonde, Louise; Jones, Trevor; Wilkinson, Diana

    2012-01-01

    In this study, an acellular modification to the alkaline comet assay to further evaluate key variables within the assay that may influence the outcome of genotoxicity studies is described. This acellular comet assay can detect differences of 0.2 Gy of (60)Co gamma-ray radiation between 0 and 1 Gy and differences of 1 Gy between 0 and 8 Gy; thus, this assay is applicable for a wide range of DNA damage levels. It is also shown that DNA damage from different radiation energies was not significantly different from (60)Co gamma-ray. This assay displayed a statistical increase in DNA damage due to uncontrolled exposure to natural light; however, the slope of the dose-response curve for light-exposed samples was similar to that for samples protected from light. A comparison of the alkaline comet assay with the acellular comet assay allowed for the intrinsic repair capacity of the alkaline comet assay to be quantified.

  8. Pertactin deficient Bordetella pertussis present a better fitness in mice immunized with an acellular pertussis vaccine.

    Science.gov (United States)

    Hegerle, N; Dore, G; Guiso, N

    2014-11-20

    Bordetella pertussis is the etiologic agent of whooping cough and has been the target of vaccination for over fifty years. The latest strategies include the use of acellular pertussis vaccines that induce specific immunity against few virulence factors amongst which pertactin is included in three and five component acellular pertussis vaccines. Recently, it has been reported that B. pertussis clinical isolates loose the production of this adhesin in regions reaching high vaccine coverage with vaccines targeting this virulence factor. We here demonstrate that isolates not producing pertactin are capable of sustaining longer infection as compared to pertactin producing isolates in an in vivo model of acellular pertussis immunization. Loosing pertactin production might thus provide a selective advantage to these isolates in this background, which could account for the upraise in prevalence of these pertactin deficient isolates in the population.

  9. Acellular biological tissues containing inherent glycosaminoglycans for loading basic fibroblast growth factor promote angiogenesis and tissue regeneration.

    Science.gov (United States)

    Lai, Po-Hong; Chang, Yen; Chen, Sung-Ching; Wang, Chung-Chi; Liang, Huang-Chien; Chang, Wei-Chun; Sung, Hsing-Wen

    2006-09-01

    It was found in our previous study that acellular tissues derived from bovine pericardia consist primarily of insoluble collagen, elastin, and tightly bound glycosaminoglycans (GAGs). It is speculated that the inherent GAGs in acellular tissues may serve as a reservoir for loading basic fibroblast growth factor (bFGF) and promote angiogenesis and tissue regeneration. This study was therefore designed to investigate effects of the content of GAGs in acellular bovine pericardia on the binding of bFGF and its release profile in vitro while its stimulation in angiogenesis and tissue regeneration in vivo were evaluated subcutaneously in a rat model. To control the content of GAGs, acellular tissues were treated additionally with hyaluronidase for 1 (Hase-D1), 3 (Hase-D3), or 5 days (Hase-D5). The in vitro results indicated that a higher content of GAGs in the acellular tissue resulted in an increase in bFGF binding and in a more gradual and sustained release of the growth factor. The in vivo results obtained at 1 week postoperatively showed that the density and the depth of neo-vessels infiltrated into the acellular tissue loaded with bFGF (acellular/bFGF) were significantly greater than the other test samples. At 1 month postoperatively, vascularized neo-connective tissues were found to fill the pores within each test sample, particularly for the acellular/bFGF tissue. These results suggested that the sustained release of bFGF from the acellular/ bFGF tissue continued to be effective in enhancing angiogenesis and generation of new tissues. In conclusion, the inherent GAGs present in acellular tissues may be used for binding and sustained release of bFGF to enhance angiogenesis and tissue regeneration.

  10. Coverage of Megaprosthesis with Human Acellular Dermal Matrix after Ewing's Sarcoma Resection: A Case Report

    Directory of Open Access Journals (Sweden)

    Robert M. Whitfield

    2011-01-01

    Full Text Available A 23-year-old female with Ewing's Sarcoma underwent tibial resection and skeletal reconstruction using proximal tibial allograft prosthetic reconstruction with distal femur endoprosthetic reconstruction and rotating hinge. Human acellular dermal matrix, (Alloderm, LifeCell, Branchburg, NJ, USA, was used to wrap the skeletal reconstruction. Soft tissue reconstruction was completed with a rotational gastrocnemius muscle flap and skin graft. Despite prolonged immobilization, the patient quickly regained full range of motion of her skeletal reconstruction. Synthetic mesh, tapes and tubes are used to perform capsule reconstruction of megaprosthesis. This paper describes the role of human acellular dermal matrix in capsule reconstruction around a megaprosthesis.

  11. Coverage of Megaprosthesis with Human Acellular Dermal Matrix after Ewing's Sarcoma Resection: A Case Report.

    Science.gov (United States)

    Whitfield, Robert M; Rinard, Jeremy; King, David

    2011-01-01

    A 23-year-old female with Ewing's Sarcoma underwent tibial resection and skeletal reconstruction using proximal tibial allograft prosthetic reconstruction with distal femur endoprosthetic reconstruction and rotating hinge. Human acellular dermal matrix, (Alloderm, LifeCell, Branchburg, NJ, USA), was used to wrap the skeletal reconstruction. Soft tissue reconstruction was completed with a rotational gastrocnemius muscle flap and skin graft. Despite prolonged immobilization, the patient quickly regained full range of motion of her skeletal reconstruction. Synthetic mesh, tapes and tubes are used to perform capsule reconstruction of megaprosthesis. This paper describes the role of human acellular dermal matrix in capsule reconstruction around a megaprosthesis.

  12. Usual interstitial pneumonia end-stage features from explants with radiologic and pathological correlations.

    Science.gov (United States)

    Rabeyrin, Maud; Thivolet, Françoise; Ferretti, Gilbert R; Chalabreysse, Lara; Jankowski, Adrien; Cottin, Vincent; Pison, Christophe; Cordier, Jean-François; Lantuejoul, Sylvie

    2015-08-01

    Idiopathic pulmonary fibrosis (IPF) is the most frequent and severe idiopathic interstitial pneumonia, with typical high-resolution computed tomography (HRCT) features and histologic pattern of usual interstitial pneumonia (UIP); its main differential diagnosis is fibrotic nonspecific interstitial pneumonia (F-NSIP). Usual interstitial pneumonia was mainly described from lung biopsies, and little is known on explants. Twenty-two UIP/IPF explants were analyzed histologically and compared with previous open lung biopsies (OLBs; n = 11) and HRCT (n = 19), when available. Temporospatial heterogeneity and subpleural and paraseptal fibrosis were similarly found in UIP/IPF explants and OLB (91%-95%). Fibroblastic foci were found in 82% of OLBs and 100% of explants, with a higher mean score in explants (P = .023). Honeycombing was present in 64% of OLBs and 95% of explants, with a higher mean score in explants (P = .005). Almost 60% of UIP/IPF explants showed NSIP areas and 41% peribronchiolar fibrosis; inflammation, bronchiolar metaplasia, and vascular changes were more frequent in UIP/IPF explants; and Desquamative Interstitial Pneumonia (DIP)-like areas were not common (18%-27%). Numerous large airspace enlargements with fibrosis were frequent in UIP/IPF explants (59%). On HRCT, honeycombing was observed in 95% of the cases and ground-glass opacities in 53%, correlating with NSIP areas or acute exacerbation at histology. Six patients had combined IPF and emphysema. Lesions were more severe in UIP/IPF explants, reflecting the worsening of the disease. Usual interstitial pneumonia/IPF explants more frequently presented with confounding lesions such as NSIP areas, peribronchiolar fibrosis, and airspace enlargements with fibrosis sometimes associated with emphysema.

  13. The influence of season collection of explants on micropropagation of peach rootstock GF-677

    Directory of Open Access Journals (Sweden)

    Elektra Spahiu

    2013-02-01

    Full Text Available The influence of season on the rate of multiplication on in vitro culture of peach rootstock GF- 677 was investigated on Murashige and Skoog (MS media, supplemented with GA3 0.1 mg/L and IAA 0.1mg/l. Benzyladenine (BAP at concentrations 1mg/l was used in the multiplication stage and 1mg/l IBA in the stage of rooting. Shoot-tip and nodal segment explants were collected from 5 years old rootstock GF-677 (Prunus persica x Prunus amygdalus in February 24th (from dormant shoots that have been sprouted in climatic room, March 22th, April 20th, May 18th and September 15th during the 2009 growing season and have been sterilized by sodium hypochlorite (NaOCl 10% for 20 min. The data on the effect of the season collection of the explants on number of shoots per explants, the mean shoot length and the percentage of rooted shoots were recorded six weeks after culture. In vitro performance of explants indicated a positive correlation between shoot proliferation and season collection The highest number of shoots per explants (3,5 was obtained on explants collected in March 22th (3,5, which was on a par with explants collected in February 24th (from shoots that have been sprouted in climatic room. Moreover, the highest shoot length was observed on explants collected on February and March (1,53cm and 1,505cm respectively. The percentage of rooted shoots from explants sampled on February was not markedly greater than those sampled on March. The number of shoots per explants, the shoot length and the percentage of rooted shoots on explants sampled in April, May and September were significantly lower than those sampled in February and March. The amount of chlorophyll a + b of the shoots coming from explants collected in March was markedly greater than those collected in February, April, May and September.

  14. Effect of peripheral nerve on the neurite growth from retinal explants in culture

    Institute of Scientific and Technical Information of China (English)

    LiuLi; SoKwokfai

    1990-01-01

    The effect of peripheral nerve (PN) on neurite outgrowth from retinal explants of adult hamsters was examined.Cultures of retinal explants,and co-cultures of retinal explants and PN were performed using chick retinal basement memebrane (BM) as substrate.The presence of PN increases the number and length of neurite outgrowth.In addition,a high proportion of neurites situated close to PN tend to grow towards it.Since there was no contact between retinal explants and PN,we suggest that PN might secete diffusible substances to attract the neurites to grow towards it.

  15. Micropropagation of Cyrtopodium paludicolum (Orchidaceae from root tip explants

    Directory of Open Access Journals (Sweden)

    Dayana Rotili Nunes Picolotto

    2017-06-01

    Full Text Available An efficient protocol for in vitro plant propagation of Cyrtopodium paludicolum has been developed using root tips dissected from well-developed seedlings. Root tips were cultured on Knudson medium supplemented with α-naphthaleneacetic acid (NAA, and/or thidiazuron (TDZ. TDZ did not induce protocorm-like bodies (PLBs in the NAA absence, indicating phytoregulators synergistic effect. Medium supplemented with 1.34 μM NAA and 2.27 μM TDZ resulted in better response on PBLs, and subsequent shoot differentiation (55.25 shoots per explant, and in better rooting number and root length responses, favoring acclimatization with 90% of survived plants. However, the medium supplemented with only NAA (1.34 μM resulted in 33.50 shoots per explant. Histological sections confirmed that only one PLB was induced per responsive root tip, and it showed numerous dispersed and extended meristemoids, or division centers that originated new PBLs. Additionally, this protocol could be an excellent model to study molecular aspects of root to shoot conversion.

  16. Microscopic and spectroscopic investigation of an explanted opacified intraocular lens

    Energy Technology Data Exchange (ETDEWEB)

    Simon, V., E-mail: viosimon@phys.ubbcluj.ro [Babeş-Bolyai University, Faculty of Physics and Interdisciplinary Research Institute on Bio-Nano-Sciences, 400084 Cluj-Napoca (Romania); Radu, T.; Vulpoi, A. [Babeş-Bolyai University, Faculty of Physics and Interdisciplinary Research Institute on Bio-Nano-Sciences, 400084 Cluj-Napoca (Romania); Rosca, C. [Optilens Clinic of Ophthalmology, 400604 Cluj-Napoca (Romania); Eniu, D. [Iuliu Haţieganu University of Medicine and Pharmacy, Department of Molecular Sciences, 400349 Cluj-Napoca (Romania)

    2015-01-15

    Highlights: • Changes on intraocular lens (IOL) surface after implantation. • Partial opacification of IOL central area. • Elemental composition on IOL surface prior to and after implantation. • First XPS depth profiling examination of the opacifying deposits. • Cell-mediated hydroxyapatite structuring. - Abstract: The investigated polymethylmethacrylate intraocular lens explanted an year after implantation presented a fine granularity consisting of ring-like grains of about 15 μm in diameter. In order to evidence the changes occurred on intraocular lens relative to morphology, elemental composition and atomic environments, microscopic and spectroscopic analyses were carried out using scanning electron microscopy (SEM), Fourier transform infrared (FTIR), energy-dispersive X-ray (EDS), and X-ray photoelectron (XPS) spectroscopies. The results revealed that the grains contain hydroxyapatite mineral phase. A protein layer covers the lens both in opacified and transparent zones. The amide II band is like in basal epithelial cells. The shape and size of the grains, and the XPS depth profiling results indicate the possibility of a cell-mediated process involving lens epithelial cells which fagocitated apoptotic epithelial cells, and in which the debris derived from cell necrosis were calcified. To the best of our knowledge, this is the first investigation on explanted intraocular lenses using XPS depth profiling in order to examine the inside of the opacifying deposits.

  17. Microscopic and spectroscopic investigation of an explanted opacified intraocular lens

    Science.gov (United States)

    Simon, V.; Radu, T.; Vulpoi, A.; Rosca, C.; Eniu, D.

    2015-01-01

    The investigated polymethylmethacrylate intraocular lens explanted an year after implantation presented a fine granularity consisting of ring-like grains of about 15 μm in diameter. In order to evidence the changes occurred on intraocular lens relative to morphology, elemental composition and atomic environments, microscopic and spectroscopic analyses were carried out using scanning electron microscopy (SEM), Fourier transform infrared (FTIR), energy-dispersive X-ray (EDS), and X-ray photoelectron (XPS) spectroscopies. The results revealed that the grains contain hydroxyapatite mineral phase. A protein layer covers the lens both in opacified and transparent zones. The amide II band is like in basal epithelial cells. The shape and size of the grains, and the XPS depth profiling results indicate the possibility of a cell-mediated process involving lens epithelial cells which fagocitated apoptotic epithelial cells, and in which the debris derived from cell necrosis were calcified. To the best of our knowledge, this is the first investigation on explanted intraocular lenses using XPS depth profiling in order to examine the inside of the opacifying deposits.

  18. Biomechanical properties of acellular sciatic nerves treated with a modified chemical method

    Institute of Scientific and Technical Information of China (English)

    Xinlong Ma; Zhao Yang; Xiaolei Sun; Jianxiong Ma; Xiulan Li; Zhenzhen Yuan; Yang Zhang; Honggang Guo

    2011-01-01

    Nerve grafts are able to adapt to surrounding biomechanical environments if the nerve graft itself exhibits appropriate biomechanical properties (load, elastic modulus, etc.). The present study was designed to determine the differences in biomechanical properties between fresh and chemically acellularized sciatic nerve grafts. Two different chemical methods were used to establish acellular nerve grafts. The nerve was chemically extracted in the Sondell method with a combination of Triton X-100 (nonionic detergent) and sodium deoxycholate (anionic detergent), and in the modified method with a combination of Triton X-200 (anionic detergent), sulfobetaine-10 (SB-10, amphoteric detergents), and sulfobetaine-16 (SB-16, amphoteric detergents). Following acellularization, hematoxylin-eosin staining and scanning electron microscopy demonstrated that the effect of acellularization via the modified method was similar to the traditional Sondell method. However, effects of demyelination and nerve fiber tube integrity were superior to the traditional Sondell method. Biomechanical testing showed that peripheral nerve graft treated using the chemical method resulted in decreased biomechanical properties (ultimate load, ultimate stress, ultimate strain, and mechanical work to fracture) compared with fresh nerves, but the differences had no statistical significance (P > 0.05). These results demonstrated no significant effect on biomechanical properties of nerves treated using the chemical method. In conclusion, nerve grafts treated via the modified method removed Schwann cells, preserved neural structures, and ensured biomechanical properties of the nerve graft, which could be more appropriate for implantation studies.

  19. Flexor tendon tissue engineering: acellularization of human flexor tendons with preservation of biomechanical properties and biocompatibility.

    Science.gov (United States)

    Pridgen, Brian C; Woon, Colin Y L; Kim, Maxwell; Thorfinn, Johan; Lindsey, Derek; Pham, Hung; Chang, James

    2011-08-01

    Acellular human tendons are a candidate scaffold for tissue engineering flexor tendons of the hand. This study compared acellularization methods and their compatibility with allogeneic human cells. Human flexor tendons were pretreated with 0.1% ethylenediaminetetracetic acid (EDTA) for 4  h followed by 24  h treatments of 1% Triton X-100, 1% tri(n-butyl)phosphate, or 0.1% or 1% sodium dodecyl sulfate (SDS) in 0.1% EDTA. Outcomes were assessed histologically by hematoxylin and eosin and SYTO green fluorescent nucleic acid stains and biochemically by a QIAGEN DNeasy kit, Sircol collagen assay, and 1,9 dimethylmethylene blue glycosaminoglycan assay. Mechanical data were collected using a Materials Testing System to pull to failure tendons acellularized with 0.1% SDS. Acellularized tendons were re-seeded in a suspension of human dermal fibroblasts. Attachment of viable cells to acellularized tendon was assessed biochemically by a cell viability assay and histologically by a live/dead stain. Data are reported as mean±standard deviation. Compared with the DNA content of fresh tendons (551±212  ng DNA/mg tendon), only SDS treatments significantly decreased DNA content (1% SDS [202.8±37.4  ng DNA/mg dry weight tendon]; 0.1% SDS [189±104  ng DNA/mg tendon]). These findings were confirmed by histology. There was no decrease in glycosaminoglycans or collagen following acellularization with SDS. There was no difference in the ultimate tensile stress (55.3±19.2 [fresh] vs. 51.5±6.9 [0.1% SDS] MPa). Re-seeded tendons demonstrated attachment of viable cells to the tendon surface using a viability assay and histology. Human flexor tendons were acellularized with 0.1% SDS in 0.1% EDTA for 24  h with preservation of mechanical properties. Preservation of collagen and glycoaminoglycans and re-seeding with human cells suggest that this scaffold is biocompatible. This will provide a promising scaffold for future human flexor tendon tissue engineering studies to

  20. [STUDY ON MODIFICATION OF BIOMATERIALS OF ACELLULAR BOVINE PERICARDIUM WITH DIFFERENT CROSSLINKING REAGENTS].

    Science.gov (United States)

    Xiao, Hongtao; Tian, Shemin; Zha, Xinjian; Wei, Ying; Huang, Hongjun; Li, Yun; Yang, Huanna; Xia, Chengde; Niu, Xihua

    2015-10-01

    To investigate the effects of modification of acellular bovine pericardium with 1-ethyl-3-(3- dinethylami-nopropyl) carbodimide (EDC)/N-hydroxysuccininide (NHS) or genipin and find out the best crosslinking reagent. The cellular components of the bovine pericardiums were removed. The effects of decellularization were tested by HE staining. The acellular bovine pericardiums were crosslinked with EDC/NHS (EDC/NHS group) or genipin (genipin group). The properties of the crosslinked acellular matrix were evaluated by scanning electron microscope (SEM), matrix thickness, crosslinking index, mechanical property, denaturation temperature, enzymatic degradation, and cytotoxicity test before and after the crosslinking. Acellular bovine pericardium (ABP group) or normal bovine pericardium (control group) were harvested as controls. SEM showed that collagen fibers were reticulated in bovine pericardial tissues after crosslinked by EDC/NHS or genipin, and relative aperture of the collagen fiber was from 10 to 20 μm. The thickness and denaturation temperature of the scaffolds were increased significantly after crosslinking with EDC/NHS or genipin (P 0.05). The difference had no statistical significance in crosslinking index between EDC/NHS group and genipin group (t = 0.205, P = 0.218). The degradation rate in EDC/NHS group and genipin group was significantly lower than that in ABP group and control group (P 0.05). The break elongation in EDC/NHS group and genipin group were significantly increased than those in ABP group and control group (P 0.05). Cytotoxicity of genipin crosslinked tissue (grade 1) were much lower than that of EDC/NHS (grade 2) at 5 days. Acellular bovine pericardium crosslinked with genipin has better biocompatibility than EDC/NHS.

  1. Organogênese direta de explantes cotiledonares e regeneração de plantas de mogango Direct organogenesis of cotyledon explants and plant regeneration of squash

    Directory of Open Access Journals (Sweden)

    André Luís Lopes da Silva

    2006-06-01

    Full Text Available Os objetivos foram induzir a organogênese direta de explantes cotiledonares de mogango e estudar a regeneração de plântulas completas a partir das brotações adventícias. Foram utilizados cotilédones como explantes, originados das plântulas de mogango com 20 dias após a semeadura. O meio basal utilizado foi o MS (MURASHIGE & SKOOG, 1962 suplementado com 30g L-1 de sacarose e 7g L-1 de agar. Foram testadas as concentrações de 6-benzilaminopurina (BAP de 0; 0,5; 1,0 e 2,0mg L-1. Explantes de ápices caulinares e segmentos nodais de brotações adventícias foram então cultivados em meio MS suplementado com 30g L-1 de sacarose e 7g L-1 de agar. Maiores concentrações de BAP no meio MS promoveram um aumento da percentagem de explantes cotiledonares com brotações adventícias e uma redução da percentagem de enraizamento. Explantes de segmentos nodais e ápices caulinares oriundos de brotações adventícias cresceram e enraizaram em meio MS sem reguladores de crescimento. Altas percentagens de enraizamento dependem do tamanho dos explantes utilizados.The objectives were to induce direct organogenesis of squash cotyledons and to study the regeneration of complete plantlets from adventitious shoot. Cotyledon explants of 20-day seedlings were cultured in MS (MURASHIGE & SKOOG, 1962 medium supplemented with 30g L-1 of sucrose and 7g L-1 of agar. The 6-benzilaminopurina (BAP concentrations of 0, 0.5, 1.0 and 2.0mg L-1 were tested. Apical and nodal explants from adventitious shoots were transferred to MS medium supplemented with 30g L-1 of sucrose and 7g L-1 of agar. Increasing BAP concentrations in the MS medium enhance the percentage of adventitious shoot and reduce the percentage of root organogenesis of squash cotyledon explants. Apical and nodal explants from adventitious shoot regenerated plantlets with roots in MS medium without growth regulators. High percentage of plantlet rooting depends upon the size of the explants.

  2. Bone tumor

    Science.gov (United States)

    Tumor - bone; Bone cancer; Primary bone tumor; Secondary bone tumor; Bone tumor - benign ... The cause of bone tumors is unknown. They often occur in areas of the bone that grow rapidly. Possible causes include: Genetic defects ...

  3. EXPLANTATION OF MESANGIAL CELL HILLOCKS - A METHOD FOR OBTAINING HUMAN MESANGIAL CELLS IN CULTURE

    NARCIS (Netherlands)

    MULLER, EW; KIM, Y; MICHAEL, AF; VERNIER, RL; VANDERHEM, GK; VANDERWOUDE, FJ

    1992-01-01

    A simple method is presented for selective cell culture of human mesangial cells using explanatation of mesangial cell hillocks. Glomeruli which had been incubated with collagenase were explanted on plastic tissue culture flasks. Three to 6 weeks after explantation, a rapidly growing multilayer of e

  4. In vitro morphogenic events in culture of Lotus corniculatus L. seedling root explants

    Directory of Open Access Journals (Sweden)

    Jan J. Rybczyński

    2011-01-01

    Full Text Available The experiments were carried out on Lotus corniculatus (L. seedling root explants of the cultivar varieties Skrzeszowicka, Caroll A10 and strain 175. Callus formation and shoot regeneration were the major explant response depended mainly on of the studied genotype and used plant growth regulators (PGRs. Primary cortex of proximal and distal end of explant was the most active tissue for callus proliferation. For shoot primordia differentiation deeper zones of cortex took a part. The process of meristematic centre initiation was not uniform and various level of shoot differentiation events were observed not earlier than 3 weeks of culture. Usually, the shoot primordia regeneration began on proximal rather than distal end of the explant. BAP rather than urea derivatives stimulated shoot proliferation in extended cultures. Increasing of BAP and TDZ concentrations brought about the explant polarity and expansion of the meristematic zones. The explant position in root did not have significant influence on the number of regenerated shoots. The cultures only had better bud formation by TDZ when compared to BAP. BAP stimulated bud formation and development of the shoots from them. Short term of TDZ treatment of explants stimulated meristem formation which developed into buds and shoots. CPPU stimulated callus proliferation and bud formation when explants pretreatment was prolonged from 12 to 36 hrs.

  5. Explant culture of rat colon: A model system for studying metabolism of chemical carcinogens

    DEFF Research Database (Denmark)

    Autrup, Herman; Stoner, G.D.; Jackson, F.

    1978-01-01

    An explant culture system has been developed for the long-term maintenance of colonic tissue from the rat. Explants of 1 cm2 in size were placed in tissue-culture dishes to which was added 2 ml of CMRL-1066 medium supplemented with glucose, hydrocortisone, beta-retinyl acetate, and either 2.5% bo...

  6. EXPLANTATION OF MESANGIAL CELL HILLOCKS - A METHOD FOR OBTAINING HUMAN MESANGIAL CELLS IN CULTURE

    NARCIS (Netherlands)

    MULLER, EW; KIM, Y; MICHAEL, AF; VERNIER, RL; VANDERHEM, GK; VANDERWOUDE, FJ

    1992-01-01

    A simple method is presented for selective cell culture of human mesangial cells using explanatation of mesangial cell hillocks. Glomeruli which had been incubated with collagenase were explanted on plastic tissue culture flasks. Three to 6 weeks after explantation, a rapidly growing multilayer of e

  7. [The effect of Solcoseryl on explant cultures of the hippocampus].

    Science.gov (United States)

    Lindner, G; Grosse, G; Goworek, K; Franz, C; Liebezeit, K

    1979-01-01

    Explants of hippocampus from fetal rats were cultivated in Maximow chambers in semisynthetic medium up to 12 days in vitro. The cultures were fixed Bouin, slided 15 micron, coloured with Klüver-Barrera and some morphological parameters were tested. 1. The nerve fiber index increased by influence of 1% Solcoseryl in relation to control cultures, which growed in minimal medium. An essential stimulation was observed by application of placentar serum and embryonal extract into the culture medium. 2. Die decrease of the number of neurons and glial cells per unit of area and a small decrease of the area of neuron nuclei was discussed in relation to the effect of the pharmacon Solcoseryl on O2- consumption. 3. Solcoseryl (firm Solco AG, Base) is an extract of calf blood. It can not substitute other tissue extracts.

  8. Glucosamine increases hyaluronic acid production in human osteoarthritic synovium explants

    Directory of Open Access Journals (Sweden)

    Uitterlinden EJ

    2008-09-01

    Full Text Available Abstract Background Glucosamine (GlcN used by patients with osteoarthritis was demonstrated to reduce pain, but the working mechanism is still not clear. Viscosupplementation with hyaluronic acid (HA is also described to reduce pain in osteoarthritis. The synthesis of HA requires GlcN as one of its main building blocks. We therefore hypothesized that addition of GlcN might increase HA production by synovium tissue. Methods Human osteoarthritic synovium explants were obtained at total knee surgery and pre-cultured for 1 day. The experimental conditions consisted of a 2 days continuation of the culture with addition of N-Acetyl-glucosamine (GlcN-Ac; 5 mM, glucosamine-hydrochloride (GlcN-HCl; 0.5 and 5 mM, glucose (Gluc; 0.5 and 5 mM. Hereafter HA production was measured in culture medium supernatant using an enzyme-linked binding protein assay. Real time RT-PCR was performed for hyaluronic acid synthase (HAS 1, 2 and 3 on RNA isolated from the explants. Results 0.5 mM and 5 mM GlcN-HCl significantly increased HA production compared to control (approximately 2 – 4-fold, whereas GlcN-Ac had no significant effect. Addition of 5 mM Gluc also increased HA production (approximately 2-fold, but 0.5 mM Gluc did not. Gene expression of the HA forming enzymes HAS 1, 2 and 3 was not altered by the addition of GlcN or Gluc. Conclusion Our data suggest that exogenous GlcN can increase HA production by synovium tissue and is more effective at lower concentrations than Gluc. This might indicate that GlcN exerts its potential analgesic properties through stimulation of synovial HA production.

  9. Plant Regeneration of Sweet Potato via Somatic Embryogenesis from Different Explants

    Institute of Scientific and Technical Information of China (English)

    Ling ZHANG; Hongxuan XU; Baifu QIN; Zhihua LIA0; Min CHEN; Chunxian YANG; Yufan FU; Qitang ZHANG

    2012-01-01

    [Objective] This study aimed to regenerate plants of sweet potato (Ipomoea batatas) cultivar Xushu22 via somatic embryogenesis, using leaf and shoot apex as explants. [Method] The leaf and shoot apex of Xushu 22 were separately cultured on MSB medium and MSD medium. The induced embryogenic calluses were then cultured on MS medium. The regeneration frequency of leaf and shoot apex ex- plants were respectively calculated. [Result] The average frequency of leaf explants developing somatic callus was 95.69% compared to 30.56% in case of shoot apex explants. There were different types of morphogenic structures in the process of so- matic embryo development. Leaf explants gave a high regeneration frequency to 60.61%, while the regeneration frequency of shoot apices was 22%. In addition, no morphological variations were observed in the regeneration plants. [Conclusion] Leaf explant was better than shoot apices in plant regeneration of Xushu22 via somatic embryogenesis.

  10. Explant analysis of the Biomet Magnum/ReCap metal-on-metal hip joint

    Science.gov (United States)

    Hunt, B. J.; Richardson, V. M.; Langton, D. J.; Smith, E.; Joyce, T. J.

    2017-01-01

    Objectives The high revision rates of the DePuy Articular Surface Replacement (ASR) and the DePuy ASR XL (the total hip arthroplasty (THA) version) have led to questions over the viability of metal-on-metal (MoM) hip joints. Some designs of MoM hip joint do, however, have reasonable mid-term performance when implanted in appropriate patients. Investigations into the reasons for implant failure are important to offer help with the choice of implants and direction for future implant designs. One way to assess the performance of explanted hip prostheses is to measure the wear (in terms of material loss) on the joint surfaces. Methods In this study, a coordinate measuring machine (CMM) was used to measure the wear on five failed cementless Biomet Magnum/ReCap/ Taperloc large head MoM THAs, along with one Biomet ReCap resurfacing joint. Surface roughness measurements were also taken. The reason for revision of these implants was pain and/or adverse reaction to metal debris (ARMD) and/or elevated blood metal ion levels. Results The mean wear rate of the articulating surfaces of the heads and acetabular components of all six joints tested was found to be 6.1 mm3/year (4.1 to 7.6). The mean wear rate of the femoral head tapers of the five THAs was 0.054 mm3/year (0.021 to 0.128) with a mean maximum wear depth of 5.7 µm (4.3 to 8.5). Conclusion Although the taper wear was relatively low, the wear from the articulating surfaces was sufficient to provide concern and was potentially large enough to have been the cause of failure of these joints. The authors believe that patients implanted with the ReCap system, whether the resurfacing prosthesis or the THA, should be closely monitored. Cite this article: S. C. Scholes, B. J. Hunt, V. M. Richardson, D. J. Langton, E. Smith, T. J. Joyce. Explant analysis of the Biomet Magnum/ReCap metal-on-metal hip joint. Bone Joint Res 2017;6:113–122. DOI: 10.1302/2046-3758.62.BJR-2016-0130.R2. PMID:28246095

  11. Characterisation of the circulating acellular proteome of healthy sheep using LC-MS/MS-based proteomics analysis of serum

    OpenAIRE

    Chemonges, Saul; Gupta, Rajesh; Mills, Paul C.; Steven R. Kopp; Sadowski, Pawel

    2017-01-01

    Background Unlike humans, there is currently no publicly available reference mass spectrometry-based circulating acellular proteome data for sheep, limiting the analysis and interpretation of a range of physiological changes and disease states. The objective of this study was to develop a robust and comprehensive method to characterise the circulating acellular proteome in ovine serum. Methods Serum samples from healthy sheep were subjected to shotgun proteomic analysis using nano liquid chro...

  12. New Insights on the Composition and the Structure of the Acellular Extrinsic Fiber Cementum by Raman Analysis

    Science.gov (United States)

    Colard, Thomas; Falgayrac, Guillaume; Bertrand, Benoit; Naji, Stephan; Devos, Olivier; Balsack, Clara; Delannoy, Yann; Penel, Guillaume

    2016-01-01

    Acellular extrinsic fiber cementum is a mineralized tissue that covers the cervical half of the tooth root surface. It contains mainly extrinsic or Sharpey’s fibers that run perpendicular to the root surface to anchor the tooth via the periodontal ligament. Acellular cementum is continuously and slowly produced throughout life and exhibits an alternating bright and dark pattern under light microscopy. However, although a better understanding of the structural background of acellular cementum is relevant to many fields, such as cementochronology, periodontology and tissue engineering, acellular cementum remains rarely studied and poorly understood. In this work, we studied the acellular cementum at the incremental line scale of five human mandibular canines using polarized Raman spectroscopy. We provided Raman imaging analysis and polarized acquisitions as a function of the angular orientation of the sample. The results showed that mineral crystals were always parallel to collagen fibrils, and at a larger scale, we proposed an organizational model in which we found radial collagen fibers, “orthogonal” to the cementum surface, and “non-orthogonal” fibers, which consist of branching and bending radial fibers. Concerning the alternating pattern, we observed that the dark lines corresponded to smaller, more mineralized and probably more organized bands, which is consistent with the zoological assumption that incremental lines are produced during a winter rest period of acellular cementum growth. PMID:27936010

  13. New Insights on the Composition and the Structure of the Acellular Extrinsic Fiber Cementum by Raman Analysis.

    Science.gov (United States)

    Colard, Thomas; Falgayrac, Guillaume; Bertrand, Benoit; Naji, Stephan; Devos, Olivier; Balsack, Clara; Delannoy, Yann; Penel, Guillaume

    2016-01-01

    Acellular extrinsic fiber cementum is a mineralized tissue that covers the cervical half of the tooth root surface. It contains mainly extrinsic or Sharpey's fibers that run perpendicular to the root surface to anchor the tooth via the periodontal ligament. Acellular cementum is continuously and slowly produced throughout life and exhibits an alternating bright and dark pattern under light microscopy. However, although a better understanding of the structural background of acellular cementum is relevant to many fields, such as cementochronology, periodontology and tissue engineering, acellular cementum remains rarely studied and poorly understood. In this work, we studied the acellular cementum at the incremental line scale of five human mandibular canines using polarized Raman spectroscopy. We provided Raman imaging analysis and polarized acquisitions as a function of the angular orientation of the sample. The results showed that mineral crystals were always parallel to collagen fibrils, and at a larger scale, we proposed an organizational model in which we found radial collagen fibers, "orthogonal" to the cementum surface, and "non-orthogonal" fibers, which consist of branching and bending radial fibers. Concerning the alternating pattern, we observed that the dark lines corresponded to smaller, more mineralized and probably more organized bands, which is consistent with the zoological assumption that incremental lines are produced during a winter rest period of acellular cementum growth.

  14. Influence of mineral phase in mineralization of a biocomposite containing chitosan, demineralized bone matrix and bone ash—in vitro study

    Indian Academy of Sciences (India)

    Krithiga Gunasekaran; Santhosh Kumar Baskar; Divya Sapphire Mohan; Thotapalli P Sastry

    2014-05-01

    A resorbable composite which acts as a active barrier in guided bone regeneration was fabricated using chitosan, demineralized bone matrix and bone ash. Its potential to form bone like apatite in simulated body fluid was assessed in this study. The mechanical strength of these composites was correlated with bone ash ratios and composites with better tensile strength were studied for their acellular bioactivity by incubating in simulated body fluid for 21 days. Composites without bone ash did not show acellular bioactivity which was confirmed by thermogravimetric analysis. In case of biocomposites with bone ash, there was an increase in residual weight indicating the mineralization of the composite. The composite containing bone ash has shown the peaks related to phosphate vibrations in its Fourier-transform infrared spectrum. Scanning micrographs revealed formation of apatite like crystals on its surface. Ca/P ratio was found to be 1.7 which is nearer to that of natural bone. Thus, prepared composites can be used as resorbable biocomposite in maxillofacial and oral defects.

  15. The metabolic dynamics of cartilage explants over a long-term culture period

    Directory of Open Access Journals (Sweden)

    E.K Moo

    2011-01-01

    Full Text Available INTRODUCTION: Although previous studies have been performed on cartilage explant cultures, the generalized dynamics of cartilage metabolism after extraction from the host are still poorly understood due to differences in the experimental setups across studies, which in turn prevent building a complete picture. METHODS: In this study, we investigated the response of cartilage to the trauma sustained during extraction and determined the time needed for the cartilage to stabilize. Explants were extracted aseptically from bovine metacarpal-phalangeal joints and cultured for up to 17 days. RESULTS: The cell viability, cell number, proteoglycan content, and collagen content of the harvested explants were analyzed at 0, 2, 10, and 17 days after explantation. A high percentage of the cartilage explants were found to be viable. The cell density initially increased significantly but stabilized after two days. The proteoglycan content decreased gradually over time, but it did not decrease to a significant level due to leakage through the distorted peripheral collagen network and into the bathing medium. The collagen content remained stable for most of the culture period until it dropped abruptly on day 17. CONCLUSION: Overall, the tested cartilage explants were sustainable over long-term culture. They were most stable from day 2 to day 10. The degradation of the collagen on day 17 did not reach diseased levels, but it indicated the potential of the cultures to develop into degenerated cartilage. These findings have implications for the application of cartilage explants in pathophysiological fields.

  16. An ex vivo porcine nasal mucosa explants model to study MRSA colonization.

    Directory of Open Access Journals (Sweden)

    Pawel Tulinski

    Full Text Available Staphylococcus aureus is an opportunistic pathogen able to colonize the upper respiratory tract and skin surfaces in mammals. Methicillin-resistant S. aureus ST398 is prevalent in pigs in Europe and North America. However, the mechanism of successful pig colonization by MRSA ST398 is poorly understood. To study MRSA colonization in pigs, an ex vivo model consisting of porcine nasal mucosa explants cultured at an air-liquid interface was evaluated. In cultured mucosa explants from the surfaces of the ventral turbinates and septum of the pig nose no changes in cell morphology and viability were observed up to 72 h. MRSA colonization on the explants was evaluated followed for three MRSA ST398 isolates for 180 minutes. The explants were incubated with 3×10(8 CFU/ml in PBS for 2 h to allow bacteria to adhere to the explants surface. Next the explants were washed and in the first 30 minutes post adhering time, a decline in the number of CFU was observed for all MRSA. Subsequently, the isolates showed either: bacterial growth, no growth, or a further reduction in bacterial numbers. The MRSA were either localized as clusters between the cilia or as single bacteria on the cilia surface. No morphological changes in the epithelium layer were observed during the incubation with MRSA. We conclude that porcine nasal mucosa explants are a valuable ex vivo model to unravel the interaction of MRSA with nasal tissue.

  17. Effect of vasoactive intestinal peptide on pulmonary surfactants phospholipid synthesis in lung explants

    Institute of Scientific and Technical Information of China (English)

    Lian LI; Zi-qiang LUO; Fu-wen ZHOU; Dan-dan FENG; Cha-xiang GUAN; Chang-qing ZHANG; Xiu-hong SUN

    2004-01-01

    AIM: To investigate the effect of vasoactive intestinal peptide (VIP) on pulmonary surfactants (PS) phospholipid synthesis in cultured lung explants. METHODS: Lung explants were cultured with serum-free medium, [methyl-3H]choline incorporation, total phospholipid, phosphatidylcholine, activity of choline-phosphate cytidylyltransferase (CCT) and CCTα mRNA level in lung explants were determined. RESULTS: (1) VIP (10-10-10-7 mol/L) for 16 h promoted [methyl-3H]choline incorporation in dose dependence and VIP (10-8 mol/L) for 2 h-16 h promoted [methylz3H]choline incorporation in time dependence. (2) VIP (10-8 mol/L) enhanced the contents of total phospholipidsand phosphatidylcholine in lung explants. (3) VIP (10-10-10-7 mol/L) elevated microsomal CCT activity of lung explants in dose dependence. (4) VIP (10-8 mol/L) increased expression of CCTα mRNA in lung explants and alveolar type Ⅱ cells (ATII). (5) [D-P-Cl-Phe(6)-Leu(17)]-VIP (10-6 mol/L), a VIP receptors antagonist, abolished the increase of [3H]choline incorporation, microsomal CCT activity and CCTα mRNA level induced by VIP (10-8 mol/L) in lung explants. CONCLUSION: VIP could enhance synthesis of phosphatidylcholine, the major component of pulmonary surfactants by enhancing microsomal CCT activity and CCTα mRNA level via VIP receptormediated pathway.

  18. Culturing Schwann Cells from Neonatal Rats by Improved Enzyme Digestion Combined with Explants-culture Method.

    Science.gov (United States)

    Liu, Di; Liang, Xiao-Chun; Zhang, Hong

    2016-08-01

    Objective To develop an improved method for culturing Schwann cells(SCs) by using both enzyme digestion and explants-culture approaches and compared with traditional explants-culture method and general hemi-explants-culture method. Methods Bilaterally sciatic nerves and brachial plexus nerves were dissected from 3 to 5-day-old neonatal SD rats and explants-culture method,general hemi-explants-culture method,and improved enzyme digestion combined with explants-culture method were adopted to culture SCs,respectively. SCs were digested and passaged after 7 days in culture and counted under the microscope. The purity of SCs was identified by S-100 immunofluorescence staining. Results The SCs of improved method group grew fastest and the total number of cells obtained was(1.85±0.13)×10(6);the SCs of the hemi-explants-culture method group grew slower than the improved method group and the total number of cells obtained was (1.10±0.10)×10(6);the SCs of the explants-culture method group grew slowest and the total number of cells obtained was (0.77±0.03)×10(6).The total number of cells obtained showed significant difference among the three groups(Pculture method group,and (74.50±4.23)% in the explants-culture method group(Pculture method can obtain sufficient amount of high-purity SCs in a short time and thus may be applied in further research on peripheral nerve regeneration.

  19. Genome-wide transcriptome and expression profile analysis of Phalaenopsis during explant browning.

    Directory of Open Access Journals (Sweden)

    Chuanjun Xu

    Full Text Available Explant browning presents a major problem for in vitro culture, and can lead to the death of the explant and failure of regeneration. Considerable work has examined the physiological mechanisms underlying Phalaenopsis leaf explant browning, but the molecular mechanisms of browning remain elusive. In this study, we used whole genome RNA sequencing to examine Phalaenopsis leaf explant browning at genome-wide level.We first used Illumina high-throughput technology to sequence the transcriptome of Phalaenopsis and then performed de novo transcriptome assembly. We assembled 79,434,350 clean reads into 31,708 isogenes and generated 26,565 annotated unigenes. We assigned Gene Ontology (GO terms, Kyoto Encyclopedia of Genes and Genomes (KEGG annotations, and potential Pfam domains to each transcript. Using the transcriptome data as a reference, we next analyzed the differential gene expression of explants cultured for 0, 3, and 6 d, respectively. We then identified differentially expressed genes (DEGs before and after Phalaenopsis explant browning. We also performed GO, KEGG functional enrichment and Pfam analysis of all DEGs. Finally, we selected 11 genes for quantitative real-time PCR (qPCR analysis to confirm the expression profile analysis.Here, we report the first comprehensive analysis of transcriptome and expression profiles during Phalaenopsis explant browning. Our results suggest that Phalaenopsis explant browning may be due in part to gene expression changes that affect the secondary metabolism, such as: phenylpropanoid pathway and flavonoid biosynthesis. Genes involved in photosynthesis and ATPase activity have been found to be changed at transcription level; these changes may perturb energy metabolism and thus lead to the decay of plant cells and tissues. This study provides comprehensive gene expression data for Phalaenopsis browning. Our data constitute an important resource for further functional studies to prevent explant browning.

  20. Genome-wide transcriptome and expression profile analysis of Phalaenopsis during explant browning.

    Science.gov (United States)

    Xu, Chuanjun; Zeng, Biyu; Huang, Junmei; Huang, Wen; Liu, Yumei

    2015-01-01

    Explant browning presents a major problem for in vitro culture, and can lead to the death of the explant and failure of regeneration. Considerable work has examined the physiological mechanisms underlying Phalaenopsis leaf explant browning, but the molecular mechanisms of browning remain elusive. In this study, we used whole genome RNA sequencing to examine Phalaenopsis leaf explant browning at genome-wide level. We first used Illumina high-throughput technology to sequence the transcriptome of Phalaenopsis and then performed de novo transcriptome assembly. We assembled 79,434,350 clean reads into 31,708 isogenes and generated 26,565 annotated unigenes. We assigned Gene Ontology (GO) terms, Kyoto Encyclopedia of Genes and Genomes (KEGG) annotations, and potential Pfam domains to each transcript. Using the transcriptome data as a reference, we next analyzed the differential gene expression of explants cultured for 0, 3, and 6 d, respectively. We then identified differentially expressed genes (DEGs) before and after Phalaenopsis explant browning. We also performed GO, KEGG functional enrichment and Pfam analysis of all DEGs. Finally, we selected 11 genes for quantitative real-time PCR (qPCR) analysis to confirm the expression profile analysis. Here, we report the first comprehensive analysis of transcriptome and expression profiles during Phalaenopsis explant browning. Our results suggest that Phalaenopsis explant browning may be due in part to gene expression changes that affect the secondary metabolism, such as: phenylpropanoid pathway and flavonoid biosynthesis. Genes involved in photosynthesis and ATPase activity have been found to be changed at transcription level; these changes may perturb energy metabolism and thus lead to the decay of plant cells and tissues. This study provides comprehensive gene expression data for Phalaenopsis browning. Our data constitute an important resource for further functional studies to prevent explant browning.

  1. Clinical evaluation of an acellular dermal allograft for increasing the zone of attached gingiva.

    Science.gov (United States)

    Shulman, J

    1996-03-01

    Grafting with autogenous tissue or freeze-dried skin is the generally accepted method for increasing and/or restoring the width of attached gingiva. This article describes the periodontal use of an acellular dermal allograft previously available for treating burn patients. When used as a gingival graft, this new dermal allograft has major potential advantages over the previously available periodontal graft materials, including improved color and contour match, elimination of multiple surgeries, and unlimited availability. The technique and results of acellular dermal grafting are presented and discussed. The learning objective of this article is to describe the principles and the clinical procedure of this technique. Several cases are used to illustrate this technique.

  2. Simulating strange attraction of acellular slime mould Physarum polycephaum to herbal tablets

    OpenAIRE

    Adamatzky, Andrew

    2012-01-01

    Plasmodium of acellular slime mould Physarum polycephalum exhibits traits of wave-like behaviour. The plasmodium's behaviour can be finely tuned in laboratory experiments by using herbal tablets. A single tablet acts as a fixed attractor: plasmodium propagates towards the tablet, envelops the tablet with its body and stays around the tablet for several days. Being presented with several tablets the plasmodium executes limit cycle like motions. The plasmodium performs sophisticated routines of...

  3. Randomized controlled trial of minimally invasive surgery using acellular dermal matrix for complex anorectal fistula

    Institute of Scientific and Technical Information of China (English)

    Ma-Mu-Ti-Jiang; A; ba-bai-ke-re; Er-Ha-Ti; Ai

    2010-01-01

    AIM: To compare the efficacy and safety of acellular dermal matrix (ADM) bioprosthetic material and endorectal advancement flap (ERAF) in treatment of complex anorectal fistula. METHODS: Ninety consecutive patients with complex anorectal fistulae admitted to Anorectal Surgical Department of First Affi liated Hospital, Xinjiang Medical University from March 2008 to July 2009, were enrolled in this study. Complex anorectal fistula was diagnosed following its clinical, radiographic, or endoscopic diagnostic cr...

  4. Porcine acellular dermis-based breast reconstruction: complications and outcomes following adjuvant radiotherapy

    OpenAIRE

    Ng, Cho Ee; Pieri, Andrew; Fasih, Tarannum

    2015-01-01

    Background Acellular dermal matrices (ADM) such as StratticeTM are increasingly used in UK during implant-based reconstruction. However, there are mixed opinions regarding the compatibility of radiotherapy treatment in pre- and post-reconstructed breasts. The aims of this study are to audit the rates of radiation induced complications in patients who underwent breast reconstruction using StratticeTM and establish whether there is an association between timing of radiotherapy and complication ...

  5. Biomimetic acellular detoxified glutaraldehyde cross-linked bovine pericardium for tissue engineering

    Energy Technology Data Exchange (ETDEWEB)

    Mathapati, Santosh; Bishi, Dillip Kumar [Stem Cell and Molecular Biology Laboratory, Department of Biotechnology, Indian Institute of Technology Madras, Chennai (India); Frontier Lifeline Pvt Ltd. and Dr. K. M. Cherian Heart Foundation, Mogappair, Chennai (India); Healthcare and Energy Materials Laboratory, NUSNNI, Faculty of Engineering, National University of Singapore (Singapore); Guhathakurta, Soma [Departmet of Engineering Design, Indian Institute of Technology Madras, Chennai (India); Cherian, Kotturathu Mammen [Frontier Lifeline Pvt Ltd. and Dr. K. M. Cherian Heart Foundation, Mogappair, Chennai (India); Venugopal, Jayarama Reddy; Ramakrishna, Seeram [Healthcare and Energy Materials Laboratory, NUSNNI, Faculty of Engineering, National University of Singapore (Singapore); Verma, Rama Shanker, E-mail: vermars@iitm.ac.in [Stem Cell and Molecular Biology Laboratory, Department of Biotechnology, Indian Institute of Technology Madras, Chennai (India)

    2013-04-01

    Glutaraldehyde (GLUT) processing, cellular antigens, calcium ions in circulation, and phospholipids present in the native tissue are predominantly responsible for calcification, degeneration, and lack of natural microenvironment for host progenitor cell migration in tissue implants. The study presents an improved methodology for adhesion and proliferation of endothelial progenitor cells (EPCs) without significant changes in biomechanical and biodegradation properties of the processed acellular bovine pericardium. The anti-calcification potential of the processed tissue was enhanced by detoxification of GLUT-cross-linked bovine pericardium by decellularization, pretreating it with ethanol or removing the free aldehydes by citric acid treatment and lyophilization. The treated tissues were assessed for biomechanical properties, GLUT ligand quantification, adhesion, proliferation of EPCs, and biodegradability. The results indicate that there was no significant change in biomechanical properties and biodegradability when enzymatic hydrolysis (p > 0.05) is employed in detoxified acellular GLUT cross-linked tissue (DBP–G–CA–ET), compared with the native detoxified GLUT cross-linked bovine pericardium (NBP–G–CA–ET). DBP–G–CA–ET exhibited a significant (p > 0.05) increase in the viability of EPCs and cell adhesion as compared to acellular GLUT cross-linked bovine pericardium (p < 0.05). Lyophilized acellular detoxified GLUT cross-linked bovine pericardium, employed in our study as an alternative to conventional GLUT cross-linked bovine pericardium, might provide longer durability and better biocompatibility, and reduce calcification. The developed bovine pericardium patches could be used in cardiac reconstruction and repair, arteriotomy, soft tissue repair, and general surgical procedures with tissue regeneration dimensions. - Highlights: ► We improved the quality of patch biomaterial for cardiovascular surgical procedures. ► Bovine pericardium was

  6. Explant culture of rat colon: A model system for studying metabolism of chemical carcinogens

    DEFF Research Database (Denmark)

    Autrup, Herman; Stoner, G.D.; Jackson, F.

    1978-01-01

    An explant culture system has been developed for the long-term maintenance of colonic tissue from the rat. Explants of 1 cm2 in size were placed in tissue-culture dishes to which was added 2 ml of CMRL-1066 medium supplemented with glucose, hydrocortisone, beta-retinyl acetate, and either 2....... The explants were incubated at 30 degrees C. The viability of the tissue was measured both by incorporation of specific precursors into cellular macromolecules and by monitoring of tissue morphology with light and electron microscopy. Cultured rat colon was able to metabolize benzo[alpha]pyrene, 7...

  7. Preparation and characterization of an acellular bovine pericardium intended for manufacture of valve bioprostheses.

    Science.gov (United States)

    Goissis, Gilberto; Giglioti, Aparecida de Fátima; Braile, Domingo Marcolino

    2011-05-01

    Major problems with biological heart valves post-implantation are associated with progressive structural deterioration and calcification attributed to glutaraldehyde processing, dead cells, and cell fragments present in the native tissue. In spite of these problems, glutaraldehyde still is the reagent of choice. The results with acellular matrix xenograft usually prepared by detergent treatment in association with enzymes are rather conflicting because while preserving mechanical properties, tissue morphology and collagen structure are process dependent. This work describes a chemical approach for the preparation of an acellular bovine pericardium matrix intended for the manufacture of heart valve bioprostheses. Cell removal was performed by an alkaline extraction in the presence of calcium salts for periods ranging from 6 to 48 h. The results showed that cell removal was achieved after 12 h, with swelling and negative charge increasing with processing time. Nevertheless, collagen fibril structure, ability to form fibrils, and stability to collagenase were progressive after 24-h processing. There was no denaturation of the collagen matrix. A process is described for the preparation of acellular bovine pericardium matrices with preserved fibril structure and morphology for the manufacture of cardiac valve bioprostheses and may be used in other applications for tissue reconstruction. © 2011, Copyright the Authors. Artificial Organs © 2011, International Center for Artificial Organs and Transplantation and Wiley Periodicals, Inc.

  8. Development and characterization of a full-thickness acellular porcine cornea matrix for tissue engineering.

    Science.gov (United States)

    Du, Liqun; Wu, Xinyi

    2011-07-01

    Our aim was to produce a natural, acellular matrix from porcine cornea for use as a scaffold in developing a tissue-engineered cornea replacement. Full-thickness, intact porcine corneas were decellularized by immersion in 0.5% (wt/vol) sodium dodecyl sulfate. The resulting acellular matrices were then characterized and examined specifically for completeness of the decellularization process. Histological analyses of decellularized corneal stromas showed that complete cell and α-Gal removal was achieved, while the major structural proteins including collagen type I and IV, laminin, and fibronectin were retained. DAPI staining did not detect any residual DNA within the matrix, and the DNA contents, which reflect the presence of cellular materials, were significantly diminished in the decellularized cornea. The collagen content of the decellularized cornea was well maintained compared with native tissues. Uniaxial tensile testing indicated that decellularization did not significantly compromise the ultimate tensile strength of the tissue (P > 0.05). In vitro cytotoxicity assays using rabbit corneal fibroblast cultures excluded the presence of soluble toxins in the biomaterial. In vivo implantation to rabbit interlamellar stromal pockets showed good biocompability. In summary, a full-thickness natural acellular matrix retaining the major structural components and strength of the cornea has been successfully developed. The matrix is biocompatible with cornea-derived cells and has potential for use in corneal transplantation and tissue-engineering applications. © 2011, Copyright the Authors. Artificial Organs © 2011, International Center for Artificial Organs and Transplantation and Wiley Periodicals, Inc.

  9. Histopathology of explanted AlphaCor due to keratoprosthesis extrusion.

    Science.gov (United States)

    Caporossi, Aldo; Mazzotta, Cosimo; Balestrazzi, Angelo; Occhini, Rossella; Toti, Paolo; Traversi, Claudio

    2006-07-01

    AlphaCor keratoprosthesis (KPro) is a new-concept poly (2-hydroxyethyl methacrylate) one-piece KPro that makes possible a two-step implantation technique easy to perform with a short learning curve. In literature an 18% incidence of AlphaCor removal due to melting complications is reported. The histopathology of corneal tissue removed during a re-operation while bearing an AlphaCor KPro has previously been described in the literature only in one report. Herein, the first histological features of an AlphaCor-corneal complex explanted because of KPro extrusion is described. The histopathology of the AlphaCor-corneal complex is characterized by mild inflammation in the corneal tissues, limited to the region surrounding the anteriorized and extruded part of the KPro. It is not possible to fully understand the mechanisms that trigger the device extrusion. One possible explanation could be a dislocation of the prosthesis in the corneal pocket due to the untied fixation stitch. Another explanation could be a foreign body reaction induced by KPro.

  10. Tropic responses of potato single-node explant cultures

    Directory of Open Access Journals (Sweden)

    Vinterhalter D.

    2012-01-01

    Full Text Available A special in-vitro protocol was elaborated enabling the production of potato single-node explant plantlets that can be used as objects for tropic studies. In light-grown plantlets, achievement of a full (90° phototropic (PT curvature required 75 to 120 min of continuous unilateral blue light irradiation or 120-135 min of gravitropic stimulation (GT. Time-lapse photography revealed that the curves describing PT and GT bending have a sigmoid shape. Continuous BL irradiation was necessary for the induction of continuous PT bending. If the BL was turned off after 30-50 min of PT stimulation, the bending gradually decreased and stopped in darkness after 25.0 ± 2.0 min. Within this period, curvature increased by 15.5 ± 1.5°. When the BL was turned off upon completion of PT bending (when the plantlets reached an angle of 90°, the plantlets entered the phase of fast straightening. The 90° PT curvature was significantly exaggerated in darkness by turning the jars from a vertical to horizontal position providing 120.74 ± 2.5° as the final curvature angle after two more hours in darkness.

  11. Estabelecimento in vitro de explantes de três cultivares de bananeira Explants establishment in vitro of three banana cultivars

    Directory of Open Access Journals (Sweden)

    Silvia Nietsche

    2006-06-01

    Full Text Available O objetivo do presente trabalho foi avaliar dois protocolos de desinfestação na micropropagação em três cultivares de bananeira. Utilizaram-se mudas das cultivares de bananeira Prata Anã, FHIA 18 e SH3640. Os protocolos 1 e 2 de desinfestação (Des. 1 e Des. 2 foram realizados utilizando-se os seguintes produtos: solução fungicida de Derosol, álcool comercial, solução de hipoclorito de sódio e de hipoclorito de cálcio e tween 20, apresentando variações na concentração dos produtos em cada um dos protocolos. As avaliações para a porcentagem de contaminação foram realizadas diariamente por meio do número total de tubos contaminados e não contaminados. O delineamento foi em blocos casualizados, com quatro repetições, em um sistema fatorial. As contaminações que ocorreram foram causadas exclusivamente por bactérias. Independentemente do tratamento utilizado, os índices de contaminação foram superiores a 29% para as três cultivares testadas. Estes resultados indicam a necessidade de adequar novas metodologias de assepsias de explantes de bananeira.The purpose of this work was to evaluate two protocols of disinfestations in the micropropagation on young plants of three banana cultivars: Prata-Anã, FHIA 18 and SH3640. Protocols 1 and 2 of disinfestations (Dis. 1 and Dis. 2 were carried out using the products: fungicidal solution of Derosol, commercial alcohol, solution of sodium hypo chlorite and calcium hypo chlorite and tween 20, presenting variations in their concentration in every one of the protocols. The evaluations for contamination rate were taken daily by means of the total number of contaminated and non-contaminated tubes. A random block design in a factorial scheme with four repetitions was used. Exclusively bacteria had caused all contamination. Independently of the used treatment the contamination levels had been up to 29% for all the three cultivars tested. These results indicate the necessity of

  12. Mechanically loaded ex vivo bone culture system 'Zetos': Systems and culture preparation

    Directory of Open Access Journals (Sweden)

    C M Davies

    2006-04-01

    Full Text Available This paper introduces the culture preparation of ovine, bovine and human cancellous bone cores to be used in an explants model Zetos. The three dimensional (3D bone cores were prepared and evaluated for all three animals. Bone cells in vivo constantly interact with each other, migratory cells, surrounding extracellular matrix (ECM and interstitial fluid in a microenvironment, which continuously responds to various endogenous and exogenous stimuli. The Zetos system was designed to culture and mechanically load viable cancellous bone explants in their near natural microenvironment. This 3D ex vivo system bridges the current gap between in vitro and in vivo methods. One aim of this work was to compare the macro and micro-architecture of ovine, bovine and human cancellous bone tissue in preparation for culture within the Zetos system in order to determine the optimal source of experimental material. A second aim was to optimise the preparations of the bone cores as well as develop techniques involved during tissue maintenance. Bone core response was visualised using histological and immunohistochemical methods. The results demonstrate that cancellous bone explants vary greatly in trabecular density and bone volume depending on species, age and location. Sheep and human samples displayed the greatest variation between bones cores when compared to bovine. Even cores taken from the same animal possessed very different characteristics. The histology demonstrated normal bone and cell structure after the core preparation. Immunohistochemistry results demonstrated antigen retention after preparation methods.

  13. Muscle acellular scaffold as a biomaterial: effects on C2C12 cell differentiation andinteraction with the murine host environment

    Directory of Open Access Journals (Sweden)

    Perniconi eBarbara

    2014-09-01

    Full Text Available The extracellular matrix (ECM of decellularized organs possesses the characteristics of the idealtissue-engineering scaffold (i.e. histocompatibility, porosity, degradability, non-toxicity. Wepreviously observed that the muscle acellular scaffold (MAS is a pro-myogenic environment invivo. In order to determine whether MAS, which is basically muscle ECM, behaves as a myogenicenvironment, regardless of its location, we analysed MAS interaction with both muscle and nonmusclecells and tissues, to assess the effects of MAS on cell differentiation. Bone morphogeneticprotein treatment of C2C12 cells cultured within MAS induced osteogenic differentiation in vitro,thus suggesting that MAS does not irreversibly commit cells to myogenesis. In vivo MAS supportedformation of nascent muscle fibres when replacing a muscle (orthotopic position. However,heterotopically grafted MAS did not give rise to muscle fibres when transplanted within the renalcapsule. Also, no muscle formation was observed when MAS was transplanted under the xiphoidprocess, in spite of the abundant presence of cells migrating along the laminin-based MASstructure. Taken together, our results suggest that MAS itself is not sufficient to induce myogenicdifferentiation. It is likely that the pro-myogenic environment of MAS is not strictly related to theintrinsic properties of the muscle scaffold (e.g. specific muscle ECM proteins. Indeed, it is morelikely that myogenic stem cells colonising MAS recognise a muscle environment that ultimatelyallows terminal myogenic differentiation. In conclusion, MAS may represent a suitableenvironment for muscle and non-muscle 3D constructs characterised by a highly organised structurewhose relative stability promotes integration with the surrounding tissues. Our work highlights theplasticity of MAS, suggesting that it may be possible to consider MAS for a wider range of tissueengineering applications than the mere replacement of volumetric muscle loss.

  14. Tissue-engineered graft constructed by self-derived cells and heterogeneous acellular matrix

    Institute of Scientific and Technical Information of China (English)

    HUANG Hui-min; WU Shao-feng; REN Hong

    2006-01-01

    Background: Endothelial and smooth muscle cells were used as seeding cells and heterogeneous acellularized matrix was used as scaffold to construct the tissue-engineered graft. Methods: A 2 weeks piglet was selected as a donor of seeding cells. Two-centimetre length of common carotid artery was dissected. Endothelial cells and smooth muscle cells were harvested by trypsin and collagenase digestion respectively. The isolated cells were cultured and expanded using routine cell culture technique.An adult sheep was used as a donor of acellularized matrix. The thoracic aorta was harvested and processed by a multi-step decellularizing technique to remove the original cells and preserve the elastic and collagen fibers. The cultured smooth muscle cells and endothelial cells were then seeded to the acellularized matrix and incubated in vitro for another 2 weeks. The cell seeded graft was then transplanted to the cell-donated piglet to substitute part of the native pulmonary artery. Results: The cultured cells from piglet were characterized as endothelial cells by the presence of specific antigens vWF and CD31, and smooth muscle cells by the presence of specific antigen α-actin on the cell surface respectively with immunohistochemical technique. After decellularizing processing for the thoracic aorta from sheep, all the cellular components were extracted and elastic and collagen fibers kept their original morphology and structure. The maximal load of acellular matrix was decreased and 20% lower than that of untreated thoracic aorta, but the maximal tensions between them were not different statistically and they had similar load-tension curves. Three months after transplantation, the animal was sacrificed and the graft was removed for observation. The results showed that the inner surfaces of the graft were smooth, without thrombosis and calcification. Under microscopy, a great number of growing cells could be seen and elastic and collagen fibers were abundant. Conclusion

  15. Callogenesis in stem explants of Eucalyptus grandis (Hill ex Maiden showing resilience in oxide calluses

    Directory of Open Access Journals (Sweden)

    Regina Paula Willemen Pereira

    Full Text Available Callogenesis was achieved using growth regulators at different concentrations to obtain the best characteristics regarding the calluses texture aspect with the main objective to produce friable calluses from stem explants of Eucalyptus grandis. Stem segments of approximately 1 cm were excised from 30-day seedlings. The treatments were prepared based on the MS medium added with TDZ alone or combined with AIA; 2,4-D; ANA. Each treatment consisted of five repetitions with five explants per plate. After 210 days of in vitro culture, the calluses were evaluated visually and by manipulation for texture. The explants that did not produce calluses were those exposed to treatments with 2,4-D 50.0 µM; TDZ 3.0 µM and in absence of a growth regulator (control.The best protocols for maintenance of friable calluses of stem explants of Eucalyptus grandis under the conditions performed in this work were those using only TDZ.

  16. In vitro direct regeneration in mint from different explants on half ...

    African Journals Online (AJOL)

    STORAGESEVER

    2009-09-15

    Sep 15, 2009 ... e.g. Limonene, cineol, polygon, piperitone in the genus mentha, have ... Each treatment had 10 replicates containing 2 explants and all ex- periments were ... plant type and exogenous level of growth regulator in the medium.

  17. Organ explant culture of neonatal rat ventricles: a new model to study gene and cell therapy.

    Directory of Open Access Journals (Sweden)

    A Dénise den Haan

    Full Text Available Testing cardiac gene and cell therapies in vitro requires a tissue substrate that survives for several days in culture while maintaining its physiological properties. The purpose of this study was to test whether culture of intact cardiac tissue of neonatal rat ventricles (organ explant culture may be used as a model to study gene and cell therapy. We compared (immuno histology and electrophysiology of organ explant cultures to both freshly isolated neonatal rat ventricular tissue and monolayers. (Immuno histologic studies showed that organ explant cultures retained their fiber orientation, and that expression patterns of α-actinin, connexin-43, and α-smooth muscle actin did not change during culture. Intracellular voltage recordings showed that spontaneous beating was rare in organ explant cultures (20% and freshly isolated tissue (17%, but common (82% in monolayers. Accordingly, resting membrane potential was -83.9±4.4 mV in organ explant cultures, -80.5±3.5 mV in freshly isolated tissue, and -60.9±4.3 mV in monolayers. Conduction velocity, measured by optical mapping, was 18.2±1.0 cm/s in organ explant cultures, 18.0±1.2 cm/s in freshly isolated tissue, and 24.3±0.7 cm/s in monolayers. We found no differences in action potential duration (APD between organ explant cultures and freshly isolated tissue, while APD of monolayers was prolonged (APD at 70% repolarization 88.8±7.8, 79.1±2.9, and 134.0±4.5 ms, respectively. Organ explant cultures and freshly isolated tissue could be paced up to frequencies within the normal range for neonatal rat (CL 150 ms, while monolayers could not. Successful lentiviral (LV transduction was shown via Egfp gene transfer. Co-culture of organ explant cultures with spontaneously beating cardiomyocytes increased the occurrence of spontaneous beating activity of organ explant cultures to 86%. We conclude that organ explant cultures of neonatal rat ventricle are structurally and electrophysiologically similar

  18. An early look at the Organ Procurement and Transplantation Network explant pathology form data.

    Science.gov (United States)

    Harper, Ann M; Edwards, Erick; Washburn, W Kenneth; Heimbach, Julie

    2016-06-01

    In April 2012, the Organ Procurement and Transplantation Network (OPTN) implemented an online explant pathology form for recipients of liver transplantation who received additional wait-list priority for their diagnosis of hepatocellular carcinoma (HCC). The purpose of the form was to standardize the data being reported to the OPTN, which had been required since 2002 but were submitted to the OPTN in a variety of formats via facsimile. From April 2012 to December 2014, over 4500 explant forms were submitted, allowing for detailed analysis of the characteristics of the explanted livers. Data from the explant pathology forms were used to assess agreement with pretransplant imaging. Explant data were also used to assess the risk of recurrence. Of those with T2 priority, 55.7% were found to be stage T2 on explant. Extrahepatic spread (odds ratio [OR] = 6.8; P < 0.01), poor tumor differentiation (OR = 2.8; P < 0.01), microvascular invasion (OR = 2.6; P < 0.01), macrovascular invasion (OR = 3.2; P < 0.01), and whether the Milan stage based on the number and size of tumors on the explant form was T4 (OR = 2.4; P < 0.01) were the strongest predictors of recurrence. In conclusion, this analysis confirms earlier findings that showed an incomplete agreement between pretransplant imaging and posttransplant pathology in terms of HCC staging, though the number of patients with both no pretransplant treatment and no tumor in the explant was reduced from 20% to <1%. In addition, several factors were identified (eg, tumor burden, age, sex, region, ablative therapy, alpha-fetoprotein, Milan stage, vascular invasion, satellite lesions, etc.) that were predictive of HCC recurrence, allowing for more targeted surveillance of high-risk recipients. Continued evaluation of these data will help shape future guidelines or policy recommendations. Liver Transplantation 22 757-764 2016 AASLD.

  19. In vitro direct shoot regeneration from proximal, middle and distal segment of Coleus forskohlii leaf explants

    OpenAIRE

    Krishna, Gaurav; P. Sairam Reddy; Anoop Nair, N.; P. W. Ramteke; Bhattacharya, P. S.

    2010-01-01

    Coleus forskohlii is an endangered multipurpose medicinal plant that has widespread applications. In spite of this, there have been few attempts to propagate its cultivation in India. The present communication presents an in vitro rapid regeneration method using leaf explants of Coleus forskohlii through direct organogenesis. Leaf explants that were excised into three different segments i.e. proximal (P), middle (M) and distal (D) were cultured on Murashige and Skoog (MS) basal medium supplem...

  20. Harms and responsibilities associated with battery-operated implants (BOI): who controls postmortem explantation?

    Science.gov (United States)

    Bramstedt, Katrina A

    2013-01-01

    The postmortem issues raised by battery-operated implants (BOI) are complex and issues of consent, setting (clinical vs research), and environmental risks have received little attention in bioethics literature. Analyzing the issues, the following are argued: (1) Patients receiving BOIs should sign a consent form that includes a requirement for postmortem explant of the device; (2) BOI consent forms should require the explanted devices be returned to their manufacturers for Returned Product Analysis; (3) Failure to explant and analyze devices from the research setting fails the research goal of generation of knowledge for the benefit of future patients; (4) Failure to explant and analyze devices from the clinical setting allows product defects to be potentially hidden from patients, families, clinicians, manufacturers, and regulatory agencies; (5) Bodies buried with BOIs potentially harm the environment; (6) Religious or philosophical objections to autopsy should not supersede the duty to explant and analyze BOIs; (7) The concepts herein for BOIs could potentially extend to non-BOI if the device has failure modes that can lead to a potentially life-threatening event or can cause permanent debilitating health issues, and the burial or cremation of the device poses environmental harm. In these situations, neither the patient (premortem) nor family (postmortem) should have the right to refuse explant.

  1. In Vitro Spermatogenesis in Explanted Adult Mouse Testis Tissues.

    Science.gov (United States)

    Sato, Takuya; Katagiri, Kumiko; Kojima, Kazuaki; Komeya, Mitsuru; Yao, Masahiro; Ogawa, Takehiko

    2015-01-01

    Research on in vitro spermatogenesis is important for elucidating the spermatogenic mechanism. We previously developed an organ culture method which can support spermatogenesis from spermatogonial stem cells up to sperm formation using immature mouse testis tissues. In this study, we examined whether it is also applicable to mature testis tissues of adult mice. We used two lines of transgenic mice, Acrosin-GFP and Gsg2-GFP, which carry the marker GFP gene specific for meiotic and haploid cells, respectively. Testis tissue fragments of adult GFP mice, aged from 4 to 29 weeks old, which express GFP at full extension, were cultured in medium supplemented with 10% KSR or AlbuMAX. GFP expression decreased rapidly and became the lowest at 7 to 14 days of culture, but then slightly increased during the following culture period. This increase reflected de novo spermatogenesis, confirmed by BrdU labeling in spermatocytes and spermatids. We also used vitamin A-deficient mice, whose testes contain only spermatogonia. The testes of those mice at 13-21 weeks old, showing no GFP expression at explantation, gained GFP expression during culturing, and spermatogenesis was confirmed histologically. In addition, the adult testis tissues of Sl/Sld mutant mice, which lack spermatogenesis due to Kit ligand mutation, were cultured with recombinant Kit ligand to induce spermatogenesis up to haploid formation. Although the efficiency of spermatogenesis was lower than that of pup, present results showed that the organ culture method is effective for the culturing of mature adult mouse testis tissue, demonstrated by the induction of spermatogenesis from spermatogonia to haploid cells.

  2. Human neural progenitor cells promote photoreceptor survival in retinal explants.

    Science.gov (United States)

    Englund-Johansson, Ulrica; Mohlin, Camilla; Liljekvist-Soltic, Ingela; Ekström, Per; Johansson, Kjell

    2010-02-01

    Different types of progenitor and stem cells have been shown to provide neuroprotection in animal models of photoreceptor degeneration. The present study was conducted to investigate whether human neural progenitor cells (HNPCs) have neuroprotective properties on retinal explants models with calpain- and caspase-3-dependent photoreceptor cell death. In the first experiments, HNPCs in a feeder layer were co-cultured for 6 days either with postnatal rd1 mouse or normal rat retinas. Retinal histological sections were used to determine outer nuclear layer (ONL) thickness, and to detect the number of photoreceptors with labeling for calpain activity, cleaved caspase-3 and TUNEL. The ONL thickness of co-cultured rat and rd1 retinas was found to be almost 10% and 40% thicker, respectively, compared to controls. Cell counts of calpain activity, cleaved caspase-3 and TUNEL labeled photoreceptors in both models revealed a 30-50% decrease when co-cultured with HNPCs. The results represent significant increases of photoreceptor survival in the co-cultured retinas. In the second experiments, for an identification of putative survival factors, or a combination of them, a growth factor profile was performed on conditioned medium. The relative levels of various growth factors were analyzed by densitometric measurements of growth factor array membranes. Following growth factors were identified as most potential survival factors; granulocyte colony stimulating factor (G-CSF), granulocyte-macrophage colony stimulating factor (GMCSF), insulin-like growth factor II (IGF-II), neurotrophic factor 3 (NT-3), placental growth factor (PIGF), transforming growth factors (TGF-beta1 and TGF-beta2) and vascular endothelial growth factor (VEGF-D). HNPCs protect both against calpain- and caspase-3-dependent photoreceptor cell death in the rd1 mouse and against caspase-3-dependent photoreceptor cell death in normal rat retinas in vitro. The protective effect is possibly achieved by a variety of

  3. TGF-ß1 enhances the BMP-2-induced chondrogenesis of bovine synovial explants and arrests downstream differentiation at an early stage of hypertrophy.

    Directory of Open Access Journals (Sweden)

    Nahoko Shintani

    Full Text Available BACKGROUND: Synovial explants furnish an in-situ population of mesenchymal stem cells for the repair of articular cartilage. Although bone morphogenetic protein 2 (BMP-2 induces the chondrogenesis of bovine synovial explants, the cartilage formed is neither homogeneously distributed nor of an exclusively hyaline type. Furthermore, the downstream differentiation of chondrocytes proceeds to the stage of terminal hypertrophy, which is inextricably coupled with undesired matrix mineralization. With a view to optimizing BMP-2-induced chondrogenesis, the modulating influences of fibroblast growth factor 2 (FGF-2 and transforming growth factor beta 1 (TGF-ß1 were investigated. METHODOLOGY/PRINCIPAL FINDINGS: Explants of bovine calf metacarpal synovium were exposed to BMP-2 (200 ng/ml for 4 (or 6 weeks. FGF-2 (10 ng/ml or TGF-ß1 (10 ng/ml was introduced at the onset of incubation and was present either during the first week of culturing alone or throughout its entire course. FGF-2 enhanced the BMP-2-induced increase in metachromatic staining for glycosaminoglycans (GAGs only when it was present during the first week of culturing alone. TGF-ß1 enhanced not only the BMP-2-induced increase in metachromasia (to a greater degree than FGF-2, but also the biochemically-assayed accumulation of GAGs, when it was present throughout the entire culturing period; in addition, it arrested the downstream differentiation of cells at an early stage of hypertrophy. These findings were corroborated by an analysis of the gene- and protein-expression levels of key cartilaginous markers and by an estimation of individual cell volume. CONCLUSIONS/SIGNIFICANCE: TGF-ß1 enhances the BMP-2-induced chondrogenesis of bovine synovial explants, improves the hyaline-like properties of the neocartilage, and arrests the downstream differentiation of cells at an early stage of hypertrophy. With the prospect of engineering a mature, truly articular type of cartilage in the context of

  4. Evaluating acellular versus cellular perfusate composition during prolonged ex vivo lung perfusion after initial cold ischaemia for 24 hours.

    Science.gov (United States)

    Becker, Simon; Steinmeyer, Jasmin; Avsar, Murat; Höffler, Klaus; Salman, Jawad; Haverich, Axel; Warnecke, Gregor; Ochs, Matthias; Schnapper, Anke

    2016-01-01

    Normothermic ex vivo lung perfusion (EVLP) has developed as a powerful technique to evaluate particularly marginal donor lungs prior to transplantation. In this study, acellular and cellular perfusate compositions were compared in an identical experimental setting as no consensus has been reached on a preferred technique yet. Porcine lungs underwent EVLP for 12 h on the basis of an acellular or a cellular perfusate composition after 24 h of cold ischaemia as defined organ stress. During perfusion, haemodynamic and respiratory parameters were monitored. After EVLP, the lung condition was assessed by light and transmission electron microscopy. Aerodynamic parameters did not show significant differences between groups and remained within the in vivo range during EVLP. Mean oxygenation indices were 491 ± 39 in the acellular group and 513 ± 53 in the cellular group. Groups only differed significantly in terms of higher pulmonary artery pressure and vascular resistance in the cellular group. Lung histology and ultrastructure were largely well preserved after prolonged EVLP and showed only minor structural alterations which were similarly present in both groups. Prolonged acellular and cellular EVLP for 12 h are both feasible with lungs prechallenged by ischaemic organ stress. Physiological and ultrastructural analysis showed no superiority of either acellular or cellular perfusate composition.

  5. Bone Biopsy

    Science.gov (United States)

    ... News Physician Resources Professions Site Index A-Z Bone Biopsy Bone biopsy uses a needle and imaging ... the limitations of Bone Biopsy? What is a Bone Biopsy? A bone biopsy is an image-guided ...

  6. Indução de calos embriogênicos em explantes de cupuaçuzeiro Induction of embryogenics calli in cupuassu explants

    Directory of Open Access Journals (Sweden)

    Maria das Graças Rodrigues Ferreira

    2004-08-01

    Full Text Available Objetivou-se a indução de calos embriogênicos em cupuaçuzeiro, em função do tipo de explante e meio de cultura. Foram testados como explantes, segmentos cotiledonares e eixos embrionários divididos em três partes: região da plúmula, radícula e hipocótilo. Os explantes foram cultivados em 2 diferentes meios de cultura: 1 MS suplementado com 2,4-D (1 mg L-1 e Cinetina (0,25 mg L-1; 2 MS acrescido de ANA (5 mg L-1 e Cinetina (0,25 mg L-1. Constatou-se que a região do hipocótilo foi a parte mais responsiva do eixo embrionário, formando calos com aspecto branco e friável. As auxinas testadas nos meios não estimularam o processo embriogênico em calos de cupuaçuzeiro.It was studied the induction of embryogenics calli in cupuassu, in function of kind of explant and culture medium. Cotyledons segments and embryonic axes were tested and divided in three parts: region of plumule, radicule and hypocotile. The explants were cultivated in two different culture medium: 1 MS supplemented with 2,4-D (1 mg L-1 and Kinetin (0,25 mg L-1; 2 MS supplemented with NAA (5 mg L-1 and Kinetin (0,25 mg L-1. The hypocotile region demonstrated to be more responsive segment of the embryonic axe, forming callus with white and friable aspect. No somatic embryogenesis was evidenced in callus of cupuassu with auxines testeds in the medium.

  7. Bone Densitometry (Bone Density Scan)

    Science.gov (United States)

    ... News Physician Resources Professions Site Index A-Z Bone Densitometry (DEXA) Bone densitometry, also called dual-energy ... limitations of DEXA Bone Densitometry? What is a Bone Density Scan (DEXA)? Bone density scanning, also called ...

  8. Reactivation of HSV-1 following explant of tree shrew brain.

    Science.gov (United States)

    Li, Lihong; Li, Zhuoran; Li, Xin; Wang, Erlin; Lang, Fengchao; Xia, Yujie; Fraser, Nigel W; Gao, Feng; Zhou, Jumin

    2016-06-01

    Herpes Simplex Virus type I (HSV-1) latently infects peripheral nervous system (PNS) sensory neurons, and its reactivation leads to recurring cold sores. The reactivated HSV-1 can travel retrograde from the PNS into the central nervous system (CNS) and is known to be causative of Herpes Simplex viral encephalitis. HSV-1 infection in the PNS is well documented, but little is known on the fate of HSV-1 once it enters the CNS. In the murine model, HSV-1 genome persists in the CNS once infected through an ocular route. To gain more details of HSV-1 infection in the CNS, we characterized HSV-1 infection of the tree shrew (Tupaia belangeri chinensis) brain following ocular inoculation. Here, we report that HSV-1 enters the tree shrew brain following ocular inoculation and HSV-1 transcripts, ICP0, ICP4, and LAT can be detected at 5 days post-infection (p.i.), peaking at 10 days p.i. After 2 weeks, ICP4 and ICP0 transcripts are reduced to a basal level, but the LAT intron region continues to be expressed. Live virus could be recovered from the olfactory bulb and brain stem tissue. Viral proteins could be detected using anti-HSV-1 antibodies and anti-ICP4 antibody, during the acute stage but not beyond. In situ hybridization could detect LAT during acute infection in most brain regions and in olfactory bulb and brain stem tissue well beyond the acute stage. Using a homogenate from these tissues' post-acute infection, we did not recover live HSV-1 virus, supporting a latent infection, but using a modified explant cocultivation technique, we were able to recover reactivated virus from these tissues, suggesting that the HSV-1 virus latently infects the tree shrew CNS. Compared to mouse, the CNS acute infection of the tree shrew is delayed and the olfactory bulb contains most latent virus. During the acute stage, a portion of the infected tree shrews exhibit symptoms similar to human viral encephalitis. These findings, together with the fact that tree shrews are closely

  9. Staphylococcus aureus induces hypoxia and cellular damage in porcine dermal explants.

    Science.gov (United States)

    Lone, Abdul G; Atci, Erhan; Renslow, Ryan; Beyenal, Haluk; Noh, Susan; Fransson, Boel; Abu-Lail, Nehal; Park, Jeong-Jin; Gang, David R; Call, Douglas R

    2015-06-01

    We developed a porcine dermal explant model to determine the extent to which Staphylococcus aureus biofilm communities deplete oxygen, change pH, and produce damage in underlying tissue. Microelectrode measurements demonstrated that dissolved oxygen (DO) in biofilm-free dermal tissue was 4.45 ± 1.17 mg/liter, while DO levels for biofilm-infected tissue declined sharply from the surface, with no measurable oxygen detectable in the underlying dermal tissue. Magnetic resonance imaging demonstrated that biofilm-free dermal tissue had a significantly lower relative effective diffusion coefficient (0.26 ± 0.09 to 0.30 ± 0.12) than biofilm-infected dermal tissue (0.40 ± 0.12 to 0.48 ± 0.12; P < 0.0001). Thus, the difference in DO level was attributable to biofilm-induced oxygen demand rather than changes in oxygen diffusivity. Microelectrode measures showed that pH within biofilm-infected explants was more alkaline than in biofilm-free explants (8.0 ± 0.17 versus 7.5 ± 0.15, respectively; P < 0.002). Cellular and nuclear details were lost in the infected explants, consistent with cell death. Quantitative label-free shotgun proteomics demonstrated that both proapoptotic programmed cell death protein 5 and antiapoptotic macrophage migration inhibitory factor accumulated in the infected-explant spent medium, compared with uninfected-explant spent media (1,351-fold and 58-fold, respectively), consistent with the cooccurrence of apoptosis and necrosis in the explants. Biofilm-origin proteins reflected an extracellular matrix-adapted lifestyle of S. aureus. S. aureus biofilms deplete oxygen, increase pH, and induce cell death, all factors that contribute to impede wound healing.

  10. Efficient culture protocol for plant regeneration from cotyledonary petiole explants of Jatropha curcas L.

    Directory of Open Access Journals (Sweden)

    Ying Liu

    2016-09-01

    Full Text Available A high-frequency and reproducible protocol for induction of adventitious shoot buds and plant regeneration from cotyledonary petiole explants of Jatropha curcas L. has been developed. The cotyledonary petiole explants of J. curcas cultured directly on medium supplemented with thidiazuron (TDZ induce regeneration of poor quality shoot buds that have a low regeneration frequency. However, treating the explants with high concentrations (10–60 mg/L of TDZ solution for certain time periods (5–80 min significantly increased the regeneration frequency and improved the quality of the regenerated shoot buds. The best shoot buds induction (88.42% and number of shoot buds (12.67 per explant were observed when in vitro explants were treated with 20 mg/L TDZ solution for 20 min before being transferred on hormone-free medium after 30 days. Regeneration was also influenced by the orientation (horizontal or vertical of the explants on the medium, and by the origin of the cotyledonary petioles (in vitro or in vivo used for the preparation of explants. We performed subsequent experiments for elongation and rooting of the regenerated shoot buds. Addition of L-arginine to the medium was conducive to the elongation of the shoot buds. A concentration of 7.5 mg/L L-arginine yielded the best results. The elongated shoots could initiate roots to become intact plantlets in half-strength Murashige and Skoog medium containing 0.1 mg/L indole-3-butyric acid. After acclimatization, these plantlets could be transplanted to the soil and the growth was normal. Therefore, application of the methods described here helped to increase plant regeneration efficiency.

  11. Lack of association between mannose binding lectin and antibody responses after acellular pertussis vaccinations.

    Directory of Open Access Journals (Sweden)

    Kirsi Gröndahl-Yli-Hannuksela

    Full Text Available BACKGROUND: Mannose-binding lectin (MBL is one of the key molecules in innate immunity and its role in human vaccine responses is poorly known. This study aimed to investigate the possible association of MBL polymorphisms with antibody production after primary and booster vaccinations with acellular pertussis vaccines in infants and adolescents. METHODOLOGY/PRINCIPAL FINDINGS: Five hundred and sixty eight subjects were included in this study. In the adolescent cohort 355 subjects received a dose of diphtheria and tetanus toxoids and acellular pertussis (dTpa vaccine ten years previously. Follow-up was performed at 3, 5 and 10 years. Infant cohort consisted of 213 subjects, who had received three primary doses of DTaP vaccine at 3, 5, and 12 months of age according to Finnish immunization program. Blood samples were collected before the vaccinations at 2,5 months of age and after the vaccinations at 13 months and 2 years of age. Concentrations of IgG antibodies to pertussis toxin, filamentous hemagglutinin, and pertactin and antibodies to diphtheria and tetanus toxoids were measured by standardized enzyme-linked immunosorbant assay. Single nucleotide polymorphisms of MBL2 gene exon1 (codons 52, 54, 57 were examined. MBL serum concentration was also measured from the adolescent cohort. No association was found with MBL2 exon 1 polymorphisms and antibody responses against vaccine antigens, after primary and booster dTpa vaccination. CONCLUSIONS: This study indicates that MBL polymorphisms do not affect the production and persistence of antibodies after acellular pertussis vaccination. Our finding also suggests that MBL might not be involved in modulating antibody responses to the vaccines made of purified bacterial proteins.

  12. [Effect of two different acellular lung matrices on α-SMA expression in A549 cells].

    Science.gov (United States)

    Chen, C; Wang, Z Y; Weng, J; Wang, Z B; Mei, J; Du, X H; Wang, L

    2017-01-24

    Objective: To explore the effect of acellular normal and fibrotic lung matrices on alpha smooth muscle actin (α-SMA) expression in human lung adenocarcinoma cell line A549. Methods: Twenty adult SD rats were randomly divided into normal group and idiopathic pulmonary fibrosis(IPF)group (n=10 each). The pulmonary fibrosis was induced by Bleomycin. Normal and fibrotic decellularized lungs were made, then sections with 500 μm thick were cut by a standard Vibratome. None scaffold was set as control group. A549 cells were seeded dropwise into different slices (normal and fibrotic scaffolds), and cultured for one week in vitro. The expression of α-SMA was measured by immunofluorescence staining and quantitative real time polymerase chain reaction (qRT-PCR). Results: In control group, the expression of α-SMA protein was positive in A549 cells by immunofluorescence staining. However, it expressed weakly both in normal and fibrotic scaffold group, and the fluorescence intensity in fibrotic scaffold group was significant lower than that in normal group (PSMA mRNA in normal and fibrotic scaffold group were (0.70±0.11) and (0.55±0.12), which were significant lower than that of control group (1.28±0.21) (PSMA mRNA in fibrotic scaffold group was decreased compared to that in normal scaffold group (PSMA in human lung adenocarcinoma cell line A549. It may inhibit the movement of A549 cells in acellular normal and fibrotic lung matrices, especially in acellular fibrotic lung scaffold.

  13. Biomimetic acellular detoxified glutaraldehyde cross-linked bovine pericardium for tissue engineering.

    Science.gov (United States)

    Mathapati, Santosh; Bishi, Dillip Kumar; Guhathakurta, Soma; Cherian, Kotturathu Mammen; Venugopal, Jayarama Reddy; Ramakrishna, Seeram; Verma, Rama Shanker

    2013-04-01

    Glutaraldehyde (GLUT) processing, cellular antigens, calcium ions in circulation, and phospholipids present in the native tissue are predominantly responsible for calcification, degeneration, and lack of natural microenvironment for host progenitor cell migration in tissue implants. The study presents an improved methodology for adhesion and proliferation of endothelial progenitor cells (EPCs) without significant changes in biomechanical and biodegradation properties of the processed acellular bovine pericardium. The anti-calcification potential of the processed tissue was enhanced by detoxification of GLUT-cross-linked bovine pericardium by decellularization, pretreating it with ethanol or removing the free aldehydes by citric acid treatment and lyophilization. The treated tissues were assessed for biomechanical properties, GLUT ligand quantification, adhesion, proliferation of EPCs, and biodegradability. The results indicate that there was no significant change in biomechanical properties and biodegradability when enzymatic hydrolysis (p>0.05) is employed in detoxified acellular GLUT cross-linked tissue (DBP-G-CA-ET), compared with the native detoxified GLUT cross-linked bovine pericardium (NBP-G-CA-ET). DBP-G-CA-ET exhibited a significant (p>0.05) increase in the viability of EPCs and cell adhesion as compared to acellular GLUT cross-linked bovine pericardium (ppericardium, employed in our study as an alternative to conventional GLUT cross-linked bovine pericardium, might provide longer durability and better biocompatibility, and reduce calcification. The developed bovine pericardium patches could be used in cardiac reconstruction and repair, arteriotomy, soft tissue repair, and general surgical procedures with tissue regeneration dimensions. Copyright © 2012 Elsevier B.V. All rights reserved.

  14. Adipose tissue regeneration in vivo using micronized acellular allogenic dermis as an injectable scaffold.

    Science.gov (United States)

    Lee, Hee Young; Yang, Hyun Jin; Rhie, Jong Won; Han, Ki Talk

    2014-10-01

    Over the past few years, the clinical use of injectable artificial materials in plastic surgery has increased. In addition, autologous lipoimplantation is being performed for volume replacement of soft tissue for aesthetic purposes. In this study, acellular allogenic dermis was utilized as a scaffold for the culturing of preadipocytes, confirming the possibility of three-dimensional proliferation of progenitor cells, the eventual differentiation of stromal cells in adipose tissue into the adipocytes, and the in vivo implantation of such adipocytes to form fat tissue. Preadipocytes, recently called ASCs (adipose tissue-derived stromal/stem cells), were cultured in acellular allogenic dermis, successfully attached to the dermal particles in a three-dimensional structure, and proliferated, differentiated, and eventually formed a cluster. For the in vivo implantation, four groups were formed: the first group was cultured within the dermal scaffold for 24 h before implantation (24-h preconditioned group), the second group was induced for differentiation for 10 days before implantation (10-day preconditioned group), the third group was implanted immediately after cell propagation (nonpreconditioned group), and the control group was implanted with only dermal scaffold. In vivo implanted preadipocytes showed great differentiation into adipocytes within the dermal scaffolds. Also, the 10-day preconditioned group showed a greater volume of fat tissue compared to the 24-h preconditioned group. From these results, we confirmed that after a three-dimensional culture in acellular allogenic dermis, implanted preadipocytes formed a greater amount of fat tissue and that this could be a possible effective method for future soft tissue restoration.

  15. Acellular pertussis booster in adolescents induces Th1 and memory CD8+ T cell immune response.

    Directory of Open Access Journals (Sweden)

    Nikolaus Rieber

    Full Text Available In a number of countries, whole cell pertussis vaccines (wcP were replaced by acellular vaccines (aP due to an improved reactogenicity profile. Pertussis immunization leads to specific antibody production with the help of CD4(+ T cells. In earlier studies in infants and young children, wcP vaccines selectively induced a Th1 dominated immune response, whereas aP vaccines led to a Th2 biased response. To obtain data on Th1 or Th2 dominance of the immune response in adolescents receiving an aP booster immunization after a wcP or aP primary immunization, we analyzed the concentration of Th1 (IL-2, TNF-α, INF-γ and Th2 (IL-4, IL-5, IL-10 cytokines in supernatants of lymphocyte cultures specifically stimulated with pertussis antigens. We also investigated the presence of cytotoxic T cell responses against the facultative intracellular bacterium Bordetella pertussis by quantifying pertussis-specific CD8(+ T cell activation following the aP booster immunization. Here we show that the adolescent aP booster vaccination predominantly leads to a Th1 immune response based on IFNgamma secretion upon stimulation with pertussis antigen, irrespective of a prior whole cell or acellular primary vaccination. The vaccination also induces an increase in peripheral CD8(+CD69(+ activated pertussis-specific memory T cells four weeks after vaccination. The Th1 bias of this immune response could play a role for the decreased local reactogenicity of this adolescent aP booster immunization when compared to the preceding childhood acellular pertussis booster. Pertussis-specific CD8(+ memory T cells may contribute to protection against clinical pertussis.

  16. Porcine vesical acellular matrix graft of tunica albuginea for penile reconstruction

    Institute of Scientific and Technical Information of China (English)

    Kwan-Joong Joo; Byung-Soo Kim; Jeong-Ho Han; Chang-Ju Kim; Chil-Hun Kwon; Heung-Jae Park

    2006-01-01

    Aim: To characterize the feasibility of the surgical replacement of the penile tunica albuginea (TA) and to evaluate the value of a porcine bladder acellular matrix (BAM) graft. Methods: Acellular matrices were constructed from pigs'bladders by cell lysis, and then examined by scanning electron microscopy (SEM). Expression levels of the mRNA of the vascular endothelial growth factor (VEGF) receptor, fibroblast growth factor (FGF)-1 receptor, neuregulin, and brain-derived neurotrophic factor (BDNF) in the acellular matrix and submucosa of the pigs' bladders were determined through the reverse transcription-polymerase chain reaction (PCR). A 5 mm × 5 mm square was excised from the penile TA of nine rabbits. The defective TA was then covered in porcine BAM. Equal numbers of animals were sacrificed and histochemically examined at 2, 4 and 6 months after implantation. Results: SEM of the BAM showed collagen fibers with many pores. VEGF receptor, FGF-1 receptor and neuregulin mRNA were expressed in the porcine BAM; BDNF mRNA was not detected. Two months after implantation, the graft sites exhibited excellent healing without contracture, and the fusion between the graft and the neighboring normal TA appeared to be well established. There were no significant histological differences between the implanted tunica and the normal control tunica at 6 months after implantation. Conclusion: The porcine BAM graft resulted in a structure which was sufficiently like that of the normal TA. This implantation might be considered applicable to the reconstruction of the TA in conditions such as trauma or Peyronie's disease.

  17. Meshed acellular dermal matrix:technique and application in implant based breast reconstruction

    Institute of Scientific and Technical Information of China (English)

    Dino Zammit; Jonathan Kanevsky; Fan-Yi Meng; Tassos Dionisopoulos

    2016-01-01

    Alloderm was the first acellular dermal matrix used and remains a popular choice among plastic surgeons. However, while the overall surgical outcome of breast reconstruction using alloderm has been a success, the economic burden on the health care system makes it a subject of frequent re-evaluations in cost-effectiveness. Prompted by the high price of $3,700 USD for a 6 cm × 16 cm area, our group proposes the meshing of AlloDerm to decrease the total amount needed for breast reconstruction, while achieving comparable surgical outcomes as using unmeshed alloderm.

  18. Delayed repair in a case of forearm fascial muscle herniation using non-cross-linked acellular porcine dermal matrix.

    Science.gov (United States)

    Hartmann, Christoph E A; Branford, Olivier A; Floyd, David

    2012-09-01

    The options for treatment of symptomatic muscle herniation in the limbs traditionally include fasciotomy, direct repair, tendon weave graft (palmaris longus), fascial graft (tensor fascia lata), and synthetic mesh (prolene). A recent case report has described the use of acellular cadaveric dermal matrix to reconstruct fascial defects in 2 cases. We describe the use of Strattice, a non-cross-linked acellular porcine dermal matrix, as a fascial underlay graft in a case of symptomatic upper limb muscle herniation. We propose that Strattice has the advantages over cadaveric dermal matrices in terms of avoiding the use of human donor tissue. It has suitable tensile properties to be used for reconstructing fascial defects.

  19. Plant regeneration from cotyledonary explants of Eucalyptus camaldulensis Regeneração de plantas de Eucalyptus camaldulensis a partir das explantes cotiledonares

    Directory of Open Access Journals (Sweden)

    Roberson Dibax

    2005-08-01

    Full Text Available Breeding methods based on genetic transformation techniques need to be implemented for Eucalyptus camaldulensis to shorten the long breeding cycles and avoid manipulation of adult trees; that requires the development of plant regeneration protocols enabling development of plants from transformed tissues. The present work aimed to optimise the regeneration process already established for the species. Cotyledonary leaves of E. camaldulensis were cultured in MS medium supplemented with naphthaleneacetic acid (NAA and 6-benzylaminopurine (BAP combinations. The most efficient treatment for bud indirect regeneration (2.7 µmol L-1 NAA and 4.44 µmol L-1 BAP was used for further experiments. When explants were kept in the dark during the first 30 days, the percentage of explants forming calluses increased and explant necrosis was reduced in comparison with light-cultured explants. Mineral medium modifications were compared and half-strength MS mineral medium turned out to be as efficient as full-strength medium, producing 54% and 47% of explants with buds, respectively. For shoot elongation, MS medium with half-strength nitrate and ammonium salts, and 0.2% activated charcoal yielded rooted shoots 1 to 8 cm high after one month. The procedure is an efficient protocol for E. camadulensis plant regeneration, reducing the stages necessary for the obtention of complete plants.A implementação, para espécies florestais, de técnicas de melhoramento baseadas em métodos de transformação genética, permitirá reduzir os longos ciclos de melhoramento e evitar a manipulação de árvores adultas. Isto implica dispor de um protocolo de regeneração que permita o desenvolvimento de plantas a partir de tecidos transformados. Este trabalho teve como objetivo otimizar este protocolo de regeneração para Eucalyptus camaldulensis. Folhas cotiledonares foram cultivadas em meio de cultura MS suplementado com combinações de ácido naftalenoacético (ANA e 6

  20. Effects of the re-innervation of organotypic skin explants on the epidermis.

    Science.gov (United States)

    Lebonvallet, Nicolas; Boulais, Nicholas; Le Gall, Christelle; Pereira, Ulysse; Gauché, Dominique; Gobin, Eric; Pers, Jacques-Olivier; Jeanmaire, Christine; Danoux, Louis; Pauly, Gilles; Misery, Laurent

    2012-02-01

    The nervous system takes part in skin homeostasis and interacts with skin cells. In in vitro organotypic skin models, these interactions are lost owing to the absence of nerve endings. We have developed an in vitro organotypic skin model based on a re-innervated human skin explant using primary sensory neurons from the dorsal root ganglia of rats. After 10 days of co-culture between skin explant and neurons, a dense network of nerve fibres was observed. The epidermis and dermis presented nerve fibres associated with cellular body from sensory neurons introduced in the co-culture. Epidermal thickness, cell density and quality of re-innervated skin explant were all higher when skin explants were re-innervated by sensory neurons at 10 days of culture. Proliferation of epidermal cell was not modified, but the apoptosis was significantly diminished. Hence, this innovative model of co-cultured skin explants and neurons allows better epidermal integrity and could be useful for studies concerning interactions between the skin and its peripheral nervous system.

  1. Development of a murine ocular posterior segment explant culture for the study of intravitreous vector delivery.

    Science.gov (United States)

    Denk, Nora; Misra, Vikram; Sandmeyer, Lynne S; Bauer, Bianca B; Singh, Jaswant; Forsyth, George W; Grahn, Bruce H

    2015-01-01

    The objective of this study was to develop a murine retinal/choroidal/scleral explant culture system to facilitate the intravitreous delivery of vectors. Posterior segment explants from adult mice of 2 different age groups (4 wk and 15 wk) were cultured in serum-free medium for variable time periods. Tissue viability was assessed by gross morphology, cell survival quantification, activated caspase-3 expression, and immunohistochemistry. To model ocular gene therapy, explants were exposed to varying transducing units of a lentiviral vector expressing the gene for green fluorescent protein for 48 h. Explant retinal cells remained viable for approximately 1 wk, although the ganglion cell layer developed apoptosis between 4 and 7 d. Following vector infusion into the posterior segment cups, viral transduction was noted in multiple retinal layers in both age groups. An age of donor mouse influence was noted and older mice did not transduce as well as younger mice. This explant offers an easily managed posterior segment ocular culture with minimum disturbance of the tissue, and may be useful for investigating methods of enhancing retinal gene therapy under controlled conditions.

  2. Ex Vivo Produced Oral Mucosa Equivalent by Using the Direct Explant Cell Culture Technique

    Directory of Open Access Journals (Sweden)

    Kamile Öztürk

    2012-09-01

    Full Text Available Objective: The aim of this study is the histological and immunohistochemical evaluation of ex vivo produced oral mucosal equivalents using keratinocytes cultured by direct explant technique.Material and Methods: Oral mucosa tissue samples were obtained from the keratinized gingival tissues of 14 healthy human subjects. Human oral mucosa keratinocytes from an oral mucosa biopsy specimen were dissociated by the explant technique. Once a sufficient population of keratinocytes was reached, they were seeded onto the type IV collagen coated “AlloDerm” and taken for histological and immunohistochemical examinations at 11 days postseeding of the keratinocytes on the cadaveric human dermal matrix.Results: Histopathologically and immunohistochemically, 12 out of 14 successful ex vivo produced oral mucosa equivalents (EVPOME that consisted of a stratified epidermis on a dermal matrix have been developed with keratinocytes cultured by the explant technique.Conclusion: The technical handling involved in the direct explant method at the beginning of the process has fewer steps than the enzymatic method and use of the direct explant technique protocol for culturing of human oral mucosa keratinocyte may be more adequate for EVPOME production.

  3. Optical and atomic force microscopy of an explanted AcrySof intraocular lens with glistenings.

    Science.gov (United States)

    Dogru, M; Tetsumoto, K; Tagami, Y; Kato, K; Nakamae, K

    2000-04-01

    To assess the surface morphology and cause of glistenings in an explanted AcrySof intraocular lens (IOL). Shakai Hoken Kobe Central Hospital, Kobe, Japan. A 63-year-old Japanese man had implantation of an AcrySof IOL in the capsular bag. One month postoperatively, he had a neodymium:YAG laser capsulotomy for posterior capsule opacification, which changed the IOL's position in the capsular bag. A few months later, the patient developed disabling night glare from intralenticular glistenings and progressive hyperopic refractive error. The IOL was explanted and then analyzed by optical microscopy and atomic force microscopy (AFM). Laboratory analysis of control AcrySof IOLs kept in a balanced salt solution at steady room and body temperature for 2 months was also performed to evaluate the cause of the glistenings observed clinically. Optical microscopy showed that the explanted AcrySof IOL had several microvacuoles; no abnormalities were observed in the control AcrySof IOLs before or after folding at the room and body temperatures. The AFM analysis showed a significant change in the surface morphology of the explanted IOL, including vacuolar formations in the posterior surface as well as numerous anterior surface irregularities. No microvacuoles or surface morphology alterations were observed in the control AcrySof IOLs by AFM analysis. The glistenings in the explanted AcrySof IOL were likely caused by temperature changes and not mechanical stress from folding.

  4. In vitro assessment of the soft tissue/implant interface using porcine gingival explants.

    Science.gov (United States)

    Abdulmajeed, Aous A; Willberg, Jaana; Syrjänen, Stina; Vallittu, Pekka K; Närhi, Timo O

    2015-01-01

    The biologic seal of peri-implant soft tissue is crucial for long-term prognosis of oral implants. This in vitro study describes a novel tissue culture model using porcine gingival explants to evaluate the soft tissue/implant interface. Two different types of substrates were investigated: (a) plain polymer: BisGMA-TEGDMA (50-50 %) and (b) unidirectional fiber-reinforced composite (FRC). Porcine gingival explants were obtained from a local slaughterhouse. The experimental implants (n = 4) were inserted into the middle of freshly excised porcine gingival explants and cultured at the air/liquid interface up to 14 days. Porcine gingival explants with no implants served as baseline controls. The specimens were fixed and processed for the preparation of undecalcified samples. Histological analysis of the soft tissue/implant interface was carried out using a light-microscope. Microscopic evaluation suggests that the gingival explants established epithelial and connective tissue attachment to both implant types over the incubation period. FRC surfaces seemed to have a favorable tissue response with a sign of an outward epithelial migration. However, tissue degeneration was observed at the end of the experiment. In conclusion, this in vitro model maintains mucosal viability and ability to histologically evaluate soft tissue attachment to biomaterials rendering it a time efficient and cost effective model that may reduce the need for animal experiments.

  5. Organogênese de explante foliar de clones de Eucalyptus grandis x E. urophylla Organogenesis of the leaf explant of Eucalyptus grandis x E. urophylla clones

    Directory of Open Access Journals (Sweden)

    Elisa Cristina Soares de Carvalho Alves

    2004-05-01

    Full Text Available O objetivo deste trabalho foi avaliar os efeitos dos reguladores de crescimento TDZ [1-fenil-3-(1,2,3-tia-diazol-5-iluréia], BAP (6-benzilaminopurina e ANA (ácido naftalenoacético no desempenho da propagação in vitro por organogênese de explante foliar de três clones híbridos de Eucalyptus grandis x Eucalyptus urophylla. Houve resposta diferenciada dos clones quanto a intensidade, textura e coloração dos calos, em razão dos tratamentos com os reguladores de crescimento. Os melhores resultados de calejamento dos três genótipos foram observados nos tratamentos com a combinação dos reguladores de crescimento TDZ (0,5 mg L-1 e ANA (0,1 mg L-1, obtendo-se 100% de calejamento no explante foliar. Os piores resultados de calejamento foram observados nos tratamentos com a combinação dos reguladores de crescimento BAP (0,1 mg L-1 e ANA (0,1 mg L-1. Em relação à regeneração, a melhor resposta foi obtida com 1,0 mg L-1 BAP em que 8% dos calos formados a partir de explantes foliares regeneraram gemas, com número médio destas formadas por calo igual a 4,2.The aim of this work was to evaluate the effects of growth regulators TDZ [1-phenil-3-(1,2,3-thiadiazol-5-yl urea], BAP (6-benzilaminopurine e NAA (Naphthalene acetic acid on the in vitro propagation by organogenesis from foliar explants of Eucalyptus grandis x E. urophylla. Depending on the clone used, there were singular responses to growth regulators treatment regarding callusing intensity, texture and color. The best results of the three genotypes used were observed with the TDZ (0.5 mg L-1 and NAA (0.1 mg L-1 treatment, where 100% of the foliar explants presented callus. The worst results were observed with the BAP (0.1 mg L-1 and NAA (0.1 mg L-1 treatment. Subsequently, considering the regeneration process, the best response was achieved with 1.0 mg L-1 BAP, in which 8% of the calli regenerated buds, with an average of 4.2 buds per explant.

  6. Histological observation on acellular nerve grafts co-cultured with Schwann cells for repairing defects of the sciatic nerve

    Institute of Scientific and Technical Information of China (English)

    Xiaohong Sun; Jiangyi Tian; Xiaojie Tong; Xu Zhang; Zheng He

    2006-01-01

    BACKGROUND: Animal experiments and clinical studies about tissue engineering method applied to repair nerve injury mainly focus on seeking ideal artificial nerve grafts, nerve conduit and seed cells. Autologous nerve, allogeneic nerve and xenogeneic nerve are used to bridge nerve defects, it is one of the methods to promote the repair of nerve injury by culturing and growing Schwann cells, which can secrete various neurotrophic factor activities, in the grafts.OBJECTIVE: To observe the effect of acellular nerve grafts co-cultured with Schwann cells in repairing defects of sciatic nerve.DESIGN: An observational comparative study.SETTING: Tissue Engineering Laboratory of China Medical University.MATERIALS: The experiment was carried out in the Tissue Engineering Laboratory of China Medical University between April 2004 and April 2005. Forty neonatal Sprague-Dawley rats of 5-8 days (either males or females) and 24 male Wistar rats of 180-220 g were provided by the experimental animal center of China Medical University.METHODS: ① Culture of Schwann cells: The bilateral sciatic nerves and branchial plexus were isolated from the 40 neonatal SD rats. The sciatic nerves were enzymatically digested with collagenase and dispase, isolatd, purified and cultured with the method of speed-difference adhersion, and identified with the SABC immunohistochemical method. ② Model establishment: In vitro Schwann cells were microinjected into 10-mm long acellular nerve grafts repairing a surgically created gap in the rat sciatic nerve.According to the different grafted methods, the animals were randomly divided into three groups: autografts (n=8), acellular nerve grafts (n=8), or acellular nerve grafts with Schwann cells (n=8). ③ The regenerated nerve fiber number and average diameter of myeline sheath after culture were statistically anlayzed.MAIN OUTCOME MEASURES: ① The regenerated nerve ultrastructure, total number and density of myelinated nerve fibers, and the thickness of

  7. Patient-specific three-dimensional explant spheroids derived from human nasal airway epithelium

    DEFF Research Database (Denmark)

    Marthin, June Kehlet; Stevens, Elizabeth Munkebjerg; Larsen, Lars Allan

    2017-01-01

    BACKGROUND: Three-dimensional explant spheroid formation is an ex vivo technique previously used in studies of airway epithelial ion and water transport. Explanted cells and sheets of nasal epithelium form fully differentiated spheroids enclosing a partly fluid-filled lumen with the ciliated apical....... Nasal ciliary cells and sheets were removed on day 1 by nasal brush biopsy and analyzed with regard to ciliary beat pattern-and frequency using high-speed video imaging for standard reference values. Three-dimensional explant spheroid formation was initiated in the same individual on the same day...... was successful in 15 out of 18 (82%) sampled individuals. Thus, formation was successful in seven healthy controls and eight PCD patients, while unsuccessful in 3 with PCD due to infection. Median (range) number of days in culture before harvesting of spheroids was 4 (1-5) in healthy versus 2 (1-5) in PCD...

  8. Embryoid Body-Explant Outgrowth Cultivation from Induced Pluripotent Stem Cells in an Automated Closed Platform

    Science.gov (United States)

    Tone, Hiroshi; Yoshioka, Saeko; Akiyama, Hirokazu; Nishimura, Akira; Ichimura, Masaki; Nakatani, Masaru; Kiyono, Tohru

    2016-01-01

    Automation of cell culture would facilitate stable cell expansion with consistent quality. In the present study, feasibility of an automated closed-cell culture system “P 4C S” for an embryoid body- (EB-) explant outgrowth culture was investigated as a model case for explant culture. After placing the induced pluripotent stem cell- (iPSC-) derived EBs into the system, the EBs successfully adhered to the culture surface and the cell outgrowth was clearly observed surrounding the adherent EBs. After confirming the outgrowth, we carried out subculture manipulation, in which the detached cells were simply dispersed by shaking the culture flask, leading to uniform cell distribution. This enabled continuous stable cell expansion, resulting in a cell yield of 3.1 × 107. There was no evidence of bacterial contamination throughout the cell culture experiments. We herewith developed the automated cultivation platform for EB-explant outgrowth cells. PMID:27648449

  9. In vitro organogenesis and somatic embryogenesis from leaf explants of Leucosceptrum canum sm.

    Science.gov (United States)

    Pal, A; Banerjee, A; Dhar, K

    1985-10-01

    Plantlets were obtained from leaf explants of a Labiatae tree - Leucosceptrum canum Sm. using plant tissue culture techniques. Two types of calli proliferated from the leaf explants when grown on different media, one of which was amenable to somatic embryogenesis. Differentiation of the embryoids started from the fourth passage of culture and continued up to the seventh passage. The number of embryoids decreased with the age of the callus. The capacity of such embryoids to form entire plantlets was studied using different nutrient mileux. Embryoids formed plantlets on Murashige and Skoog's (MS) medium fortified with benzylaminopurine plus indolebutyric acid. Organogenesis was observed in shoot-buds derived from explants of in vitro regenerated plantlets on MS basal medium supplemented with benzylaminopurine. Culture regenerated plantlets were transferred to MS medium without sucrose and growth hormones; finally transferred to pots containing sterile vermiculite where they are growing.

  10. Sterile Acellular Dermal Collagen as a Treatment for Rippling Deformity of Breast

    Directory of Open Access Journals (Sweden)

    Brittany Busse

    2014-01-01

    Full Text Available Prosthetic implants are frequently used for breast augmentation and breast reconstruction following mastectomy. Unfortunately, long-term aesthetic results of prosthetic breast restoration may be hindered by complications such as rippling, capsular contracture, and implant malposition. The advent of use of acellular dermal matrices has greatly improved the outcomes of prosthetic breast reconstruction. We describe a case of rippling deformity of breast that was treated using an acellular dermal matrix product, AlloMax. The patient presented with visible rippling of bilateral prosthetic breast implants as well as significant asymmetry of the breasts after multiple excisional biopsies for right breast ductal carcinoma in situ. A 6×10 cm piece of AlloMax was placed on the medial aspect of each breast between the implant and the skin flap. Follow-up was performed at 1 week, 3 months, and 1 year following the procedure. The patient recovered well from the surgery and there were no complications. At her first postoperative follow-up the patient was extremely satisfied with the result. At her 3-month and 1-year follow-up she had no recurrence of her previous deformity and no new deformity.

  11. Tissue engineering of rat bladder using marrow-derived mesenchymal stem cells and bladder acellular matrix.

    Directory of Open Access Journals (Sweden)

    Daniel L Coutu

    Full Text Available Bladder replacement or augmentation is required in congenital malformations or following trauma or cancer. The current surgical solution involves enterocystoplasty but is associated with high complication rates. Strategies for bladder tissue engineering are thus actively sought to address this unmet clinical need. Because of the poor efficacy of synthetic polymers, the use of bladder acellular matrix (BAM has been proposed. Indeed when cellular components are removed from xenogenic or allogeneic bladders, the extracellular matrix scaffold thus obtained can be used alone or in combination with stem cells. In this study, we propose the use of BAM seeded with marrow-derived mesenchymal stem cells (MSCs for bladder tissue engineering. We optimized a protocol for decellularization of bladder tissue from different species including rat, rabbit and swine. We demonstrate the use of non-ionic detergents followed by nuclease digestion results in efficient decellularization while preserving the extracellular matrix. When MSCs were seeded on acellular matrix scaffold, they remained viable and proliferative while adopting a cellular phenotype consistent with their microenvironment. Upon transplantation in rats after partial cystectomy, MSC-seeded BAM proved superior to unseeded BAM with animals recovering nearly 100% normal bladder capacity for up to six months. Histological analyses also demonstrated increased muscle regeneration.

  12. The acellular matrix (ACM) for bladder tissue engineering: A quantitative magnetic resonance imaging study.

    Science.gov (United States)

    Cheng, Hai-Ling Margaret; Loai, Yasir; Beaumont, Marine; Farhat, Walid A

    2010-08-01

    Bladder acellular matrices (ACMs) derived from natural tissue are gaining increasing attention for their role in tissue engineering and regeneration. Unlike conventional scaffolds based on biodegradable polymers or gels, ACMs possess native biomechanical and many acquired biologic properties. Efforts to optimize ACM-based scaffolds are ongoing and would be greatly assisted by a noninvasive means to characterize scaffold properties and monitor interaction with cells. MRI is well suited to this role, but research with MRI for scaffold characterization has been limited. This study presents initial results from quantitative MRI measurements for bladder ACM characterization and investigates the effects of incorporating hyaluronic acid, a natural biomaterial useful in tissue-engineering and regeneration. Measured MR relaxation times (T(1), T(2)) and diffusion coefficient were consistent with increased water uptake and glycosaminoglycan content observed on biochemistry in hyaluronic acid ACMs. Multicomponent MRI provided greater specificity, with diffusion data showing an acellular environment and T(2) components distinguishing the separate effects of increased glycosaminoglycans and hydration. These results suggest that quantitative MRI may provide useful information on matrix composition and structure, which is valuable in guiding further development using bladder ACMs for organ regeneration and in strategies involving the use of hyaluronic acid.

  13. Conjunctival structural and functional reconstruction using acellular bovine pericardium graft (Normal GEN®) in rabbits.

    Science.gov (United States)

    Huang, Danping; Xu, Bing; Yang, Xiaonan; Xu, Binbin; Zhao, Jing

    2016-04-01

    To evaluate the effectiveness of acellular bovine pericardium grafts (Normal GEN®) used as scaffolds for conjunctival reconstruction. The acellular bovine pericardium graft and the amnion graft were implanted into the bulbar conjunctival defects of adult rabbits. Conjunctival samples of implanted materials and blank defect controls were observed at day 3, 7, 14, 21, 28, and 56 postoperatively. Histological examination was observed at day 14, 28, and 56 of surgery, including hematoxylin-eosin staining, periodic acid-Schiff staining, and Masson's trichrome staining, while immunofluorescent microscopy was observed at 14 days and 28 days after surgery. Results were compared among the Normal GEN®, amnion, and blank defect controls. All three groups showed complete conjunctival reconstruction. Wounds that were not grafted closed by formation of conjunctival scar characterized by a linear array of densely packed collagen fibers in Tenon's capsule. Subepithelial tissue in the grafted groups comprised a loosely organized network of randomly oriented collagen that resembled that of the normal bulbar conjunctiva. However, there was a dense layer of aligned collagen between the conjunctival Tenon's capsule and the sclera in the NormalGEN® group, about 250 μm in thickness. Implantation of the NormalGEN® graft promoted the formation of conjunctiva as a kind of scaffold both in structure and in function. It had more advantageous mechanical properties than the amnion, strong and elastic, during the period of conjunctival reconstruction.

  14. Results of Acellular Dermis Matrix Graft Used for Tympanoplasty in Guinea Pig Model

    Directory of Open Access Journals (Sweden)

    Farhad Farahani

    2015-03-01

    Full Text Available Introduction: To describe the underlay tympanoplasty technique using an acellular dermal graft(AlloDerm for tympanic membrane (TM reconstruction in a guinea pig model and to demonstrate the feasibility of the technique using AlloDerm tissue harvested from the prepuce as a source of tissue for future grafting in human TM reconstruction.   Materials and Methods: The prepuce was divided during circumcision and the acellular dermis was prepared using a number of standard processes. Two groups of guinea pigs were prepared. In the case group (20 guinea pigs and 40 ears removal of TM was performed with tympanoplasty using AlloDerm, and in the control group (eight guinea pigs and 16 ears, removal of TM was performed without tympanoplasty. In each group, the TM was completely removed in one ear and partially removed on the other side, and the integrity of the TMs was re-evaluated after 8 weeks.   Results: In the case group, the healing rates in the completely and partially removed TMs were 83.3% and 94.4%, respectively. The difference in healing rate (0% and 66.7%, respectively was statistically significant (P

  15. Combination of acellular nerve graft and schwann cells-like cells for rat sciatic nerve regeneration.

    Science.gov (United States)

    Gao, Songtao; Zheng, Yan; Cai, Qiqing; Deng, Zhansheng; Yao, Weitao; Wang, Jiaqiang; Wang, Xin; Zhang, Peng

    2014-01-01

    To investigate the effect of tissue engineering nerve on repair of rat sciatic nerve defect. Forty-five rats with defective sciatic nerve were randomly divided into three groups. Rats in group A were repaired by acellular nerve grafts only. Rats in group B were repaired by tissue engineering nerve. In group C, rats were repaired by autogenous nerve grafts. After six and twelve weeks, sciatic nerve functional index (SFI), neural electrophysiology (NEP), histological and transmission electron microscope observation, recovery ratio of wet weight of gastrocnemius muscle, regenerated myelinated nerve fibers number, nerve fiber diameter, and thickness of the myelin sheath were measured to assess the effect. After six and twelve weeks, the recovery ratio of SFI and wet weight of gastrocnemius muscle, NEP, and the result of regenerated myelinated nerve fibers in groups B and C were superior to that of group A (P 0.05). The tissue engineering nerve composed of acellular allogenic nerve scaffold and Schwann cells-like cells can effectively repair the nerve defect in rats and its effect was similar to that of the autogenous nerve grafts.

  16. Xenogeneic acellular conjunctiva matrix as a scaffold of tissue-engineered corneal epithelium.

    Directory of Open Access Journals (Sweden)

    Haifeng Zhao

    Full Text Available Amniotic membrane-based tissue-engineered corneal epithelium has been widely used in the reconstruction of the ocular surface. However, it often degrades too early to ensure the success of the transplanted corneal epithelium when treating patients with severe ocular surface disorders. In the present study, we investigated the preparation of xenogeneic acellular conjunctiva matrix (aCM and evaluated its efficacy and safety as a scaffold of tissue-engineered corneal epithelium. Native porcine conjunctiva was decellularized with 0.1% sodium dodecyl sulfate (SDS for 12 h at 37°C and sterilized via γ-irradiation. Compared with native conjunctiva, more than 92% of the DNA was removed, and more than 90% of the extracellular matrix components (glycosaminoglycan and collagen remained after the decellularization treatment. Compared with denuded amniotic membrane (dAM, the aCM possessed favorable optical transmittance, tensile strength, stability and biocompatibility as well as stronger resistance to degradation both in vitro and in vivo. The corneal epithelial cells seeded on aCM formed a multilayered epithelial structure and endured longer than did those on dAM. The aCM-based tissue-engineered corneal epithelium was more effective in the reconstruction of the ocular surface in rabbits with limbal stem cell deficiency. These findings support the application of xenogeneic acellular conjunctiva matrix as a scaffold for reconstructing the ocular surface.

  17. Micropropagation of Araucaria excelsa R. Br. var. glauca Carrière from orthotropic stem explants.

    Science.gov (United States)

    Sarmast, Mostafa Khoshhal; Salehi, Hassan; Khosh-Khui, Morteza

    2012-07-01

    The objectives of the present work were in vitro propagation of Araucaria excelsa R. Br. var. glauca Carrière (Norfolk Island pine) with focus on the evaluation of the mean number of shoots per explant (MNS/E) and mean length of shoots per explants (MLS/E) produced by different parts of the orthotropic stem of A. excelsa R. Br. var. glauca in response to plant growth regulators. Norfolk Island pine axillary meristems responded very well to the 2-iso-pentenyl adenine (2iP) and thidiazuron (TDZ) levels. Explants taken from stem upper segments in the media containing 2iP had a higher MNS/E (3.47) and MLS/E (6.27 mm) in comparison to those taken from stem lower segments, which were 0.71 and 0.51 mm, respectively. Using 0.045 μM TDZ in the MS medium not only resulted in 4.60 MNS/E with 7.08 mm MLS/E but proliferated shoots showed a good performance as well. Investigating the best position of stem explant on mother plant as well as the best concentrations of growth regulators were performed which were useful for efficient micropropagation of this plant. Thirty three percent of explants were rooted in the MS medium containing 3 % sucrose, supplemented with 7.5 μM of both NAA and IBA for 2 weeks before transferring to a half strength MS medium without any growth regulator. Plantlets obtained were acclimatized and transferred to the greenhouse with less than 20 % mortality. This procedure considered the first successful report for regeneration and acclimatization of A. excelsa R. Br. var. glauca plantlet through main stem explants.

  18. Acellular dermal matrix and negative pressure wound therapy: a tissue-engineered alternative to free tissue transfer in the compromised host.

    Science.gov (United States)

    Menn, Zachary K; Lee, Edward; Klebuc, Michael J

    2012-02-01

    Free tissue transfer has revolutionized lower extremity reconstruction; however, its use in elderly patients with multiple medical problems can be associated with elevated rate s of perioperative morbidity and mortality. This study evaluates the use of acellular dermal matrix (ADM) in conjunction with negative pressure wound therapy (NPWT) and delayed skin graft application as an alternative to free tissue transfer in this compromised population. Bilayer, ADM (Integra, Plainsboro, NJ) was used in conjunction with NPWT (Wound V.A.C, Kinetic Concepts Inc., San Antonio, TX) to achieve vascularized coverage of complex lower extremity wounds with denuded tendon and bone in elderly, medically compromised patients. Following incorporation, the matrix was covered with split-thickness skin graft. Four patients (age range, 50 to 76 years) with multiple medical comorbidities were treated with the above protocol. The average time to complete vascularization of the matrix was 29 days. Definitive closure with split-thickness skin graft was achieved in three patients and one wound healed by secondary intention. No medical or surgical complications were encountered and stable soft tissue coverage was achieved in all patients. This early experience suggests that dermal substitute and NPWT with delayed skin graft application can provide a reasonable tissue-engineered alternative to free tissue transfer in the medically compromised individual.

  19. In Vitro Propagation of Desmodium gangeticum (L. DC. from Cotyledonary Nodal Explants

    Directory of Open Access Journals (Sweden)

    U R Vishwakarma

    2009-01-01

    Full Text Available An in vitro procedure for rapid multiplication of medicinally important plant Desmodium gangeticum (L. DC. (Fabaceae, has been developed using cotyledonary nodal explant. An average of 9.2 shoots per explant were obtained by culturing cotyledonary nodal explaint on Murashige and Skoog′s medium containing 8.8 μM BAP and 21.2 μM NAA, in combination, within 28 days. These shoots were rooted on half strength MS medium supplemented with IAA 17.1 μM. Rooted plantlets were hardened using 1:1:1 mixture of soil, river sand and vermiculite under green house conditions.

  20. Mechanical Stimulus Inhibits the Growth of a Bone Tissue Model Cultured In Vitro

    Institute of Scientific and Technical Information of China (English)

    Zong-ming Wan; Lu Liu; Jian-yu Li; Rui-xin Li; Yong Guo; Hao Li; Jian-ming Zhang; Xi-zheng Zhang

    2013-01-01

    Objectives To construct the cancellous bone explant model and a method of culturing these bone tissues in vitro, and to investigate the effect of mechanical load on growth of cancellous bone tissue in vitro. Methods Cancellous bone were extracted from rabbit femoral head and cut into 1-mm-thick and 8-mm-diameter slices under sterile conditions. HE staining and scanning electron microscopy were employed to identify the histomorphology of the model after being cultured with a new dynamic load and circulating perfusion bioreactor system for 0, 3, 5, and 7 days, respectively. We built a three-dimensional model using microCT and analyzed the loading effects using finite element analysis. The model was subjected to mechanical load of 1000, 2000, 3000, and 4000μεrespectively for 30 minutes per day. After 5 days of continuous stimuli, the activities of alkaline phosphatase (AKP) and tartrate-resistant acid phosphatase (TRAP) were detected. Apoptosis was analyzed by DNA ladder detection and caspase-3/8/9 activity detection. Results After being cultured for 3, 5, and 7 days, the bone explant model grew well. HE staining showed the apparent nucleus in cells at the each indicated time, and electron microscope revealed the living cells in the bone tissue. The activities of AKP and TRAP in the bone explant model under mechanical load of 3000 and 4000μεwere significantly lower than those in the unstressed bone tissues (all P Conclusions The cancellous bone explant model extracted from the rabbit femoral head could be alive at least for 7 days in the dynamic load and circulating perfusion bioreactor system, however, pathological mechanical load could affect the bone tissue growth by apoptosis in vitro. The differentiation of osteoblasts and osteoclasts might be inhibited after the model is stimulated by mechanical load of 3000 and 4000με.

  1. Bone within a bone

    Energy Technology Data Exchange (ETDEWEB)

    Williams, H.J.; Davies, A.M. E-mail: wendy.turner@roh.nhs.uk; Chapman, S

    2004-02-01

    The 'bone within a bone' appearance is a well-recognized radiological term with a variety of causes. It is important to recognize this appearance and also to be aware of the differential diagnosis. A number of common conditions infrequently cause this appearance. Other causes are rare and some remain primarily of historical interest, as they are no longer encountered in clinical practice. In this review we illustrate some of the conditions that can give the bone within a bone appearance and discuss the physiological and pathological aetiology of each where known.

  2. T-Cell Responses before and after the Fifth Consecutive Acellular Pertussis Vaccination in 4-Year-Old Dutch Children

    NARCIS (Netherlands)

    Schure, Rose-Minke; Hendrikx, Lotte H.; de Rond, Lia G. H.; Ozturk, Kemal; Sanders, Elisabeth A. M.; Berbers, Guy A. M.; Buisman, Anne-Marie

    2012-01-01

    Immunization with acellular pertussis vaccine (aP) induces higher specific antibody levels and fewer adverse reactions than does immunization with the whole-cell vaccine (wP). However, antibody levels in infants induced by both types of pertussis vaccines wane already after 1 year. Therefore, long-t

  3. Effect of formaldehyde on acellular-nuclear DNA%甲醛对脱细胞DNA的影响

    Institute of Scientific and Technical Information of China (English)

    杨丽娟; 王越越; 朱海燕

    2013-01-01

    目的 探索甲醛对脱细胞DNA的影响,并初步建立脱细胞-核DNA检测加合物的新模型.方法 用染毒缓冲液配制浓度分别为4%、1%、0.25%和0%的甲醛,用羟自由基损伤的脱细胞-核DNA作为检测加合物的试验模型,每组6张脱细胞-核DNA板,用彗星实验检测各组脱细胞-核DNA损伤情况,组间差异用SPSS 11.0软件统计分析.结果 甲醛染毒的DNA损伤顺序为:0%=0.25%>1%组>4%,呈现明显的剂量-反应关系.结论 甲醛可直接和脱细胞DNA片段形成加合物和/或DNA-DNA交联,脱细胞-核DNA模型可用于DNA加合物和/或交联物的检测.%Objective To explore the impact of formaldehyde on acellular-nuclear DNA, and initially to establish a new model of acellular nuclear DNA to test adducts. Methods The acellular-nuclear DNA used as a test model, sealed boxes pre-positioned slides fixed with acellular-nuclear DNA. Different concentrations (4% , 1% , 0. 25% and 0% ) of formaldehyde were added to boxes. Six acellular-nuclear DNA slides were used in each group. DNA damage was measured with comet assay, and statistical analysis of the difference between the groups with SPSS 11.0 software. Results DNA damage exposed to formaldehyde in such sequences; 0% =0.25% > 1% > 4% with significant dose-response relationship. Conclusion Formaldehyde can directly react with acellular DNA, and form adducts and/or DNA-DNA crosslinking, acellular-Nuclear DNA model can be used to detect DNA adducts and cross-linked.

  4. In vitro white spot syndrome virus (WSSV) replication in explants of the heart of freshwater crab, Paratelphusa hydrodomous.

    Science.gov (United States)

    Nathiga Nambi, K S; Abdul Majeed, S; Sundar Raj, N; Taju, G; Madan, N; Vimal, S; Sahul Hameed, A S

    2012-08-01

    Explants from different organs of freshwater crab, Paratelphusa hydrodomous were prepared to establish an in vitro system for replication of white spot syndrome virus (WSSV) of shrimp. Heart explants were maintained for 53 days without any morphological changes in EX-CELL™ 405 medium with and without serum whereas the explants of eye muscle, gill, shell membrane and appendage muscle died within 15 days of culture period. The heart explants on different days of culture were exposed to WSSV for 10 days to study the viral replication. The infection of WSSV in explants of the heart was confirmed by PCR, RT-PCR, Western blot, histology, immunohistochemistry, bioassay and transmission electron microscopy. The WSSV was quantified by real-time PCR and indirect ELISA. The WSSV inoculum prepared from the heart explants of crab caused significant mortality in Penaeus monodon in challenge experiments and the results indicate that the WSSV which replicated in the heart explants of freshwater crab maintains its infectivity as in marine shrimp. The results indicate that the heart explants of P. hydrodomous would be a good alternative to whole animals for production of WSSV.

  5. Induction of bulb organogenesis in in vitro cultures of tarda tulip (Tulipa tarda Stapf.) from seed-derived explants.

    Science.gov (United States)

    Maślanka, Małgorzata; Bach, Anna

    2014-01-01

    A protocol for obtaining bulbs via in vitro organogenesis was developed for tarda tulip (Tulipa tarda Stapf). Scale explants were obtained from bulbs formed at the base of seedlings or from adventitious bulbs that developed from callus tissue forming on stolons or on germinating seeds. Some explants were subjected to chilling at 5°C for 12 wk. The culture media contained 3 or 6% sucrose and was supplemented with either no growth regulators, either 0.5 μM 6-benzyl-aminopurine (BAP) or 18.9 or 94.6 μM abscisic acid (ABA). Cultures were maintained in the dark at 20°C. Callus tissue developed mainly on media without growth regulators or with BAP. Callus was formed from up to 96% of explants derived from non-chilled adventitious bulbs that were treated with 3% sucrose and 0.5 μM BAP. Less callus was formed from chilled explants compared with non-chilled explants. Newly formed adventitious bulbs appeared on the explants via direct and indirect organogenesis. The media with BAP promoted the formation of adventitious bulbs at a rate of 56-92% from non-chilled explants, whereas a maximum rate of 36% was observed from chilled explants. ABA inhibited the induction of adventitious bulbs and callus. The adventitious bulbs obtained in these experiments contained a meristem, which was evidence that they had developed properly.

  6. Low-intensity pulsed ultrasound (LIPUS) and pulsed electromagnetic field (PEMF) treatments affect degeneration of cultured articular cartilage explants

    NARCIS (Netherlands)

    Tan, Lijun; Ren, Yijin; Kooten, van Theo G.; Grijpma, Dirk W.; Kuijer, Roel

    2015-01-01

    Purpose: Articular cartilage has some capacity for self-repair. Clinically used low-intensity pulsed ultrasound (LIPUS) and pulsed electromagnetic field (PEMF) treatments were compared in their potency to prevent degeneration using an explant model of porcine cartilage. Methods: Explants of porcine

  7. CALLUS INDUCTION AND PLANT REGENERATION IN PUNICA GRANATUM L. ?NANA' FROM LEAF EXPLANTS

    Directory of Open Access Journals (Sweden)

    Alireza Bonyanpour

    2013-09-01

    Full Text Available ABSTRACT In this investigation, leaf explants of a local cultivar of dwarf pomegranate were placed on Murashige and Skoog (1962 (MS medium supplemented with various concentrations of 6-benzyl adenin (BA and naphthalene acetic acid (NAA for callus induction. After 40 days, maximum callus induction was observed on a media containing 1 mg L-1 BA and 0.2 to 0.4 mg L-1 NAA. However, the highest callus growth was obtained on a medium containing 1 mg L-1 BA and 1 mg L-1 NAA. The highest number of shoots (7 shoots per explants was obtained by transferring the calli to the media containing 5 mg L-1 BA with 0.1 mg L-1 NAA. Maximum shoot proliferation was observed when shoots were cultured on woody plant medium (WPM supplemented with 5 mg L-1 kinetin (Kin. In this treatment, after 4 subcultures, 36 shoots were produced from one original explant. Among treatments used in rooting experiments, shoots cultured on WPM medium containing 0.2 mg L-1 indol butyric acid (IBA had the maximum root percentage (100% and good root growth (2.06 cm mean length and 2 roots in each explants. Rooted plantlets were cultured in a soil mixture containing vermiculite (60%, perlite (30% and coco peat (10% v/v. After 2 months, 80% of plants survived and transferred to the greenhouse.

  8. Culture of human adipose tissue explants leads to profound alteration of adipocyte gene expression.

    Science.gov (United States)

    Gesta, S; Lolmède, K; Daviaud, D; Berlan, M; Bouloumié, A; Lafontan, M; Valet, P; Saulnier-Blache, J S

    2003-03-01

    Primary culture of adipose tissue has often been used to investigate pharmacological and nutritional regulation of adipocyte gene expression. Possible alteration of adipocyte gene expression by primary culture on its own has not been explored in detail. In order to address this issue, explants were prepared from human subcutaneous adipose tissue recovered from plastic surgery and maintained for 0 to 48 h in DMEM supplemented with 10 % serum. At different time points, adipocytes were isolated from the explants by collagenase digestion, and mRNA expression and lipolysis were studied. Culture was associated with an accumulation of tumor necrosis factor-alpha (TNFalpha) in the culture medium, an increase in anaerobic glycolysis, and an increase in the basal lipolysis. In parallel, a rapid and dramatic decrease in the level of mRNA encoding for several adipocyte-specific proteins such as adipocyte lipid-binding protein, hormone-sensitive lipase, lipoprotein lipase, and peroxisome proliferation activating receptor-gamma2 was observed in isolated adipocytes. These downregulations were reminiscent of a dedifferentiation process. In parallel, primary culture was associated with an increase in adipocyte beta-actin, TNFalpha, glucose transporter-1 and hypoxia-induced factor-1alpha mRNAs. Treatment of explants with agents that increase cAMP (isobutylmethylxanthine and forskolin) prevented TNFalpha production and expression and culture-induced alterations of adipocyte gene expression. These data show that primary culture of human adipose tissue explants dramatically alters adipocyte gene expression.

  9. Re-use of explanted DDD pacemakers as VDD- clinical utility and cost effectiveness.

    Science.gov (United States)

    Namboodiri, K K N; Sharma, Y P; Bali, H K; Grover, A

    2004-01-01

    Re-use of DDD pulse generators explanted from patients died of unrelated causes is associated with an additional cost of two transvenous leads if implanted as DDD itself, and high rate of infection according to some studies. We studied the clinical and economical aspects of reutilization of explanted DDD pacemakers programmed to VDD mode. Out of 28 patients who received VDD pacemaker during the period, October 2000- September 2001 in the Department of Cardiology, PGIMER, Chandigarh, 5 poor patients were implanted with explanted DDD pulse generators programmed to VDD mode. Each implantation was planned and carried out according to a standard protocol. The age ranged from 45 to 75 (mean-61) years. The indications for pacing were complete heart block (4) and second degree AV block (1). The clinical profile, costs and complications, if any were noted and followed up at regular intervals. The results were compared with patients who received new DDD pulse generators during this period. The additional cost for the atrial lead was not required in these patients. None of these patients had any local site infection. Compared to the two-lead system, the single lead system provided more rapid implantation and minimized complications associated with placement of an atrial lead. The explanted DDD pacemaker can be safely reused as VDD mode with same efficacy in selected patient population. This is associated with lower cost and complications compared to reimplantation as DDD itself.

  10. Investigating the Skoog-Miller Model for Organogenesis Using Sweet Potato Root Explants.

    Science.gov (United States)

    Delany, William; And Others

    1994-01-01

    Describes an experiment in which groups of students in a plant tissue culture course worked together to test application of the Skoog-Miller model (developed by Skoog and Miller in regeneration of tobacco experiments to demonstrate organogenesis) to sweet potato root explants. (ZWH)

  11. In vitro regeneration of Centaurium erythraea Rafn from shoot tips and other seedling explants

    Directory of Open Access Journals (Sweden)

    Ewelina Piątczak

    2011-01-01

    Full Text Available Various explants from 30-day-old seedlings of Centaurium erythraea Rafn were evaluated for their morphogenetic capacity under in vitro culture conditions. Shoot formation from shoot tip explants was achieved mainly through adventitious bud differentiation. The highest number of shoots (up to 43.3 ± 2.2 from a single shoot tip was obtained on Murashige and Skoog medium (MS supplemented with indole-3-acetic acid (IAA (0.57 μM and 6-benzylaminopurine (BAP (4.4 μM. Adventitious shoot regeneration was also achieved through organogenesis from calluses obtained from hypocotyls, cotyledons, roots and leaves on MS medium containing IAA (2.85 μM and BAP (0.88 μM. Significant differences were noted between explant types in their effects on shoot regeneration. In the primary culture, the best response was obtained either from calluses derived from roots or leaves (44.4 ± 4.5 and 40.2 ± 6.0 shoots per callus, respectively. The number of subcultures of inoculated calluses affected both the multiplication rate (the number of shoots/explant and shoot morphology (the frequency of shoot hyperhydricity. Shoots rooted with the frequency of 94-100% after culture on MS medium without growth regulators. Plantlets were successfully acclimatized (97% under high relative humidity and then moved to the greenhouse.

  12. Detection of basal and potassium-evoked acetylcholine release from embryonic DRG explants.

    Science.gov (United States)

    Bernardini, Nadia; Tomassy, Giulio Srubek; Tata, Ada Maria; Augusti-Tocco, Gabriella; Biagioni, Stefano

    2004-03-01

    Spontaneous and potassium-induced acetylcholine release from embryonic (E12 and E18) chick dorsal root ganglia explants at 3 and 7 days in culture was investigated using a chemiluminescent procedure. A basal release ranging from 2.4 to 13.8 pm/ganglion/5 min was detected. Potassium application always induced a significant increase over the basal release. The acetylcholine levels measured in E12 explants were 6.3 and 38.4 pm/ganglion/5 min at 3 and 7 days in culture, respectively, while in E18 explant cultures they were 10.7 and 15.5 pm/ganglion/5 min. In experiments performed in the absence of extracellular Ca2+ ions, acetylcholine release, both basal and potassium-induced, was abolished and it was reduced by cholinergic antagonists. A morphometric analysis of explant fibre length suggested that acetylcholine release was directly correlated to neurite extension. Moreover, treatment of E12 dorsal root ganglion-dissociated cell cultures with carbachol as cholinergic receptor agonist was shown to induce a higher neurite outgrowth compared with untreated cultures. The concomitant treatment with carbachol and the antagonists at muscarinic receptors atropine and at nicotinic receptors mecamylamine counteracted the increase in fibre outgrowth. Although the present data have not established whether acetylcholine is released by neurones or glial cells, these observations provide the first evidence of a regulated release of acetylcholine in dorsal root ganglia.

  13. Production of immunoglobulins in gingival tissue explant cultures from juvenile periodontitis patients

    Energy Technology Data Exchange (ETDEWEB)

    Hall, E.R.; Falkler, W.A. Jr.; Suzuki, J.B. (Univ. of Maryland Dental School, Baltimore (USA))

    1990-10-01

    B lymphocytes and plasma cells are histologically observed in granulomatous periodontal tissues of juvenile periodontitis (JP) patients. Local immune processes may participate in protective or immunopathologic roles in the pathogenesis of this disease. An in vitro explant culture system was utilized to demonstrate the production of immunoglobulins by diseased JP tissues. Immunodiffusion studies using goat anti-human gamma, alpha, or mu chain serum revealed IgG to be the major immunoglobulin present in 92% of the day 1 supernatant fluids (SF) of the 47 JP gingival tissue explant cultures. IgA was present in 15% of the SF; however, no IgM was detected. Staph Protein A isolated 14C-labeled IgG from the SF, when allowed to react with goat anti-human gamma chain serum, formed lines of precipitation. Positive autoradiographs confirmed the biosynthesis of IgG by the explant cultures. The in vitro gingival tissue explant culture system described provides a useful model for the study of localized immunoglobulins produced by diseased tissues of JP patients.

  14. Investigating the Skoog-Miller Model for Organogenesis Using Sweet Potato Root Explants.

    Science.gov (United States)

    Delany, William; And Others

    1994-01-01

    Describes an experiment in which groups of students in a plant tissue culture course worked together to test application of the Skoog-Miller model (developed by Skoog and Miller in regeneration of tobacco experiments to demonstrate organogenesis) to sweet potato root explants. (ZWH)

  15. Callus induction and plant regeneration from different explant types of Miscanthus x ogiformis Honda 'Giganteus'

    DEFF Research Database (Denmark)

    Holme, Inger Bæksted; Petersen, Karen Koefoed

    1996-01-01

    . The explants were cultured on urashige and Skoog medium supplemented with 4.5, 13.6, 22.6 or 31.7 μM 2,4-dichlorophenoxyacetic acid. Three types of callus were formed but only one was embryogenic and regenerated plants. Callus induction and formation of embryogenic callus depended on the type and developmental...

  16. Staphylococcus aureus induces hypoxia and cellular damage in porcine dermal explants

    Science.gov (United States)

    Methicillin-resistant Staphylococcus aureus (MRSA) can infect wounds and produce difficult-to- treat biofilms. To determine the extent that MRSA biofilms can deplete oxygen, change pH and damage host tissue, we developed a porcine dermal explant model on which we cultured GFP-labeled MRSA biofilms. ...

  17. Adventitious shoot regeneration from leaf explants of southern highbush blueberry cultivars

    Science.gov (United States)

    Protocols were developed to optimize adventitious shoot regeneration from four southern highbush blueberry cultivars. Leaf explants from six-week-old shoots of the four cultivars were excised and cultured on ten WPM (woody plant medium)-based regeneration media each containing thidiazuron (TDZ) (4.5...

  18. Effect of explants, hormonal combination and genotype on micropropagation of pepper

    Directory of Open Access Journals (Sweden)

    Z. Moheb Mohamadi

    2014-07-01

    Full Text Available Pepper (Capsicum annuum is generally propagated from seeds. In Iran, imported hybrid seeds are used for this purpose. Germination of the greenhouse pepper seeds and stabilization of their seedlings takes a long time. But, tissue culture technique could rapidly propagate this plant with a homogeneous genetic structure in a large scale. An attempt was made in this study to evaluate the effects of different genotypes, explants and growth regulators on callus production and regeneration and introduce the appropriate protocol for micropropagation of commercial varieties of pepper in Iran. Analysis of variance indicated that effect of different genotypes, interaction of various explants and combination of hormones and interaction between genotypes, explants and hormones were significant on the evaluated traits at 1% level. Based on the results, for micropropagation of pepper with leaves and cotyledons, combination of 1 mg/l IAA and 5 mg/l BAP was selected. It seems that using leaf explants for micropropagation of pepper in large scale in greenhouses would be suitable.

  19. Effect of clinorotation on in vitro cultured explants of Mentha piperita L.

    Science.gov (United States)

    Paolicchi, Fabio; Mensuali-Sodi, Anna; Tognoni, Franco

    2002-02-14

    An in vitro culture system was used to study the influence of gravity on axillary shoot formation and adventitious root regeneration in Mentha piperita L. The direction of the gravity vector was altered by displacing stem node explants in different orientations. Also, microgravity conditions were simulated by rotating the explants on a horizontal clinostat so that the main axis of nodes was either parallel (Cpa) or perpendicular to the clinostat axis (Ccp and Ccf, centripetally and centrifugally oriented, respectively). Mint nodes were cultured on solidified Linsmaier and Skoog's medium [Physiol. Plant. 18 (1965) 100] adding a filter-sterilized aqueous solution of 2 mg/l benzyladenine (BA) in half of the cultures. The proliferation of axillary shoots as well as adventitious root formation were not affected by altering upright explant orientation. On the contrary clinorotation was able to modify plantlet development. In absence of BA, leaf width was hindered by Cpa treatment and penultimate internode length was enhanced by Ccp. Furthermore, a negative effect of Cpa treatment was observed in root length parameter, while Ccp increased the root number both in absence and in presence of BA. An effect strictly connected to clinorotation in presence of BA was the occurrence of hyperhydricity. Moreover, explants under clinorotation treatments switched their gravitropic response modifying shoot curvature.

  20. Fumonisin B₁ (FB₁) Induces Lamellar Separation and Alters Sphingolipid Metabolism of In Vitro Cultured Hoof Explants.

    Science.gov (United States)

    Reisinger, Nicole; Dohnal, Ilse; Nagl, Veronika; Schaumberger, Simone; Schatzmayr, Gerd; Mayer, Elisabeth

    2016-03-24

    One of the most important hoof diseases is laminitis. Yet, the pathology of laminitis is not fully understood. Different bacterial toxins, e.g. endotoxins or exotoxins, seem to play an important role. Additionally, ingestion of mycotoxins, toxic secondary metabolites of fungi, might contribute to the onset of laminitis. In this respect, fumonsins are of special interest since horses are regarded as species most susceptible to this group of mycotoxins. The aim of our study was to investigate the influence of fumonisin B₁ (FB₁) on primary isolated epidermal and dermal hoof cells, as well as on the lamellar tissue integrity and sphingolipid metabolism of hoof explants in vitro. There was no effect of FB₁ at any concentration on dermal or epidermal cells. However, FB₁ significantly reduced the separation force of explants after 24 h of incubation. The Sa/So ratio was significantly increased in supernatants of explants incubated with FB₁ (2.5-10 µg/mL) after 24 h. Observed effects on Sa/So ratio were linked to significantly increased sphinganine concentrations. Our study showed that FB₁ impairs the sphingolipid metabolism of explants and reduces lamellar integrity at non-cytotoxic concentrations. FB₁ might, therefore, affect hoof health. Further in vitro and in vivo studies are necessary to elucidate the effects of FB₁ on the equine hoof in more detail.

  1. Induction of in vitro shoots of Billbergia euphemiae E. Morren (Bromeliaceae from leaf explants

    Directory of Open Access Journals (Sweden)

    Mariela Justiniano Simão

    2016-10-01

    Full Text Available Bromeliads are an important group for the maintenance of the Atlantic Forest, with many threatened species due to exacerbated extraction and destruction of their natural habitats. Considering the need of developing protocols for the conservation of these species, the aim of this work was to evaluate the effect of different growth regulators in the in vitro induction of shoots of Billbergia euphemiae. Leaf explants were excised from seedlings derived from in vitro germination and grown on MS medium supplemented with NAA (0, 1 or 2 μM and BA (0, 2, 4 or 6 μM combinations. The evaluation of the number of shoots per explant, shoot length, number of leaves per shoot and longest leaf length average was carried out after 30 and 60 days of culture. The best in vitro responses were observed in the presence of 1 μM NAA after 60 days of culture, which induced the best production of shoots per explant (16.39, as well as the highest rates of shoot length (1.08 cm, number of leaves per shoot (5.00 and the longest leaf length (0.56 cm. This work determined the best conditions for shoot production from leaf explants of B. euphemiae, being the first report on micropropagation of this species.

  2. A RIFAMPICINA NA DESCONTAMINAÇÃO BACTERIANA DE EXPLANTES DE MAMOEIRO PROVENIENTES DO CAMPO

    Directory of Open Access Journals (Sweden)

    GIOVANNI RODRIGUES VIANNA

    1997-01-01

    Full Text Available Observou-se alta contaminação bacteriana nos explantes de mamoeiro introduzidos in vitro, a partir de plantas matrizes desenvolvidas no campo, independentemente da época do ano em que se realizaram as coletas. O uso de desinfestantes superficiais, como álcool e hipoclorito de sódio, garantiram níveis aceitáveis de controle apenas para fungos, não para bactérias. A rifampicina, por tratamento de imersão ou introdução em meio de cultura, controlou satisfatoriamente as contaminações de caráter endofítico, obtendo-se 70% de explantes sadios, sem sinais de fitotoxicidade.High contamination by bacteria was observed in papaya tissue cuttings introduced in vitro from plants grown in the field, independent of the period of the year that samples were collected. The use of alcohol and sodium hypoclorite did not guarantee good bacteria control. Rifampicin, added as an immersion solution treatment or in the culture media, controlled the internal contamination of explants, without damaging the cuttings. Up to 70% of healthy tissue explants were obtained by the use of rifampicin.

  3. Explant culture of human peripheral lung. I. Metabolism of benzo[alpha]pyrene

    DEFF Research Database (Denmark)

    Stoner, G.D.; Harris, C.C.; Autrup, Herman

    1978-01-01

    Human lung explants have been maintained in vitro for a period of 25 days. Autoradiographic studies indicated that the broncholar epithelial cells, type 2 alveolar epithelial cells, and stromal fibroblasts incorporated 3H-thymidine during the culture. After 7 to 10 days, type 2 cells were the pre...

  4. Effect of explant type on the rooting and acclimatization of Dianthus serotinus Waldst. & Kit.

    Directory of Open Access Journals (Sweden)

    Marković Marija

    2014-01-01

    Full Text Available The effect of the concentration of MS salts and explant type on D. serotinus rooting and acclimatization was investigated in order to optimize a protocol for the micropropagation of this species. The obtained results showed that explant type as well as the concentration of MS salts had a significant effect on rooting, and the highest rooting rate (85-86,7% was achieved when culturing single-node cuttings and terminal buds on a half-strength MS medium supplemented with 0,5 mgL-1 NAA. Nevertheless, mean number of roots per explant was higher on the MS media (15,3-18,6 than on the half-strength MS media (11,8-13,4. The best acclimatization rate was obtained in a 4:1 mixture of peat and sand (83,3-86,7%. The explant type from which in vitro plantlets developed had no effect on the acclimatization rate. [Projekat Ministarstva nauke Republike Srbije, br. 43007

  5. Wear study of Total Ankle Replacement explants by microstructural analysis.

    Science.gov (United States)

    Cottrino, S; Fabrègue, D; Cowie, A P; Besse, J-L; Tadier, S; Gremillard, L; Hartmann, D J

    2016-08-01

    The implantation of Total Ankle Replacement (TAR) prostheses generally gives satisfactory results. However, a high revision rate is associated with the Ankle Evolutive System (AES) implant, due to periprosthetic osteolysis that generates significant cortical lesions and bone cysts in the periprosthetic region. Radioclinical and histological analyses of peri-implant tissues show the presence of numerous foreign particles that may come from the implant. It is known that a precocious wear of materials may lead to an important rate of foreign body in tissues and may generate osteolysis lesions and inflammatory reactions. Thus the objectives of this retrospective study of 10 AES TAR implants (recovered after revision surgeries) are to understand how the prostheses wear out, which part is the most stressed and to determine the nature and size of foreign body particles. A better understanding of friction mechanisms between the three parts of the implant and of the nature and morphology of foreign particles generated was needed to explain the in vivo behavior of the implant. This was achieved using microstuctural and tomographic analysis of both implants parts and periprosthetic tissues.

  6. Morphological and functional characteristics of three-dimensional engineered bone-ligament-bone constructs following implantation.

    Science.gov (United States)

    Ma, Jinjin; Goble, Kristen; Smietana, Michael; Kostrominova, Tatiana; Larkin, Lisa; Arruda, Ellen M

    2009-10-01

    The incidence of ligament injury has recently been estimated at 400,000/year. The preferred treatment is reconstruction using an allograft, but outcomes are limited by donor availability, biomechanical incompatibility, and immune rejection. The creation of an engineered ligament in vitro solely from patient bone marrow stromal cells (has the potential to greatly enhance outcomes in knee reconstructions. Our laboratory has developed a scaffoldless method to engineer three-dimensional (3D) ligament and bone constructs from rat bone marrow stem cells in vitro. Coculture of these two engineered constructs results in a 3D bone-ligament-bone (BLB) construct with viable entheses, which was successfully used for medial collateral ligament (MCL) replacement in a rat model. 1 month and 2 month implantations were applied to the engineered BLBs. Implantation of 3D BLBs in a MCL replacement application demonstrated that our in vitro engineered tissues grew and remodeled quickly in vivo to an advanced phenotype and partially restored function of the knee. The explanted 3D BLB ligament region stained positively for type I collagen and elastin and was well vascularized after 1 and 2 months in vivo. Tangent moduli of the ligament portion of the 3D BLB 1 month explants increased by a factor of 2.4 over in vitro controls, to a value equivalent to those observed in 14-day-old neonatal rat MCLs. The 3D BLB 1 month explants also exhibited a functionally graded response that closely matched native MCL inhomogeneity, indicating the constructs functionally adapted in vivo.

  7. Increased angiogenic response in aortic explants of collagen XVIII/endostatin-null mice.

    Science.gov (United States)

    Li, Qing; Olsen, Bjorn R

    2004-08-01

    Endostatin, a proteolytic fragment of basement membrane-associated collagen XVIII, has been shown to be a potent angiogenesis inhibitor both in vivo and in vitro when given at high concentrations. The precise molecular mechanisms by which it functions and whether or not it plays a role in physiological regulation of angiogenesis are not clear. In mice with targeted null alleles of Col18a1, there appears to be no major abnormality in vascular patterns or capillary density in most organs. Furthermore, the growth of experimental tumors is not increased. However, a detailed analysis of induced angiogenesis in these mice has not been performed. Therefore, we compared the angiogenic responses induced by in vitro culture of aortic explants from collagen XVIII/endostatin-null mice (ko) to wild-type (wt) littermates. We found a twofold increase in microvessel outgrowth in explants from ko mice, relative to wt explants. This increased angiogenesis was reduced to the wt level by the addition of low levels (0.1 microg/ml) of recombinant mouse or human endostatin during the culture period. To address cellular/molecular mechanisms underlying this difference in angiogenic response between ko and wt mice, we isolated endothelial cells from both strains and compared their biological behavior. Proliferation assays showed no difference between the two types of endothelial cells. In contrast, adhesion assays showed a striking difference in their ability to adhere to fibronectin suggesting that collagen XVIII/endostatin may regulate interactions between endothelial cells and underlying basement membrane-associated components, including fibronectin, such that in the absence of collagen XVIII/endostatin, endothelial cells are more adhesive to fibronectin. In the aortic explant assay, characterized by dynamic processes of microvessel elongation and regression, this may result in stabilization of newly formed vessels, reduced regression, and a net increase in microvessel outgrowth in

  8. Milk Thistle Extract and Silymarin Inhibit Lipopolysaccharide Induced Lamellar Separation of Hoof Explants in Vitro

    Directory of Open Access Journals (Sweden)

    Nicole Reisinger

    2014-10-01

    Full Text Available The pathogenesis of laminitis is not completely identified and the role of endotoxins (lipopolysaccharides, LPS in this process remains unclear. Phytogenic substances, like milk thistle (MT and silymarin, are known for their anti-inflammatory and antioxidant properties and might therefore have the potential to counteract endotoxin induced effects on the hoof lamellar tissue. The aim of our study was to investigate the influence of endotoxins on lamellar tissue integrity and to test if MT and silymarin are capable of inhibiting LPS-induced effects in an in vitro/ex vivo model. In preliminary tests, LPS neutralization efficiency of these phytogenics was determined in an in vitro neutralization assay. Furthermore, tissue explants gained from hooves of slaughter horses were tested for lamellar separation after incubation with different concentrations of LPS. By combined incubation of explants with LPS and either Polymyxin B (PMB; positive control, MT or silymarin, the influence of these substances on LPS-induced effects was assessed. In the in vitro neutralization assay, MT and silymarin reduced LPS concentrations by 64% and 75%, respectively, in comparison PMB reduced 98% of the LPS concentration. In hoof explants, LPS led to a concentration dependent separation. Accordantly, separation force was significantly decreased by 10 µg/mL LPS. PMB, MT and silymarin could significantly improve tissue integrity of explants incubated with 10 µg/mL LPS. This study showed that LPS had a negative influence on the structure of hoof explants in vitro. MT and silymarin reduced endotoxin activity and inhibited LPS-induced effects on the lamellar tissue. Hence, MT and silymarin might be used to support the prevention of laminitis and should be further evaluated for this application.

  9. Induction of cardiac muscle differentiation in isolated animal pole explants of Xenopus laevis embryos.

    Science.gov (United States)

    Logan, M; Mohun, T

    1993-07-01

    We have isolated a cDNA fragment encoding a portion of the myosin heavy chain alpha-isoform (XMHC alpha) in the amphibian, Xenopus laevis. The XMHC alpha transcript is highly enriched in adult heart RNA and is expressed exclusively in embryonic heart tissue. It therefore provides a tissue-specific marker for cardiac muscle differentiation during early embryogenesis. Using an RNAase protection assay, we can detect the onset of cardiac muscle differentiation in an anterior, ventral region of tailbud embryos, many hours before the appearance of a beating heart. Whole-mount in situ RNA hybridisation indicates that expression of the XMHC alpha gene is restricted to the developing heart primordium. XMHC alpha gene expression can also be induced in isolated animal pole explants of blastulae by treatment with the growth factor, activin A. Induction is dose-dependent, requiring high doses of the growth factor compared with that required for myotomal (skeletal) muscle differentiation. In contrast, no XMHC alpha transcripts are detected in explants incubated with basic FGF, despite the induction of myotomal muscle differentiation. Activin-induced explants show a similar temporal pattern of XMHC alpha gene expression to that found in normal embryogenesis. Furthermore, cells expressing this gene appear clustered in one or two foci within fused explant aggregates, which often show regular, spontaneous contractions after several days in culture. These results show that terminal differentiation of cardiac muscle can occur in growth factor-induced explants and may be distinguished from skeletal muscle differentiation by the dose and nature of the inducing factor.

  10. Dissection and culture of mouse dopaminergic and striatal explants in three-dimensional collagen matrix assays.

    Science.gov (United States)

    Schmidt, Ewoud R E; Morello, Francesca; Pasterkamp, R Jeroen

    2012-03-23

    Midbrain dopamine (mdDA) neurons project via the medial forebrain bundle towards several areas in the telencephalon, including the striatum(1). Reciprocally, medium spiny neurons in the striatum that give rise to the striatonigral (direct) pathway innervate the substantia nigra(2). The development of these axon tracts is dependent upon the combinatorial actions of a plethora of axon growth and guidance cues including molecules that are released by neurites or by (intermediate) target regions(3,4). These soluble factors can be studied in vitro by culturing mdDA and/or striatal explants in a collagen matrix which provides a three-dimensional substrate for the axons mimicking the extracellular environment. In addition, the collagen matrix allows for the formation of relatively stable gradients of proteins released by other explants or cells placed in the vicinity (e.g. see references 5 and 6). Here we describe methods for the purification of rat tail collagen, microdissection of dopaminergic and striatal explants, their culture in collagen gels and subsequent immunohistochemical and quantitative analysis. First, the brains of E14.5 mouse embryos are isolated and dopaminergic and striatal explants are microdissected. These explants are then (co)cultured in collagen gels on coverslips for 48 to 72 hours in vitro. Subsequently, axonal projections are visualized using neuronal markers (e.g. tyrosine hydroxylase, DARPP32, or βIII tubulin) and axon growth and attractive or repulsive axon responses are quantified. This neuronal preparation is a useful tool for in vitro studies of the cellular and molecular mechanisms of mesostriatal and striatonigral axon growth and guidance during development. Using this assay, it is also possible to assess other (intermediate) targets for dopaminergic and striatal axons or to test specific molecular cues.

  11. Tetanus-diphtheria-acellular pertussis vaccination of adults in the USA.

    Science.gov (United States)

    Gidengil, Courtney A; Sandora, Thomas J; Lee, Grace M

    2008-07-01

    Pertussis is an important cause of morbidity and mortality, and its incidence has been increasing in adolescents and adults over the past two decades. Waning immunity in adolescents and adults may be partially responsible. Adults can suffer significant illness from pertussis and its complications, such as pneumonia, rib fractures and syncope. Moreover, adults serve as a source of disease for infants, who are more vulnerable to severe complications and even death. The economic burden of pertussis is substantial, in terms of both medical and nonmedical costs. Fortunately, the burden of pertussis disease can now be safely and effectively reduced by vaccinating adults with tetanus-diphtheria-acellular pertussis (Tdap) vaccine. Further research is needed to elucidate the role of vaccination in pregnant women and those over 65 years of age, and also to determine whether further booster doses of Tdap are needed.

  12. Gravity related behavior of the acellular slime mold Physarum polycephalum (7-IML-1)

    Science.gov (United States)

    Block, I.

    1992-01-01

    The objective of the experiment is to investigate the effect of near weightlessness on a single cell. The test object is the acellular slime mold Physarum polycephalum. This cell is composed of a network of protoplastic strands which perform rhythmic contractions in the minute range. These contractions of the strands' ectoplastic walls generate the force to drive the vigorous shuttle streaming of fluid protoplasm inside the strands (hydrostatic pressure flow). A net transport of protoplasm in one direction determines the direction of the cell's locomotion itself. In this way, gravity modifies the contraction rhythm of the strands, the streaming velocity of protoplasm in the strands, and the direction of locomotion of the whole slime mold (geotaxis). The other parts of this experiment will address the major question of how this cell, which does not possess any specialized gravireceptors, gets the information about the direction of the gravity vector. Details of the experimental setup are given.

  13. A novel porcine acellular dermal matrix scaffold used in periodontal regeneration

    Institute of Scientific and Technical Information of China (English)

    Jing Guo; Hui Chen; Ying Wang; Cheng-Bo Cao; Guo-Qiang Guan

    2013-01-01

    Regeneration of periodontal tissue is the most promising method for restoring periodontal structures. To find a suitable bioactive three- dimensional scaffold promoting cell proliferation and differentiation is critical in periodontal tissue engineering. The objective of this study was to evaluate the biocompatibility of a novel porcine acellular dermal matrix as periodontal tissue scaffolds both in vitroand in vivo. The scaffolds in this study were purified porcine acellular dermal matrix (PADM) and hydroxyapatite-treated PADM (HA-PADM). The biodegradation patterns of the scaffolds were evaluated in vitro. The biocompatibility of the scaffolds in vivo was assessed by implanting them into the sacrospinal muscle of 20 New Zealand white rabbits. The hPDL cells were cultured with PADM or HA-PADM scaffolds for 3, 7, 14, 21 and 28 days. Cell viability assay, scanning electron microscopy (SEM), hematoxylin and eosin (H&E) staining, immunohistochemistry and confocal microscopy were used to evaluate the biocompatibility of the scaffolds. In vitro, both PADM and HA-PADM scaffolds displayed appropriate biodegradation pattern, and also, demonstrated favorable tissue compatibility without tissue necrosis, fibrosis and other abnormal response. The absorbance readings of the WST-1 assay were increased with the time course, suggesting the cell proliferation in the scaffolds. The hPDL cells attaching, spreading and morphology on the surface of the scaffold were visualized by SEM, H&E staining, immnuohistochemistry and confocal microscopy, demonstrated that hPDL cells were able to grow into the HA-PADM scaffolds and the amount of cells were growing up in the course of time. This study proved that HA-PADM scaffold had good biocompatibility in animals in vivoand appropriate biodegrading characteristics in vitro. The hPDL cells were able to proliferate and migrate into the scaffold. These observations may suggest that HA-PADM scaffold is a potential cell carrier

  14. Comparative effectiveness of acellular versus whole-cell pertussis vaccines in teenagers.

    Science.gov (United States)

    Klein, Nicola P; Bartlett, Joan; Fireman, Bruce; Rowhani-Rahbar, Ali; Baxter, Roger

    2013-06-01

    During the 1990s, the United States switched from combined diphtheria, tetanus toxoids, whole-cell pertussis (DTwP) vaccines to combined acellular pertussis (DTaP) vaccines because of safety concerns. After a 2010-2011 pertussis outbreak, we sought to evaluate whether disease risk in 10 to 17 year olds differed between those who previously received DTwP from those who received DTaP. A case-control study among individuals born from 1994 to 1999 who received 4 pertussis-containing vaccines during the first 2 years of life at Kaiser Permanente Northern California (KPNC). We separately compared pertussis polymerase chain reaction (PCR)-positive cases with PCR-negative and KPNC-matched controls. We assessed risk of pertussis relative to vaccine type in early childhood (4 DTwPs, mixed DTwP/DTaP, or 4 DTaPs) by using conditional logistic regression stratified for calendar time and adjusted for gender, race, medical clinic, and receipt of reduced antigen content acellular pertussis (Tdap) vaccine. We compared 138 PCR-positive cases with 899 PCR-negative and 54 339 KPNC-matched controls. Teenagers who had received 4 DTwPs were much less likely to be pertussis PCR-positive than those who had received 4 DTaPs (odds ratio 5.63, 95% confidence interval 2.55-12.46) or mixed DTwP/DTaP vaccines (odds ratio 3.77, 95% confidence interval 1.57-9.07). Decreasing number of DTwP doses was significantly associated with increased pertussis risk (P vaccines in childhood were more protected during a pertussis outbreak than were those who received DTaP vaccines.

  15. Effect of schedule on reactogenicity and antibody persistence of acellular and whole-cell pertussis vaccines: value of laboratory tests as predictors of clinical performance.

    Science.gov (United States)

    Miller, E; Ashworth, L A; Redhead, K; Thornton, C; Waight, P A; Coleman, T

    1997-01-01

    The performance of four acellular pertussis vaccines containing between two and five pertussis antigens combined with diphtheria and tetanus toxoids was compared with that of British whole-cell diphtheria/tetanus/pertussis (DTP) vaccine both in laboratory assays for potency, toxicity and immunogenicity, and for reactogenicity and immunogenicity in infants. Clinical responses were evaluated in double blind randomized Phase II trials using 3/5/9 month and 2/3/4 month schedules. The acellular DTPs had much lower toxicity than whole-cell DTP in laboratory tests and were significantly less pyrogenic than whole-cell DTP under both schedules. Local reactions were not consistently lower in acellular than whole-cell vaccinees and varied with the source of the diphtheria and tetanus antigens used. Differences in endotoxin level and content of active pertussis toxin (PT) between acellular DTP vaccines were not clinically significant. The reactogenicity advantage of the acellular vaccines was substantially reduced under the 2/3/4 month schedule due to the reduced reactogenicity of the whole-cell DTP vaccine when given at a younger age. There was no relationship between antigen content measured in micrograms per dose and ELISA antibody responses to filamentous haemagglutinin (FHA) and PT in infants, nor was murine immunogenicity predictive of immunogenicity in humans. Antibody response to PT was attenuated in the whole-cell group under the 2/3/4 month schedule but was unaffected in the group receiving acellular vaccines with individually purified components; antibody response to pertactin (69 kDa antigen) was similar in recipients of the whole-cell and component acellular vaccines under the 2/3/4 month schedule. PT antibody persistence until 4-5 years of age was significantly better in recipients of the component acellular than either the whole-cell vaccine or the co-purified acellular vaccine under the 3/5/9 month schedule. However, diphtheria antitoxin levels were reduced in

  16. Acellularization-Induced Changes in Tensile Properties Are Organ Specific - An In-Vitro Mechanical and Structural Analysis of Porcine Soft Tissues.

    Directory of Open Access Journals (Sweden)

    Stefan Schleifenbaum

    Full Text Available Though xenogeneic acellular scaffolds are frequently used for surgical reconstruction, knowledge of their mechanical properties is lacking. This study compared the mechanical, histological and ultrastructural properties of various native and acellular specimens.Porcine esophagi, ureters and skin were tested mechanically in a native or acellular condition, focusing on the elastic modulus, ultimate tensile stress and maximum strain. The testing protocol for soft tissues was standardized, including the adaption of the tissue's water content and partial plastination to minimize material slippage as well as templates for normed sample dimensions and precise cross-section measurements. The native and acellular tissues were compared at the microscopic and ultrastructural level with a focus on type I collagens.Increased elastic modulus and ultimate tensile stress values were quantified in acellular esophagi and ureters compared to the native condition. In contrast, these values were strongly decreased in the skin after acellularization. Acellularization-related decreases in maximum strain were found in all tissues. Type I collagens were well-preserved in these samples; however, clotting and a loss of cross-linking type I collagens was observed ultrastructurally. Elastins and fibronectins were preserved in the esophagi and ureters. A loss of the epidermal layer and decreased fibronectin content was present in the skin.Acellularization induces changes in the tensile properties of soft tissues. Some of these changes appear to be organ specific. Loss of cross-linking type I collagen may indicate increased mechanical strength due to decreasing transverse forces acting upon the scaffolds, whereas fibronectin loss may be related to decreased load-bearing capacity. Potentially, the alterations in tissue mechanics are linked to organ function and to the interplay of cells and the extracellular matrix, which is different in hollow organs when compared to skin.

  17. Side effects of cellular and acellular DPT vaccine in children aged from 3 months to 5 years

    Directory of Open Access Journals (Sweden)

    Durmišević Smajil

    2004-01-01

    Full Text Available Introduction Both mild and severe local and systemic postvaccination reactions are seen more rarely in infants immunized with DTPa than in those immunized with DTPw vaccine. Material and methods By analysis of medical records and follow-up of patients, the authors searched for sings of adverse effects of DPT vaccines, comparing cellular and acellular vaccines in children aged from three months to five years. The results of investigation were analyzed using X2. Results Out of the total number of 940 applied vaccines, 329 were cellular and 611 were acellular. Body temperature over 38.5oC occurred in 3% of children immunized with cellular DTPw, and vomiting occurred in 0.8% of those immunized with acellular DTPa vaccine. Vomiting occurred (more than five times in 0.9% of children immunized with DPTw and in 0.32% of children immunized with DPTa. Other undesirable symptoms like swelling, redness and pain in the arm were found in 0.6% of children immunized with DPTw, and in 0.32% of children immunized with DPTa; prolonged crying (three hours or longer was registered in 0.3% of cases immunized with DPTw, and in 0.16% of immunized with DPTa vaccine. Convulsions and collapse appeared only in 0.3% of children immunized with DPTw. Discussion Our investigation shows that local and generalized undesirable postvaccination reactions occurred in 5.4% of children immunized with DPTw and in 1.64 of children immunized with DPTa. The latest clinical investigations show that acellular pertussis vaccines are successful in prevention of pertussis and that they are quite safe for infants; in our investigations, local and generalized reactions were markedly rare in children immunized with DPTa. Conclusion Undesirable postvaccination reactions after application of acellular DPT vaccines are less frequent than it is described in relevant references. The most frequent postvaccination reactions was raised body temperature (38.5oC. Convulsions and collapses were not

  18. Influencing micropropagation in Clitoria ternatea L. through the manipulation of TDZ levels and use of different explant types.

    Science.gov (United States)

    Mukhtar, Seemab; Ahmad, Naseem; Khan, Md Imran; Anis, Mohammad; Aref, Ibrahim M

    2012-10-01

    A comparative performance of two explants types (CN and Nodal) for their efficiency to induce multiple shoot regeneration in Clitoria ternatea has been carried out. Thidiazuron (TDZ) in different concentrations (0.05-2.5 μM) was used as a supplement to the Murashige and Skoog's (MS) basal media. Explant type apart, two factors viz. concentration and exposure duration to TDZ played an important role in affecting multiple shoot regeneration. Cotyledonary node explants produced the best results at 0.1 μM TDZ, while in nodal explants the highest rate of shoot formation was achieved on MS medium supplemented with 1.0 μM TDZ. In both the explants, shoot multiplication increased when the regenerated shoots were subcultured on hormone free MS medium after 4 weeks of exposure to TDZ. Among the two, cotyledonary node explants produced considerably higher number of shoots at a comparatively lower concentration of TDZ than nodal explants. The regenerated shoots rooted best on MS medium containing 1.0 μM indole-3-butyric acid (IBA) and were successfully established in pots containing garden soil with 88 % survival rate. All the regenerated plants showed normal morphology and growth characteristics.

  19. Bone scan

    Science.gov (United States)

    ... legs, or spine fractures) Diagnose a bone infection (osteomyelitis) Diagnose or determine the cause of bone pain, ... 2015:chap 43. Read More Broken bone Metabolism Osteomyelitis Review Date 12/10/2015 Updated by: Jatin ...

  20. Bone Cancer

    Science.gov (United States)

    Cancer that starts in a bone is uncommon. Cancer that has spread to the bone from another ... more common. There are three types of bone cancer: Osteosarcoma - occurs most often between ages 10 and ...

  1. Bone Diseases

    Science.gov (United States)

    Your bones help you move, give you shape and support your body. They are living tissues that rebuild constantly ... childhood and your teens, your body adds new bone faster than it removes old bone. After about ...

  2. Bone Markers

    Science.gov (United States)

    ... markers may be seen in conditions such as: Osteoporosis Paget disease Cancer that has spread to the bone (metastatic bone disease) Hyperparathyroidism Hyperthyroidism Osteomalacia in adults and rickets in children—lack of bone mineralization, ...

  3. A Novel 3D Skin Explant Model to Study Anaerobic Bacterial Infection

    Directory of Open Access Journals (Sweden)

    Grazieli Maboni

    2017-09-01

    Full Text Available Skin infection studies are often limited by financial and ethical constraints, and alternatives, such as monolayer cell culture, do not reflect many cellular processes limiting their application. For a more functional replacement, 3D skin culture models offer many advantages such as the maintenance of the tissue structure and the cell types present in the host environment. A 3D skin culture model can be set up using tissues acquired from surgical procedures or post slaughter, making it a cost effective and attractive alternative to animal experimentation. The majority of 3D culture models have been established for aerobic pathogens, but currently there are no models for anaerobic skin infections. Footrot is an anaerobic bacterial infection which affects the ovine interdigital skin causing a substantial animal welfare and financial impact worldwide. Dichelobacter nodosus is a Gram-negative anaerobic bacterium and the causative agent of footrot. The mechanism of infection and host immune response to D. nodosus is poorly understood. Here we present a novel 3D skin ex vivo model to study anaerobic bacterial infections using ovine skin explants infected with D. nodosus. Our results demonstrate that D. nodosus can invade the skin explant, and that altered expression of key inflammatory markers could be quantified in the culture media. The viability of explants was assessed by tissue integrity (histopathological features and cell death (DNA fragmentation over 76 h showing the model was stable for 28 h. D. nodosus was quantified in all infected skin explants by qPCR and the bacterium was visualized invading the epidermis by Fluorescent in situ Hybridization. Measurement of pro-inflammatory cytokines/chemokines in the culture media revealed that the explants released IL1β in response to bacteria. In contrast, levels of CXCL8 production were no different to mock-infected explants. The 3D skin model realistically simulates the interdigital skin and has

  4. Acellular dermal allograft for vestibuloplasty--an alternative to autogenous soft tissue grafts in preprosthetic surgical procedures: a clinical report.

    Science.gov (United States)

    Bhola, Monish; Newell, Donald H; Hancock, Everett B

    2003-06-01

    Various vestibular extension procedures have been described in the literature over the past 6 decades, including the use of free gingival grafts. An acellular dermal allograft has been used as a substitute for autogenous soft tissue grafts in root coverage procedures. This clinical report describes the use of such an allograft as a substitute for palatal donor tissue in the vestibular extension of an edentulous maxillary arch with multiple frenum attachments before fabrication of a complete denture. In this patient, healing was uneventful, and 6-month clinical observations demonstrated an apical positioning of the mucogingival junction with an increase in vestibular depth, and the absence of multiple frenae. The acellular dermal allograft appears to be a useful substitute for autogenous palatal grafts in preprosthetic surgery. This approach has many advantages over the free gingival graft, including no donor site morbidity, unlimited availability, and better color match.

  5. Human acellular dermal matrix for repair of abdominal wall defects: review of clinical experience and experimental data.

    Science.gov (United States)

    Holton, Luther H; Kim, Daniel; Silverman, Ronald P; Rodriguez, Eduardo D; Singh, Navin; Goldberg, Nelson H

    2005-01-01

    The use of prosthetic mesh for the tension-free repair of incisional hernias has been shown to be more effective than primary suture repair. Unfortunately, prosthetic materials can be a suboptimal choice in a variety of clinical scenarios. In general, prosthetic materials should not be implanted into sites with known contamination or infection because they lack an endogenous vascular network and are thus incapable of clearing bacteria. This is of particular relevance to the repair of recurrent hernias, which are often refractory to repair because of indolent bacterial colonization that weakens the site and retards appropriate healing. Although fascia lata grafts and muscle flaps can be employed for tension-free hernia repairs, they carry the potential for significant donor site morbidity. Recently, a growing number of clinicians have used human acellular dermal matrix as a graft material for the tension-free repair of ventral hernias. This material has been shown to become revascularized in both animal and human subjects. Once repopulated with a vascular network, this graft material is theoretically capable of clearing bacteria, a property not found in prosthetic graft materials. Unlike autologous materials such as fascial grafts and muscle flaps, acellular dermal matrix can be used without subjecting the patient to additional morbidity in the form of donor site complications. This article presents a thorough review of the current literature, describing the properties of human acellular dermal matrix and discussing both animal and human studies of its clinical performance. In addition to the review of previously published clinical experiences, we discuss our own preliminary results with the use of acellular dermal matrix for ventral hernia repair in 46 patients.

  6. Early escharectomy and concurrent composite skin grafting over human acellular dermal matrix scaffold for covering deep facial burns.

    Science.gov (United States)

    Tang, Bing; Zhu, Bin; Liang, Yue-Ying; Bi, Liang-Kuan; Chen, Bin; Hu, Zhi-Cheng; Zhang, Kai; Zhu, Jia-Yuan

    2011-04-01

    Although escharectomy and full-thickness skin autografting have been widely used to treat deep facial burns, the clinical outcomes remain unacceptable. Composite razor-thin skin grafting over acellular dermal matrix scaffold has been used successfully in repairing burns of the trunk and limbs, but its use in covering deep facial burns has rarely been reported. In this study, the authors investigated the clinical outcomes of early escharectomy and concurrent composite razor-thin skin autografting and acellular dermal matrix scaffold for treating deep facial burns. Patients with deep facial burns (n = 16) involving 8 to 30 percent of the total body surface area received early escharectomy by postburn day 3 and concurrent, one-stage, large, razor-thin skin autografting on top of human acellular dermal matrix scaffold. Wound dressings were changed on postoperative days 7, 9, and 12 to examine the survival of skin autografts. Patients were followed up for 12 months to evaluate their facial profiles. The take rate of composite skin autografts was 97.3 percent at postoperative day 12. At the follow-up visit, the skin autografts appeared normal in color, with soft texture and good elasticity. The skin junctures showed little scarring. The patients exhibited a chubby facial appearance and abundant expression, except for one patient with microstomia and two patients with ectropion who required further plastic surgical interventions. Early escharectomy and concurrent composite razor-thin skin autografting on top of acellular dermal matrix scaffold constitute an effective and favorable option for covering deep facial burns, especially for patients with limited donor sites.

  7. Long-Term Followup of Dermal Substitution with Acellular Dermal Implant in Burns and Postburn Scar Corrections

    OpenAIRE

    Juhasz, I.; Kiss, B.; Lukacs, L.; Erdei, I.; Peter, Z.; Remenyik, E.

    2010-01-01

    Full-thickness burn and other types of deep skin loss will result in scar formation. For at least partial replacement of the lost dermal layer, there are several options to use biotechnologically derived extracellular matrix components or tissue scaffolds of cadaver skin origin. In a survey, we have collected data on 18 pts who have previously received acellular dermal implant Alloderm. The age of these patients at the injury varied between 16 months and 84 years. The average area of the impl...

  8. Effect of explant origin and different growth regulators on micropropagation of Pistacia atlantica ssp. mutica

    Directory of Open Access Journals (Sweden)

    Ali-Ashraf Mehrabi

    2015-06-01

    Full Text Available Propagation of wild pistachio as a multipurpose woody species is a hard and tedious task. In this research, an effective in vitro protocol was developed for rapid proliferation of wild pistachio (Pistacia atlantica ssp. mutica in MS medium supplemented with B5 vitamins and different growth regulators. Rooting of plantlets was tested by two treatments containing Rhizopon and IBA in ex vitro. With respect to the results, the nodal segments explants, produced the highest shoot frequency, leaf frequency and the tallest shoots. On the other hand, the tallest shoots were generated from shoot tip explant and medium containing of TDZ plus IAA. Both treatments (Rhizopon and IBA led to a remarkable increase in the number of roots, root length and rooting percentage compared to the control. These results may be applied for rapid proliferation to spread the pistachio trees and shrubs that are difficult and time consuming.

  9. Acetylcholine causes rooting in leaf explants of in vitro raised tomato (Lycopersicon esculentum Miller) seedlings.

    Science.gov (United States)

    Bamel, Kiran; Gupta, Shrish Chandra; Gupta, Rajendra

    2007-05-30

    The animal neurotransmitter acetylcholine (ACh) induces rooting and promotes secondary root formation in leaf explants of tomato (Lycopersicon esculentum Miller var. Pusa Ruby), cultured in vitro on Murashige and Skoog's medium. The roots originate from the midrib of leaf explants and resemble taproot. ACh at 10(-5) M was found to be the optimum over a wide range of effective concentrations between 10(-7) and 10(-3) M. The breakdown products, choline and acetate were ineffective even at 10(-3) M concentration. ACh appears to have a natural role in tomato rhizogenesis because exogenous application of neostigmine, an inhibitor of ACh hydrolysis, could mimic the effect of ACh. Neostigmine, if applied in combination with ACh, potentiated the ACh effect.

  10. One-stage explant-implant procedure of exposed porous orbital implants

    DEFF Research Database (Denmark)

    Toft, Peter B; Rasmussen, Marie L Roed; Prause, Jan Ulrik

    2011-01-01

    ) orbital implant from January 2000 to February 2009 were included. The patient records were reviewed; patients were interviewed by telephone and invited for a clinical examination. Histopathological examination was carried out on the removed implants. Main outcome measures were: presence of exposure......Purpose:  To investigate the risks of implant exposure after a combined explant-implant procedure in patients with an exposed porous orbital implant. Methods:  Twenty-four consecutive patients who had a combined explant-implant procedure of an exposed hydroxyapatite (21) or porous polyethylene (3...... of the new implant or not, patient graded satisfaction with the cosmetic result, and presence of poor motility. Results:  None of the new implants became exposed or infected in the follow-up period of 25 [3-94] months (median [range]). The patients scored their satisfaction with the cosmetic result...

  11. Efficient plant regeneration from cotyledon explants of bottle gourd (Lagenaria siceraria Standl.).

    Science.gov (United States)

    Han, J-S; Oh, D-G; Mok, I-G; Park, H-G; Kim, C K

    2004-11-01

    Using cotyledon explants excised from seedlings germinated in vitro, an efficient plant regeneration system via organogenesis was established for bottle gourd (Lagenaria siceraria Standl.). Maximum shoot regeneration was obtained when the proximal parts of cotyledons from 4-day-old seedlings were cultured on MS medium with 3 mg/l BA and 0.5 mg/l AgNO(3) under a 16-h photoperiod. After 3-4 weeks of culture, 21.9-80.7% of explants from the five cultivars regenerated shoots. Adventitious shoots were successfully rooted on a half-strength MS medium with 0.1 mg/l IAA for 2-3 weeks. Flow cytometric analysis revealed that most of the regenerated plants derived from culture on medium with AgNO(3) were diploid.

  12. Activity of MMP-9 after repair of abdominal wall defects with acellular and crosslinked bovine pericardium in rabbit.

    Science.gov (United States)

    Singh, Himani; Kumar, Naveen; Sharma, A K; Kataria, Meena; Munjal, Ashok; Kumar, Amit; Dewangan, Rukmani; Kumar, Vineet; Devarathnam, J; Kumar, Sachin

    2014-02-01

    This study was undertaken for the identification of matrix metalloproteinases (MMPs) in extracts obtained from native, acellular and crosslinked bovine pericardium (in vitro), as well as in the plasma after implantation of these biomaterials in rabbits (in vivo). Native pericardium (NP) expressed a 72 kDa (MMP-2) band; whereas, in acellular pericardium (AP) two bands (10 kDa and 92 kDa) of MMPs were observed of which, 92 kDa band was very faint. AP crosslinked with glutaraldehyde did not show any gelatinase activity and thus reflects the creation of new additional chemical bonds between the collagen molecules which has been effectively removed. Gelatin zymography showed only one major band of 92 kDa in all the implanted and untreated rabbit plasma, but the relative amount of 92 kDa was 1-2 times higher in acellular bovine pericardium implanted rabbits as compared to crosslinked and native groups. In NP group, the 92 kDa band was the dullest among the three groups. This indicated that the level of MMP-9 corresponds to the degree of collagen degradation. © 2012 The Authors. International Wound Journal © 2012 Medicalhelplines.com Inc and John Wiley & Sons Ltd.

  13. Characteristic of c-Kit+ progenitor cells in explanted human hearts

    OpenAIRE

    Matuszczak, Sybilla; Czapla, Justyna; Jarosz-Biej, Magdalena; Wiśniewska, Ewa; Cichoń, Tomasz; Smolarczyk, Ryszard; Kobusińska, Magdalena; Gajda, Karolina; Wilczek, Piotr; Śliwka, Joanna; Zembala, Michał; Zembala, Marian; Szala, Stanisław

    2014-01-01

    According to literature data, self-renewing, multipotent, and clonogenic cardiac c-Kit+ progenitor cells occur within human myocardium. The aim of this study was to isolate and characterize c-Kit+ progenitor cells from explanted human hearts. Experimental material was obtained from 19 adult and 7 pediatric patients. Successful isolation and culture was achieved for 95 samples (84.1 %) derived from five different regions of the heart: right and left ventricles, atrium, intraventricular septum,...

  14. Stent explantation from an arteriovenous fistula for hemodialysis: a case report.

    Science.gov (United States)

    Nano, Giovanni; Dalainas, Ilias; Casana, Renato; Bianchi, Paolo; Lupattelli, Tommaso; Malacrida, Giovanni; Tealdi, Domenico G

    2006-01-01

    Percutaneous transluminal angioplasty is the first treatment of a dysfunctional vascular access for hemodialysis. A case of stenting of a native arteriovenous hemodialysis fistula is reported that was treated with a stent placement at the anastomosis level, with explantation of the stent after complete thrombosis of the fistula 48 hours after the procedure. It is preferable to treat arteriovenous fistulas with simple balloon dilatation, avoiding stenting of the fistula, especially in the anastomosis site.

  15. Concurrent Connection of Embryonic Chick Heart Using a Microfluidic Device for Organ-Explant-Chip

    OpenAIRE

    Owaki, Hirofumi; Masuda, Taisuke; Kawahara, Tomohiro; Miyasaka, Kota; Ogura, Toshihiko; Arai, Fumihito

    2013-01-01

    We propose a concurrent microvascular connection method called suction-induced vascular fixation (SVF) method for the achievement of Organ-Explant-Chip which is a biologically-designed simulator having biological materials such as cells, tissues, and organs. The advantages of proposed method with using a microfluidic device are as follows: (1) operation of flexible objects (blood vessels), (2) alignment the blood vessels concurrently, and (3) reduction of the DOFs of the blood vessels. From t...

  16. Protoplast Isolation in Lupin ( Lupinus mutabilis Sweet): Determination of Optimum Explant Sources and Isolation Conditions

    OpenAIRE

    BABAOĞLU, Mehmet

    2000-01-01

    Effects of cultural factors on the yield, viability and division of protoplasts were investigated in Lupinus mutabilis Sweet containing a high protein content as well as a reasonable oil content which may make this species an alternative crop to soybean in Turkey. Explants from different in vitro seedling parts were evaluated on the suitability of protoplast isolation and viability. Leaf mesophyll was the most suitable tissue as a protoplast source. Pectinases as well as cellulases were es...

  17. Ni2+ treatment causes cement gland formation in ectoderm explants of Xenopus laevis embryo

    Institute of Scientific and Technical Information of China (English)

    HUANGYONG; XIAOYANDING

    1999-01-01

    We found T-type calcium channel blocker Ni2+ can efficiently induce the formation of cement gland in Xenopus laevis animal cap explants.Nother T-typer specific calcium channel blocker Amiloride can also induce the formation of cement gland,while L-type specific calcium channel blocker Nifedipine as no inductive effect.These results may offer us an new approach to study the differentiation of cement gland through the change of intracelluar calcium concentration.

  18. Bordetella avium Causes Induction of Apoptosis and Nitric Oxide Synthase in Turkey Tracheal Explant Cultures

    OpenAIRE

    Miyamoto, David M.; Ruff, Kristin; Beach, Nathan M.; Dorsey-Oresto, Angella; Masters, Isaac; Temple, Louise M.

    2011-01-01

    Bordetellosis is an upper respiratory disease of turkeys caused by Bordetella avium in which the bacteria attach specifically to ciliated respiratory epithelial cells. Little is known about the mechanisms of pathogenesis of this disease, which has a negative impact in the commercial turkey industry. In this study, we produced a novel explant organ culture system that was able to successfully reproduce pathogenesis of B. avium in vitro, using tracheal tissue derived from 26 day-old turkey embr...

  19. Aescin Content in Embryogenic Callus and in Embryoids from Leaf Explants of Aesculus hippocastanum.

    Science.gov (United States)

    Profumo, P; Caviglia, A M; Gastaldo, P; Dameri, R M

    1991-02-01

    HPLC determinations of the aescin contents in calli and embryoids from leaf explants of AESCULUS HIPPOCASTANUM L. were carried out in order to determine whether it was possible to obtain aescin-forming proliferations IN VITRO. The results indicate that embryogenic calli and embryoids derived from them produce an amount of active principle higher than that of horse-chestnut seeds. The data are discussed in terms of the relation between tissue differentiation and secondary metabolites biosynthesis.

  20. Aescin formation in calli and embryoids from cotyledon and stem explants of Aesculus hippocastanum L.

    Science.gov (United States)

    Profumo, P; Caviglia, A M; Gastaldo, P

    1994-11-01

    Aescin in calli and embryoids obtained from both cotyledon and stem explants of Aesculus hippocastanum were investigated by HPLC. Determinations were carried out on tissues cultured in agarized medium supplemented with growth substances (2,4-dichlorophenoxyacetic acid; kinetin; 1-naphthaleneacetic acid). The results indicate that aescin was produced in all the analysed samples. The amount of active principle present in some samples was higher than that found in horse-chestnut seeds.

  1. In vitro mutagenesis in citrus breeding: I. Gamma rays sensitivity of cultivar pera explants; Uso de mutagenese in vitro no melhoramento de citros: I. sensibilidade a raios gama de explantes do cultivar pera

    Energy Technology Data Exchange (ETDEWEB)

    Tulmann Neto, A.; Mendes, B.M.J.; Ando, A. [Centro de Energia Nuclear na Agricultura (CENA), Piracicaba, SP (Brazil); Cristofani, M. [Instituto Agronomico de Campinas, SP (Brazil)

    1994-12-31

    In order to obtain basic data for future research in mutation breeding through in vitro technology in sweet orange, gamma rays sensitivity of different types of plants were analysed. After irradiation the following observations were carried out: response of explants, number of embryoids and adventitious buds, callus weight and rooting formation. Different results were obtained accordingly to the explant used and are presented. (author). 21 refs., 6 tabs.

  2. Optimization of Agrobacterium mediated genetic transformation of cotyledonary node explants of Vigna radiata.

    Science.gov (United States)

    Yadav, Sushil Kumar; Katikala, Sweety; Yellisetty, Varalaxmi; Kannepalle, Annapurna; Narayana, Jyothi Lakshmi; Maddi, Vanaja; Mandapaka, Maheswari; Shanker, Arun Kumar; Bandi, Venkateswarlu; Bharadwaja, Kirti Pulugurtha

    2012-12-01

    A reproducible and highly efficient protocol for genetic transformation mediated by Agrobacterium has been established for greengram (Vigna radiata L. Wilczek). Double cotyledonary node (DCN) explants were inoculated with Agrobacterium tumefaciens strain LBA 4404 harboring a binary vector pCAMBIA 2301 containing neomycin phosphotransferase (npt II) gene as selectable marker, β-glucuronidase (GUS) as a reporter (uidA) gene and annexin 1 bj gene. Important parameters like optical density of Agrobacterium culture, culture quantity, infection medium, infection and co-cultivation time and acetosyringone concentration were standardized to optimize the transformation frequency. Kanamycin at a concentration of 100 mg/l was used to select transformed cells. Transient and stable GUS expressions were studied in transformed explants and regenerated putative plants, respectively. Transformed shoot were produced on regeneration medium containing 100 mg/l kanamycin and 250 mg/l cefotaxime and rooted on ½ MS medium. Transient and constitutive GUS expression was observed in DCN explants and different tissues of T(0) and T(1) plants. Rooted T(0) and T(1) shoots confirming Polymerase Chain Reaction (PCR) positive for npt II and annexin 1bj genes were taken to maturity to collect the seeds. Integration of annexin gene into the greengram genome was confirmed by Southern blotting.

  3. Further assessment of neuropathology in retinal explants and neuroprotection by human neural progenitor cells

    Science.gov (United States)

    Mohlin, Camilla; Liljekvist-Soltic, Ingela; Johansson, Kjell

    2011-10-01

    Explanted rat retinas show progressive photoreceptor degeneration that appears to be caspase-12-dependent. Decrease in photoreceptor density eventually affects the inner retina, particularly in the bipolar cell population. Explantation and the induced photoreceptor degeneration are accompanied by activation of Müller and microglia cells. The goal of this study was to determine whether the presence of a feeder layer of human neural progenitor cells (hNPCs) could suppress the degenerative and reactive changes in the explants. Immunohistochemical analyses showed considerable sprouting of rod photoreceptor axon terminals into the inner retina and reduced densities of cone and rod bipolar cells. Both sprouting and bipolar cell degenerations were significantly lower in retinas cultured with feeder layer cells compared to cultured controls. A tendency toward reduced microglia activation in the retinal layers was also noted in the presence of feeder layer cells. These results indicate that hNPCs or factors produced by them can limit the loss of photoreceptors and secondary injuries in the inner retina. The latter may be a consequence of disrupted synaptic arrangement.

  4. In vitro regeneration from petiole explants of non-toxic Jatropha curcas

    KAUST Repository

    Kumar, Nitish

    2011-01-01

    Jatropha curcas, a multipurpose shrub has acquired significant economic potential as biodiesel plant. The seeds or pressed cake is toxic due to the presence of toxic substances and is not useful as food/fodder despite having the best protein composition. A simple, efficient, and reproducible method for plant regeneration through direct organogenesis from petiole explants of non-toxic J. curcas was developed using Murashige and Skoog (MS) medium supplemented with different concentrations of thidiazuron (TDZ). The best induction of shoot buds (57.61%), and number of shoot buds (4.98) per explant were obtained when in vitro petiole explants were placed horizontally on MS medium supplemented with 2.27 mu M TDZ. The Induced shoot buds were transferred to MS medium containing 10 mu M kinetin (Kn), 4.5 mu M 6-benzyl aminopurine (BA), and 5.5 mu M alpha-naphthaleneacetic acid (NAA) for shoot proliferation and subsequent elongation was achieved on MS medium supplemented with 2.25 mu M BA and 8.5 mu M IAA. The elongated shoots could be rooted on half-strength MS medium with 15 mu M IBA, 11.4 mu M IAA and 5.5 mu M NAA with more than 90% survival rate. (C) 2010 Elsevier B.V. All rights reserved.

  5. Phytic acid decreases deoxynivalenol and fumonisin B1-induced changes on swine jejunal explants

    Directory of Open Access Journals (Sweden)

    Elisângela Olegário da Silva

    2014-01-01

    Full Text Available The purpose of the present study was to investigate the effects of phytic acid (IP6 on morphological and immunohistochemical parameters on intestinal explants exposed to deoxynivalenol (DON and fumonisin B1 (FB1. The jejunal explants were exposed for 4 h to different treatments: control, DON (10 μM, DON plus 2.5 mM or 5 mM IP6, FB1 (70 μM, and FB1 plus 2.5 mM or 5 mM IP6. Both mycotoxins induced significant intestinal lesions and decreased villi height. The presence of 2.5 mM and 5 mM IP6 significantly inhibited the morphological changes caused by the mycotoxins. DON induced a significant increase in caspase-3 (83% and cyclooxygenase-2 (71.3% expression compared with the control. The presence of 5 mM IP6 induced a significant decrease in caspase-3 (43.7% and Cox-2 (48% expression compared with the DON group. FB1 induced a significant increase in caspase-3 expression (47% compared to the control, whereas IP6 induced no significant change in this expression. A significant decrease in cell proliferation was observed when explants were exposed to 5 mM of IP6 in comparison with the DON and FB1 groups. The present data provide evidence that phytic acid modulates the toxic effects induced by DON and FB1 on intestinal tissue.

  6. Phytic acid decreases deoxynivalenol and fumonisin B1-induced changes on swine jejunal explants.

    Science.gov (United States)

    da Silva, Elisângela Olegário; Gerez, Juliana Rubira; do Carmo Drape, Thalisie; Bracarense, Ana Paula F R L

    2014-01-01

    The purpose of the present study was to investigate the effects of phytic acid (IP6) on morphological and immunohistochemical parameters on intestinal explants exposed to deoxynivalenol (DON) and fumonisin B1 (FB1). The jejunal explants were exposed for 4 h to different treatments: control, DON (10 μM), DON plus 2.5 mM or 5 mM IP6, FB1 (70 μM), and FB1 plus 2.5 mM or 5 mM IP6. Both mycotoxins induced significant intestinal lesions and decreased villi height. The presence of 2.5 mM and 5 mM IP6 significantly inhibited the morphological changes caused by the mycotoxins. DON induced a significant increase in caspase-3 (83%) and cyclooxygenase-2 (71.3%) expression compared with the control. The presence of 5 mM IP6 induced a significant decrease in caspase-3 (43.7%) and Cox-2 (48%) expression compared with the DON group. FB1 induced a significant increase in caspase-3 expression (47%) compared to the control, whereas IP6 induced no significant change in this expression. A significant decrease in cell proliferation was observed when explants were exposed to 5 mM of IP6 in comparison with the DON and FB1 groups. The present data provide evidence that phytic acid modulates the toxic effects induced by DON and FB1 on intestinal tissue.

  7. Pericardium Plug in the Repair of the Corneoscleral Fistula After Ahmed Glaucoma Valve Explantation

    Science.gov (United States)

    Yoo, Chungkwon; Kwon, Sung Wook

    2008-01-01

    We report four cases in which a pericardium (Tutoplast®) plug was used to repair a corneoscleral fistula after Ahmed Glaucoma Valve (AGV) explantation. In four cases in which the AGV tube had been exposed, AGV explantation was performed using a pericardium (Tutoplast®) plug to seal the defect previously occupied by the tube. After debridement of the fistula, a piece of processed pericardium (Tutoplast®), measured 1 mm in width, was plugged into the fistula and secured with two interrupted 10-0 nylon sutures. To control intraocular pressure, a new AGV was implanted elsewhere in case 1, phaco-trabeculectomy was performed concurrently in case 2, cyclophotocoagulation was performed postoperatively in case 3 and anti-glaucomatous medication was added in case 4. No complication related to the fistula developed at the latest follow-up (range: 12~26 months). The pericardium (Tutoplast®) plug seems to be an effective method in the repair of corneoscleral fistulas resulting from explantation of glaucoma drainage implants. PMID:19096247

  8. Bordetella avium causes induction of apoptosis and nitric oxide synthase in turkey tracheal explant cultures.

    Science.gov (United States)

    Miyamoto, David M; Ruff, Kristin; Beach, Nathan M; Stockwell, Stephanie B; Dorsey-Oresto, Angella; Masters, Isaac; Temple, Louise M

    2011-09-01

    Bordetellosis is an upper respiratory disease of turkeys caused by Bordetella avium in which the bacteria attach specifically to ciliated respiratory epithelial cells. Little is known about the mechanisms of pathogenesis of this disease, which has a negative impact in the commercial turkey industry. In this study, we produced a novel explant organ culture system that was able to successfully reproduce pathogenesis of B. avium in vitro, using tracheal tissue derived from 26 day-old turkey embryos. Treatment of the explants with whole cells of B. avium virulent strain 197N and culture supernatant, but not lipopolysaccharide (LPS) or tracheal cytotoxin (TCT), specifically induced apoptosis in ciliated cells, as shown by annexin V and TUNEL staining. LPS and TCT are known virulence factors of Bordetella pertussis, the causative agent of whooping cough. Treatment with whole cells of B. avium and LPS specifically induced NO response in ciliated cells, shown by uNOS staining and diaphorase activity. The explant system is being used as a model to elucidate specific molecules responsible for the symptoms of bordetellosis.

  9. Rapid and simple method for in vivo ex utero development of mouse embryo explants.

    Science.gov (United States)

    Gonçalves, André B; Thorsteinsdóttir, Sólveig; Deries, Marianne

    2016-01-01

    The in utero development of mammals drastically reduces the accessibility of the mammalian embryo and therefore limits the range of experimental manipulation that can be done to study functions of genes or signaling pathways during embryo development. Over the past decades, tissue and organ-like culture methods have been developed with the intention of reproducing in vivo situations. Developing accessible and simple techniques to study and manipulate embryos is an everlasting challenge. Herein, we describe a reliable and quick technique to culture mid-gestation explanted mouse embryos on top of a floating membrane filter in a defined medium. Viability of the cultured tissues was assessed by apoptosis and proliferation analysis showing that cell proliferation is normal and there is only a slight increase in apoptosis after 12h of culture compared to embryos developing in utero. Moreover, differentiation and morphogenesis proceed normally as assessed by 3D imaging of the transformation of the myotome into deep back muscles. Not only does muscle cell differentiation occur as expected, but so do extracellular matrix organization and the characteristic splitting of the myotome into the three epaxial muscle groups. Our culture method allows for the culture and manipulation of mammalian embryo explants in a very efficient way, and it permits the manipulation of in vivo developmental events in a controlled environment. Explants grown under these ex utero conditions simulate real developmental events that occur in utero.

  10. Pistacia lentiscus fruit oil reduces oxidative stress in human skin explants caused by hydrogen peroxide.

    Science.gov (United States)

    Ben Khedir, S; Moalla, D; Jardak, N; Mzid, M; Sahnoun, Z; Rebai, T

    2016-10-01

    We investigated the efficacy of Pistacia lentiscus fruit oil (PLFO) for protecting human skin from damage due to oxidative stress. PLFO contains natural antioxidants including polyphenols, sterols and tocopherols. We compared the antioxidant potential of PLFO with extra virgin olive oil (EVOO). Explants of healthy adult human skin were grown in culture with either PLFO or EVOO before adding hydrogen peroxide (H2O2). We also used cultured skin explants to investigate the effects of PLFO on lipid oxidation and depletion of endogenous antioxidant defense enzymes including glutathione peroxidase (GPx), superoxide dismutase (SOD) and catalase (CAT) one day after 2 h exposure to H2O2. We found that PLFO scavenged radicals and protected skin against oxidative injury. PLFO exhibited greater antioxidant and free radical scavenging activity than EVOO. Skin explants treated with PLFO inhibited H2O2 induced MDA formation by inhibition of lipid oxidation. In addition, the oil inhibited H2O2 induced depletion of antioxidant defense enzymes including GPx, SOD and CAT. We found that treatment with PLFO repaired skin damage owing to its antioxidant properties.

  11. Collaborative study on a Guinea pig serological method for the assay of acellular pertussis vaccines.

    Science.gov (United States)

    Winsnes, R; Sesardic, D; Daas, A; Terao, E; Behr-Gross, M-E

    2009-10-01

    An international collaborative study (coded BSP083) was performed under the aegis of the Biological Standardisation Programme supported by the Council of Europe and the European Commission, with the aim of replacing the in vivo challenge assays for potency determination of combined acellular pertussis (aP) vaccines by a refined procedure also allowing reduction of animal use. This study investigates whether the immunogenicity of aP vaccine components could be assayed in a guinea pig (gp) serology model, using the same vaccine immunising doses as for D and T components potency testing, instead of using separate animals as is currently done. The BSP83 project is a follow up of 3 former collaborative studies (coded BSP019, BSP034 and BSP035) on serological methods for the potency testing of tetanus (T) and diphtheria (D) vaccines for human use. The use of gp instead of mice serology has the advantage of providing a larger volume of good quality antiserum for the assay of several vaccine components in the same sample, hence providing the opportunity for animal sparing. The results of Phase I of the study demonstrated that gp serology may be a useful method for the immunogenicity assay of acellular pertussis vaccines. This was confirmed in Phase II of the study, using 7 different combined aP vaccines in an international collaborative study involving 17 laboratories from both public and private sectors. Clear dose-response relationships were observed for different vaccines by ELISA, for antibodies against aP antigens, i.e. pertussis toxin (PT), filamentous haemagglutinin (FHA), fimbrial agglutinogens-2/3 (Fim 2/3) and pertactin (PRN). Intra- and inter-laboratory variations of aP ELISA results were found to be within an acceptable range. For some combined vaccines, however, the range of vaccine dilutions for immunisation confirmed to be optimal for D and T potency testing may not provide optimal dose-response for all aP components. Method adjustments may thus be required

  12. Bone Tissue Engineering with Adipose-Derived Stem Cells in Bioactive Composites of Laser-Sintered Porous Polycaprolactone Scaffolds and Platelet-Rich Plasma

    Directory of Open Access Journals (Sweden)

    Han-Tsung Liao

    2013-10-01

    Full Text Available Three-dimensional porous polycaprolactone (PCL scaffolds with consistent inter-pore channels, 83% porosity and 300–400 μm pore size were fabricated via selective laser sintering. The PCL scaffold was combined with platelet-rich plasma (PRP to form a bioactive composite and studied for potential application in bone tissue engineering using porcine adipose-derived stem cells (PASCs. The PCL/PRP/PASCs construct showed enhanced cell seeding efficiency and synergistically increased the differentiation capability of PASCs in osteogenic medium toward the osteoblast lineage, judging from elevated alkaline phosphatase activity and up-regulated osteogenic genes expression. For in vivo study, a 3 cm × 3 cm mandible defect was created in pigs and reconstructed by implanting acellular PCL scaffolds or PCL/PRP/PASCs constructs. Both groups showed new bone formation, however, the new bone volume was 5.1 times higher for PCL/PRP/PASCs 6 months post-operation. The bone density was less and loose in the acellular PCL group and the Young’s modulus was only 29% of normal bone. In contrast, continued and compact bone formation was found in PCL/PRP/PASCs and the Young’s modulus was 81% that of normal bone. Masson’s trichrome stain, immunohistochemical analysis of osteocalcin and collagen type I also confirmed new bone formation.

  13. Differential antiepileptic effects of the organic calcium antagonists verapamil and flunarizine in neurons of organotypic neocortical explants from newborn rats

    NARCIS (Netherlands)

    Bingmann, D; Speckmann, E J; Baker, R E; Ruijter, J; de Jong, B. M.

    1988-01-01

    Effects of the organic calcium antagonists verapamil and flunarizine on pentylenetetrazol induced paroxysmal depolarizations were tested in organotypic neocortical explants taken from neonatal rats. In these in vitro experiments the papaverin derivative verapamil depressed, and finally abolished, ep

  14. Reduced inflammatory cell recruitment and tissue damage in spinal cord injury by acellular spinal cord scaffold seeded with mesenchymal stem cells.

    Science.gov (United States)

    Wang, Yu-Hai; Chen, Jian; Zhou, Jing; Nong, Feng; Lv, Jin-Han; Liu, Jia

    2017-01-01

    Therapy using acellular spinal cord (ASC) scaffolds seeded with bone marrow stromal cells (BMSCs) has previously been shown to restore function of the damaged spinal cord and improve functional recovery in a rat model of acute hemisected spinal cord injury (SCI). The aim of the present study was to determine whether BMSCs and ASC scaffolds promote the functional recovery of the damaged spinal cord in a rat SCI model through regulation of apoptosis and immune responses. Whether this strategy regulates secondary inflammation, which is characterized by the infiltration of immune cells and inflammatory mediators to the lesion site, in SCI repair was investigated. Basso, Beattie, and Bresnahan scores revealed that treatment with BMSCs seeded into an ASC scaffold led to a significant improvement in motor function recovery compared with treatment with an ASC scaffold alone or untreated controls at 2 and 8 weeks after surgery (Pscaffold significantly decreased the number of terminal deoxynucleotidyl transferase dUTP nick end labeling-positive cells, as compared with the ASC scaffold only and control groups. These results suggested that the use of BMSCs decreased the apoptosis of neural cells and thereby limited tissue damage at the lesion site. Notably, the use of BMSCs with an ASC scaffold also decreased the recruitment of macrophages (microglia; Pscaffold only groups. BMSCs regulated inflammatory cell recruitment to promote functional recovery. However, there was no significant difference in IgM-positive expression among the three groups (P>0.05). The results of this study demonstrated that BMSCs seeded into ASC scaffolds for repair of spinal cord hemisection defects promoted functional recovery through the early regulation of inflammatory cell recruitment with inhibition of apoptosis and secondary inflammation.

  15. Healing rates for challenging rotator cuff tears utilizing an acellular human dermal reinforcement graft

    Science.gov (United States)

    Agrawal, Vivek

    2012-01-01

    Purpose: This study presents a retrospective case series of the clinical and structural outcomes (1.5 T MRI) of arthroscopic rotator cuff repair with acellular human dermal graft reinforcement performed by a single surgeon in patients with large, massive, and previously repaired rotator cuff tears. Materials and Methods: Fourteen patients with mean anterior to posterior tear size 3.87 ± 0.99 cm (median 4 cm, range 2.5–6 cm) were enrolled in the study and were evaluated for structural integrity using a high-field (1.5 T) MRI at an average of 16.8 months after surgery. The Constant-Murley scores, the Flexilevel Scale of Shoulder Function (Flex SF), scapular plane abduction, and strength were analyzed. Results: MRI results showed that the rotator cuff repair was intact in 85.7% (12/14) of the patients studied. Two patients had a Sugaya Type IV recurrent tear (2 of 14; 14.3%), which were both less than 1 cm. The Constant score increased from a preoperative mean of 49.72 (range 13–74) to a postoperative mean of 81.07 (range 45–92) (P value = 0.009). Flexilevel Scale of Shoulder Function (Flex SF) Score normalized to a 100-point scale improved from a preoperative mean of 53.69 to a postoperative mean of 79.71 (P value = 0.003). The Pain Score improved from a preoperative mean of 7.73 to a postoperative mean of 13.57 (P value = 0.008). Scapular plane abduction improved from a preoperative mean of 113.64° to a postoperative mean of 166.43° (P value = 0.010). The strength subset score improved from a preoperative mean of 1.73 kg to a postoperative mean of 7.52 kg (P value = 0.006). Conclusions: This study presents a safe and effective technique that may help improve the healing rates of large, massive, and revision rotator cuff tears with the use of an acellular human dermal allograft. This technique demonstrated favorable structural healing rates and statistically improved functional outcomes in the near term. Level of Evidence: 4. Retrospective case series. PMID

  16. Response of cotyledon explants of Capsicum annuum L. cv. kujawianka to chosen plant growth regulators in in vitro culture

    Directory of Open Access Journals (Sweden)

    Alicja Fraś

    2014-02-01

    Full Text Available Shoot buds originated directly on cotyledon explants of Capsicum annuum L. cv. Kujawianka, when Linsmaier and Skoog medium was enriched with BAP (2 mg/l. Kinetin (2 mg/l or kinetin with IAA (1 mg/l + 1 mg/l induced indirect shoot buds regeneration from callus. Rooting was obtained with explants cultivated on a medium containing NAA (0,5 mg/l. Occurrence of the early stages of differentiation was proved at the histological level.

  17. Structures of benzo(a)pyrene-nucleic acid adducts formed in human and bovine bronchial explants

    DEFF Research Database (Denmark)

    1977-01-01

    obtained evidence that the same derivative is involved in the binding of BP to the DNA of human bronchial explants, although details of the specific isomer involved and of the structure of the adduct were not reported. We describe here studies on RNA and DNA adducts formed by human bronchial explants...... and provide evidence that the structures of the major adducts are similar to those formed in the analogous bovine system....

  18. In vitro clonal propagation of Achyranthes aspera L. and Achyranthes bidentata Blume using nodal explants

    Institute of Scientific and Technical Information of China (English)

    Wesely Edward Gnanaraj; Johnson MarimuthuAntonisamy; Mohanamathi RB

    2012-01-01

    Objective: To develop the reproducible in vitro propagation protocols for the medicinally important plants viz., Achyranthes aspera (A. aspera) L. and Achyranthes bidentata (A. bidentata) Blume using nodal segments as explants. Methods: Young shoots of A. aspera and A. bidentata were harvested and washed with running tap water and treated with 0.1% bavistin and rinsed twice with distilled water. Then the explants were surface sterilized with 0.1% (w/v) HgCl2 solutions for 1 min. After rinsing with sterile distilled water for 3-4 times, nodal segments were cut into smaller segments (1 cm) and used as the explants. The explants were placed horizontally as well as vertically on solid basal Murashige and Skoog (MS) medium supplemented with 3% sucrose, 0.6% (w/v) agar (Hi-Media, Mumbai) and different concentration and combination of 6-benzyl amino purine (BAP), kinetin (Kin), naphthalene acetic acid (NAA) and indole acetic acid (IAA) for direct regeneration.Results:Adventitious proliferation was obtained from A. aspera and A. bidentata nodal segments inoculated on MS basal medium with 3% sucrose and augmented with BAP and Kin with varied frequency. MS medium augmented with 3.0 mg/L of BAP showed the highest percentage (93.60±0.71) of shootlets formation for A. aspera and (94.70±0.53) percentages for A. bidentata. Maximum number of shoots/explants (10.60±0.36) for A. aspera and (9.50±0.56) for A. bidentata was observed in MS medium fortified with 5.0 mg/L of BAP. For A. aspera, maximum mean length (5.50±0.34) of shootlets was obtained in MS medium augmented with 3.0 mg/L of Kin and for A. bidentata (5.40±0.61) was observed in the very same concentration. The highest percentage, maximum number of rootlets/shootlet and mean length of rootlets were observed in 1/2 MS medium supplemented with 1.0 mg/L of IBA. Seventy percentages of plants were successfully established in polycups. Sixty eight percentages of plants were well established in the green house condition

  19. Re-use of explanted osteosynthesis devices: a reliable and inexpensive reprocessing protocol.

    Science.gov (United States)

    Danesi, Valentina; Cristofolini, Luca; Stea, Susanna; Traina, Francesco; Beraudi, Alina; Tersi, Luca; Harman, Melinda; Viceconti, Marco

    2011-10-01

    Orthopaedic surgical treatments emphasizing immobilization using open reduction and internal fixation with osteosynthesis devices are widely accepted for their efficacy in treating complex fractures and reducing permanent musculoskeletal deformity. However, such treatments are profoundly underutilized in low- and middle-income countries (LMIC), partially due to inadequate availability of the costly osteosynthesis devices. Orthopaedic surgeons in some LMIC regularly re-use osteosynthesis devices in an effort to meet treatment demands, even though such devices typically are regulated for single-use only. The purpose of this study is to report a reprocessing protocol applied to explanted osteosynthesis devices obtained at a leading trauma care hospital. Explanted osteosynthesis devices were identified through a Register of Explanted Orthopaedic Prostheses. Guidelines to handle ethical issues were approved by the local Ethical Committee and informed patient consent was obtained at the time of explant surgery. Primary acceptance criteria were established and applied to osteosynthesis devices explanted between 2005 and 2008. A rigorous protocol for conducting decontamination and visual inspection based on specific screening criteria was implemented using simple equipment that is readily available in LMIC. A total of 2050 osteosynthesis devices, including a large variety of plates, screws and staples, were reprocessed using the decontamination and inspection protocols. The acceptance rate was 66%. Estimated labour time and implementation time of the protocol to reprocess a typical osteosynthesis unit (1 plate and 5 screws) was 25 min, with an estimated fixed cost (in Italy) of €10 per unit for implementing the protocol, plus an additional €5 for final sterilization at the end-user hospital site. This study was motivated by the treatment demands encountered by orthopaedic surgeons providing medical treatment in several different LMIC and their need for access to basic

  20. IN VITRO REGENERATION FROM SHOOT TIP AND NODAL EXPLANTS OF SIMAROUBA GLAUCA DC, A PROMISING BIODIESEL TREE

    Directory of Open Access Journals (Sweden)

    Shastri P. Shukla

    2013-03-01

    Full Text Available An efficient regeneration protocol was developed from shoot tip and nodal explants of Simarouba glauca DC, a promising biodiesel plant. Nodal explants appeared to have better regeneration capacity than shoot tip explants (40% in the tested media. The highest regeneration frequency (90% and shoot number (7.00 ± 1.00 shoots per explants were obtained in nodal explants in Murashige and Skoog’s (MS medium supplemented with 6-benzylaminopurine (BAP 4.43 μM and α-naphthalene acetic acid (NAA 5.36 μM.Induced shoot buds were multiplied and elongated on the MS medium supplemented with BAP (4.44 μM, NAA (5.36 μM and TDZ (Thidiazuron 2.27 μM with 9.66±0.33 (mean length 5.35±0.32 cm and 9.00±0.57 (mean length 4.51±0.15cm shoots using nodal segments and shoot tip explants, respectively. Halfstrength woody plant medium (WPM containing 2.46μM indole-3-butyric acid (IBA produced the maximum number of roots (6.00±1.15. The rooted plantlets were hardened on MS basal liquid medium and subsequently in polycups containing sterile soil and vermiculite (1:1 and successfully established in pots.

  1. Bone Densitometry (Bone Density Scan)

    Science.gov (United States)

    ... of DXA Bone Densitometry? What is a Bone Density Scan (DXA)? Bone density scanning, also called dual-energy x-ray absorptiometry ( ... is today's established standard for measuring bone mineral density (BMD). An x-ray (radiograph) is a noninvasive ...

  2. Response of root explants to in vitro cultivation of marketable garlic cultivars Resposta dos explantes radiculares ao cultivo in vitro de cultivares comerciais de alho

    Directory of Open Access Journals (Sweden)

    Danielle C Scotton

    2013-03-01

    Full Text Available Garlic cultivars are sexually sterile under standard growth conditions, with direct implications for commercial production costs as well as breeding programs. Garlic is propagated commercially via bulblets, which facilitates disease transmission and virus load accumulation over vegetative generations. Tissue culture produces virus-free clones that are more productive, while keeping the desired traits of the cultivar. Consequently, this technique allows studies of garlic genetics as well as guarantees genetic conservation of varieties. We aimed at analyzing the in vitro regeneration of eight marketable cultivars of garlic using root segments as explants. For each genotype, bulblet-derived explants were isolated and introduced into MS medium supplemented with 2,4-D and 2-iP. Calli were transferred to MS medium supplemented with 8.8 mM BAP and 0.1 mM NAA (regeneration medium A, or with 4.6 mM kinetin alone (regeneration medium B. The calli were then evaluated for regeneration frequency after sixty days of in vitro cultivation. The noble cultivar 'Jonas' presented the highest rates of plant regeneration among the cultivars tested. The medium A, which contained auxin and cytokinin, induced the highest regeneration rates of all cultivars. The process described herein is simple, reproducible and can potentially be used as a tool in molecular breeding strategies for other marketable cultivars and genotypes of garlic.Cultivares de alho são sexualmente estéreis sob condições padrão de cultivo, com implicações diretas nos custos de produção comercial, bem como em programas de melhoramento. O alho é comercialmente propagado por meio de bulbilhos, o que facilita a transmissão de doenças e leva ao acúmulo de cargas virais ao longo das gerações. A cultura de tecidos produz clones livres de vírus que são mais produtivos, mantendo as características desejadas da cultivar. Consequentemente, esta técnica permite estudar a genética do alho, bem

  3. Sealing and explant types on the mangaba micropropagation Tipo de vedação e explantes na micropropagação de mangabeira

    Directory of Open Access Journals (Sweden)

    Aline de Jesus Sá

    2012-08-01

    Full Text Available In micropropagation, especially for mangaba tree botanical variety of Northeastern Brazil, limiting aspects such as ethylene accumulation in the cultivation flask and loss of vigor in subcultures have been observed. This study was aimed at assessing the technical and scientific knowledge of the in vitro propagation of botanical mangaba tree variety and at improving the micropropagation protocol, establishing the in vitro cultivation time, the best type of flask sealing and explant at different micropropagation stages. For the establishment phase and for the first and second subcultures, the MS medium with 3% sucrose and 0.6% agar, supplemented with 1 mg L-1 IAA and 1 mg L-1 BA was used. Evaluations were performed at 30, 50 and 65 days of in vitro cultivation. The best types of flask sealing for the establishment phase were the PVC film and Para-film® and for the first subculture the Para-film® seal. In the second subculture the PVC film and Para-film® seals promoted the best growth. The median and basal nodal segments presented the best performance in the first subculture. No significant effect of explant type was observed in the second subculture. The ideal subculture interval in the establishment phase and the first and second subcultures is 50 days.Na micropropagação, especialmente para mangaba, variedade botânica da árvore do Nordeste do Brasil, aspectos limitantes, como acúmulo de etileno no recipiente de cultivo e perda de vigor em subculturas têm sido observados. Neste estudo, objetivou-se avaliar o conhecimento técnico e científico da propagação in vitro de mangabeira, variedade botânica do Nordeste do Brasil, e melhorar o protocolo de micropropagação, o melhor tipo de vedação frasco e explante em diferentes etapas. Para a fase de estabelecimento e para as subculturas primeiro e segundo, foi utlizado o meio MS com 3% de sacarose e agar 0,6%, suplementado com 1 mg L-1 de AIA e 1 mg L-1 de BAP. As avaliações foram

  4. Bone marrow aspiration

    Science.gov (United States)

    Iliac crest tap; Sternal tap; Leukemia - bone marrow aspiration; Aplastic anemia - bone marrow aspiration; Myelodysplastic syndrome - bone marrow aspiration; Thrombocytopenia - bone marrow aspiration; Myelofibrosis - bone marrow aspiration

  5. Human Keratinocyte Growth and Differentiation on Acellular Porcine Dermal Matrix in relation to Wound Healing Potential

    Directory of Open Access Journals (Sweden)

    Robert Zajicek

    2012-01-01

    Full Text Available A number of implantable biomaterials derived from animal tissues are now used in modern surgery. Xe-Derma is a dry, sterile, acellular porcine dermis. It has a remarkable healing effect on burns and other wounds. Our hypothesis was that the natural biological structure of Xe-Derma plays an important role in keratinocyte proliferation and formation of epidermal architecture in vitro as well as in vivo. The bioactivity of Xe-Derma was studied by a cell culture assay. We analyzed growth and differentiation of human keratinocytes cultured in vitro on Xe-Derma, and we compared the results with formation of neoepidermis in the deep dermal wounds treated with Xe-Derma. Keratinocytes cultured on Xe-Derma submerged in the culture medium achieved confluence in 7–10 days. After lifting the cultures to the air-liquid interface, the keratinocytes were stratified and differentiated within one week, forming an epidermis with basal, spinous, granular, and stratum corneum layers. Immunohistochemical detection of high-molecular weight cytokeratins (HMW CKs, CD29, p63, and involucrin confirmed the similarity of organization and differentiation of the cultured epidermal cells to the normal epidermis. The results suggest that the firm natural structure of Xe-Derma stimulates proliferation and differentiation of human primary keratinocytes and by this way improves wound healing.

  6. Does Acellular Dermal Matrix Thickness Affect Complication Rate in Tissue Expander Based Breast Reconstruction?

    Directory of Open Access Journals (Sweden)

    Jessica F. Rose

    2016-01-01

    Full Text Available Background. While the benefits of using acellular dermal matrices (ADMs in breast reconstruction are well described, their use has been associated with additional complications. The purpose of this study was to determine if ADM thickness affects complications in breast reconstruction. Methods. A retrospective chart review was performed including all tissue expander based breast reconstructions with AlloDerm (LifeCell, Branchburg, NJ over 4 years. We evaluated preoperative characteristics and assessed postoperative complications including seroma, hematoma, infection, skin necrosis, and need for reintervention. We reviewed ADM thickness and time to Jackson-Pratt (JP drain removal. Results. Fifty-five patients underwent 77 ADM-associated tissue expander based breast reconstructions, with average age of 48.1 years and average BMI of 25.9. Average ADM thickness was 1.21 mm. We found higher complication rates in the thick ADM group. Significant associations were found between smokers and skin necrosis (p<0.0001 and seroma and prolonged JP drainage (p=0.0004; radiated reconstructed breasts were more likely to suffer infections (p=0.0085, and elevated BMI is a significant predictor for increased infection rate (p=0.0037. Conclusion. We found a trend toward increased complication rates with thicker ADMs. In the future, larger prospective studies evaluating thickness may provide more information.

  7. Xenogeneic acellular dermal matrix in combination with pectoralis major myocutaneous flap reconstructs hypopharynx and cervical esophagus.

    Science.gov (United States)

    Yin, Danhui; Tang, Qinglai; Wang, Shuang; Li, Shisheng; He, Xiangbo; Liu, Jiajia; Liu, Bingbing; Yang, Mi; Yang, Xinming

    2015-11-01

    The aim of this study was to explore xenogeneic acellular dermal matrix (ADM) in combination with pectoralis major myocutaneous flap in hypopharynx and cervical esophagus reconstruction. A total of five patients were treated with this surgical method to reconstruct hypopharynx and cervical esophagus in Second Xiangya Hospital between January 2012 and April 2013. Four of them had hypopharyngeal carcinoma with laryngeal and cervical esophageal invasion, while the fifth patient with hypopharyngeal cancer had developed scars and atresia after postoperative radiotherapy. The defect length after hypopharyngeal and cervical esophageal resection was 6-8 cm, and was repaired by a combination of ADM and pectoralis major myocutaneous flap by our team. Interestingly, the four patients had primary healing and regained their eating function about 2-3 weeks after surgery, the fifth individual suffered from pharyngeal fistula, but recovered after dressing change about 2 months. Postoperative esophageal barium meals revealed that the pharynx and esophagus were unobstructed in all five patients. Xenogeneic ADM in combination with pectoralis major myocutaneous flap for hypopharynx and cervical esophagus reconstruction is a simple, safe and effective method with fewer complications. Nevertheless, according to the defect length of the cervical esophagus, the patients need to strictly follow the medical advice.

  8. Tetanus, diphtheria, and acellular pertussis vaccination among women of childbearing age-United States, 2013.

    Science.gov (United States)

    O'Halloran, Alissa C; Lu, Peng-Jun; Williams, Walter W; Ding, Helen; Meyer, Sarah A

    2016-07-01

    The incidence of pertussis in the United States has increased since the 1990s. Tetanus, diphtheria, and acellular pertussis (Tdap) vaccination of pregnant women provides passive protection to infants. Tdap vaccination is currently recommended for pregnant women during each pregnancy, but coverage among pregnant women and women of childbearing age has been suboptimal. Data from the 2013 Behavioral Risk Factor Surveillance System (BRFSS) and 2013 National Health Interview Survey (NHIS) were used to determine national and state-specific Tdap vaccination coverage among women of childbearing age by self-reported pregnancy status at the time of the survey. Although this study could not assess coverage of Tdap vaccination received during pregnancy because questions on whether Tdap vaccination was received during pregnancy were not asked in BRFSS and NHIS, demographic and access-to-care factors associated with Tdap vaccination coverage in this population were assessed. Tdap vaccination coverage among all women 18-44 years old was 38.4% based on the BRFSS and 23.3% based on the NHIS. Overall, coverage did not differ by pregnancy status at the time of the survey. Coverage among all women 18-44 years old varied widely by state. Age, race and ethnicity, education, number of children in the household, and access-to-care characteristics were independently associated with Tdap vaccination in both surveys. We identified associations of demographic and access-to-care characteristics with Tdap vaccination that can guide strategies to improve vaccination rates in women during pregnancy.

  9. Brainless but Multi-Headed: Decision Making by the Acellular Slime Mould Physarum polycephalum.

    Science.gov (United States)

    Beekman, Madeleine; Latty, Tanya

    2015-11-20

    Because of its peculiar biology and the ease with which it can be cultured, the acellular slime mould Physarum polycephalum has long been a model organism in a range of disciplines. Due to its macroscopic, syncytial nature, it is no surprise that it has been a favourite amongst cell biologists. Its inclusion in the experimental tool kit of behavioural ecologists is much more recent. These recent studies have certainly paid off. They have shown that, for an organism that lacks a brain or central nervous system, P. polycephalum shows rather complex behaviour. For example, it is capable of finding the shortest path through a maze, it can construct networks as efficient as those designed by humans, it can solve computationally difficult puzzles, it makes multi-objective foraging decisions, it balances its nutrient intake and it even behaves irrationally. Are the slime mould's achievements simply "cute", worthy of mentioning in passing but nothing to take too seriously? Or do they hint at the fundamental processes underlying all decision making? We will address this question after reviewing the decision-making abilities of the slime mould. Copyright © 2015 Elsevier Ltd. All rights reserved.

  10. Identification of substrates for transglutaminase in Physarum polycephalum, an acellular slime mold, upon cellular mechanical damage.

    Science.gov (United States)

    Wada, Fumitaka; Hasegawa, Hiroki; Nakamura, Akio; Sugimura, Yoshiaki; Kawai, Yoshiki; Sasaki, Narie; Shibata, Hideki; Maki, Masatoshi; Hitomi, Kiyotaka

    2007-06-01

    Transglutaminases are Ca(2+)-dependent enzymes that post-translationally modify proteins by crosslinking or polyamination at specific polypeptide-bound glutamine residues. Physarum polycephalum, an acellular slime mold, is the evolutionarily lowest organism expressing a transglutimase whose primary structure is similar to that of mammalian transglutimases. We observed transglutimase reaction products at injured sites in Physarum macroplasmodia upon mechanical damage. With use of a biotin-labeled primary amine, three major proteins constituting possible transglutimase substrates were affinity-purified from the damaged slime mold. The purified proteins were Physarum actin, a 40 kDa Ca(2+)-binding protein with four EF-hand motifs (CBP40), and a novel 33 kDa protein highly homologous to the eukaryotic adenine nucleotide translocator, which is expressed in mitochondria. Immunochemical analysis of extracts from the damaged macroplasmodia indicated that CBP40 is partly dimerized, whereas the other proteins migrated as monomers on SDS/PAGE. Of the three proteins, CBP40 accumulated most significantly around injured areas, as observed by immunofluoresence. These results suggested that transglutimase reactions function in the response to mechanical injury.

  11. Speed-accuracy trade-offs during foraging decisions in the acellular slime mould Physarum polycephalum.

    Science.gov (United States)

    Latty, Tanya; Beekman, Madeleine

    2011-02-22

    Speed-accuracy trade-offs (SATs) are thought to be a fundamental feature of biological information processing, yet most evidence of SATs comes from animals. Here, we examine SATs in the foraging decisions of an acellular, amoeboid organism: the slime mould Physarum polycephalum. Slime moulds were given a simple discrimination task: selecting the highest-quality food item from a set of three options. We investigated the effect of two stressors, light exposure and hunger, on the speed and accuracy of decision-making. We also examined the effect of task difficulty. When given a difficult discrimination task, stressed individuals tend to make faster decisions than non-stressed individuals. This effect was reversed in plasmodia given easy discrimination tasks, where stressed individuals made slower decisions than non-stressed individuals. We found evidence of SATs, such that individuals who made fast decisions were more likely to make costly errors by selecting the worst possible food option. Our results suggest that SATs occur in a wider range of taxa than previously considered.

  12. Speed–accuracy trade-offs during foraging decisions in the acellular slime mould Physarum polycephalum

    Science.gov (United States)

    Latty, Tanya; Beekman, Madeleine

    2011-01-01

    Speed–accuracy trade-offs (SATs) are thought to be a fundamental feature of biological information processing, yet most evidence of SATs comes from animals. Here, we examine SATs in the foraging decisions of an acellular, amoeboid organism: the slime mould Physarum polycephalum. Slime moulds were given a simple discrimination task: selecting the highest-quality food item from a set of three options. We investigated the effect of two stressors, light exposure and hunger, on the speed and accuracy of decision-making. We also examined the effect of task difficulty. When given a difficult discrimination task, stressed individuals tend to make faster decisions than non-stressed individuals. This effect was reversed in plasmodia given easy discrimination tasks, where stressed individuals made slower decisions than non-stressed individuals. We found evidence of SATs, such that individuals who made fast decisions were more likely to make costly errors by selecting the worst possible food option. Our results suggest that SATs occur in a wider range of taxa than previously considered. PMID:20826487

  13. Nanopatterned acellular valve conduits drive the commitment of blood-derived multipotent cells

    Science.gov (United States)

    Di Liddo, Rosa; Aguiari, Paola; Barbon, Silvia; Bertalot, Thomas; Mandoli, Amit; Tasso, Alessia; Schrenk, Sandra; Iop, Laura; Gandaglia, Alessandro; Parnigotto, Pier Paolo; Conconi, Maria Teresa; Gerosa, Gino

    2016-01-01

    Considerable progress has been made in recent years toward elucidating the correlation among nanoscale topography, mechanical properties, and biological behavior of cardiac valve substitutes. Porcine TriCol scaffolds are promising valve tissue engineering matrices with demonstrated self-repopulation potentiality. In order to define an in vitro model for investigating the influence of extracellular matrix signaling on the growth pattern of colonizing blood-derived cells, we cultured circulating multipotent cells (CMC) on acellular aortic (AVL) and pulmonary (PVL) valve conduits prepared with TriCol method and under no-flow condition. Isolated by our group from Vietnamese pigs before heart valve prosthetic implantation, porcine CMC revealed high proliferative abilities, three-lineage differentiative potential, and distinct hematopoietic/endothelial and mesenchymal properties. Their interaction with valve extracellular matrix nanostructures boosted differential messenger RNA expression pattern and morphologic features on AVL compared to PVL, while promoting on both matrices the commitment to valvular and endothelial cell-like phenotypes. Based on their origin from peripheral blood, porcine CMC are hypothesized in vivo to exert a pivotal role to homeostatically replenish valve cells and contribute to hetero- or allograft colonization. Furthermore, due to their high responsivity to extracellular matrix nanostructure signaling, porcine CMC could be useful for a preliminary evaluation of heart valve prosthetic functionality. PMID:27789941

  14. Biopolymer gel matrix as acellular scaffold for enhanced dermal tissue regeneration.

    Science.gov (United States)

    Judith, Rangasamy; Nithya, Mariappan; Rose, Chellan; Mandal, Asit Baran

    2012-07-01

    Biological grafts have drawbacks such as donor scarcity, disease transmission, tissue infection, while the scaffolds of either collagen or chitosan fabrics fail to become part of the tissue at the wound site, though they favor the formation of connective tissue matrix. This study developed a novel composite consisting of the combination of atelocollagen and chitosan in order to provide a biodegradable molecular matrix in gel form as a biomimetic surface for cell attachment, to promote the wound healing in excision wounds. We found that the topical application of biopolymer composite on the wound promoted cell proliferation, migration and collagen deposition overtime. The enhanced cellular activity in the collagen-chitosan treated wound tissue was also assed by increased levels of Platelet derived growth factor (PDGF) and Nerve growth factor (NGF) associated with elevated levels of antioxidants and decreased level of lipid peroxidation. The acellular matrix-like topical application material is designed to guide the eventual re-establishment of an anatomically normal skin. The results of this study demonstrate the feasibility of multi-cell regeneration on a molecular system that mimics tissue engineering in vivo.

  15. A anorectal fistula treatment with acellular extracellular matrix: A new technique

    Institute of Scientific and Technical Information of China (English)

    Wei-Liang Song; Zhen-Jun Wang; Yi Zheng; Xin-Qing Yang; Ya-Ping Peng

    2008-01-01

    AIM:To investigate a new technique of the anorectal fistula treatment with acellular extracellular matrix (AEM).METHODS: Thirty patients with anorectal fistula were treated with AEM.All fistula tracts and primary openings were identified using conventional fistula probe.All tracts were curetted with curet and irrigated with hydrogen peroxide and metronidazole.The AEM was pulled into the fistula tract from secondary to primary opening.The material was secured at the level of the primary opening.The excess AEM was trimmed at skin level at the secondary opening.RESULTS: All of the 30 patients had successful closure of their fistula after a 7-14 d follow-up.The healing rate of anal fistula in treatment group was 100%.The ache time,healing time and anal deformation of treatment group were obviously superior to traditional surgical methods.CONCLUSION: Using AEM anal fistula plug in treatment that causes the anorectal fistula is safe and successful in 100% of patients.It can reduce pain,shorten disease course and protect anal function.

  16. A new candidate substrate for cell-matrix adhesion study: the acellular human amniotic matrix.

    Science.gov (United States)

    Guo, Qianchen; Lu, Xuya; Xue, Yuan; Zheng, Hong; Zhao, Xiaotao; Zhao, Huajian

    2012-01-01

    In vivo adhesions between cells and the extracellular matrix play a crucial role in cell differentiation, proliferation, and migration as well as tissue remodeling. Natural three-dimensional (3D) matrices, such as self-assembling matrices and Matrigel, have limitations in terms of their biomechanical properties. Here, we present a simple method to produce an acellular human amniotic matrix (AHAM) with preserved biomechanical properties and a favorable adhesion potential. On the stromal side of the AHAM, human foreskin fibroblasts (HFFs) attached and extended with bipolar spindle-shaped morphology proliferated to multilayer networks, invaded into the AHAM, and migrated in a straight line. Moreover, αV integrin, paxillin, and fibronectin were observed to colocalize after 24 h of HFF culture on the stromal side of the AHAM. Our results indicate that the AHAM may be an ideal candidate as a cell-matrix adhesion substrate to study cell adhesion and invasion as well as other functions in vitro under a tensile force that mimics the in vivo environment.

  17. Production of an acellular matrix from amniotic membrane for the synthesis of a human skin equivalent.

    Science.gov (United States)

    Sanluis-Verdes, Anahí; Yebra-Pimentel Vilar, Maria Teresa; García-Barreiro, Juan Javier; García-Camba, Marta; Ibáñez, Jacinto Sánchez; Doménech, Nieves; Rendal-Vázquez, Maria Esther

    2015-09-01

    Human amniotic membrane (HAM) has useful properties as a dermal matrix substitute. The objective of our work was to obtain, using different enzymatic or chemical treatments to eliminate cells, a scaffold of acellular HAM for later use as a support for the development of a skin equivalent. The HAM was separated from the chorion, incubated and cryopreserved. The membrane underwent different enzymatic and chemical treatments to eliminate the cells. Fibroblasts and keratinocytes were separately obtained from skin biopsies of patients following a sequential double digestion with first collagenase and then trypsin-EDTA (T/E). A skin equivalent was then constructed by seeding keratinocytes on the epithelial side and fibroblasts on the chorionic side of the decellularizated HAM. Histological, immunohistochemical, inmunofluorescent and molecular biology studies were performed. Treatment with 1% T/E at 37 °C for 30 min totally removed epithelial and mesenchymal cells. The HAM thus treated proved to be a good matrix to support adherence of cells and allowed the achievement of an integral and intact scaffold for development of a skin equivalent, which could be useful as a skin substitute for clinical use.

  18. A New Candidate Substrate for Cell-Matrix Adhesion Study: The Acellular Human Amniotic Matrix

    Directory of Open Access Journals (Sweden)

    Qianchen Guo

    2012-01-01

    Full Text Available In vivo adhesions between cells and the extracellular matrix play a crucial role in cell differentiation, proliferation, and migration as well as tissue remodeling. Natural three-dimensional (3D matrices, such as self-assembling matrices and Matrigel, have limitations in terms of their biomechanical properties. Here, we present a simple method to produce an acellular human amniotic matrix (AHAM with preserved biomechanical properties and a favorable adhesion potential. On the stromal side of the AHAM, human foreskin fibroblasts (HFFs attached and extended with bipolar spindle-shaped morphology proliferated to multilayer networks, invaded into the AHAM, and migrated in a straight line. Moreover, αV integrin, paxillin, and fibronectin were observed to colocalize after 24 h of HFF culture on the stromal side of the AHAM. Our results indicate that the AHAM may be an ideal candidate as a cell-matrix adhesion substrate to study cell adhesion and invasion as well as other functions in vitro under a tensile force that mimics the in vivo environment.

  19. Three-dimensional Reconstruction of the Microstructure of Human Acellular Nerve Allograft

    Science.gov (United States)

    Zhu, Shuang; Zhu, Qingtang; Liu, Xiaolin; Yang, Weihong; Jian, Yutao; Zhou, Xiang; He, Bo; Gu, Liqiang; Yan, Liwei; Lin, Tao; Xiang, Jianping; Qi, Jian

    2016-01-01

    The exact inner 3D microstructure of the human peripheral nerve has been a mystery for decades. Therefore, it has been difficult to solve several problems regarding peripheral nerve injury and repair. We used high-resolution X-ray computed microtomography (microCT) to scan a freeze-dried human acellular nerve allograft (hANA). The microCT images were then used to reconstruct a 3D digital model, which was used to print a 3D resin model of the nerve graft. The 3D digital model of the hANA allowed visualization of all planes. The magnified 3D resin model clearly showed the nerve bundles and basement membrane tubes of the hANA. Scanning electron microscopy (SEM) was used to analyse the microstructure of the hANA. Compared to the SEM images, the microCT image clearly demonstrated the microstructure of the hANA cross section at a resolution of up to 1.2 μm. The 3D digital model of the hANA facilitates a clear and easy understanding of peripheral nerve microstructure. Furthermore, the enlarged 3D resin model duplicates the unique inner structure of each individual hANA. This is a crucial step towards achieving 3D printing of a hANA or nerve that can be used as a nerve graft. PMID:27476584

  20. Does tetanus-diphtheria-acellular pertussis vaccination interfere with serodiagnosis of pertussis infection?

    Science.gov (United States)

    Pawloski, Lucia C; Kirkland, Kathryn B; Baughman, Andrew L; Martin, Monte D; Talbot, Elizabeth A; Messonnier, Nancy E; Tondella, Maria Lucia

    2012-06-01

    An anti-pertussis toxin (PT) IgG enzyme-linked immunosorbent assay (ELISA) was analytically validated for the diagnosis of pertussis at a cutoff of 94 ELISA units (EU)/ml. Little was known about the performance of this ELISA in the diagnosis of adults recently vaccinated with tetanus-diphtheria-acellular pertussis (Tdap) vaccine, which contains PT. The goal of this study was to determine when the assay can be used following Tdap vaccination. A cohort of 102 asymptomatic health care personnel (HCP) vaccinated with Tdap (Adacel; Sanofi Pasteur) were aged 19 to 79 years (median, 47 years) at vaccination. For each HCP, specimens were available for evaluation at 2 to 10 time points (prevaccination to 24 months postvaccination), and geometric mean concentrations (GMC) for the cohort were calculated at each time point. Among 97 HCP who responded to vaccination, a mixed-model analysis with prediction and tolerance intervals was performed to estimate the time at which serodiagnosis can be used following vaccination. The GMCs were 8, 21, and 9 EU/ml at prevaccination and 4 and 12 months postvaccination, respectively. Eight (8%) of the 102 HCP reached antibody titers of ≥94 EU/ml during their peak response, but none had these titers by 6 months postvaccination. The calculated prediction and tolerance intervals were <94 EU/ml by 45 and 75 days postvaccination, respectively. Tdap vaccination 6 months prior to testing did not confound result interpretation. This seroassay remains a valuable diagnostic tool for adult pertussis.

  1. Degradation and erosion mechanisms of bioresorbable porous acellular vascular grafts: an in vitro investigation.

    Science.gov (United States)

    Gade, Piyusha S; Lee, Keewon; Pfaff, Blaise N; Wang, Yadong; Robertson, Anne M

    2017-07-01

    A fundamental mechanism of in situ tissue regeneration from biodegradable synthetic acellular vascular grafts is the effective interplay between graft degradation, erosion and the production of extracellular matrix. In order to understand this crucial process of graft erosion and degradation, we conducted an in vitro investigation of grafts (n = 4 at days 1, 4, 7, 10 each) exposed to enzymatic degradation. Herein, we provide constitutive relationships for mass loss and mechanical properties based on much-needed experimental data. Furthermore, we formulate a mathematical model to provide a physics-based framework for understanding graft erosion. A novel finding is that despite their porous nature, grafts lost mass exponentially via surface erosion demonstrating a 20% reduction in outer diameter and no significant change in apparent density. A diffusion based, concentration gradient-driven mechanistic model of mass loss through surface erosion was introduced which can be extended to an in vivo setting through the use of two degradation parameters. Furthermore, notably, mechanical properties of degrading grafts did not scale with mass loss. Thus, we introduced a damage function scaling a neo-Hookean model to describe mechanical properties of the degrading graft; a refinement to existing mass-dependent growth and remodelling (G&R) models. This framework can be used to improve accuracy of well-established G&R theories in biomechanics; tools that predict evolving structure-function relationships of neotissues and guide graft design. © 2017 The Author(s).

  2. Purification design and practice for pertactin, the third component of acellular pertussis vaccine, from Bordetella pertussis.

    Science.gov (United States)

    Li, Zenglan; Zhang, Yan; Wang, Qi; Li, Zhengjun; Liu, Yongdong; Zhang, Songping; Zhang, Guifeng; Ma, Guanghui; Luo, Jian; Su, Zhiguo

    2016-07-25

    Development of acellular pertussis vaccine (aPV) requires purification of several components from Bordetella pertussis. While the components pertussis toxin (PT) and filamentous hemagglutinin (FHA) have been successfully purified, the third component, pertactin, proves to be a difficult target due to its very low concentration. In order to solve its purification problem, we performed the surface potential analysis with GRASP2 program. The results demonstrated that there are two major charge patches, one negative and one positive, which are located separately on this linear protein. For this special feature, we designed a dual ion exchange chromatography strategy including an anionic exchange and a cationic exchange process for separation of pertactin from the heat extract of B. pertussis. The initial anionic exchange chromatography concentrated the product from 1.7% to 14.6%, with recovery of 80%. The second cationic exchange chromatography increased the purity to 33%, with recovery of 83%. The final purification was accomplished by hydrophobic interaction chromatography, yielding a purity of 96%. The total recovery of the three columns was 61%. Characterization of the purified antigen was performed with CD, intrinsic fluorescence, HP-SEC and western-blot, showing that the purified protein kept its natural conformation and immune-reactivity. The rationally designed process proved to be feasible, and it is suitable for large-scale preparation of the third aPV component pertactin.

  3. Acellular Dermal Matrix in Reconstructive Breast Surgery: Survey of Current Practice among Plastic Surgeons

    Science.gov (United States)

    Ibrahim, Ahmed M. S.; Koolen, Pieter G. L.; Ashraf, Azra A.; Kim, Kuylhee; Mureau, Marc A. M.; Lee, Bernard T.

    2015-01-01

    Background: Acellular dermal matrices (ADMs) in plastic surgery have become increasingly popular particularly for breast reconstruction. Despite their advantages, questions exist regarding their association with a possible increased incidence of complications. We describe a collective experience of plastic surgeons’ use of ADMs in reconstructive breast surgery using an internet-based survey. Methods: Members of the American Society of Plastic Surgeons were recruited through voluntary, anonymous participation in an online survey. The web-based survey garnered information about participant demographics and their experience with ADM use in breast reconstruction procedures. After responses were collected, all data were anonymously processed. Results: Data were ascertained through 365 physician responses of which 99% (n = 361) completed the survey. The majority of participants were men (84.5%) between 51 and 60 years (37.4%); 84.2% used ADM in breast reconstruction, including radiated patients (79.7%). ADM use was not favored for nipple reconstruction (81.5%); 94.6% of participants used drains, and 87.8% administered antibiotics postoperatively. The most common complications were seroma (70.9%) and infection (16%), although 57.4% claimed anecdotally that overall complication rate was unchanged after incorporating ADM into their practice. High cost was a deterrent for ADM use (37.5%). Conclusions: Plastic surgeons currently use ADM in breast reconstruction for both immediate and staged procedures. Of those responding, a majority of plastic surgeons will incorporate drains and use postoperative antibiotics for more than 48 hours. PMID:25973359

  4. A anorectal fistula treatment with acellular extracellular matrix: A new technique

    Science.gov (United States)

    Song, Wei-Liang; Wang, Zhen-Jun; Zheng, Yi; Yang, Xin-Qing; Peng, Ya-Ping

    2008-01-01

    AIM: To investigate a new technique of the anorectal fistula treatment with acellular extracellular matrix (AEM). METHODS: Thirty patients with anorectal fistula were treated with AEM. All fistula tracts and primary openings were identified using conventional fistula probe. All tracts were curetted with curet and irrigated with hydrogen peroxide and metronidazole. The AEM was pulled into the fistula tract from secondary to primary opening. The material was secured at the level of the primary opening. The excess AEM was trimmed at skin level at the secondary opening. RESULTS: All of the 30 patients had successful closure of their fistula after a 7-14 d follow-up. The healing rate of anal fistula in treatment group was 100%. The ache time, healing time and anal deformation of treatment group were obviously superior to traditional surgical methods. CONCLUSION: Using AEM anal fistula plug in treatment that causes the anorectal fistula is safe and successful in 100% of patients. It can reduce pain, shorten disease course and protect anal function. PMID:18720541

  5. Hormonal regulation of medullary bone metabolism in the laying hen

    Energy Technology Data Exchange (ETDEWEB)

    Harrison, J.R.

    1987-01-01

    A new organ culture system for the study of bone formation has been developed using medullary bone, a non-structural, metabolically active form of bone which is found in the marrow cavities of egg-laying birds. In the presence of fetal calf serum, bone explants were viable in culture by morphological criteria, and retained large numbers of osteoblasts and osteoclasts. Incorporation of /sup 3/H-proline into collagenase-digestible protein (CDP) and non-collagen protein (NCP) was determined using purified bacterial collagenase. Collagen accounted for over 10% of the total protein labeled. The calcium-regulating hormones, parathyroid hormone and 1,25-dihydroxyvitamin D3 (1,25(OH)2D3), caused a dose-dependent inhibition of /sup 3/H-proline incorporation into CDP. The effective dose range of 1,25(OH)2D3 was 0.1 nM to 100 nM, while that of PTH was 1.0 nM to 100 nM. The effect of both hormones was specific for collagen, since /sup 3/H-proline incorporation into NCP was unaffected. Hydroxyproline analysis of bone explants and culture medium revealed that both hormones decreased the total hydroxyroline content of the cultures, suggesting that the inhibition of /sup 3/H-proline incorporation into DCP is due to inhibition of collagen synthesis.

  6. Platelet-rich plasma-induced bone marrow mesenchymal stem cells versus autologous nerve grafting for sciatic nerve repair

    Institute of Scientific and Technical Information of China (English)

    Changsuo Xia; Yajuan Li; Wen Cao; Zhaohua Yu

    2010-01-01

    Autologous nerve grafting is the gold standard of peripheral nerve repair.We previously showed that autologous platelet-rich plasma(PRP)contains high concentrations of growth factors and can induce in vitro cultured bone marrow mesenchymal stem cells(BMSCs)to differentiate into Schwann cells.Here we used PRP-induced BMSCs combined with chemically extracted acellular nerves to repair sciatic nerve defects and compared the effect with autologous nerve grafting.The BMSCs and chemically extracted acellular nerve promoted target muscle wet weight restoration,motor nerve conduction velocity,and axonal and myelin sheath regeneration,with similar effectiveness to autologous nerve grafting.This finding suggests that PRP induced BMSCs can be used to repair peripheral nerve defects.

  7. "High-grade" central acellular carcinoma and matrix-producing carcinoma of the breast: correlation between ultrasonographic findings and pathological features.

    Science.gov (United States)

    Yamaguchi, Rin; Tanaka, Maki; Mizushima, Yasuko; Hirai, Yoshitake; Yamaguchi, Miki; Terasaki, Hiroshi; Yokoyama, Toshiro; Tsuchiya, Shin-ichi; Nakashima, Osamu; Yano, Hirohisa

    2011-09-01

    High-grade carcinoma with a large central acellular zone (central acellular carcinoma, CAC) and matrixproducing carcinoma (MPC) are aggressive tumors that both have a central myxomatous acellular zone. Their characteristic morphology may be useful in diagnostic imaging. Ultrasonographic findings based on the Breast Imaging Recording and Data System (BI-RADS) and detailed histological features were evaluated in 11 cases of CAC and 2 cases of MPC to characterize their features. Safranin-O staining was undertaken for the evaluation of central acellular zones in these tumors. Overall, ultrasonography demonstrated heterogeneous hyperechoic lesions in the center of the hypoechoic mass. Posterior echo enhancement was observed in all but 1 case. One case was classified as malignant and the others as "borderline." Histologically, cancer tissue was located in the periphery of the tumor with a ring-like structure and fewer cellular central areas comprising hyaline cartilage myxoid material such as those stained by safranin-O. The present study showed that the pathological findings of CACs and MPCs accurately reflect the ultrasonographic findings. Tumors that showed hyperechoic areas in the center of the hypoechoic mass, with posterior echo enhancement indicating acellular zones composed by myxochondroid material, and that were also relatively round on ultrasonography may be benign, but evaluation is required to exclude CAC and MPC.

  8. Studies on Callus Induction and Regeneration of Medicinal Plant Chicory (Cichorium intybus L. from Leaf and Petiole Explants

    Directory of Open Access Journals (Sweden)

    H. Hadizadeh

    2016-07-01

    Full Text Available Introduction: Chicory (Cichorium intybus L. belongs to Asteraceae family is commonly known as witloof chicory. The leaves and the roots of this medicinal plant are edible and commonly used as salad. Some varieties are also cultivated as coffee substitute after roasting the roots. All parts of the plant contain these volatile oils, with the majority of the toxic components concentrated in the plant's root. In folk medicine, the plant is used for the treatment of diarrhea, spleen enlargement, fever, and vomiting. Antihepatotoxic activity on damaged rat’s liver sections and anti-bacterial activity of this crop has been recently reported. In vitro regeneration from leaf explants with various hormonal combinations has been reported previously. Moreover, in vitro regeneration of Chicory from cotyledon explants using different combinations of plant growth regulators has been studied. Also, a protocol for the regeneration of plantlets from leaf and petiole explants of witloof chicory has been developed. The aim of the present investigation was optimization of callus induction and shoot regeneration from leaf and petiole tissues of Chicory (Esfahan genotype. Materials and Methods: In this investigation, Esfahan genotype was used for callus induction and direct shoot regeneration. Seeds were first washed with running tap water for 30 min then seeds were surface sterilized by dipping in 70% ethanol for 90 s and rinsed with sterile distilled water, followed by immersing in 5% sodium hypochlorite solution for 25 min and thereafter rinsed for 30 min with sterile distilled water. The basal medium used in this investigation was MS. For shoot regeneration, leaf and petiole explants (5 mm segments were excised from 4-week-old sterile seedlings and cultured on MS medium containing different combinations of NAA / BA and KIN / BA in two separate experiments. Experiments were performed factorial based on completely randomized design. Cultures were incubated at 25

  9. Effects of thidiazuron and paclobutrazol on regeneration potential of tulip flower stalk explants in vitro and subsequent shoot multiplication

    Directory of Open Access Journals (Sweden)

    Małgorzata Podwyszyńska

    2011-01-01

    Full Text Available The effects of TDZ and paclobutrazol on the primary regeneration on tulip flower stalk explants of six cultivars and subsequent shoot multiplication were examined. Explants, flower stalk slices, were excised from cooled and subsequently forced bulbs. The explants were incubated for two months in darkness on medium containing NAA and cytokinins, 2iP and BAP, as control, or TDZ (0.5-4 mg l-1 and paclobutrazol (0.05-0.4 mg l-1. Then, the regenerating explants were subcultured on medium with TDZ and NAA applied at low concentrations. Different regeneration capabilities were found depending on cultivar and growth regulators. The percentage of explants forming leaf-like structures ranged, on the control medium, from 80% in 'Blue Parrot' and 'Prominence' to below 30% in 'Apeldoorn' and 'Mirjoran'. TDZ, applied at optimum for each cultivar concentration, greatly increased the regeneration potential up to 70-100%. Paclobutrazol, added to the TDZ-containing medium, significantly enhanced the response of explants, resulting in high numbers of leaf-like structures formed per explant (13.7-22.8. The structures developed gradually into characteristic forms: the growing up cotyledonary leaf, the probable root primordium formed at its base, the growing downwards stolon and the shoot meristem developed finely on its tip. It is suggested that such primary regeneration may have a nature of somatic embryogenesis. Then, the adventitious shoots developed and formed clusters, which were divided into 2-3 smaller ones every two months. The growth regulators, used at initial stage, markedly influenced subsequent shoot multiplication. Thus, the most intensive shoot formation was noted with TDZ at concentrations of 0.5-2 mg l-1 and paclobutrazol of 0.05-0.1 mg l-1.

  10. Simulação da soja geneticamente modificada tolerante ao glyphosate por meio do cultivo de explantes Simulation of the transgenic soybean tolerant to glyphosate through explant cultivation

    Directory of Open Access Journals (Sweden)

    Sérgio C. Siqueira

    1999-04-01

    Full Text Available O objetivo do experimento consistiu na simulação in vitro da soja transgênica tolerante ao glyphosate, através do cultivo de explantes em meios de cultura contendo aminoácidos aromáticos. As avaliações basearam-se nos efeitos do glyphosate sobre sementes oriundas de explantes de soja (Glycine max (L. Merr. cv. UFV-16. Para tanto, explantes de soja foram cultivados em meios de cultura líquidos com pH em torno de 5,0. Cada explante constou de um legume completamente expandido contendo duas sementes de aproximadamente 100 mg, conectada a um segmento de caule de 45 mm de comprimento. Os tratamentos testados foram: A = glutamina (Gln; B = Gln + fenilalanina (Phe + tirosina (Tyr + triptofano (Trp; C = Gln + glyphosate; D = Gln + Phe + Tyr + Trp + glyphosate. O experimento foi conduzido sob irradiância de 80 mmol-2s-1 a 25oC por 204 horas. Nos tratamentos que receberam aminoácidos aromáticos e glutamina, o herbicida não afetou as massas fresca e seca das sementes, como também, não afetou seus constituintes bioquímicos (proteínas, óleo, ácidos graxos, carboidratos e clorofilas. Portanto, a suplementação exógena de aminoácidos aromáticos suprime os efeitos fitotóxicos do glyphosate sobre explantes de soja, permitindo estudos sobre o seu modo de ação e metabolismo nas sementes, uma vez que os explantes se comportaram analogamente à soja transgênica não suscetível ao herbicida.The objective of this experiment consisted in simulation in vitro of the transgenic soybean tolerant to glyphosate through explant cultivation in culture medium containing aromatic amino acids. The effects of glyphosate on soybean (Glycine max (L. Merrill were evaluated in seeds harvested from explants of cv UFV-16. The soybean explants were cultivated in liquid medium culture with pH about 5.0. Each explant consisted of one fruit completely expanded, containing two seeds of 100 mg approximately, and connected to a stem segment of 45 mm length. The

  11. T cells fail to develop in the human skin-cell explants system; an inconvenient truth

    Directory of Open Access Journals (Sweden)

    Vanderlocht Joris

    2011-02-01

    Full Text Available Abstract Background Haplo-identical hematopoietic stem cell (HSC transplantation is very successful in eradicating haematological tumours, but the long post-transplant T-lymphopenic phase is responsible for high morbidity and mortality rates. Clark et al. have described a skin-explant system capable of producing host-tolerant donor-HSC derived T-cells. Because this T-cell production platform has the potential to replenish the T-cell levels following transplantation, we set out to validate the skin-explant system. Results Following the published procedures, while using the same commercial components, it was impossible to reproduce the skin-explant conditions required for HSC differentiation towards mature T-cells. The keratinocyte maturation procedure resulted in fragile cells with minimum expression of delta-like ligand (DLL. In most experiments the generated cells failed to adhere to carriers or were quickly outcompeted by fibroblasts. Consequently it was not possible to reproduce cell-culture conditions required for HSC differentiation into functional T-cells. Using cell-lines over-expressing DLL, we showed that the antibodies used by Clark et al. were unable to detect native DLL, but instead stained 7AAD+ cells. Therefore, it is unlikely that the observed T-lineage commitment from HSC is mediated by DLL expressed on keratinocytes. In addition, we did confirm expression of the Notch-ligand Jagged-1 by keratinocytes. Conclusions Currently, and unfortunately, it remains difficult to explain the development or growth of T-cells described by Clark et al., but for the fate of patients suffering from lymphopenia it is essential to both reproduce and understand how these co-cultures really "work". Fortunately, alternative procedures to speed-up T-cell reconstitution are being established and validated and may become available for patients in the near future.

  12. Agrobacterium-mediated transformation of oat (Avena sativa L.) cultivars via immature embryo and leaf explants.

    Science.gov (United States)

    Gasparis, Sebastian; Bregier, Cezary; Orczyk, Waclaw; Nadolska-Orczyk, Anna

    2008-11-01

    This paper reports on the successful Agrobacterium-mediated transformation of oat, and on some factors influencing this process. In the first step of the experiments, three cultivars, two types of explant, and three combinations of strain/vectors, which were successfully used for transformation of other cereals were tested. Transgenic plants were obtained from the immature embryos of cvs. Bajka, Slawko and Akt and from leaf base explants of cv. Bajka after transformation with A. thumefaciens strain LBA4404(pTOK233). The highest transformation rate (12.3%) was obtained for immature embryos of cv. Bajka. About 79% of the selected plants proved to be transgenic; however, only 14.3% of the T(0) plants and 27.5% of the T(1) showed GUS expression. Cell competence of both types of explant differed in terms of their transformation ability and transgene expression. The next step of the study was to test the suitability for oat transformation of the pGreen binary vector combined with different selection cassettes: nptII or bar under the nos or 35S promoter. Transgenic plants were selected in combinations transformed with nos::nptII, 35S::nptII and nos::bar. The highest transformation efficiency (5.3%) was obtained for cv. Akt transformed with nos::nptII. A detailed analysis of the T(0) plants selected from a given callus line and their progeny revealed that they were the mixture of transgenic, chimeric-transgenic and non-transgenic individuals. Southern blot analysis of T(0) and T(1) showed simple integration pattern with the low copy number of the introduced transgenes.

  13. Plant regeneration from single-nodal-stem explants of legume tree Prosopis alba Griseb.

    Science.gov (United States)

    Castillo de Meier, G; Bovo, O A

    2000-08-01

    Seeds of Prosopis alba were scarified with abrasive paper and placed to germinate on MS (Murashige and Skoog 1962) nutrient medium. After 7 days of culture, the basal part of cotyledons was removed and pieces of 4 mm" from distal parts were cultured on Murashige and Skoog (1962) mineral salts and vitamins (MS) (3% sucrose) supplemented with growth regulators. Callus proliferation took place in the majority of the media tested. A low percentage of calluses with green buds that developed on MS basal medium containing 0.1 mg.L-1 2,4-D alone or supplemented with BAP at 0.1 mg.L-1 was observed. Neither cotyledonary segments in any medium assayed regenerated the whole plants. Bud elongation (near 70%) was achieved when single-nodal-stem segments cut from 20 days old seedlings were cultured on MS salts supplemented with 3 mg.L-1 NAA or 3 mg.L-1 IBA combined with 0.05 mg.L-1 KIN after 60 days in culture. Multiple shoots per bud were also observed. Single-nodal-stem segments from five-year-old plants were also cultured on the same media used for seedling explants. Maximal frequency of explants with bud elongation (near 70%) was found on MS with 0.1 mg.L-1 NAA plus 1 mg.L-1 BAP after 60 days of culture. Single-nodal-stem explants cut from adult trees (more than 20 years) were also employed, but the number of bud elongation was lesser. For rooting, the elongated shoots were transferred to a semisolid or liquid MS culture medium employing a paper bridge, supplemented with 0.5 mg.L-1 IBA or 0.1 mg.L-1 NAA.

  14. Glucocorticoids affect 24 h clock genes expression in human adipose tissue explant cultures.

    Directory of Open Access Journals (Sweden)

    Purificación Gómez-Abellán

    Full Text Available AIMS: to examine firstly whether CLOCK exhibits a circadian expression in human visceral (V and subcutaneous (S adipose tissue (AT in vitro as compared with BMAL1 and PER2, and secondly to investigate the possible effect of the glucocorticoid analogue dexamethasone (DEX on positive and negative clock genes expression. SUBJECTS AND METHODS: VAT and SAT biopsies were obtained from morbid obese women (body mass index ≥ 40 kg/m(2 (n = 6. In order to investigate rhythmic expression pattern of clock genes and the effect of DEX on CLOCK, PER2 and BMAL1 expression, control AT (without DEX and AT explants treated with DEX (2 hours were cultured during 24 h and gene expression was analyzed at the following times: 10:00 h, 14:00 h, 18:00 h, 22:00 h, 02:00 h and 06:00 h, using qRT-PCR. RESULTS: CLOCK, BMAL1 and PER2 expression exhibited circadian patterns in both VAT and SAT explants that were adjusted to a typical 24 h sinusoidal curve. PER2 expression (negative element was in antiphase with respect to CLOCK and in phase with BMAL1 expression (both positive elements in the SAT (situation not present in VAT. A marked effect of DEX exposure on both positive and negative clock genes expression patterns was observed. Indeed, DEX treatment modified the rhythmicity pattern towards altered patterns with a period lower than 24 hours in all genes and in both tissues. CONCLUSIONS: 24 h patterns in CLOCK and BMAL1 (positive clock elements and PER2 (negative element mRNA levels were observed in human adipose explants. These patterns were altered by dexamethasone exposure.

  15. Propagation of goldenrod (Solidago canadensis L. from leaf and nodal explants

    Directory of Open Access Journals (Sweden)

    Jun Li

    2012-02-01

    Full Text Available Goldenrod (Solidago canadensis L. is an invasive plant species in many countries except North America but a cut-flower species worldwide. There is a need to generate and propagate goldenrod clones efficiently for research and commercial purposes. A callus induction and plantlet regeneration system was developed by studying the influence of explant type and different concentrations of plant growth regulators. The highest callus production from leaf segments was obtained on Murashige and Skoog’s medium (MS medium supplemented with 1.0 mg/L naphthalene acetic acid (NAA and 1.0 mg/L 6-benzylaminopurine (BA. Adventitious shoots could be regenerated directly from leaf explants without an intermediate callus phase with the highest shoot induction percentage of 87.2%. The largest number of adventitious shoots per leaf explant (3.2 was obtained on MS medium supplemented with 0.4 mg/L NAA and 2.0 mg/L BA. MS medium supplemented with 0.1 mg/L NAA and 1.0 mg/L BA was the best medium for axillary shoot regeneration from nodal segments. The highest root number and longest roots occurred on half-strength MS without the addition of any growth regulator. Rooted plantlets were then transferred to a soil-based growth medium, placed in a greenhouse, and acclimatized with 100% success. All surviving plants grew normally without showing any morphological varia­tion when compared to those grow from seed. This regeneration protocol may be used to produce certain biotypes of goldenrod suitable for genetic transformation rapid propagation of goldenrod for commercial purposes or for screening fungi and toxins as potential biocontrol agents against this weed.

  16. Agrobacterium-mediated transformation of finger millet (Eleusine coracana (L.) Gaertn.) using shoot apex explants.

    Science.gov (United States)

    Ceasar, S Antony; Ignacimuthu, S

    2011-09-01

    A new Agrobacterium-mediated transformation system was developed for finger millet using shoot apex explants. The Agrobacterium strain LBA4404 harboring binary vector pCAMBIA1301, which contained hygromycin phosphotransferase (hptII) as selectable marker gene and β-glucuronidase (GUS) as reporter gene, was used for optimization of transformation conditions. Two finger millet genotypes, GPU 45 and CO 14, were used in this study. The optimal conditions for the Agrobacterium-mediated transformation of finger millet were found to be the co-cultivation of explants obtained on the 16th day after callus induction (DACI), exposure of explants for 30 min to agrobacterial inoculum and 3 days of co-cultivation on filter paper placed on medium supplemented with 100 μM acetosyringone (AS). Addition of 100 μM L: -cysteine in the selection medium enhanced the frequency of transformation and transgenic plant recovery. Both finger millet genotypes were transformed by Agrobacterium. A frequency of 19% transient expression with 3.8% stable transformation was achieved in genotype GPU 45 using optimal conditions. Five stably transformed plants were fully characterized by Southern blot analysis. A segregation analysis was also performed in four R(1) progenies, which showed normal Mendelian pattern of transgene segregation. The inheritance of transgenes in R(1) progenies was also confirmed by Southern blot analysis. This is the first report on Agrobacterium-mediated transformation of finger millet. This study underpins the introduction of numerous agronomically important genes into the genome of finger millet in the future.

  17. Osteogenesis effect of guided bone regeneration combined with alveolar cleft grafting: assessment by cone beam computed tomography.

    Science.gov (United States)

    Xiao, W-L; Zhang, D-Z; Chen, X-J; Yuan, C; Xue, L-F

    2016-06-01

    Cone beam computed tomography (CBCT) allows for a significantly lower radiation dose than conventional computed tomography (CT) scans and provides accurate images of the alveolar cleft area. The osteogenic effect of guided bone regeneration (GBR) vs. conventional alveolar bone grafting alone for alveolar cleft defects was evaluated in this study. Sixty alveolar cleft patients were divided randomly into two groups. One group underwent GBR using acellular dermal matrix film combined with alveolar bone grafting using iliac crest bone grafts (GBR group), while the other group underwent alveolar bone grafting only (non-GBR group). CBCT images were obtained at 1 week and at 3 months following the procedure. Using Simplant 11.04 software, the bone resorption rate was calculated and compared between the two groups. The bone resorption rate from 1 week to 3 months following bone grafting without the GBR technique was 36.50±5.04%, whereas the bone resorption rate using the GBR technique was 31.69±5.50% (P=0.017). The application of autogenous iliac bone combined with the GBR technique for alveolar bone grafting of alveolar cleft patients can reduce bone resorption and result in better osteogenesis.

  18. Outcome of total knee replacement following explantation and cemented spacer therapy

    Directory of Open Access Journals (Sweden)

    Ghanem, Mohamed

    2016-03-01

    Full Text Available Background: Infection after total knee replacement (TKR is one of the serious complications which must be pursued with a very effective therapeutic concept. In most cases this means revision arthroplasty, in which one-setting and two-setting procedures are distinguished. Healing of infection is the conditio sine qua non for re-implantation. This retrospective work presents an assessment of the success rate after a two-setting revision arthroplasty of the knee following periprosthetic infection. It further considers drawing conclusions concerning the optimal timing of re-implantation.Patients and methods: A total of 34 patients have been enclosed in this study from September 2005 to December 2013. 35 re-implantations were carried out following explantation of total knee and implantation of cemented spacer. The patient’s group comprised of 53% (18 males and 47% (16 females. The average age at re-implantation time was 72.2 years (ranging from 54 to 85 years. We particularly evaluated the microbial spectrum, the interval between explantation and re-implantation, the number of surgeries that were necessary prior to re-implantation as well as the postoperative course. Results: We reported 31.4% (11 reinfections following re-implantation surgeries. The number of the reinfections declined with increasing time interval between explantation and re-implantation. Patients who developed reinfections were operated on (re-implantation after an average of 4.47 months. Those patients with uncomplicated course were operated on (re-implantation after an average of 6.79 months. Nevertheless, we noticed no essential differences in outcome with regard to the number of surgeries carried out prior to re-implantation. Mobile spacers proved better outcome than temporary arthrodesis with intramedullary fixation.Conclusion: No uniform strategy of treatment exists after peri-prosthetic infections. In particular, no optimal timing can be stated concerning re

  19. In situ fiber-optical monitoring of cytosolic calcium in tissue explant cultures

    CERN Document Server

    Ryser, Manuel; Geiser, Marianne; Frenz, Martin; Rička, Jaro

    2014-01-01

    We present a fluorescence-lifetime based method for monitoring cell and tissue activity in situ, during cell culturing and in the presence of a strong autofluorescence background. The miniature fiber-optic probes are easily incorporated in the tight space of a cell culture chamber or in an endoscope. As a first application we monitored the cytosolic calcium levels in porcine tracheal explant cultures using the Calcium Green-5N (CG5N) indicator. Despite the simplicity of the optical setup we are able to detect changes of calcium concentration as small as 2.5 nM, with a monitoring time resolution of less than 1 s.

  20. Stable genetic transformation of Jatropha curcas via Agrobacterium tumefaciens-mediated gene transfer using leaf explants

    KAUST Repository

    Kumar, Nitish

    2010-07-01

    Jatropha curcas is an oil bearing species with multiple uses and considerable economic potential as a biofuel crop. A simple and reproducible protocol was developed for Agrobacterium tumefaciens-mediated stable genetic transformation of J. curcas using leaf explains. Agrobacterium strain LBA 4404 harbouring the binary vector pCAMBIA 1304 having sense-dehydration responsive element binding (S-DREB2A), beta-glucuronidase (gus), and hygromycin-phosphotransferase (hpt) genes were used for gene transfer. A number of parameters such as preculture of explains, wounding of leaf explants, Agrobacterium growth phase (OD), infection duration, co-cultivation period, co-cultivation medium pH, and acetosyringone, were studied to optimized transformation efficiency. The highest transformation efficiency was achieved using 4-day precultured, non-wounded leaf explants infected with Agrobacterium culture corresponding to OD(600)=0.6 for 20 min, followed by co-cultivation for 4 days in a co-cultivation medium containing 100 mu M acetosyringone, pH 5.7. Co-cultivated leaf explants were initially cultured on Murashige and Skoog (MS) medium supplemented with 2.27 mu M thidiazuron (TDZ) for regeneration of shoot buds, followed by selection on same medium with 5 mu g ml(-1) hygromycin. Selected shoot buds were transferred to MS medium containing 10 mu M kinetin (Kn), 4.5 mu M 6-benzyl aminopurine (BA), and 5.5 mu M alpha-naphthaleneacetic acid (NAA) for proliferation. The proliferated shoots were elongated on MS medium supplemented with 2.25 mu M BA and 8.5 mu M indole-3-acetic acid (IAA). The elongated shoots were rooted on half strength MS medium supplemented with 15 mu M indole-3-butyric acid (IBA), 5.7 mu M IAA, 5.5 mu M NAA, and 0.25 mg l(-1) activated charcoal. GUS histochemical analysis of the transgenic tissues further confirmed the transformation event. PCR and DNA gel blot hybridization were performed to confirm the presence of transgene. A transformation efficiency of 29% was

  1. In Vitro Shoot Bud Differentiation from Hypocotyl Explants of Chili Peppers (Capsicum annuum L.)

    OpenAIRE

    Owk ANIEL KUMAR; Sape SUBBA TATA

    2010-01-01

    Chili pepper (Capsicum annuum L.) is an economically important spice crop in tropical and subtropical countries. In vitro plant regeneration was obtained from 15th day old hypocotyl explants of three chili pepper cultivars (Capsicum annuum L., var. �X-235�, var. �PC-1� and var. �Pusa Jwala�). Among the genotypes of Capsicum L. var. �X-235� responded better than the var. �PC-1� and var. �Pusa Jwala�. MS medium containing BAP (4.0 mg/l) and IAA (0.5 mg/l) was found to be the best medium for the...

  2. Variation in phytate accumulation in common bean (Phaseolus vulgaris L. fruit explants

    Directory of Open Access Journals (Sweden)

    Cileide Maria Medeiros Coelho

    2008-02-01

    Full Text Available The in vitro synthesis of phytate was studied in common bean fruit explants. Different concentrations of sucrose; phosphorus (P; myo-inositol; abscisic acid (ABA; glutamine and methionine, were tested. Fixed concentrations of these compounds were tested at different periods (0, 3, 6 and 9 days. Variation in phytate coincided with different concentrations of sucrose, myo-inositol, P and ABA for the duration tested. These compounds caused an accumulation of phytate and were more effective in the presence of myo-inositol and P. The accumulation of P varied less than phytate for the different treatments tested in vitro. In conclusion, P, sucrose, ABA, and myo-inositol caused an increase in the phytate of bean seed, showing that it could be possible to alter its content by culturing bean fruit explants in vitro.O fósforo é armazenado na forma de fitato nas sementes, o qual forma complexos estáveis e insolúveis com minerais e proteínas, conferindo efeito antinutriente. A síntese de fitato foi estudada em cultivo de explantes de fruto de feijão in vitro sob diferentes concentrações de sacarose, fósforo (P, mio-inositol, ácido abscísico (ABA, glutamina e metionina. Fixada a concentração destes compostos, testou-se os diferentes tempos de cultivo (0, 3, 6 e 9 dias. A variação no acúmulo de fitato ocorreu na presença de sacarose, mio-inositol, P e ABA nas diferentes concentrações e tempos testados. O acúmulo mais efetivo de fitato ocorreu na presença de mio-inositol e P. O acúmulo de P variou menos do que fitato em todos os tratamentos. Em conclusão, P, sacarose, ABA e mio-inositol causaram aumento no fitato acumulado nas sementes, mostrando que foi possível alterar a síntese de fitato em cultivo de explantes de fruto de feijão.

  3. In situ osteogenesis: regeneration of 10-cm mandibular defect in porcine model using recombinant human bone morphogenetic protein-2 (rhBMP-2) and Helistat absorbable collagen sponge.

    Science.gov (United States)

    Carstens, Michael H; Chin, Martin; Li, X Jian

    2005-11-01

    Traditional bone grafting relies upon the incorporation of a bone-cell bearing structure into a recipient site. The graft serves as a scaffold that is eventually replaced and remodeled. This process is known as osteoconduction. Recombinant human bone morphogenetic protein-2 (rhBMP-2) is commercially available as an acellular implant in which the protein is bound to an absorbable collagen sponge (ACS). The rhBMP-2/ACS implant converts undifferentiated mesenchymal stem cells into osteoblasts and promotes an intense local neovascular response. This process, known as osteoinduction, produces bone via membranous, chondroid, or endochondral ossification. The type of bone synthesis depends upon the mesenchymal substrate and the local cellular environment. Using this simple technique, bone defects can be resynthesized with good outcomes and a significant reduction in donor site morbidity. Repair of a critical-sized mandibular resection defect with ISO is described. Basic science concepts of rhBMP-2, relevant histopathologic findings, and clinical application are described.

  4. Structural and redox behavior of OxyVita, a zero-linked polymeric hemoglobin: comparison with natural acellular polymeric hemoglobins.

    Science.gov (United States)

    Harrington, John P; Orlik, Kseniya; Orlig, Kseniya; Zito, Samantha L; Wollocko, Jacek; Wollocko, Hanna

    2010-04-01

    A zero-linked polymeric hemoglobin (OxyVita Hb) has been developed for application as an acellular therapeutic hemoglobin-based-oxygen-carrier (HBOC). For effective and safe oxygen binding, transport and delivery, an HBOC must meet essential molecular requirements related to its structural integrity and redox stability. OxyVita is a super polymer possessing an average M.wt. of 17 x 10(6) Da. Structural integrity was determined by unfolding studies of OxyVita in the presence of increasing concentrations of urea. The unfolding midpoints (D(1/2)) of different preparations of OxyVita (solution and powder forms) were compared to Lumbricus Hb (LtHb) and Arenicola Hb (ArHb), natural acellular polymeric hemoglobins, which are serving as models for an effective and safe acellular HBOC. Reduction studies of OxyVita Hb using endogenous reducing agents were also investigated. Results from these studies indicate that: 1) OxyVita Hb exhibits greater resistance to conformational change than either LtHb or ArHb in the reduced (oxyHb) state; and 2) the reduction of met OxyVita Hb to oxyHb occurs slowly in the presence of either ascorbic acid (70% reduction in 560 min.) or beta-NADH (40% reduction in 90 min.). These studies provide consistent evidence that OxyVita Hb possesses physiochemical properties that exhibit structural integrity and redox behavior necessary for functioning as an effective and safe HBOC within clinical applications. These results are in agreement with observations made by other investigators as to the reduction in heme-loss of OxyVita Hb, essential for the reversible binding/release of molecular oxygen within the circulatory system.

  5. Characterisation of the circulating acellular proteome of healthy sheep using LC-MS/MS-based proteomics analysis of serum.

    Science.gov (United States)

    Chemonges, Saul; Gupta, Rajesh; Mills, Paul C; Kopp, Steven R; Sadowski, Pawel

    2016-01-01

    Unlike humans, there is currently no publicly available reference mass spectrometry-based circulating acellular proteome data for sheep, limiting the analysis and interpretation of a range of physiological changes and disease states. The objective of this study was to develop a robust and comprehensive method to characterise the circulating acellular proteome in ovine serum. Serum samples from healthy sheep were subjected to shotgun proteomic analysis using nano liquid chromatography nano electrospray ionisation tandem mass spectrometry (nanoLC-nanoESI-MS/MS) on a quadrupole time-of-flight instrument (TripleTOF® 5600+, SCIEX). Proteins were identified using ProteinPilot™ (SCIEX) and Mascot (Matrix Science) software based on a minimum of two unmodified highly scoring unique peptides per protein at a false discovery rate (FDR) of 1% software by searching a subset of the Universal Protein Resource Knowledgebase (UniProtKB) database (http://www.uniprot.org). PeptideShaker (CompOmics, VIB-UGent) searches were used to validate protein identifications from ProteinPilot™ and Mascot. ProteinPilot™ and Mascot identified 245 and 379 protein groups (IDs), respectively, and PeptideShaker validated 133 protein IDs from the entire dataset. Since Mascot software is considered the industry standard and identified the most proteins, these were analysed using the Protein ANalysis THrough Evolutionary Relationships (PANTHER) classification tool revealing the association of 349 genes with 127 protein pathway hits. These data are available via ProteomeXchange with identifier PXD004989. These results demonstrated for the first time the feasibility of characterising the ovine circulating acellular proteome using nanoLC-nanoESI-MS/MS. This peptide spectral data contributes to a protein library that can be used to identify a wide range of proteins in ovine serum.

  6. Ascorbic acid inhibits the squamous metaplasia that results from treatment of tracheal explants with asbestos or benzo[a]pyrene-coated asbestos.

    Science.gov (United States)

    Holtz, G; Bresnick, E

    1988-01-01

    Hamster tracheal explants were maintained in culture in the presence or absence of benzo[a]pyrene (BP), crocidolite asbestos, or BP-coated crocidolite. Dose-dependent squamous metaplasia was observed in the treated samples. L-Ascorbic acid and DL-alpha-tocopherol were able to partially protect the tracheal explants from the metaplastic response induced by crocidolite. Furthermore, ascorbic acid reduced the extent of metaplasia observed in hamster tracheal explants that were exposed to BP-crocidolite.

  7. Daily Serum Collection after Acellular Dermal Matrix-Assisted Breast Reconstruction

    Directory of Open Access Journals (Sweden)

    Glenda Giorgia Caputo

    2015-05-01

    Full Text Available BackgroundThe acellular dermal matrix (ADM-assisted breast reconstruction technique is widely known, but discouraging results due to early postoperative complications have been reported. As the literature identifies seroma as the most common issue after breast surgery without identifying its pathogenesis, we aimed to report the trend of postoperative daily serum collection after ADM-assisted breast reconstruction and compare it with data in the literature in order to discover more about this little-known topic.MethodsA retrospective study on 28 consecutive patients who received ADM-assisted breast reconstruction between February 2013 and February 2014 was performed. In order to reduce the number of variables that could affect serum production, only one brand of ADM was used and all tissues were handled gently and precisely. The daily drainage volume was recorded per patient during the first four days of hospitalization. Likewise, postoperative complications were noted during routine follow-up.ResultsIn total, five (17.9% bilateral and 23 (82.1% unilateral ADM-assisted breast reconstructions (33 implants were performed. The mean age, body mass index, and length of hospital stay were 53.6 years, 21.3 kg/m2, and 4.5 days, respectively. One major complication led to implant loss (3.0%, and nine minor complications were successfully treated with ambulatory surgery (27.3%. Serum collection linearly decreased after 24 hours postoperatively.ConclusionsDaily drainage decreased following the theoretical decline of acute inflammation. In concordance with the literature, daily serum production may not be related to the use of ADM.

  8. Plastic Surgery and Acellular Dermal Matrix: Highlighting Trends from 1999 to 2013.

    Science.gov (United States)

    Daar, David A; Gandy, Jessica R; Clark, Emily G; Mowlds, Donald S; Paydar, Keyianoosh Z; Wirth, Garrett A

    2016-05-01

    The last decade has ushered in a rapidly expanding global discussion regarding acellular dermal matrix (ADM) applications, economic analyses, technical considerations, benefits, and risks, with recent emphasis on ADM use in breast surgery. This study aims to evaluate global trends in ADM research using bibliometric analysis. The top nine Plastic Surgery journals were determined by impact factor (IF). Each issue of the nine journals between 1999 and 2013 was accessed to compile a database of articles discussing ADM. Publications were further classified by IF, authors' geographic location, study design, and level of evidence (LOE, I-V). Productivity index and productivity share were calculated for each region. In total, 256 ADM articles were accessed. The annual global publication volume increased significantly by 4.2 (0.87) articles per year (p<0.001), with a mean productivity index of 36.3 (59.0). The mean impact factor of the nine journals increased significantly from 0.61 (0.11) to 2.47 (0.99) from 1993 to 2013 (p<0.001). Despite this increase in the global ADM literature, the majority of research was of weaker LOE (level I: 2.29% and level II: 9.17%). USA contributed the most research (87%), followed by Asia (4.76%) and Western Europe (4.71%). USA contributed the greatest volume of research. Regarding clinical application of ADM, the majority of publications focused on ADM use in breast surgery, specifically breast reconstruction (154 articles, 60.2%). The majority of research was of lower LOE; thus, efforts should be made to strengthen the body of literature, particularly with regard to cost analysis.

  9. Potential sites for the perception of gravity in the acellular slime mold Physarum polycephalum

    Science.gov (United States)

    Block, I.; Briegleb, W.

    Recently a gravisensitivity of the acellular slime mold Physarum polycephalum, which possesses no specialized gravireceptor, could be established by conducting experiments under simulated and under real near weightlessness. In these experiments macroplasmodia showed a modulation of their contraction rhythm followed by regulation phenomena. Until now the perception mechanism for the gravistimulus is unknown, but several findings indicate the involvement of mitochondria: A) During the impediment of respiration the Og-reaction is inhibited and the regulation is reduced. B) The response to a light stimulus and the following regulation phenomena strongly resemble the behavior during exposure to Og, the only difference is that the two reactions are directed into opposite directions. In the blue-light reaction a flavin of the mitochondrial matrix seems to be involved in the light perception. C) The contraction rhythm as well as its modulations are coupled to rhythmic changes in the levels of ATP and calcium ions, involving the mitochondria as sites of energy production and of Ca++-storage. - So the mitochondria could be the site of the regulation and they possibly are the receptor sites for the light and gravity stimuli. - Also the observation of a morphologic polarity of the slime mold's plasmodial strands has to be considered: Cross-sections reveal that the ectoplasmic wall surrounding the streaming endoplasm is much thinner on the physically lower side than on the upper side of the strand - this applies to strands lying on or hanging on a horizontal surface. So, in addition to the mitochondria, also the morphologic polarity may be involved in the perception mechanism of the observed gravisensitivity and of the recently established geotaxis. - The potential role of the nuclei and of the contractile elements in the perception of gravity is also discussed.

  10. Using porcine acellular collagen matrix (Pelvicol® in bladder augmentation: experimental study

    Directory of Open Access Journals (Sweden)

    Ayyildiz Ali

    2006-01-01

    Full Text Available PURPOSE: Evaluate the rabbit augmented bladder with PelvicolÒ. MATERIALS AND METHODS: Twenty New Zealand rabbits were divided into 4 groups. Bladder augmentation was performed using a 10 x 10 mm sized porcine acellular collagen matrix. The material was placed on the dome of the bladder wall as a patch with 5-0 polyglycolic sutures. The bladder was resected on the 7th, 14th day, 30th and 90th days, and processed for histological analysis. RESULTS: No stone formation was found in the first, second and fourth weeks. In the first week, there was inflammatory appearance and roughness in the reconstructed area when compared to other sites on the bladder wall. The material could not be seen in some bladders because of acute inflammatory reaction. The normal bladder epithelium was found on the part of the bladder wall that follows the surface of the eroded material. In the second week, edema was observed through the bladder wall. Perivesical fat tissue increased and it was not easy to distinguish it from the surrounding area. In the fourth week, the bladder wall was thickened and there was a sensation of hardness present. The inner and outer surface of the material was darker than in the other bladders. In the third month, there was no inflammatory reaction; however, there was micro calcification and irregular detrusor regeneration. CONCLUSIONS: PelvicolÒ cannot be suitable material for bladder augmentation because of the resultant micro calcification, thickening of the bladder wall and irregular development of detrusor regeneration.

  11. EFEITO DE REGULADORES DE CRESCIMENTO E TIPO DE EXPLANTES NA MORFOGÊNESE in vitro DE Capsicum annuum L.

    Directory of Open Access Journals (Sweden)

    Márcio Henrique Pereira Barbosa

    1994-01-01

    Full Text Available RESUMO Explantes apicais, cotiledonares, hipocotiledonares e brotações regeneradas in vitro de Capsicum annuum cv. Agronômico 10 foram cultivados em meio (MURASHIGE & SKOOG, 1962-MS suplementado com diferentes concentrações e combinações de BAP, adenina e tidiazuron. De acordo com o tipo de explante e reguladores de crescimento, foram obtidas diferentes respostas morfogenéticas. De modo geral, meios contendo concentrações mais elevadas de BAP e tidiazuron, promoveram a indução e produção de calos nos explantes provenientes de "seedlings", enquanto que a rizogênese foi favorecida em meios com baixas concentrações de BAP. Múltiplos brotos axilares foram formados somente quando utilizou-se como explante, brotações regeneradas in vitro. Os níveis de 26,64 e 39,96 µM de BAP possiblitaram a maximização da proliferação em aproximadamente 2,08 e 2,17 novas brotações por explante, respectivamente.

  12. Organotypic Culture of Breast Tumor Explants as a Multicellular System for the Screening of Natural Compounds with Antineoplastic Potential

    Science.gov (United States)

    Carranza-Torres, Irma Edith; Guzmán-Delgado, Nancy Elena; Coronado-Martínez, Consuelo; Bañuelos-García, José Inocente; Viveros-Valdez, Ezequiel; Morán-Martínez, Javier; Carranza-Rosales, Pilar

    2015-01-01

    Breast cancer is the leading cause of death in women worldwide. The search for novel compounds with antitumor activity, with less adverse effects and higher efficacy, and the development of methods to evaluate their toxicity is an area of intense research. In this study we implemented the preparation and culture of breast tumor explants, which were obtained from precision-cut breast tumor slices. In order to validate the model we are proposing to screen antineoplastic effect of natural compounds, we selected caffeic acid, ursolic acid, and rosmarinic acid. Using the Krumdieck tissue slicer, precision-cut tissue slices were prepared from breast cancer samples; from these slices, 4 mm explants were obtained and incubated with the selected compounds. Viability was assessed by Alamar Blue assay, LDH release, and histopathological criteria. Results showed that the viability of the explants cultured in the presence of paclitaxel (positive control) decreased significantly (P < 0.05); however, tumor samples responded differently to each compound. When the explants were coincubated with paclitaxel and compounds, a synergic effect was observed. This study shows that ex vivo culture of breast cancer explants offers a suitable alternative model for evaluating natural or synthetic compounds with antitumor properties within the complex microenvironment of the tumor. PMID:26075250

  13. Millicurrent stimulation of human articular chondrocytes cultivated in a collagen type-I gel and of human osteochondral explants

    Directory of Open Access Journals (Sweden)

    Silny Jiri

    2010-08-01

    Full Text Available Abstract Background Here we investigate the effect of millicurrent treatment on human chondrocytes cultivated in a collagen gel matrix and on human osteochondral explants. Methods Human chondrocytes from osteoarthritic knee joints were enzymatically released and transferred into a collagen type-I gel. Osteochondral explants and cell-seeded gel samples were cultivated in-vitro for three weeks. Samples of the verum groups were stimulated every two days by millicurrent treatment (3 mA, sinusoidal signal of 312 Hz amplitude modulated by two super-imposed signals of 0.28 Hz, while control samples remained unaffected. After recovery, collagen type-I, type-II, aggrecan, interleukin-1β, IL-6, TNFα and MMP13 were examined by immunohistochemistry and by real time PCR. Results With regard to the immunostainings 3 D gel samples and osteochondral explants did not show any differences between treatment and control group. The expression of all investigated genes of the 3 D gel samples was elevated following millicurrent treatment. While osteochondral explant gene expression of col-I, col-II and Il-1β was nearly unaffected, aggrecan gene expression was elevated. Following millicurrent treatment, IL-6, TNFα, and MMP13 gene expression decreased. In general, the standard deviations of the gene expression data were high, resulting in rarely significant results. Conclusions We conclude that millicurrent stimulation of human osteoarthritic chondrocytes cultivated in a 3 D collagen gel and of osteochondral explants directly influences cell metabolism.

  14. Organotypic Culture of Breast Tumor Explants as a Multicellular System for the Screening of Natural Compounds with Antineoplastic Potential

    Directory of Open Access Journals (Sweden)

    Irma Edith Carranza-Torres

    2015-01-01

    Full Text Available Breast cancer is the leading cause of death in women worldwide. The search for novel compounds with antitumor activity, with less adverse effects and higher efficacy, and the development of methods to evaluate their toxicity is an area of ​​intense research. In this study we implemented the preparation and culture of breast tumor explants, which were obtained from precision-cut breast tumor slices. In order to validate the model we are proposing to screen antineoplastic effect of natural compounds, we selected caffeic acid, ursolic acid, and rosmarinic acid. Using the Krumdieck tissue slicer, precision-cut tissue slices were prepared from breast cancer samples; from these slices, 4 mm explants were obtained and incubated with the selected compounds. Viability was assessed by Alamar Blue assay, LDH release, and histopathological criteria. Results showed that the viability of the explants cultured in the presence of paclitaxel (positive control decreased significantly (P<0.05; however, tumor samples responded differently to each compound. When the explants were coincubated with paclitaxel and compounds, a synergic effect was observed. This study shows that ex vivo culture of breast cancer explants offers a suitable alternative model for evaluating natural or synthetic compounds with antitumor properties within the complex microenvironment of the tumor.

  15. Biochemical investigations during in vitro adventitious shoot regeneration in leaflet explants from nodal segments of a mature Albizia procera tree

    Institute of Scientific and Technical Information of China (English)

    Ekta Rai; Sulochna Bouddha; Shamim Akhtar Ansari

    2016-01-01

    The in vitro adventitious shoot differentiation in leaflet explants of an adult tree differed from that of leaflet explants of seedlings of Albizia procera (Roxb.) Benth. reported previously elsewhere. The leaflet explants from an adult tree passed through an initial callus phase for 30 days on MS medium supplemented with 3% sucrose, 2.5 lM 2,4-D followed by a subsequent adventitious shoot differentiation phase for another 30 days on half MS medium supplemented with 0.25 lM each of BA and IBA. The regeneration rate of in vitro adventitious shoots in explants from the adult tree, i.e.1.66 shoots/callus, was lower than that from seedlings, i.e. [10 shoots/callus, which was reported elsewhere. Correspondingly, the activities of nitrate reductase and peroxidase, and endogenous phenol content remained very low during in vitro adventitious shoot differentiation in leaflet explants of an adult tree possibly due to lower availability of competent stem (juvenile) cells for the process.

  16. Plant Regeneration from In Vitro Cultured Hypocotyl Explants of Euonymus japonicus Cu zhi

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    Adventitious shoots were successfully regenerated from hypocotyl explants of in vitro cultures of Euonymus japonicus Cu zhi. Hypocotyl slices were cultured on Murashige and Skoog (MS) and B5 basal medium supplemented with varied concentration of different plant growth-regulators, e.g., α-naphthaleneacetic acid (NAA) and indole-3-butyric acid (IBA) in combination with 6-benzylaminopurine (6-BA) and kinetin. The study showed that shoots could be directly regenerated from hypocotyl explants without the intervening callus phase; MS medium was more suitable for adventitious shoots regeneration. The ability of hypocotyls segments to produce shoots varied depending upon their position on the seedlings. The highest regeneration rate was obtained with hypocotyl segments near to the cotyledon cultured on MS basal medium supplemented with 1.5 mg L-1 6-BA and 0.05 mg L-1 NAA (63.64%). The regenerated shoots were readily elongated on the same medium as used for multiplication and rooted on half-strength MS basal medium supplemented with 1.0 mg L-1 IBA and 100 mg L-1 activated carbon. After being transferred to greenhouse conditions, 96% of the plantlets were successfully acclimatized. This regeneration system is applied for genetic transformation now.

  17. Clonal propagation of guava (Psidium guajava L on nodal explants of mature elite cultivar

    Directory of Open Access Journals (Sweden)

    Guochen Yang

    2011-04-01

    Full Text Available Guava (Psidium guajava L. is a very valuable tropical and subtropical fruit. However, guava micro-propagation are genotypes dependent, there are several problems associated with in vitro cultures of guava including browning or blackening of culture medium due to leaching of phenolics, microbial contamination, and in vitro tissue recalcitrance. A micro-propagation system using Murashige and Skoog (MS medium with 6-benzylaminopurine (BA, kinetin and naphthaleneacetic acid (NAA was developed for guava (Psidium guajava L from mature cultivar. As part of this research various disinfection methods and plant growth regulators were tested in vitro. The most effective method involved treating explants in a 15% bleach solution for 20 mins followed by culturing them in MS medium with 250 mg/L polyvinylpyrrolidone (PVP. This method maximized the percentage of bud breakage (53.3%, while producing the minimum browning rate (18.3% for the explants. The best observed proliferation rate (71.2% occurred on the MS medium supplemented with 4.44 μM BA, 4.65 μM kinetin (KT and 0.54 μM NAA. It produced the highest mean number of shoots (2.2. Shoots were then rooted (65% when dipped in 4.9 mM Indole-3-butyric acid (IBA solution for 1 min and rooted plantlets survived (100% after acclimatization to the greenhouse.

  18. Effects of the mesonephros and insulin-like growth factor I on chondrogenesis of limb explants.

    Science.gov (United States)

    Geduspan, J S; Solursh, M

    1993-04-01

    The mesonephros has been shown to have a growth-promoting influence in vivo on limb outgrowth. This influence has been studied in detail using an organ culture system. The results show that in the presence of the mesonephros limb explants formed larger cartilages than cultures without mesonephros. Furthermore, with mesonephros, morphology of the cartilages is comparable to that of skeletal elements in vivo while cartilages formed in cultures lacking mesonephros were amorphous. The mesonephric influence also promoted the formation of a well-organized extracellular matrix in the cartilage while cartilage in cultures without mesonephros formed an abnormal appearing matrix. Cartilage matrices in cultures with or without mesonephros were immunoreactive to type IX and type II collagens, cartilage proteoglycan PGH, and link protein although cultures lacking mesonephros had a very restricted distribution of type IX collagen immunoreactivity. Despite the different distribution of type IX collagen, long-form-type IX collagen transcripts appeared similar in both types of culture based on in situ hybridization. The mesonephric effect on limb explants could be partially duplicated by the addition of insulin-like growth factor I (IGF-I) to cultures without mesonephros. Furthermore, the mesonephric influence on cartilage growth and morphological differentiation could be blocked by the addition of a blocking antibody to IGF-I to cultures with mesonephros. The results support the hypothesis that IGF-I is one of the growth factors produced by the mesonephros which may play a role in early limb development and chondrogenesis.

  19. Establishing axenic cultures from mature pecan embryo explants on media with low water availability.

    Science.gov (United States)

    Obeidy, A A; Smith, M A

    1990-12-01

    Endophytic fungi associated with mature pecan (Carya illinoensis (Wangenh.) C. Koch) nuts prevented successful, contaminant-free in vitro culture of embryo expiants, even after rigorous surface disinfestation of the nuts and careful aseptic shelling. Disinfestation with sodium hypochlorite after shell removal was also unsuccessful, because even dilute concentrations which were ineffective against the fungal contaminants prevented subsequent growth from the embryo. Explanting media with low water availability which would not sustain growth of fungal contaminants, but supported growth from mature pecan embryos, were developed as an alternative disinfestation method. The explanting media were supplemented with 0.9-1.5% agar, and other media components were selectively omitted to test their influence on water availability and fungal growth. Disinfestation of up to 65% of the cultures was accomplished, depending on the medium formulation, compared to 100% loss to contamination on control medium (0.5% agar). A complete medium (containing sucrose, salts, vitamins, 18 μM BAP, and 5 μM IBA) with 1.5% agar provided control of contamination, and encouraged subsequent regeneration from the embryo expiants, which remained free of contaminant growth through subsequent subcultures.

  20. Scanning Electron Microscopy Evaluation of an EX-PRESS Mini Glaucoma Shunt After Explantation.

    Science.gov (United States)

    Tognetto, Daniele; Cecchini, Paolo; D'Aloisio, Rossella; Vattovani, Odilla; Turco, Gianluca

    2017-01-01

    We report a case of an explanted stainless steel miniature glaucoma drainage device (EX-PRESS) implanted under a scleral flap for pseudoexfoliation open-angle glaucoma surgical treatment. The glaucoma shunt was implanted in a 75-year-old white man with medically refractive glaucoma. Cataract surgery was performed simultaneously. After 2 years, the shunt extruded through the scleral flap and the conjunctiva and it was, therefore, explanted. Scanning electron microscopy images of the EX-PRESS mini glaucoma shunt were acquired to verify the patency of the device lumen and the presence of fibrosis or cellular adhesion on the device. Energy dispersive spectroscopy for chemical surface characterization of the EX-PRESS shunt was performed. Scanning electron microscopy-acquired images showed minimal extracellular material proliferation on the lumen device. The energy dispersive spectroscopy analysis revealed a high peak of carbon suggesting the organic nature of the residuals found on the shunt lumen. The surface showed few superficial pits, likely due to an initial corrosion process.

  1. Induction of Tetraploids from Petiole Explants through Colchicine Treatments in Echinacea purpurea L.

    Directory of Open Access Journals (Sweden)

    Dahanayake Nilanthi

    2009-01-01

    Full Text Available Petiole explants were obtained from in vitro grown diploid (2x=22 Echinacea purpurea plantlets. Shoots were regenerated by culturing the explants on MS basal medium containing 0.3 mg/L benzyladenine (BA, 0.01 mg/L naphthaleneacetic acid (NAA and four concentrations (30, 60, 120, and 240 mg/L of colchicine for 30 days, or 120 mg/L of colchicine for various durations (7, 14, 21, and 28 days. The regenerated shoots were induced to root on MS basal medium with 0.01 mg/L NAA, and then the root-tips of the regenerated shoots were sampled for count of chromosome number. It was found that a treatment duration of >7 days was necessary for induction of tetraploid (4x=44 shoots, and treatment with 120 mg/L colchicine for 28 days was the most efficient for induction of tetraploids, yielding 23.5% of tetraploids among all the regenerated shoots. Chimeras were observed in almost all the treatments. However, the ratio of tetraploid to diploid cells in a chimeric plant was usually low. In comparison with diploid plants, tetraploid plants in vitro had larger stomata and thicker roots with more root branches, and had prominently shorter inflorescence stalk when mature.

  2. An efficient regeneration and rapid micropropagation protocol for Almond using dormant axillary buds as explants.

    Science.gov (United States)

    Choudhary, Ravish; Chaudhury, Rekha; Malik, Surendra Kumar; Sharma, Kailash Chandra

    2015-07-01

    An efficient in vitro protocol was standardized for Almond (Prunus dulcis) propagation using dormant axillary buds as explants. Explants were cultured on Murashige and Skoog (MS) and woody plant medium (WPM) supplemented with different concentration/combination(s) of phytohormones. MS basal medium showed lowest shoot induction and took longest duration for shoot initiation. Multiple shoots were induced in MS medium supplemented with the combination of BAP (0.5 mgL(-1)). Cultures showed poor response for rooting in all combinations of plant growth regulators (PGRs) and took 90 days for initiation. Rooting was higher in half strength of MS than in full-strength. The highest root induction (33.33%) was recorded in half MS medium supplemented with 0.1 mgL(-1) IBA (indole-3-butyric acid) followed by full strength of MS medium (20%) supplemented with IBA (0.1 mgL(-1)). α-Naphthalene acetic acid (NAA) was less effective for rooting than IBA. The highest root induction (25%) was found in half strength of MS medium supplemented with 0.1 mgL(-1) NAA followed by full strength of MS medium (20%). The protocol developed would be of use in mass propagation of almond and also support in vitro conservation.

  3. Histological and molecular evaluation of patient-derived colorectal cancer explants.

    Directory of Open Access Journals (Sweden)

    Joshua M Uronis

    Full Text Available Mouse models have been developed to investigate colorectal cancer etiology and evaluate new anti-cancer therapies. While genetically engineered and carcinogen-induced mouse models have provided important information with regard to the mechanisms underlying the oncogenic process, tumor xenograft models remain the standard for the evaluation of new chemotherapy and targeted drug treatments for clinical use. However, it remains unclear to what extent explanted colorectal tumor tissues retain inherent pathological features over time. In this study, we have generated a panel of 27 patient-derived colorectal cancer explants (PDCCEs by direct transplantation of human colorectal cancer tissues into NOD-SCID mice. Using this panel, we performed a comparison of histology, gene expression and mutation status between PDCCEs and the original human tissues from which they were derived. Our findings demonstrate that PDCCEs maintain key histological features, basic gene expression patterns and KRAS/BRAF mutation status through multiple passages. Altogether, these findings suggest that PDCCEs maintain similarity to the patient tumor from which they are derived and may have the potential to serve as a reliable preclinical model that can be incorporated into future strategies to optimize individual therapy for patients with colorectal cancer.

  4. Two-Stage Explantation of a Magnetic Lower Esophageal Sphincter Augmentation Device Due to Esophageal Erosion.

    Science.gov (United States)

    Parmar, Abhishek D; Tessler, Robert A; Chang, Howard Y; Svahn, Jonathan D

    2017-08-01

    Implanting a magnetic lower esophageal sphincter augmentation device (LINX, Torax Medical) has become an increasingly common option in the surgical management of gastroesophageal reflux disease. As the enthusiasm for placing this device increases, experience in the management of device-related complications-including erosion-is necessary. We report a staged approach to LINX removal in a 64-year-old female with symptoms of odynophagia secondary to partial erosion of a LINX device into the esophagus. The patient had a 12-bead LINX device placed in 2011 at an outside, international facility. In late 2013, she began experiencing symptoms of odynophagia. An esophagogastroduodenoscopy at our institution in October 2015 demonstrated two metallic beads eroding through the distal esophageal lumen. An elective endoscopic removal of the two visible beads was performed. A postoperative esophagram confirmed that there was no resulting esophageal perforation. The patient noted mild improvement in her symptoms. After a 12-week period to allow for complete healing, the remaining 10 beads of the LINX device were explanted laparoscopically without complication. No further procedures were undertaken. At 2 months' follow-up, the patient noted complete resolution of her symptoms. Transmural erosion of the LINX device into the esophageal lumen is a rare occurrence, with only five such complications reported in the published literature. We present the first account of LINX explantation for esophageal erosion in the United States. We demonstrated that a staged laparoendoscopic approach to LINX removal in these cases is feasible with minimal morbidity.

  5. A comparative study of candidal invasion in rabbit tongue mucosal explants and reconstituted human oral epithelium.

    Science.gov (United States)

    Jayatilake, J A M S; Samaranayake, Y H; Samaranayake, L P

    2008-06-01

    The purpose of this study is to compare the light and scanning electron microscopic (SEM) features of tissue invasion by three Candida species (C. albicans, C. tropicalis, and C. dubliniensis) in two different tissue culture models: rabbit tongue mucosal explants (RTME) and reconstituted human oral epithelium (RHOE). Tongue mucosal biopsies of healthy New Zealand rabbits were maintained in explant culture using a transwell system. RHOE was obtained from Skinethic Laboratory (Nice, France). RTME and RHOE were inoculated with C. albicans, C. tropicalis, and C. dubliniensis separately and incubated at 37 degrees C, 5% CO(2), and 100% humidity up to 48 h. Light microscopic and SEM examinations of uninfected (controls) and infected tissues were performed at 24 and 48 h. C. albicans produced characteristic hallmarks of pathological tissue invasion in both tissue models over a period of 48 h. Hyphae penetrated through epithelial cells and intercellular gaps latter resembling thigmotropism. SEM showed cavitations on the epithelial cell surfaces particularly pronounced at sites of hyphal invasion. Some hyphae on RTME showed several clusters of blastospores attached in regular arrangements resembling "appareil sporifere". C. tropicalis and C. dubliniensis produced few hyphae mainly on RTME but they did not penetrate either model. Our findings indicate that multiple host-fungal interactions such as cavitations, thigmotropism, and morphogenesis take place during candidal tissue invasion. RTME described here appears to be useful in investigations of such pathogenic processes of Candida active at the epithelial front.

  6. Thermal neutron irradiation field design for boron neutron capture therapy of human explanted liver.

    Science.gov (United States)

    Bortolussi, S; Altieri, S

    2007-12-01

    The selective uptake of boron by tumors compared to that by healthy tissue makes boron neutron capture therapy (BNCT) an extremely advantageous technique for the treatment of tumors that affect a whole vital organ. An example is represented by colon adenocarcinoma metastases invading the liver, often resulting in a fatal outcome, even if surgical resection of the primary tumor is successful. BNCT can be performed by irradiating the explanted organ in a suitable neutron field. In the thermal column of the Triga Mark II reactor at Pavia University, a facility was created for this purpose and used for the irradiation of explanted human livers. The neutron field distribution inside the organ was studied both experimentally and by means of the Monte Carlo N-particle transport code (MCNP). The liver was modeled as a spherical segment in MCNP and a hepatic-equivalent solution was used as an experimental phantom. In the as-built facility, the ratio between maximum and minimum flux values inside the phantom ((phi(max)/phi(min)) was 3.8; this value can be lowered to 2.3 by rotating the liver during the irradiation. In this study, the authors proposed a new facility configuration to achieve a uniform thermal neutron flux distribution in the liver. They showed that a phi(max)/phi(min) ratio of 1.4 could be obtained without the need for organ rotation. Flux distributions and dose volume histograms were reported for different graphite configurations.

  7. In vitro direct shoot regeneration from proximal, middle and distal segment of Coleus forskohlii leaf explants.

    Science.gov (United States)

    Krishna, Gaurav; Sairam Reddy, P; Anoop Nair, N; Ramteke, P W; Bhattacharya, P S

    2010-04-01

    Coleus forskohlii is an endangered multipurpose medicinal plant that has widespread applications. In spite of this, there have been few attempts to propagate its cultivation in India. The present communication presents an in vitro rapid regeneration method using leaf explants of Coleus forskohlii through direct organogenesis. Leaf explants that were excised into three different segments i.e. proximal (P), middle (M) and distal (D) were cultured on Murashige and Skoog (MS) basal medium supplemented with cytokinins. MS Media containing 5.0 mg L(-1) BAP (6-Benzylaminopurine) promoted regeneration of multiple shoots through direct organogenesis from the leaf, which were further elongated on MS media augmented with 0.1 mg L-1 BAP and 0.1 mg L(-1) IAA (Indole-3-acetic acid), cytokinin and auxin combination. Regenerated and elongated shoots, when transferred to ose resulted in profuse rooting plants that were transferred to soil after acclimatization and maintained in a green house. The current protocol offers a direct, mass propagation method bypassing the callus phase of C. forskohlii and is suitable for conservation, large-scale commercial cultivation, and genetic transformation with agronomically desirable traits.

  8. Induction of tetraploids from petiole explants through colchicine treatments in Echinacea purpurea L.

    Science.gov (United States)

    Nilanthi, Dahanayake; Chen, Xiao-Lu; Zhao, Fu-Cheng; Yang, Yue-Sheng; Wu, Hong

    2009-01-01

    Petiole explants were obtained from in vitro grown diploid (2x = 22) Echinacea purpurea plantlets. Shoots were regenerated by culturing the explants on MS basal medium containing 0.3 mg/L benzyladenine (BA), 0.01 mg/L naphthaleneacetic acid (NAA) and four concentrations (30, 60, 120, and 240 mg/L) of colchicine for 30 days, or 120 mg/L of colchicine for various durations (7, 14, 21, and 28 days). The regenerated shoots were induced to root on MS basal medium with 0.01 mg/L NAA, and then the root-tips of the regenerated shoots were sampled for count of chromosome number. It was found that a treatment duration of >7 days was necessary for induction of tetraploid (4x = 44) shoots, and treatment with 120 mg/L colchicine for 28 days was the most efficient for induction of tetraploids, yielding 23.5% of tetraploids among all the regenerated shoots. Chimeras were observed in almost all the treatments. However, the ratio of tetraploid to diploid cells in a chimeric plant was usually low. In comparison with diploid plants, tetraploid plants in vitro had larger stomata and thicker roots with more root branches, and had prominently shorter inflorescence stalk when mature.

  9. Ultrastructural Localization of Polygalacturonase in Ethylene-Stimulated Abscission of Tomato Pedicel Explants

    Directory of Open Access Journals (Sweden)

    Ming-Fang Qi

    2014-01-01

    Full Text Available Polygalacturonase (PG is crucial in plant organ abscission process. This paper investigated the cellular and subcellular localization of PG in ethylene-stimulated abscission of tomato pedicel explants. Confocal laser scanning microscopy of abscission zone sections with the fluorescent probe Cy3 revealed that PG was initially accumulated in parenchyma cells in cortical and vascular tissues after 8 h of ethylene treatment and then extended throughout the abscission zone when the abscission zone separated at 24 h after ethylene treatment. At the subcellular level, transmission electron microscopy with immunogold staining showed that PG showed abundant accumulation in the cortical and vascular tissues at 8 h after ethylene treatment, and the distribution area extended to the central parenchyma cells at 16 h after ethylene treatment. In addition, PGs were observed in the distal and proximal parts of the tomato pedicel explants throughout the abscission process. The results provided a visualized distribution of PG in the pedicel abscission zone and proved that PG was closely related to abscission.

  10. In vitro isolation and cultivation of rabbit tracheal epithelial cells using tissue explant technique.

    Science.gov (United States)

    Shi, Hong-Can; Lu, Dan; Li, Hai-Jia; Han, Shi; Zeng, Yan-Jun

    2013-04-01

    Epithelial cells from tracheal mucosa offer significant potential as a cell source in development of tissue-engineered trachea. The purpose of this study was to investigate and optimize a suitable culture system for tracheal epithelial cells, including the methods of primary culture, passage, identification, and cryopreservation. Epithelial cells were isolated from rabbit tracheal mucosa using tissue explant technique and were subjected to immunohistochemistry, immunofluorescence, and cryopreservation after purification. Epithelial cells reached confluency at 14-15 d. Immunohistochemical staining for cytokeratin showed brown yellow-positive cytoplasm and blue-counterstained nuclei, while immunofluorescence staining for cytokeratin showed green-positive cytoplasm and clear cell outline, indicating that the cultured cells had properties of epithelial cells. After recovery, epithelial cells exhibited high survival and viability. The results demonstrated that in vitro isolation and cultivation model was successfully established to provide high proliferative capacity, typical morphology and characteristics of tracheal epithelial cells from trachea mucosa by the use of the tissue explant technique.

  11. [Growth behavior of spiral ganglion explants on cochlear implant electrodes and their materials].

    Science.gov (United States)

    Hansen, S; Mlynski, R; Volkenstein, S; Stark, T; Schwaab, M; Dazert, S; Brors, D

    2009-04-01

    With the increasing use of cochlear implants (CIs), the insertion of alloplastic material into the inner ear is nowadays an established treatment for severe to profound hearing loss in children and adults. Beyond its widespread use, the biocompatibility of the CI electrode and its interaction with the neural structures of the cochlea is not yet established. To investigate the survival and growth behavior of spiral ganglion neurons on different CI materials, spiral ganglion explants from newborn rats were cultured on silicone and platinum, on a surface combination of silicone and platinum, and, finally, on a CI electrode. The results of this study indicate that the growth of spiral ganglion neurons in vitro is strongly influenced by the different materials and their arrangement, with platinum exhibiting the highest degree of biocompatibility with respect to neurite extension. Level differences in the surface structure between silicone and platinum lead to inhibition of neurite outgrowth. Furthermore, the culturing of spiral ganglion explants on a CI electrode leads to neurite sprouting toward the electrodes made of platinum. The biocompatibility of CI materials with spiral ganglion neurons was shown in this study, but it differs with different CI materials. Besides the material itself, the arrangement of the materials can affect the neurite extension.

  12. Increased proliferation of explanted vascular smooth muscle cells: a marker presaging atherogenesis.

    Science.gov (United States)

    Absher, P M; Schneider, D J; Baldor, L C; Russell, J C; Sobel, B E

    1997-06-01

    The JCR:LA-cp homozygous cp/cp corpulent rat is genetically predisposed to develop atherosclerosis evident after 9 and 18 months of age in males and females and to manifest metabolic derangements resembling those seen in type II diabetes in humans (hyperinsulinemia, insulin resistance, hyperglycemia and hypertriglyceridemia). The present study was undertaken to determine whether vascular smooth muscle cells (SMCs) explanted from vessels destined to become atherosclerotic later in life exhibit intrinsic properties ex vivo that presage atherogenesis to provide a means for evaluating promptly intervention designed to modify it. SMCs were cultured from aortic explants of JCR:LA-cp corpulent (cp/cp) and lean control (+/+) rats of 4, 5, 6, and 9 months of age. Compared with SMCs from controls, SMCs from cp/cp rats exhibited increased proliferation, higher saturation density, increased augmentation of proliferation in response to selected mitogens and greater adherence to extracellular matrix proteins. The increased proliferative activity ex vivo anteceded by several months the development of atherosclerotic lesions in vivo. Thus, it is a promising marker in assessments of the efficacy of interventions designed to retard or prevent atherosclerosis.

  13. Neisseria gonorrhoeae Challenge Increases Matrix Metalloproteinase-8 Expression in Fallopian Tube Explants

    Science.gov (United States)

    Juica, Natalia E.; Rodas, Paula I.; Solar, Paula; Borda, Paula; Vargas, Renato; Muñoz, Cristobal; Paredes, Rodolfo; Christodoulides, Myron; Velasquez, Luis A.

    2017-01-01

    Background: Neisseria gonorrhoeae (Ngo) is the etiological agent of gonorrhea, a sexually transmitted infection that initially infects the female lower genital tract. In untreated women, the bacteria can ascend to the upper genital reproductive tract and infect the fallopian tube (FTs), which is associated with salpingitis and can lead to impaired FT function and infertility. The extracellular matrix (ECM) plays an important role in cell migration and differentiation in the female genital tract, and some pathogens modify the ECM to establish successful infections. The ECM is regulated by matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs), their endogenous inhibitors; MMP deregulation causes pathological conditions in a variety of tissues. Results: The aim of this work was to analyze the expression and localization of MMP-3, MMP-8, MMP-9, and TIMP-1 in FT explants during Ngo infection using real-time PCR, immunohistochemistry, zymography and ELISA. No significant variations in MMP-3, MMP-9, and TIMP-1 transcript levels were observed. In contrast, a significant increase (p < 0.05) was observed for MMP-8 expression and was accompanied by stromal immunoreactivity in infected explants. ELISA results supported these findings and showed that MMP-8 release increased upon gonococcal infection. Conclusions: Our results indicate that gonococcal infection induces increased MMP-8 expression, which might contribute to FT damage during infection. PMID:28932707

  14. Neisseria gonorrhoeae Challenge Increases Matrix Metalloproteinase-8 Expression in Fallopian Tube Explants

    Directory of Open Access Journals (Sweden)

    Natalia E. Juica

    2017-09-01

    Full Text Available Background:Neisseria gonorrhoeae (Ngo is the etiological agent of gonorrhea, a sexually transmitted infection that initially infects the female lower genital tract. In untreated women, the bacteria can ascend to the upper genital reproductive tract and infect the fallopian tube (FTs, which is associated with salpingitis and can lead to impaired FT function and infertility. The extracellular matrix (ECM plays an important role in cell migration and differentiation in the female genital tract, and some pathogens modify the ECM to establish successful infections. The ECM is regulated by matrix metalloproteinases (MMPs and tissue inhibitors of metalloproteinases (TIMPs, their endogenous inhibitors; MMP deregulation causes pathological conditions in a variety of tissues.Results: The aim of this work was to analyze the expression and localization of MMP-3, MMP-8, MMP-9, and TIMP-1 in FT explants during Ngo infection using real-time PCR, immunohistochemistry, zymography and ELISA. No significant variations in MMP-3, MMP-9, and TIMP-1 transcript levels were observed. In contrast, a significant increase (p < 0.05 was observed for MMP-8 expression and was accompanied by stromal immunoreactivity in infected explants. ELISA results supported these findings and showed that MMP-8 release increased upon gonococcal infection.Conclusions: Our results indicate that gonococcal infection induces increased MMP-8 expression, which might contribute to FT damage during infection.

  15. Utilization of Aseptic Seedling Explants for In vitro Propagation of Indian Red Wood

    Directory of Open Access Journals (Sweden)

    Kishore Kumar CHIRUVELLA

    2013-12-01

    Full Text Available Micropropagation has been advocated as one of the most viable biotechnological tool for ex situ conservation of rare, endangered endemic medicinal plants germplasm. Rapid clonal micropropagation protocol for large-scale multiplication of an endemic medicinal plant Soymida febrifuga (Meliaceae was established from 15-day aseptic seedling cotyledonary node and shoot tip explants. High frequency of sprouting and shoot differentiation was observed from cotyledonary node explants compared to shoot tip, on Murashige and Skoog (MS medium fortified with BA, KN, 2-iP and CM. Of the cytokinins used, BA (3.0 mgl-1 supported highest average number and maximum multiple shoot differentiation (16.6. In vitro proliferated shoots were multiplied rapidly by culturing nodal segments as microcuttings, further subcultured on the same media for elongation. Elongated shoots upon transfer to MS medium fortified with IBA showed rooting within two weeks of culture. Rooted plantlets were successfully hardened and 75% of rooted shoots successfully survived on establishment to the soil. Plants looked healthy with no visually detectable phenotypic variations. This protocol provides a successful and rapid technique that can be used for ex situ conservation minimizing the pressure on wild populations and contributes to the conservation of this endemic medicinally potent flora.

  16. Comparative effects of plant growth regulators on leaf and stem explants of Labisia pumila var.alata

    Institute of Scientific and Technical Information of China (English)

    Anna Pick Kiong LING; Kinn Poay TAN; Sobri HUSSEIN

    2013-01-01

    Objective:Labisia pumi/a var.a/ata,commonly known as ‘Kacip Fatimah’ or ‘Selusuh Fatimah’ in Southeast Asia,is traditionally used by members of the Malay community because of its post-partum medicinal properties.Its various pharmaceutical applications cause an excessive harvesting and lead to serious shortage in natural habitat.Thus,this in vitro propagation study investigated the effects of different plant growth regulators (PGRs) on in vitro leaf and stem explants of L.pumila.Methods:The capabilities of callus,shoot,and root formation were evaluated by culturing both explants on Murashige and Skoog (MS) medium supplemented with various PGRs at the concentrations of 0,1,3,5,and 7 mg/L.Results:Medium supplemented with 3 mg/L indole-3-butyric acid (IBA) showed the optimal callogenesis from both leaf and stem explants with (72.34± 19.55)% and (70.40± 14.14)% efficacy,respectively.IBA was also found to be the most efficient PGR for root induction.A total of (50.00±7.07)% and (77.78±16.47)% of root formation were obtained from the in vitro stem and leaf explants after being cultured for (26.5±5.0) and (30.0±8.5) d in the medium supplemented with 1 and 3 mg/L of IBA,respectively.Shoot formation was only observed in stem explant,with the maximum percentage of formation ((100.00±0.00)%) that was obtained in 1 mg/L zeatin after (11.0±2.8) d of culture.Conclusions:Callus,roots,and shoots can be induced from in vitro leaf and stem explants of L.pumila through the manipulation of types and concentrations of PGRs.

  17. Analysis of Calcium Content,Hormones,and Degrading Enzymes in Tomato Pedicel Explants During Calcium-Inhibited Abscission

    Institute of Scientific and Technical Information of China (English)

    XU Tao; LI Tian-lai; QI Ming-fang

    2009-01-01

    This study was designed to analyze the changes of phytohormone concentrations,calcium fraction,and the activities of degrading enzymes during calcium-inhibited and ethyleneglycol-bis-(β-aminoethyl ether)N,N'-tetraacetic acid (EGTA)induced abscission of tomato pedicel explants.Added calcium caused an increase in the total calcium content within the abscission zone and produced a corresponding reduction (20%) in pedicel explant abscission.As expected,EGTA treatment produced the opposite effect and resulted in a decrease in the total calcium content,while accelerating abscission of pedicel explants.Hormone analysis revealed that indole-3-acetic acid (IAA) concentrations in the abscission zone first decreased and then increased before the occurrence of abscission in all treatments,with the greatest effect produced by addition of EGTA.Similarly,abscisic acid (ABA),and gibberellin (GA1+3) concentrations,and ethylene production were elevated in the abscission zone during the first 16 h before abscission when explants imbibed EGTA.With calcium treatment,the concentrations of ABA,ethylene,and GA1+3 also increased in pedicels throughout the first 16 h following exposure,but the increase was slower and less dramatic than with EGTA.Both cellulase and polygalacturonase were induced in the explants during abscission and the activities were also strengthened by treatment with EGTA.Calciumtreated explants produced lower hydrolysing enzyme activities than controls throughout abscission.Calcium played a role of mediating hormone balance and degrading enzymes activities and affected on abscission.

  18. Comparison of human nasal epithelial cells grown as explant outgrowth cultures or dissociated tissue cultures in vitro.

    Science.gov (United States)

    Jiao, Jian; Meng, Na; Wang, Hong; Zhang, Luo

    2013-12-01

    The purpose of this study was to compare cell growth characteristics, ciliated cell differentiation, and function of human nasal epithelial cells established as explant outgrowth cultures or dissociated tissue cultures. Human nasal mucosa of the uncinate process was obtained by endoscopy and epithelial cell cultures were established by explant outgrowth or dissociated tissue culture methods. Epithelial cell growth characteristics were observed by inverted phase contrast microscopy. Ciliated cell differentiation was detected by β-tubulin IVand ZO-1 immunocytochemistry. Basal and ATP-stimulated ciliary beat frequency (CBF) was measured using a highspeed digital microscopic imaging system. Both the explant and dissociated tissue cultures established as monolayers with tight junctions and differentiated cell composition, with both types of cultures comprising ciliated and non-ciliated epithelial cells. Fibroblasts were also frequently found in explant cultures but rarely seen in dissociated tissue cultures. In both culture systems, the highest ciliated cell density appeared at 7th-10th culture day and declined with time, with the lifespan of ciliated cells ranging from 14 to 21 days. Overall, 10% of the cells in explant cultures and 20% of the cells in the dissociated tissue cultures were ciliated. These two cultures demonstrated similar ciliary beat frequency values at baseline (7.78 ± 1.99 Hz and 7.91 ± 2.52 Hz, respectively) and reacted equivalently following stimulation with 100 μM ATP. The results of this study indicate that both the explant outgrowth and dissociated tissue culture techniques are suitable for growing well-differentiated nasal ciliated and non-ciliated cells, which have growth characteristics and ciliary activity similar to those of nasal epithelial cells in vivo.

  19. In vitro Regeneration of Plantlets from Leaf and Nodal explants of Aristolochia indica L.- An Important Threatened Medicinal Plant

    Institute of Scientific and Technical Information of China (English)

    Pramod V. Pattar; M.Jayaraj

    2012-01-01

    Objective: An efficient reproducible protocol has been developed for in vitro regeneration of plantlets from leaf and nodal explants of Aristolochia indica L. Methods: Wild grown plants Aristolochia indica L. were collected and grown in the departmental garden. Leaf and nodal segments (0.5-1.0 cm) from young healthy plants were first washed thoroughly under running tap water for 15 - 20 minutes and then treated with liquid detergent [5% (v/v) Tween-20] for 5-10 minutes. Later these explants were washed with double-distilled water for 5 minutes. Subsequently, explants were immersed in 70% (v/v) ethanol for 2 - 3 minutes and washed with sterile glass double distilled water for 2-3 times. Eventually, the explants were treated with an aqueous solution of 0.1% (w/v) HgCl2 for 1 - 2 minutes and rinsed for two-to-three times in sterile ddH2O to remove all traces of HgCl2. The sterilized explants were inoculated aseptically onto solid basal Murashige and Skoog’s medium with different concentrations and combinations of BAP and NAA for in vitro regeneration of plants. Results: Both leaf and nodal explants cultured on MS medium supplemented with 0.8 mg/L BAP developed into mass of callus. These calli were subcultured for the induction of shoots and roots. Shoots were induced from both calli on MS medium supplemented with 0.8 mg/L BAP+0.5 mg/L NAA. Roots were induced from in vitro shoots on MS medium supplemented with 0.8 mg/L NAA for 4 weeks. Nodal explants were more regenerative with 95 % response compared to leaf explants with 85%. Finally, these in vitro regenerated plantlets were hardened, acclimatised and successfully transferred to the field. Conclusions: The present protocol for in vitro regeneration of Aristolochia indica L. can be used to make this plant available throughout the year for traditional healers, pharmaceutical usages, germplasm conservation, commercial cultivation, and also for the production of secondary metabolites.

  20. Chitinase, beta-1,3-glucanase, osmotin, and extensin are expressed in tobacco explants during flower formation

    DEFF Research Database (Denmark)

    Neale, A D; Wahleithner, J A; Lund, Marianne;

    1990-01-01

    encodes the cell wall protein extensin, which also accumulates during pathogen attack. Another sequence family encodes the water stress-induced protein osmotin [Singh et al. (1989). Plant Physiol. 90, 1096-1101]. We found that osmotin was also induced by viral infection and wounding and, hence, could...... be considered a pathogenesis-related protein. These genes, which were highly expressed in explants during de novo flower formation but not in explants forming vegetative shoots [Meeks-Wagner et al. (1989). Plant Cell 1, 25-35], were also regulated developmentally in day-neutral and photoresponsive tobacco...

  1. Somatic embryogenesis and plant regeneration from immature embryo explant of papaya (Carica papaya L. cv. washington and honey dew).

    Science.gov (United States)

    Bhattacharya, J; Khuspe, S S; Renukdas, N N; Rawal, S K

    2002-05-01

    Immature zygotic embryo explants of Carica papaya were cultured on MS medium supplemented with 2,4-D (2.0 mg/l) and formed globular embryos on explants without callus formation in 4-6 weeks. Maturation and conversion of somatic embryos was also achieved on the same medium. Cotyledonary stage embryos germinated to 63.66 and 68.33% in cv. honey dew and washington respectively in MS basal medium supplemented ABA (0.5 microm/l). Robust development and proliferation of plantlet roots in vitro was obtained on MS basal medium. Hardened plantlets have 60% survival rate.

  2. Interleukin-10 prevents epithelial cell apoptosis by regulating IFNγ and TNFα expression in rhesus macaque colon explants

    Science.gov (United States)

    Pan, Diganta; Das, Arpita; Lala, Wendy; Kenway-Lynch, Carys S.; Liu, David X.; Veazey, Ronald S.; Pahar, Bapi

    2013-01-01

    Interleukin-10 (IL-10) is an important immunomodulatory cytokine that plays an obligate role in regulating inflammatory responses. Here we demonstrated the role of IL-10 in regulating crypts length and breadth as well as maintaining the survival of epithelial cells using rhesus colon explant cultures. Anti-IL-10 antibody treatment of colon explant cultures induced increased production of inflammatory cytokines/molecules like IFNγ, TNFα, CD107a and perforin as well as increased epithelial cell apoptosis compared to media controls tested. Our results suggest that IL-10 plays a crucial role in maintaining mucosal homeostasis by regulating mucosal IFNγ and TNFα cytokine production. PMID:23867612

  3. Sustained release of VEGF from PLGA nanoparticles embedded thermo-sensitive hydrogel in full-thickness porcine bladder acellular matrix

    Directory of Open Access Journals (Sweden)

    Song Hua

    2011-01-01

    Full Text Available Abstract We fabricated a novel vascular endothelial growth factor (VEGF-loaded poly(lactic-co-glycolic acid (PLGA-nanoparticles (NPs-embedded thermo-sensitive hydrogel in porcine bladder acellular matrix allograft (BAMA system, which is designed for achieving a sustained release of VEGF protein, and embedding the protein carrier into the BAMA. We identified and optimized various formulations and process parameters to get the preferred particle size, entrapment, and polydispersibility of the VEGF-NPs, and incorporated the VEGF-NPs into the (poly(ethylene oxide-poly(propylene oxide-poly(ethylene oxide (Pluronic® F127 to achieve the preferred VEGF-NPs thermo-sensitive gel system. Then the thermal behavior of the system was proven by in vitro and in vivo study, and the kinetic-sustained release profile of the system embedded in porcine bladder acellular matrix was investigated. Results indicated that the bioactivity of the encapsulated VEGF released from the NPs was reserved, and the VEGF-NPs thermo-sensitive gel system can achieve sol-gel transmission successfully at appropriate temperature. Furthermore, the system can create a satisfactory tissue-compatible environment and an effective VEGF-sustained release approach. In conclusion, a novel VEGF-loaded PLGA NPs-embedded thermo-sensitive hydrogel in porcine BAMA system is successfully prepared, to provide a promising way for deficient bladder reconstruction therapy.

  4. Surgical Outcomes of Deep Superior Sulcus Augmentation Using Acellular Human Dermal Matrix in Anophthalmic or Phthisis Socket.

    Science.gov (United States)

    Cho, Won-Kyung; Jung, Su-Kyung; Paik, Ji-Sun; Yang, Suk-Woo

    2016-07-01

    Patients with anophthalmic or phthisis socket suffer from cosmetic problems. To resolve those problems, the authors present the surgical outcomes of deep superior sulcus (DSS) augmentation using acellular dermal matrix in patients with anophthalmic or phthisis socket. The authors retrospectively reviewed anophthalmic or phthisis patients who underwent surgery for DSS augmentation using acellular dermal matrix. To evaluate surgical outcomes, the authors focused on 3 aspects: the possibility of wearing contact prosthesis, the degree of correction of the DSS, and any surgical complications. The degree of correction of DSS was classified as excellent: restoration of superior sulcus enough to remove sunken sulcus shadow; fair: gain of correction effect but sunken shadow remained; or fail: no effect of correction at all. Ten eyes of 10 patients were included. There was a mean 21.3 ± 37.1-month period from evisceration or enucleation to the operation for DSS augmentation. All patients could wear contact prosthesis after the operation (100%). The degree of correction was excellent in 8 patients (80%) and fair in 2. Three of 10 (30%) showed complications: eyelid entropion, upper eyelid multiple creases, and spontaneous wound dehiscence followed by inflammation after stitch removal. Uneven skin surface and paresthesia in the forehead area of the affected eye may be observed after surgery. The overall surgical outcomes were favorable, showing an excellent degree of correction of DSS and low surgical complication rates. This procedure is effective for patients who have DSS in the absence or atrophy of the eyeball.

  5. Three-dimensional scaffolds of acellular human and porcine lungs for high throughput studies of lung disease and regeneration.

    Science.gov (United States)

    Wagner, Darcy E; Bonenfant, Nicholas R; Sokocevic, Dino; DeSarno, Michael J; Borg, Zachary D; Parsons, Charles S; Brooks, Elice M; Platz, Joseph J; Khalpey, Zain I; Hoganson, David M; Deng, Bin; Lam, Ying W; Oldinski, Rachael A; Ashikaga, Takamaru; Weiss, Daniel J

    2014-03-01

    Acellular scaffolds from complex whole organs such as lung are being increasingly studied for ex vivo organ generation and for in vitro studies of cell-extracellular matrix interactions. We have established effective methods for efficient de and recellularization of large animal and human lungs including techniques which allow multiple small segments (∼ 1-3 cm(3)) to be excised that retain 3-dimensional lung structure. Coupled with the use of a synthetic pleural coating, cells can be selectively physiologically inoculated via preserved vascular and airway conduits. Inoculated segments can be further sliced for high throughput studies. Further, we demonstrate thermography as a powerful noninvasive technique for monitoring perfusion decellularization and for evaluating preservation of vascular and airway networks following human and porcine lung decellularization. Collectively, these techniques are a significant step forward as they allow high throughput in vitro studies from a single lung or lobe in a more biologically relevant, three-dimensional acellular scaffold. Copyright © 2013 Elsevier Ltd. All rights reserved.

  6. Long-Term Followup of Dermal Substitution with Acellular Dermal Implant in Burns and Postburn Scar Corrections

    Directory of Open Access Journals (Sweden)

    I. Juhasz

    2010-01-01

    Full Text Available Full-thickness burn and other types of deep skin loss will result in scar formation. For at least partial replacement of the lost dermal layer, there are several options to use biotechnologically derived extracellular matrix components or tissue scaffolds of cadaver skin origin. In a survey, we have collected data on 18 pts who have previously received acellular dermal implant Alloderm. The age of these patients at the injury varied between 16 months and 84 years. The average area of the implants was 185 cm2. Among those, 15 implant sites of 14 patients were assessed at an average of 50 months after surgery. The scar function was assessed by using the modified Vancouver Scar Scale. We have found that the overall scar quality and function was significantly better over the implanted areas than over the surrounding skin. Also these areas received a better score for scar height and pliability. Our findings suggest that acellular dermal implants are especially useful tools in the treatment of full-thickness burns as well as postburn scar contractures.

  7. Mortality and morbidity from invasive bacterial infections during a clinical trial of acellular pertussis vaccines in Sweden.

    Science.gov (United States)

    Storsaeter, J; Olin, P; Renemar, B; Lagergård, T; Norberg, R; Romanus, V; Tiru, M

    1988-09-01

    A double blind placebo-controlled efficacy trial of two acellular pertussis vaccines was conducted in 3801 6- to 11-month-old children. Four vaccinated children died during 7 to 9 months follow-up as a result of Haemophilus influenzae type b meningitis, heroin intoxication with concomitant pneumonia, suspected septicemia, and Neisseria meningitidis Group B septicemia. From the actual death rate in children belonging to the same birth cohort in Sweden that could have been eligible for the trial, one death was expected among vaccinated children. Several investigations were carried out to examine the possibility that the deaths could be causally related to the vaccination. The relative risk for hospitalization due to systemic or respiratory infections was 1.07 (95% confidence interval, 0.95 to 1.20) and 0.83 (95% confidence interval, 0.64 to 1.08) in the vaccine groups as compared with the placebo group. Subsets of the population were studied for signs of immunosuppression. There was no indication of immunoglobulin deficiency or any sign of clinically significant leukopenia or lymphocytosis in vaccine recipients. The results of this analysis provide no evidence for a causal relation between vaccination with the studied acellular pertussis vaccines and altered resistance to invasive disease caused by encapsulated bacteria. The hypothesis that the two variables are related, however, cannot be refuted from these data.

  8. Acellular Dermal Matrices and Radiotherapy in Breast Reconstruction: A Systematic Review and Meta-Analysis of the Literature

    Directory of Open Access Journals (Sweden)

    Luigi Valdatta

    2014-01-01

    Full Text Available The increasing use of commercially available acellular dermis matrices for postmastectomy breast reconstruction seems to have simplified the surgical procedure and enhanced the outcome. These materials, generally considered to be highly safe or with only minor contraindications due to the necessary manipulation in preparatory phases, allow an easier one-phase surgical procedure, in comparison with autologous flaps, offering a high patient satisfaction. Unfortunately, the claim for a higher rate of complications associated with irradiation at the implant site, especially when the radiation therapy was given before the reconstructive surgery, suggested a careful behaviour when this technique is preferred. However, this hypothesis was never submitted to a crucial test, and data supporting it are often discordant or incomplete. To provide a comprehensive analysis of the field, we searched and systematically reviewed papers published after year 2005 and registered clinical trials. On the basis of a meta-analysis of data, we conclude that the negative effect of the radiotherapy on the breast reconstruction seems to be evident even in the case of acellular dermis matrices aided surgery. However, more trials are needed to make solid conclusions and clarify the poor comprehension of all the factors negatively influencing outcome.

  9. Development and preclinical evaluation of acellular collagen scaffolding and autologous artificial connective tissue in the regeneration of oral mucosa wounds.

    Science.gov (United States)

    Espinosa, Lady; Sosnik, Alejandro; Fontanilla, Marta R

    2010-05-01

    This work assessed wound healing response in rabbit oral lesions grafted with autologous artificial connective tissue or acellular collagen scaffolds. Autologous artificial oral connective tissue (AACT) was produced using rabbit fibroblasts and collagen I scaffolds. Before implantation, AACT grafts were assayed to demonstrate the presence of fibroblasts and extracellular matrix components, as well as the expression of characteristic genes and secretion of chemokines, cytokines, and growth factors. AACT grafts were tested in the rabbits from which the fibroblasts were obtained, whereas acellular collagen type I scaffolds (CS) were evaluated in a separate group of rabbits. In both cases, contralateral wounds closed by secondary intention were used as controls. In a separate experiment, AACT-grafted wounds were directly compared with contralateral CS-grafted wounds in the same animals. Wound contraction and histological parameters were examined to evaluate closure differences between the treatments in the three animal experiments performed. Contraction of wounds grafted with AACT and CS was significantly lower than in their controls (p oral mucosa.

  10. Synthetic bone substitute engineered with amniotic epithelial cells enhances bone regeneration after maxillary sinus augmentation.

    Directory of Open Access Journals (Sweden)

    Barbara Barboni

    Full Text Available BACKGROUND: Evidence has been provided that a cell-based therapy combined with the use of bioactive materials may significantly improve bone regeneration prior to dental implant, although the identification of an ideal source of progenitor/stem cells remains to be determined. AIM: In the present research, the bone regenerative property of an emerging source of progenitor cells, the amniotic epithelial cells (AEC, loaded on a calcium-phosphate synthetic bone substitute, made by direct rapid prototyping (rPT technique, was evaluated in an animal study. MATERIAL AND METHODS: Two blocks of synthetic bone substitute (∼0.14 cm(3, alone or engineered with 1×10(6 ovine AEC (oAEC, were grafted bilaterally into maxillary sinuses of six adult sheep, an animal model chosen for its high translational value in dentistry. The sheep were then randomly divided into two groups and sacrificed at 45 and 90 days post implantation (p.i.. Tissue regeneration was evaluated in the sinus explants by micro-computer tomography (micro-CT, morphological, morphometric and biochemical analyses. RESULTS AND CONCLUSIONS: The obtained data suggest that scaffold integration and bone deposition are positively influenced by allotransplantated oAEC. Sinus explants derived from sheep grafted with oAEC engineered scaffolds displayed a reduced fibrotic reaction, a limited inflammatory response and an accelerated process of angiogenesis. In addition, the presence of oAEC significantly stimulated osteogenesis either by enhancing bone deposition or making more extent the foci of bone nucleation. Besides the modulatory role played by oAEC in the crucial events successfully guiding tissue regeneration (angiogenesis, vascular endothelial growth factor expression and inflammation, data provided herein show that oAEC were also able to directly participate in the process of bone deposition, as suggested by the presence of oAEC entrapped within the newly deposited osteoid matrix and by their

  11. Explanted diseased livers - a possible source of metabolic competent primary human hepatocytes.

    Science.gov (United States)

    Kleine, Moritz; Riemer, Marc; Krech, Till; DeTemple, Daphne; Jäger, Mark D; Lehner, Frank; Manns, Michael P; Klempnauer, Jürgen; Borlak, Jürgen; Bektas, Hueseyin; Vondran, Florian W R

    2014-01-01

    Being an integral part of basic, translational and clinical research, the demand for primary human hepatocytes (PHH) is continuously growing while the availability of tissue resection material for the isolation of metabolically competent PHH remains limited. To overcome current shortcomings, this study evaluated the use of explanted diseased organs from liver transplantation patients as a potential source of PHH. Therefore, PHH were isolated from resected surgical specimens (Rx-group; n = 60) and explanted diseased livers obtained from graft recipients with low labMELD-score (Ex-group; n = 5). Using established protocols PHH were subsequently cultured for a period of 7 days. The viability and metabolic competence of cultured PHH was assessed by the following parameters: morphology and cell count (CyQuant assay), albumin synthesis, urea production, AST-leakage, and phase I and II metabolism. Both groups were compared in terms of cell yield and metabolic function, and results were correlated with clinical parameters of tissue donors. Notably, cellular yields and viabilities were comparable between the Rx- and Ex-group and were 5.3±0.5 and 2.9±0.7×106 cells/g liver tissue with 84.3±1.3 and 76.0±8.6% viability, respectively. Moreover, PHH isolated from the Rx- or Ex-group did not differ in regards to loss of cell number in culture, albumin synthesis, urea production, AST-leakage, and phase I and II metabolism (measured by the 7-ethoxycoumarin-O-deethylase and uracil-5'-diphosphate-glucuronyltransferase activity). Likewise, basal transcript expressions of the CYP monooxygenases 1A1, 2C8 and 3A4 were comparable as was their induction when treated with a cocktail that consisted of 3-methylcholantren, rifampicin and phenobarbital, with increased expression of CYP 1A1 and 3A4 mRNA while transcript expression of CYP 2C8 was only marginally changed. In conclusion, the use of explanted diseased livers obtained from recipients with low labMELD-score might represent

  12. Combined application of acellular bovine pericardium and hyaluronic acid in prevention of postoperative pericardial adhesion.

    Science.gov (United States)

    Shen, Jia; Xu, Zhi Wei

    2014-03-01

    An experiment was designed to find the suitable acellular bovine pericardium (ABP) patch in pericardial cavity reconstruction and to evaluate the effect of sodium hyaluronic acid (NaHA) on inflammatory reaction in prevention of pericardial adhesions. The pericardial adhesion model was established in 20 rabbits, weighing from 3.2 to 3.6 kg. Groups were classified as follows: Group A (n = 5), the control group, the pericardium was directly closed; Group B (n = 5), 0.15% glutaraldehyde-treated ABP (low cross-link degree); Group C, 0.3% glutaraldehyde-treated ABP (middle cross-link degree); Group D, 0.15% glutaraldehyde-treated ABP + NaHA solution. Blood samples were collected at 6 h, 24 h, 3 days, and 5 days, to assay postoperative inflammatory reaction. The tenacity and severity of adhesions were evaluated 2 months after operation, by macroscopic and microscopic examinations, and Q-PCR (real-time quantitative polymerase chain reaction) test was used to quantitatively analyze the associated genes with adhesion. Pericardium regeneration was demonstrated by immunohistochemical technique to identify mesothelial cells. In Group D, the serum concentration of tumor necrosis factor-α (TNF-α) was significantly lower in the early postoperative period, and the mean adhesion score (adhesion between the epicardium and ABP) was significantly lower compared with the control group (Groups D vs. A: 0.20 ± 0.45 vs. 2.00 ± 0.71, P = 0.009*). The signs of degradation of the ABPs were observed 2 months postoperation in Groups D and B. Immunohistochemically, the positive cytokeratin AE1 staining results demonstrated the relatively total regeneration of the pericardium in Group D. Signs of regeneration were observed in Group D. Compared with the control group, the level of TGF-β2 in Group D was significantly lower (0.00132 ± 0.00114, P = 0.022*). The TGF-β3 level was statistically significant, being highest in Group D (0.00805 ± 0.00136, P = 0.029*). The mean quantity of Smad6 in

  13. Biological function evaluation and effects of laser micro-pore burn-denatured acellular dermal matrix.

    Science.gov (United States)

    Zhang, Youlai; Zeng, Yuanlin; Xin, Guohua; Zou, Lijin; Ding, Yuewei; Duyin, Jiang

    2017-08-18

    In the field of burns repairs, many problems exist in the shortage of donor skin, the expense of allograft or xenograft skin, temporary substitution and unsatisfactory extremity function after wound healing. Previous studies showed that burn-denatured skin could return to normal dermis formation and function. This study investigates the application of laser micro-pore burn-denatured acellular dermis matrix (DADM) from an escharotomy in the repair of burn wounds and evaluates the biological properties and wound repair effects of DADM in implantation experiments in Kunming mice. Specific-pathogen-free (SPF) Kunming mice were used in this study. A deep II° burn wound was created on the dorsum of the mice by an electric heated water bath. The full-thickness wound tissue was harvested. The necrotic tissue and subcutaneous tissue were removed. The denatured dermis was preserved and treated with 0.25% trypsin, 0.5% Triton X-100. The DADM was drilled by laser micro-pore. The biological properties and grafting effects of laser micro-pore burn-DADM were evaluated by morphology, cytokine expression levels and subcutaneous implantation experiments in Kunming mice. We found statistical significance (Plaser micro-pore burn-DADM (experimental group) compared to the control group (no laser micro-pore burn-DADM). Cytokine expression level was different in the dermal matrixes harvested at various time points after burn (24h, 48h, 72h and infected wound group). Comparing the dermal matrix from 24h burn tissue to infected wound tissue, the expression level of IL-6, MMP-24, VE-cadherin and VEGF were decreased. We found no inflammatory cells infiltration in the dermal matrix were observed in both experimental and control groups (24h burn group), while the obviously vascular infiltration and fiber fusion were observed in the experimental group after subcutaneous implantation experiments. There was better bio-performance, low immunogenicity and better dermal incorporation after treated

  14. Paracrine Engineering of Human Explant-Derived Cardiac Stem Cells to Over-Express Stromal-Cell Derived Factor 1α Enhances Myocardial Repair.

    Science.gov (United States)

    Tilokee, Everad L; Latham, Nicholas; Jackson, Robyn; Mayfield, Audrey E; Ye, Bin; Mount, Seth; Lam, Buu-Khanh; Suuronen, Erik J; Ruel, Marc; Stewart, Duncan J; Davis, Darryl R

    2016-07-01

    First generation cardiac stem cell products provide indirect cardiac repair but variably produce key cardioprotective cytokines, such as stromal-cell derived factor 1α, which opens the prospect of maximizing up-front paracrine-mediated repair. The mesenchymal subpopulation within explant derived human cardiac stem cells underwent lentiviral mediated gene transfer of stromal-cell derived factor 1α. Unlike previous unsuccessful attempts to increase efficacy by boosting the paracrine signature of cardiac stem cells, cytokine profiling revealed that stromal-cell derived factor 1α over-expression prevented lv-mediated "loss of cytokines" through autocrine stimulation of CXCR4+ cardiac stem cells. Stromal-cell derived factor 1α enhanced angiogenesis and stem cell recruitment while priming cardiac stem cells to readily adopt a cardiac identity. As compared to injection with unmodified cardiac stem cells, transplant of stromal-cell derived factor 1α enhanced cells into immunodeficient mice improved myocardial function and angiogenesis while reducing scarring. Increases in myocardial stromal-cell derived factor 1α content paralleled reductions in myocyte apoptosis but did not influence long-term engraftment or the fate of transplanted cells. Transplantation of stromal-cell derived factor 1α transduced cardiac stem cells increased the generation of new myocytes, recruitment of bone marrow cells, new myocyte/vessel formation and the salvage of reversibly damaged myocardium to enhance cardiac repair after experimental infarction. Stem Cells 2016;34:1826-1835.

  15. 脱细胞真皮基质对扩张器/假体乳房再造并发症影响的Meta分析%The impact of acellular dermal matrix on complications of breast reconstruction using tissue expander/implant: a Meta-analysis

    Institute of Scientific and Technical Information of China (English)

    董洁; 吴小蔚; 田方兴

    2013-01-01

    Objective To analyze the effect of acellular dermal matrix (ADM) on complications of breast reconstruction using tissue expander/implant,and to offer preliminary evidences for ADM clinical application.Methods Articles published from Jan.2010 to Oct.2012 were searched in Pubmed,EMbase,Science Direct and CNKI database.Literatures were filtrated according to inclusive criteria.Values were extracted from included literatures; factors regarding complications were collected.Metaanalysis was performed with Stata 12.0.Results 10 researches were included.Comparing to control group,the pooled odds ratio (OR) of overall complications,infections,hematomas/seromas,explantations are 1.51(P=0.038),1.91(P=0.032),1.80(P=0.005) and 2.37 (P=0.138) in ADM group respectively.Conclusions In breast reconstruction using tissue expander/implant,ADM increases the occurrence of hematomas/saromas as well as risks of infections and overall complications.%目的 对在扩张器/假体乳房再造术中应用脱细胞真皮基质(acellular dermal matrix,ADM)是否增加术后并发症进行探讨,以为临床应用提供初步依据.方法 计算机检索2010年1月至2012年10月Pubmed、EMbase、Science Direct、中国生物医学文献数据库和中国期刊网全文数据库中发表的文献,设定文献纳入条件,对符合条件的文献进行并发症相关数据导出,然后使用Stata 12.0进行Meta分析.结果 共有10篇文献被纳入分析.与对照组相比,ADM组总并发症、感染,血肿/血清肿及扩张器/假体取出发生率的比值比(OR)分别为1.51(P =0.038)、1.91(P=0.032)、1.80(P =0.005)和2.37(P =0.138).结论 在扩张器/假体的乳房再造术中,使用ADM对血肿/血清肿的发生有促进作用,并有增加感染率及总并发症发生率的趋势.

  16. Bone cutting.

    Science.gov (United States)

    Giraud, J Y; Villemin, S; Darmana, R; Cahuzac, J P; Autefage, A; Morucci, J P

    1991-02-01

    Bone cutting has always been a problem for surgeons because bone is a hard living material, and many osteotomes are still very crude tools. Technical improvement of these surgical tools has first been their motorization. Studies of the bone cutting process have indicated better features for conventional tools. Several non-conventional osteotomes, particularly ultrasonic osteotomes are described. Some studies on the possible use of lasers for bone cutting are also reported. Use of a pressurised water jet is also briefly examined. Despite their advantages, non-conventional tools still require improvement if they are to be used by surgeons.

  17. Dermal fat graft from simultaneous abdominoplasty as an adjunct to revision aesthetic and reconstructive breast surgery: A poor man's acellular dermal matrix?

    Directory of Open Access Journals (Sweden)

    F. Xie

    2014-01-01

    CONCLUSION: We herein report the use of free dermal fat graft in revision aesthetic and reconstructive surgery in a manner akin to recent acellular dermal matrix use. The comparable enhanced aesthetic outcomes, minimal complication rate and substantial cost savings merit dissemination to a global audience and encourage surgeons to consider this economic alternative.

  18. Primary vaccination of adults with reduced antigen-content diphtheria-tetanus-acellular pertussis or dTpa-inactivated poliovirus vaccines compared to diphtheria-tetanus-toxoid vaccines.

    NARCIS (Netherlands)

    Theeten, H.; Rumke, H.C.; Hoppener, F.J.; Vilatimo, R.; Narejos, S.; Damme, P. van; Hoet, B.

    2007-01-01

    OBJECTIVE: To evaluate immunogenicity and reactogenicity of primary vaccination with reduced-antigen-content diphtheria-tetanus-acellular pertussis (dTpa) or dTpa-inactivated poliovirus (dTpa-IPV) vaccine compared to diphtheria-tetanus-toxoid vaccines (Td) in adults > or = 40 years of age without

  19. Associations between three specific a-cellular measures of the oxidative potential of particulate matter and markers of acute airway and nasal inflammation in healthy volunteers

    NARCIS (Netherlands)

    Janssen, Nicole A H; Strak, Maciej; Yang, Aileen; Hellack, Bryan; Kelly, Frank J; Kuhlbusch, Thomas A J; Harrison, Roy M; Brunekreef, Bert; Cassee, Flemming R; Steenhof, Maaike; Hoek, Gerard

    2015-01-01

    INTRODUCTION: We evaluated associations between three a-cellular measures of the oxidative potential (OP) of particulate matter (PM) and acute health effects. METHODS: We exposed 31 volunteers for 5 h to ambient air pollution at five locations: an underground train station, two traffic sites, a farm

  20. Primary vaccination of adults with reduced antigen-content diphtheria-tetanus-acellular pertussis or dTpa-inactivated poliovirus vaccines compared to diphtheria-tetanus-toxoid vaccines.

    NARCIS (Netherlands)

    Theeten, H.; Rumke, H.C.; Hoppener, F.J.; Vilatimo, R.; Narejos, S.; Damme, P. van; Hoet, B.

    2007-01-01

    OBJECTIVE: To evaluate immunogenicity and reactogenicity of primary vaccination with reduced-antigen-content diphtheria-tetanus-acellular pertussis (dTpa) or dTpa-inactivated poliovirus (dTpa-IPV) vaccine compared to diphtheria-tetanus-toxoid vaccines (Td) in adults > or = 40 years of age without

  1. Different IgG-subclass distributions after whole-cell and acellular pertussis infant primary vaccinations in healthy and pertussis infected children

    NARCIS (Netherlands)

    Hendrikx, Lotte H.; Schure, Rose-Minke; Ozturk, Kemal; de Rond, Lia G. H.; de Greeff, S. C.; Sanders, Elisabeth A. M.; Berbers, Guy A. M.; Buisman, Anne-Marie

    2011-01-01

    The distribution of IgG-subclasses provides insight in the immunological mechanisms of protection against whooping cough. We investigated the effect of Dutch whole-cell pertussis and acellular pertussis vaccines administered in infancy on the IgG-subclass distributions in healthy children aged 12 mo

  2. Regenerative and Antibacterial Properties of Acellular Fish Skin Grafts and Human Amnion/Chorion Membrane: Implications for Tissue Preservation in Combat Casualty Care.

    Science.gov (United States)

    Magnusson, Skuli; Baldursson, Baldur Tumi; Kjartansson, Hilmar; Rolfsson, Ottar; Sigurjonsson, Gudmundur Fertram

    2017-03-01

    Improvised explosive devices and new directed energy weapons are changing warfare injuries from penetrating wounds to large surface area thermal and blast injuries. Acellular fish skin is used for tissue repair and during manufacturing subjected to gentle processing compared to biologic materials derived from mammals. This is due to the absence of viral and prion disease transmission risk, preserving natural structure and composition of the fish skin graft. The aim of this study was to assess properties of acellular fish skin relevant for severe battlefield injuries and to compare those properties with those of dehydrated human amnion/chorion membrane. We evaluated cell ingrowth capabilities of the biological materials with microscopy techniques. Bacterial barrier properties were tested with a 2-chamber model. The microstructure of the acellular fish skin is highly porous, whereas the microstructure of dehydrated human amnion/chorion membrane is mostly nonporous. The fish skin grafts show superior ability to support 3-dimensional ingrowth of cells compared to dehydrated human amnion/chorion membrane (p < 0.0001) and the fish skin is a bacterial barrier for 24 to 48 hours. The unique biomechanical properties of the acellular fish skin graft make it ideal to be used as a conformal cover for severe trauma and burn wounds in the battlefield. Reprint & Copyright © 2017 Association of Military Surgeons of the U.S.

  3. Indução de calos em explantes foliares de Murici-pequeno (Byrsonima intermedia A. Juss. Calli induction from leaf explants of murici-pequeno (Byrsonima intermedia A. Juss.

    Directory of Open Access Journals (Sweden)

    Raírys Cravo Nogueira

    2007-04-01

    Full Text Available O murici-pequeno (Byrsonima intermedia A. Juss. é um arbusto do cerrado cujo chá da casca do caule apresenta atividade adstringente nas diarréias e disenterias. O gênero Byrsonima apresenta taxa de germinação baixa e emergência lenta da plântula, dificultando a propagação sexuada. Objetivou-se com este trabalho obter calos friáveis em explantes foliares de murici-pequeno para estudos futuros em suspensão celular e metabolismo secundário, assim como embriogênese somática. Avaliou-se o efeito de diferentes concentrações da auxina 2,4-D e a sua interação com as citocininas TDZ e BAP na calogênese. Os resultados demonstraram que na ausência de 2,4-D não ocorre formação de calo em explantes foliares. A adição de BAP ou TDZ não influencia o processo de calogênese. Para a indução e proliferação de calo em explantes foliares de murici-pequeno, os resultados recomendam o uso de meio MS, acrescido de 1,0 mg L-1 de 2,4-D, mantendo os explantes em condição de escuro por 30 dias.Byrsonima intermedia A. Juss is a shrub of cerrado whose husk presents medicinal activities in diarrheas and dysenteries. The Byrsonima genus present low germination rate and slow plantlet emergency which makes difficult sexual propagation. The objective of this work was to obtain friable callus in leaf explants for future studies with suspension cells and secondary metabolites as well as somatic embryogenesis. The effect of different concentrations of 2,4-D and its interaction with TDZ and BAP in callus formation was evaluated. The results demonstrated that there is no formation of callus in leaf explants maintained in absence of 2,4-D. The addition of TDZ or BAP had no influence in the calogenesis process. For callus induction and proliferation, the results suggest the use of MS medium supplemented with 1.0 mg L-1 2,4-D maintaining the explants in the dark.

  4. Chemical and Structural Characterization of Several Mid-Term Explanted Breast Prostheses

    Directory of Open Access Journals (Sweden)

    Angela Amoresano

    2016-08-01

    Full Text Available The recent scandal of poly implant prostheses (PIP, which were found in some cases to be made of non-medical grade silicone (as reported by the European Scientific Committee on Emerging and Newly Identified Health Risks, had a great social impact. Thousands of patients asked for implant removal with significant costs for public health care systems. We analysed, by a multidisciplinary approach, sixteen different breast implants after explantation by using several analytical and structural techniques, such as Fourier Transform infrared spectroscopy (FT-IR, mass spectrometry equipped by ion coupled plasma (ICP-MS, gas-chromatography (GC-MS, and tensile testing. Traces of organic (fatty acid and inorganic (Fe, Cr, Pt, Na, and other metals substances were found in all samples, and, even if these values are under danger threshold levels, our study results highlight the possibility of bioaccumulation and tissue contamination, implying the need for continuous medical surveillance and monitoring of material aging.

  5. Immunohistochemical localization of IAA in graft union of explanted internode grafting

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    The dynamic changes of IAA in graft union of explanted internode autografting of Cucumber (Cucumis sativus Linn.) have been investigated using the immunohistochemical localization technique.It is shown that the efficiency of IAA fixation using lyophilization-gas fixation is higher than that using liquid chemical fixation.In contrast to few silver particles and no significant changes during the development of graft union cultured in hormone-free medium,more silver particles in graft union and significant changes of IAA related to graft union development have been found when graft union was cultured in medium supplemented with appropriate hormones.The fixation procedure of plant hormones and the roles of IAA in graft union are discussed.

  6. Micropropagation of Origanum acutidens (HAND.-MAZZ.) IETSWAART using stem node explants.

    Science.gov (United States)

    Yildirim, Mehmet Ugur

    2013-01-01

    Origanum acutidens (HAND.-MAZZ.) IETSWAART is a promising ornamental plant that can be widely used in landscape management. It is endemic to Eastern Anatolian region of Turkey. Tissue culture has not been used to micropropagate it. The study reports stem node explants from one-week-old seedlings of the plant for successful micropropagation. The stem nodes were cultured on MS medium containing 0.6, 1.2, 1.8, and 2.4 mg/L BAP with 0.2 mg/L NAA. Visible effects of culture media on shoot proliferation were recorded. Shoot regeneration rate was maximum on MS medium containing 1.80 mg/L BAP-0.2 mg/L NAA. The micropropagated shoots were rooted on MS medium containing 0.2 mg/L NAA. All microrooted plantlets survived during acclimatisation on peat moss. It was concluded that O. acutidens can be successfully micropropagated under in vitro conditions.

  7. Micropropagation from cultured nodal explants of rose (Rosa hybrida L. cv. ‘Perfume Delight’

    Directory of Open Access Journals (Sweden)

    Kamnoon Kanchanapoom

    2010-01-01

    Full Text Available A method for the micropropagation of rose (Rosa hybrida L. cv. ‘Perfume Delight’ was developed. First to fifth nodal explants from young healthy shoots were excised and cultured on basal medium of Murashige and Skoog (1962, MS containing several concentrations of BA and NAA. Multiple shoot formation of up to 3 shoots was obtained on MS medium supplemented with 3 mg/l BA and 0.003 mg/l NAA. Shoot readily rooted on ¼MS medium devoid of growth regulators.Rooted plantlets were hardened and established in pots at 100% survival. In vitro flowering was observed on rose plantscultured on MS medium containing 3 mg/l BA and 0.003 mg/l NAA.

  8. Depot-dependent effects of adipose tissue explants on co-cultured hepatocytes

    DEFF Research Database (Denmark)

    Du, Zhen-Yu; Ma, Tao; Lock, Erik-Jan;

    2011-01-01

    We have developed an in vitro hepatocyte-adipose tissue explant (ATE) co-culture model enabling examination of the effect of visceral and subcutaneous adipose tissues on primary rat hepatocytes. Initial analyses of inflammatory marker genes were performed in fractionated epididymal or inguinal...... levels of IL-6, TNF-a and PGE(2) in the media from inguinal ATEs co-cultured with primary rat hepatocytes were higher than that in the media from epididymal ATEs co-cultured with hepatocytes, although the significant difference was only seen in PGE(2). Lipolysis, measured as glycerol release, was similar...... in the ATEs isolated from inguinal and epididymal adipose tissues when cultured alone, but the glycerol release was higher in the ATEs isolated from epididymal than from inguinal adipose tissue when co-cultured with hepatocytes. Compared to epididymal ATEs, the ATEs from inguinal adipose tissue elicited...

  9. Identification of a benzyladenine disaccharide conjugate produced during shoot organogenesis in Petunia leaf explants

    Energy Technology Data Exchange (ETDEWEB)

    Auer, C.A. (Univ. of Connecticute, Storrs, CT (United States)); Cohen, J.D. (Beltsville Agricultural Research Ctr., MD (United States))

    1993-06-01

    Prior studies of benzyladenine (BA) metabolism in Petunia hybrida Vilm. leaf explants during shoot organogenesis revealed the presence of an abundant unidentified BA conjugate. The BA conjugate, compound C, made up to 39% of the total pool of BA conjugates in two Petunia lines and was associated with an increased shoot organogenic response when compared with a third Petunia line that did not produce any compound C. Structural analysis of compound C utilizing fast atom bombardment mass spectrometry, two methods of carbohydrate analysis, ultraviolet absorption spectra, and Fourier transform infrared spectra identified it as a new cytokinin conjugate, 6-benzylamino-9-[O-glucopyranosyl-(1[yields]3)-ribofuranosyl]-purine. Based on our prior biological studies and the similarity of this compound to related cytokinin conjugates, this disaccharide cytokinin conjugate may be part of the interconvertible pool of cytokinins active in Petunia shoot organogenesis. 17 refs., 2 figs., 2 tabs.

  10. An animal explant model for the study of human cutaneous squamous cell carcinoma.

    Directory of Open Access Journals (Sweden)

    Daniel A Belkin

    Full Text Available We established a human tissue explant model to facilitate study of cutaneous squamous cell carcinoma. We accomplished this by implanting debulked SCC, from surgical discard, into nude rats. Human SCC remained viable and continued to proliferate for at least 4 weeks and showed evidence of neovascularization. At 4 weeks, SCC implants showed a trend toward increased PCNA positive cells compared to fresh SCC cells/mm(2 tissue supporting continued proliferation throughout engraftment. Von Willebrand's Factor (VWF positive cells were found within implants and likely represented rat vessel neovascularization. Human Langerhans' (Langerin+ cells, but no T cells (CD3+, CD8+, FoxP3+, macrophages (CD163, or NK cells (NKp46, were present in SCC implants at 4 weeks. These findings support the possibility that LCs fail to migrate from cutaneous SCC and thus contribute to lack of effective antitumor response. Our findings also provide a novel model system for further study of primary cutaneous SCC.

  11. Characterisation of an accelerator-based neutron source for BNCT of explanted livers

    Energy Technology Data Exchange (ETDEWEB)

    Agosteo, S. [Politecnico di Milano (Italy). Dipartimento di Ingeneria Nucleare; Colautti, P. [INFN, Padova (Italy). Laboratori Nazionali di Legnaro; Corrado, M.G. [Universita degli Studi di Milano (Italy). Dipartimento di Fisica; d`Errico, F. [Pisa Univ. (Italy). Dipt. di Costruzioni Meccaniche e Nucleari; Matzke, M. [Physikalisch-Technische Bundesanstalt, Braunschweig (Germany); Monti, S.; Tinti, R. [ENEA-ERG-FIRE, Bologna (Italy); Silari, M. [Consiglio Nazionale delle Ricerche, Milan (Italy)

    1997-09-01

    An accelerator-based thermal neutron source for BNCT of the explanted liver was designed using the MCNP code. Neutrons are generated via (d,n) reactions by 7 MeV deuterons bombarding a beryllium target. The therapy constraints were approached by simulating an irradiation cavity placed inside a graphite reflector parallelepiped containing a heavy-water moderator in turn enclosing the beryllium target. The experimental verification was performed at the Laboratori Nazionali di Legnaro (Italy). The thermal and epithermal neutron flux was measured at various positions in the irradiation cavity by means of activation techniques employing bare and cadmium covered indium foils. Further measurements were performed with BF{sub 3} detectors. The fast neutron component of the dose equivalent and the energy spectrum above 100keV were assessed by means of a recently developed technique employing variable threshold superheated drop detectors. The prompt gamma ray dose was measured with {sup 7}LiF TLDs. (author).

  12. Light scattering and light transmittance in intraocular lenses explanted because of optic opacification.

    Science.gov (United States)

    Michelson, Jennifer; Werner, Liliana; Ollerton, Andrew; Leishman, Lisa; Bodnar, Zachary

    2012-08-01

    To assess light scattering and light transmittance in intraocular lenses (IOLs) explanted because of optic opacification. John A. Moran Eye Center, University of Utah, Salt Lake City, Utah, USA. Experimental study. Poly(methyl methacrylate) (PMMA) IOLs with snowflake degeneration, hydrophilic acrylic IOLs with different calcification patterns, and 1 calcified silicone IOL explanted from an eye with asteroid hyalosis were studied with gross and light microscopy. Light scattering was measured with an EAS-1000 Scheimpflug camera. Light transmittance was measured with a Lambda 35 UV/Vis spectrophotometer (single-beam configuration with RSA-PE-20 integrating sphere). Analyses were performed at room temperature in the hydrated state and compared with controls. The study evaluated 8 PMMA IOLs, 22 hydrophilic acrylic IOLs, and 1 silicone IOL. Light scattering was as follows: 208 to 223 computer-compatible tapes (CCTs) for PMMA IOLs with snowflake degeneration (control = 9 CCTs); 90 to 227 CCTs for calcified hydrophilic acrylic IOLs (controls = 12 to 23 CCTs); 223 CCTs for the calcified silicone IOL (control = 5 CCTs). The mean light transmittance in the visible light spectrum was 81.08% to 97.10% for PMMA IOLs (control = 98.80%); 78.94% to 97.32% for hydrophilic acrylic IOLs (controls = 97.32% to 98.66%); 94.68% for the silicone IOL (control = 97.74%). Intraocular lens opacification led to very high levels of light scattering and a potential for decreased light transmittance, which play a role in the development of symptoms such as glare and halos, decreased contrast sensitivity, and eventually decreased visual acuity. No author has a financial or proprietary interest in any material or method mentioned. Copyright © 2012 ASCRS and ESCRS. Published by Elsevier Inc. All rights reserved.

  13. Callus induction and biomass accumulation in vitro in explants from chokeberry (Aronia melanocarpa (Michx. Elliot fruit

    Directory of Open Access Journals (Sweden)

    Tatiana I. Calalb

    2014-09-01

    Full Text Available In this study, the following features were determined: biological (the optimal histogen as explant and the optimal age of donor fruit, biotechnological (type, dosage and combination of growth regulators supplements in culture medium Murashige and Skoog as well as sucrose dosage, and physical (light regime, to induce callusing and biomass accumulation in vitro from the succulent chokeberry (Aronia melanocarpa (Michx. Elliot fruit. It turned out that it was much easier to induce callus from explants composed of the epicarp and hypoderm cut from fruits at 50–60 days after flowering. The role of light regime and varied supplementation of the basic MS medium with different doses of growth regulators was established; they resulted in four pigmented carpomass: violet, cream-pink, cream-white and green. The best combinations for the proliferation of fruit callus were culture media with 0.2–2.5 mg × dm-3 2,4-D+0.5 mg × dm-3 KIN +60 g × dm-3sucrose, while for fruit biomass accumulation enriched with phenolic substances – 2.5–3.5 mg × dm-3 NAA+0.5 mg × dm-3 KIN+60 g × dm-3sucrose. The chemical study of phenolic compounds by HPLC coupled with the mass spectrometry method identified chlorogenic acid, hiperozide, quercetrin, isoquercitrin and rutozide quantitatively and qualitatively in all pigmented carpomass and fruits; an exception is p-coumaric present only qualitatively in green carpomass and absent in fruit and quercetol absent in green carpomass.

  14. Postnatal exposure history and airways: oxidant stress responses in airway explants.

    Science.gov (United States)

    Murphy, Shannon R; Schelegle, Edward S; Edwards, Patricia C; Miller, Lisa A; Hyde, Dallas M; Van Winkle, Laura S

    2012-12-01

    Postnatally, the lung continues to grow and differentiate while interacting with the environment. Exposure to ozone (O(3)) and allergens during postnatal lung development alters structural elements of conducting airways, including innervation and neurokinin abundance. These changes have been linked with development of asthma in a rhesus monkey model. We hypothesized that O(3) exposure resets the ability of the airways to respond to oxidant stress and that this is mediated by changes in the neurokinin-1 receptor (NK-1R). Infant rhesus monkeys received episodic exposure to O(3) biweekly with or without house dust mite antigen (HDMA) from 6 to 12 months of age. Age-matched monkeys were exposed to filtered air (FA). Microdissected airway explants from midlevel airways (intrapulmonary generations 5-8) for four to six animals in each of four groups (FA, O(3), HDMA, and HDMA+O(3)) were tested for NK-1R gene responses to acute oxidant stress using exposure to hydrogen peroxide (1.2 mM), a lipid ozonide (10 μM), or sham treatment for 4 hours in vitro. Airway responses were measured using real-time quantitative RT-PCR of NK-1R and IL-8 gene expression. Basal NK-1R gene expression levels were not different between the exposure groups. Treatment with ozonide or hydrogen peroxide did not change NK-1R gene expression in animals exposed to FA, HDMA, or HDMA+O(3). However, treatment in vitro with lipid ozonide significantly increased NK-1R gene expression in explants from O(3)-exposed animals. We conclude that a history of prior O(3) exposure resets the steady state of the airways to increase the NK-1R response to subsequent acute oxidant stresses.

  15. Effects of high hydrostatic pressure on bacterial growth on human ossicles explanted from cholesteatoma patients.

    Science.gov (United States)

    Dommerich, Steffen; Frickmann, Hagen; Ostwald, Jürgen; Lindner, Tobias; Zautner, Andreas Erich; Arndt, Kathleen; Pau, Hans Wilhelm; Podbielski, Andreas

    2012-01-01

    High hydrostatic pressure (HHP) treatment can eliminate cholesteatoma cells from explanted human ossicles prior to re-insertion. We analyzed the effects of HHP treatment on the microbial flora on ossicles and on the planktonic and biofilm states of selected isolates. Twenty-six ossicles were explanted from cholesteatoma patients. Five ossicles were directly analyzed for microbial growth without further treatment. Fifteen ossicles were cut into two pieces. One piece was exposed to HHP of 350 MPa for 10 minutes. Both the treated and untreated (control) pieces were then assessed semi-quantitatively. Three ossicles were cut into two pieces and exposed to identical pressure conditions with or without the addition of one of two different combinations of antibiotics to the medium. Differential effects of 10-minute in vitro exposure of planktonic and biofilm bacteria to pressures of 100 MPa, 250 MPa, 400 MPa and 540 MPa in isotonic and hypotonic media were analyzed using two patient isolates of Staphylococcus epidermidis and Neisseria subflava. Bacterial cell inactivation and biofilm destruction were assessed by colony counting and electron microscopy. A variety of microorganisms were isolated from the ossicles. Irrespective of the medium, HHP treatment at 350 MPa for 10 minutes led to satisfying but incomplete inactivation especially of gram-negative bacteria. The addition of antibiotics increased the efficacy of elimination. A comparison of HHP treatment of planktonic and biofilm cells showed that the effects of HPP were reduced by about one decadic logarithmic unit when HPP was applied to biofilms. High hydrostatic pressure conditions that are suitable to inactivate cholesteatoma cells fail to completely sterilize ossicles even if antibiotics are added. As a result of the reduced microbial load and the viability loss of surviving bacteria, however, there is a lower risk of re-infection after re-insertion.

  16. The effects of ascorbic acid on cartilage metabolism in guinea pig articular cartilage explants.

    Science.gov (United States)

    Clark, Amy G; Rohrbaugh, Amy L; Otterness, Ivan; Kraus, Virginia B

    2002-03-01

    Ascorbic acid has been associated with the slowing of osteoarthritis progression in guinea pig and man. The goal of this study was to evaluate transcriptional and translational regulation of cartilage matrix components by ascorbic acid. Guinea pig articular cartilage explants were grown in the presence of L-ascorbic acid (L-Asc), D-isoascorbic acid (D-Asc), sodium L-ascorbate (Na L-Asc), sodium D-isoascorbate (Na D-Asc), or ascorbyl-2-phosphate (A2P) to isolate and analyze the acidic and nutrient effects of ascorbic acid. Transcription of type II collagen, prolyl 4-hydroxylase (alpha subunit), and aggrecan increased in response to the antiscorbutic forms of ascorbic acid (L-Asc, Na L-Asc, and A2P) and was stereospecific to the L-forms. Collagen and aggrecan synthesis also increased in response to the antiscorbutic forms but only in the absence of acidity. All ascorbic acid forms tended to increase oxidative damage over control. This was especially true for the non-nutrient D-forms and the high dose L-Asc. Finally, we investigated the ability of chondrocytes to express the newly described sodium-dependent vitamin C transporters (SVCTs). We identified transcripts for SVCT2 but not SVCT1 in guinea pig cartilage explants. This represents the first characterization of SVCTs in chondrocytes. This study confirms that ascorbic acid stimulates collagen synthesis and in addition modestly stimulates aggrecan synthesis. These effects are exerted at both transcriptional and post-transcriptional levels. The stereospecificity of these effects is consistent with chondrocyte expression of SVCT2, shown previously to transport L-Asc more efficiently than D-Asc. Therefore, this transporter may be the primary mechanism by which the L-forms of ascorbic acid enter the chondrocyte to control matrix gene activity.

  17. Evaluating suitable internal control genes for transcriptional studies in heat-stressed mammary explants of buffaloes.

    Science.gov (United States)

    Sodhi, M; Kishore, A; Khate, K; Kapila, N; Mishra, B P; Kataria, R S; Mohanty, A K; Varshney, N; Mukesh, M

    2013-04-01

    It is now a well-accepted notion that each new experimental design requires proper evaluation of internal control genes (ICGs) for accurate normalization of expression data. In riverine buffaloes, till date no appropriate ICG has been reported for studying transcriptional response under any of the physiological stressful condition. The objective here was to test 16 well-known reference genes from different functional categories that could serve as suitable ICG during heat stress studies in buffalo mammary tissue. Briefly, the mammary explants were exposed to 45°C for 1 h and subsequently allowed to recover at 37°C for different time points (2-24 h). Three software programs, geNorm, Normfinder and BestKeeper, were used to measure gene transcript stability. RPL22 was excluded because of weak amplification and unacceptable PCR efficiency. Except GAPDH, all other genes showed expression stability within the acceptable range (<1.5). RPL4, B2M, RPS23 and EEF1A1 genes were found to be most stably expressed while GAPDH and ACTB showed least stability. The BestKeeper analysis identified high correlation for RPL4 (r=0.953) and EEF1A1 (r=0.914) with BestKeeper index. Based on the present findings, it could be suggested that geometric average of RPL4, B2M, RPS23 and EEF1A1 would provide accurate normalization to transcriptional data of buffalo mammary explant in response to heat stress. © 2012 Blackwell Verlag GmbH.

  18. Bone acidic glycoprotein-75 is a major synthetic product of osteoblastic cells and localized as 75- and/or 50-kDa forms in mineralized phases of bone and growth plate and in serum.

    Science.gov (United States)

    Gorski, J P; Griffin, D; Dudley, G; Stanford, C; Thomas, R; Huang, C; Lai, E; Karr, B; Solursh, M

    1990-09-05

    Anti-peptide and anti-protein antisera were produced which both recognize bone acidic glycoprotein-75 (Mr = 75,000) and an apparent fragment or biosynthetic intermediate (Mr = 50,000) in calcified tissues and/or serum. A fragment-precursor relationship is suggested from the fact that closely spaced doublet polypeptides of Mr = 50,000 could be produced by proteolysis of the purified protein upon long term storage. No reactivity was detected with osteopontin, bone sialoprotein, or small bone proteoglycans. Bone acidic glycoprotein-75 represents 0.5-1% of the total radiolabeled proteins synthesized by explant cultures of neonatal calvaria or growth plate, by calvarial outgrowth cultures, and by rat osteosarcoma cells. Amounts produced by explant cultures and calvarial outgrowth cultures were similar to that for osteopontin, a major product of osteoblasts. In osteosarcoma cultures, 80% of labeled antigens were associated with the cell layer fraction wherein specific immunoprecipitation pelleted Mr = 50,000 and 75,000 sized antigens. Bone acidic glycoprotein-75 (Mr = 75,000) is enriched in 4 M guanidine HCl/0.5 EDTA extracts of neonatal rat bone and growth plate tissues, whereas largely absent from heart, lung, spleen, liver, brain, and kidney. Explant cultures of these noncalcifying tissues also synthesized bone acidic glycoprotein-75 antigen, but the quantities produced were only 5% or less that obtained with calvaria. By immunohistochemistry, antigenicity is associated with the bony shaft and calcified cartilage of long bones, but is absent from associated soft tissues. These finding demonstrate that bone acidic glycoprotein-75 is antigenically distinct, predominantly localized to calcified tissues, represents a major product of normal osteoblastic cells and may undergo a characteristic fragmentation in vivo and in vitro.

  19. Uso de mutagênese "in vitro" no melhoramento de citros: I. Sensibilidade a raios gama de explantes do cultivar pêra "In vitro" mutagenesis in citrus breeding: gamma rays sensitivity of cultivar pêra explants

    Directory of Open Access Journals (Sweden)

    Augusto Tulmann Neto

    1994-01-01

    Full Text Available Para futuros trabalhos com indução de mutação "in vitro", determinou-se a sensibilidade de diferentes explantes do cultivar de laranja Pêra (Citrus sinensis Osbeck a raios gama visando a escolha das doses a utilizar. Para isso, após a irradiação, avaliou-se o número de explantes responsivos, de embrióides e de brotações adventícias, a massa do calo e a formação de enraizamento. Resultados diferentes foram obtidos de acordo com o tecido empregado. Assim, para a irradiação de nucelos, recomenda-se o uso de doses entre 20 e 40 Gy; em calos embriogênicos, de 120 Gy e, para a irradiação de cotilédones de embriões nucelares, a dose situa-se ao redor de 40 Gy.In order to obtain basic data for future research in mutation breeding through "in vitro" technology in sweet orange cv. Pêra, gamma rays sensitivity of different types of explants was analyzed. After irradiation the following observations were carried out: response of explants, number of embryoids and adventitious buds, callus weight and rooting formation. Different results were obtained according the explant used. For irradiated nucellus, the dose was 20-40 Gy and for embryogenic callus 120 Gy is indicated. For irradiation of cotyledons originated from nucellar embryoids the use of 40 Gy of gamma rays was recommended.

  20. The impact of acellular dermal matrix on tissue expander/implant loss in breast reconstruction: an analysis of the tracking outcomes and operations in plastic surgery database.

    Science.gov (United States)

    Pannucci, Christopher J; Antony, Anuja K; Wilkins, Edwin G

    2013-07-01

    Use of acellular dermal matrix in breast reconstruction has been associated with increased complications. However, existing studies are generally small, from single centers, and underpowered to control for confounding using regression techniques. Here, the Tracking Outcomes and Operations in Plastic Surgery database was used to examine the effect of acellular dermal matrix on expander/implant loss when controlling for other confounders. Analysis was limited to patients having tissue expander or implant-based breast reconstruction. Surgeon-reported data, International Classification of Diseases, Ninth Edition codes, and Current Procedural Terminology codes were used to identify independent variables. The dependent variable of interest was 30-day rates of tissue expander or implant loss. Bivariate statistics were performed. Multivariable logistic regression identified independent predictors of expander/implant loss when controlling for other confounders. Data were available for 14,249 patients. The overall rate of expander/implant loss was 2.05 percent. Bivariate analysis demonstrated acellular dermal matrix was associated with an absolute increase in expander/implant loss of 0.7 percent (1.88 percent versus 2.58 percent, p = 0.012). The regression model demonstrated that rising body mass index, current smoking, and presence of diabetes were each independent predictors of expander/implant loss. When controlling for all other identified confounders, use of acellular dermal matrix was associated with a significant increase in expander/implant loss (odds ratio, 1.42; 95 percent confidence interval, 1.04 to 1.94; p = 0.026). Thirty-day risk for expander/implant loss after tissue expander or implant-based breast reconstruction was 2.05 percent. Use of acellular dermal matrix was associated with a 0.7 percent absolute risk increase for expander/implant loss. Risk, III.

  1. Human low density lipoprotein as a substrate for in vitro steroidogenesis assays with fathead minnow ovary explants

    Science.gov (United States)

    Gonad explant in vitro steroidogenesis assays are used as part of a multifaceted strategy to detect endocrine active chemicals capable of altering steroid hormone synthesis. An in vitro steroidogenesis assay used in our laboratory involves exposing fathead minnow (FHM) gonad exp...

  2. Interference in adhesion of bacteria and yeasts isolated from explanted voice prostheses to silicone rubber by rhamnolipid biosurfactants

    NARCIS (Netherlands)

    Rodrigues, LR; Banat, IM; van der Mei, HC; Teixeira, JA; Oliveira, R

    2006-01-01

    Aims: The effects and extent of adhesion of four different bacterial and two yeast strains isolated from explanted voice prostheses to silicone rubber with and without an adsorbed rhamnolipid biosurfactant layer obtained from Pseudomonasaeruginosa DS10-129 was studied. Methods and Results: The abili

  3. Serial explant culture provides novel insights into the potential location and phenotype of corneal endothelial progenitor cells.

    Science.gov (United States)

    Walshe, Jennifer; Harkin, Damien G

    2014-10-01

    The routine cultivation of human corneal endothelial cells, with the view to treating patients with endothelial dysfunction, remains a challenging task. While progress in this field has been buoyed by the proposed existence of progenitor cells for the corneal endothelium at the corneal limbus, strategies for exploiting this concept remain unclear. In the course of evaluating methods for growing corneal endothelial cells, we have noted a case where remarkable growth was achieved using a serial explant culture technique. Over the course of 7 months, a single explant of corneal endothelium, acquired from cadaveric human tissue, was sequentially seeded into 7 culture plates and on each occasion produced a confluent cell monolayer. Sample cultures were confirmed as endothelial in origin by positive staining for glypican-4. On each occasion, small cells, closest to the tissue explant, developed into a highly compact layer with an almost homogenous structure. This layer was resistant to removal with trypsin and produced continuous cell outgrowth during multiple culture periods. The small cells gave rise to larger cells with phase-bright cell boundaries and prominent immunostaining for both nestin and telomerase. Nestin and telomerase were also strongly expressed in small cells immediately adjacent to the wound site, following transfer of the explant to another culture plate. These findings are consistent with the theory that progenitor cells for the corneal endothelium reside within the limbus and provide new insights into expected expression patterns for nestin and telomerase within the differentiation pathway.

  4. Human low density lipoprotein as a substrate for in vitro steroidogenesis assays with fathead minnow ovary explants

    Science.gov (United States)

    Gonad explant in vitro steroidogenesis assays are used as part of a multifaceted strategy to detect endocrine active chemicals capable of altering steroid hormone synthesis. An in vitro steroidogenesis assay used in our laboratory involves exposing fathead minnow (FHM) gonad exp...

  5. Conversion of neural plate explants to pre-placodal ectoderm-like tissue in vitro.

    Science.gov (United States)

    Shigetani, Yasuyo; Wakamatsu, Yoshio; Tachibana, Toshiaki; Okabe, Masataka

    2016-09-02

    Neural crest and cranial sensory placodes arise from ectodermal epithelium lying between the neural plate and non-neural ectoderm (neural border). BMP signaling is important for both an induction of the neural border and a subsequent induction of the neural crest within the neural border. In contrast, FGF signaling is important for the neural border induction and the following induction of the pre-placodal ectoderm (PPE), which later gives rise to the cranial sensory placodes. While previous studies have demonstrated that the neural plate explants could be converted to the neural crest cells by adding BMP4 in a culture medium, there is no report showing a similar conversion of the neural plate to the PPE. We therefore examined the effect of FGF2 along with BMP4 on the rostral neural plate explants and found that the explants became the simple squamous epithelia, which were characterized by the desmosomes/tonofilaments in membranes of adjacent cells. Such epithelia expressed sets of neural border markers and the PPE genes, suggesting that the neural plate explants were converted to a PPE-like tissue. This method will be useful for further studying mechanisms of PPE induction and subsequent specifications of the cranial placodes.

  6. Calogênese e rizogênese em explantes de mogno (Swietenia macrophylla King cultivados in vitro.

    Directory of Open Access Journals (Sweden)

    Silvana Cruz da Rocha

    2010-08-01

    Full Text Available A exploração de árvores tropicais realizada de forma indiscriminada, buscando espécies de alto valor econômico, tem levado várias espécies, como o mogno (Swietenia macrophylla King, ao perigo de extinção. O desenvolvimento de uma metodologia de regeneração de gemas, direta ou indireta, poderia auxiliar na obtenção de um grande número de mudas e constituir uma perspectiva à propagação sexuada. Essa última é limitada pelo fato das sementes perderem rapidamente a capacidade germinativa. No presente trabalho, foram utilizados dois tipos de explantes: fragmentos foliares e de raízes de plantas cultivadas in vitro. Após desinfestação, os explantes foram colocados em meio de cultura de Murashige e Skoog (1962 contendo três quartos da concentração de sais, vitaminas do mesmo meio, 30g.L-1 de sacarose, auxina (ácido naftaleno-acético, ANA, 0,11 µM e 0,54 µM, citocinina (cinetina, CIN, 1,2 µM, 2,3 µM, 4,7 µM e 9,3 µM; 6-benziladenina, BA, 2,2 µM, 4,4 µM e 8,8 µM ou 2-isopenteniladenina, 2-iP, 2,5 µM e 7g.L-1 de ágar. As variáveis testadas foram a concentração e o tipo de regulador de crescimento e a origem dos explantes. A cada 30 dias, os explantes foram avaliados pela contagem do número de explantes formando calos ou raízes e a consistência dos calos. Foram obtidos calos a com base nos dois tipos de explantes. Nos explantes foliares, 90% deles formaram calos em meios de cultura contendo BA 4,4 µM com ANA 0,54 µM e BA 8,9 µM com ANA 0,11 ou 0,54 µM. Nos explantes de raízes, a maior percentagem de explantes com calos foi de 55%, no meio de cultura com BA 2,2 µM e ANA 0,54 µM. Raízes adventícias foram obtidas partindo de calos e do limbo dos explantes foliares, em meios de cultura com CIN e ANA. Não foi observada a formação de gemas adventícias.

  7. CALOGÊNESE E RIZOGÊNESE EM EXPLANTES DE MOGNO (Swietenia macrophylla King CULTIVADOS IN VITRO

    Directory of Open Access Journals (Sweden)

    Marguerite Quoirin

    2004-04-01

    Full Text Available A exploração de árvores tropicais realizada de forma indiscriminada, buscando espécies de alto valor econômico, tem levado várias espécies, como o mogno (Swietenia macrophylla King, ao perigo de extinção. O desenvolvimento de uma metodologia de regeneração de gemas, direta ou indireta, poderia auxiliar na obtenção de um grande número de mudas e constituir uma perspectiva à propagação sexuada. Essa última é limitada pelo fato das sementes perderem rapidamente a capacidade germinativa. No presente trabalho, foram utilizados dois tipos de explantes: fragmentos foliares e de raízes de plantas cultivadas in vitro. Após desinfestação, os explantes foram colocados em meio de cultura de Murashige e Skoog (1962 contendo três quartos da concentração de sais, vitaminas do mesmo meio, 30g.L-1 de sacarose, auxina (ácido naftaleno-acético, ANA, 0,11 M e 0,54 M, citocinina (cinetina, CIN, 1,2 M, 2,3 M, 4,7 M e 9,3 M; 6-benziladenina, BA, 2,2 M, 4,4 M e 8,8 M ou 2-isopenteniladenina, 2-iP, 2,5 M e 7g.L-1 de ágar. As variáveis testadas foram a concentração e o tipo de regulador de crescimento e a origem dos explantes. A cada 30 dias, os explantes foram avaliados pela contagem do número de explantes formando calos ou raízes e a consistência dos calos. Foram obtidos calos a com base nos dois tipos de explantes. Nos explantes foliares, 90% deles formaram calos em meios de cultura contendo BA 4,4 M com ANA 0,54 M e BA 8,9 M com ANA 0,11 ou 0,54 M. Nos explantes de raízes, a maior percentagem de explantes com calos foi de 55%, no meio de cultura com BA 2,2 M e ANA 0,54 M. Raízes adventícias foram obtidas partindo de calos e do limbo dos explantes foliares, em meios de cultura com CIN e ANA. Não foi observada a formação de gemas adventícias.

  8. Blockade of Toll-like receptor 2 prevents spontaneous cytokine release from rheumatoid arthritis ex vivo synovial explant cultures

    LENUS (Irish Health Repository)

    Nic An Ultaigh, Sinead

    2011-02-23

    Abstract Introduction The aim of this study was to examine the effect of blocking Toll-like receptor 2 (TLR2) in rheumatoid arthritis (RA) synovial cells. Methods RA synovial tissue biopsies, obtained under direct visualization at arthroscopy, were established as synovial explant cultures ex vivo or snap frozen for immunohistology. Mononuclear cell cultures were isolated from peripheral blood and synovial fluid of RA patients. Cultures were incubated with the TLR1\\/2 ligand, Pam3CSK4 (200 ng, 1 and 10 μg\\/ml), an anti-TLR2 antibody (OPN301, 1 μg\\/ml) or an immunoglobulin G (IgG) (1 μg\\/ml) matched control. The comparative effect of OPN301 and adalimumab (anti-tumour necrosis factor alpha) on spontaneous release of proinflammatory cytokines from RA synovial explants was determined using quantitative cytokine MSD multiplex assays or ELISA. OPN301 penetration into RA synovial tissue explants cultures was assessed by immunohistology. Results Pam3CSK4 significantly upregulated interleukin (IL)-6 and IL-8 in RA peripheral blood mononuclear cells (PBMCs), RA synovial fluid mononuclear cells (SFMCs) and RA synovial explant cultures (P < 0.05). OPN301 significantly decreased Pam3CSK4-induced cytokine production of tumour necrosis factor alpha (TNF-α), IL-1β, IL-6, interferon (IFN)-γ and IL-8 compared to IgG control in RA PBMCs and SFMCs cultures (all P < 0.05). OPN301 penetration of RA synovial tissue cultures was detected in the lining layer and perivascular regions. OPN301 significantly decreased spontaneous cytokine production of TNF-α, IL-1β, IFN-γ and IL-8 from RA synovial tissue explant cultures (all P < 0.05). Importantly, the inhibitory effect of OPN on spontaneous cytokine secretion was comparable to inhibition by anti-TNFα monoclonal antibody adalimumab. Conclusions These findings further support targeting TLR2 as a potential therapeutic agent for the treatment of RA.

  9. Effects of donor plant age and explants on in vitro culture of Cedrela montana Moritz ex Turcz

    Directory of Open Access Journals (Sweden)

    Sofía Basto

    2012-12-01

    Full Text Available To evaluate the organogenic potential of Cedrela montana Moritz ex Turcz. Explants from mature (10-20 year-old and juvenile (7-18 month-old trees were collected. The first grouping included buds, leaves, and nodes derived from juvenile basal offshoots and rejuvenated shoots from cuttings. The second, included leaves, petioles, nodes, internodes and nodes of in vitro elongated shoots. The highest organogenic potential was observed in nodes from juvenile trees: 45.8% of explants presented axillary bud elongation, while 56.2% presented rooting in a growth regulator free culture medium. Fifty-one percent of elongated shoots produced adventitious shoots with 0.5 μM NAA and 0.5 μM BA; 30% with 0.5 μM NAA and 1 μM BA; and 30% with 1 μM BA. Twenty percent presented roots with 0.5 μM NAA. Root formation was stimulated in a medium supplemented with activated charcoal (5 gL-1. The acclimatization of eighty percent of plantlets regenerated from nodes, and of 72.5% in vitro generated shoots was successful. On the contrary, mature trees material presented low organogenic response. Axillary bud elongation was recorded just in 10.7% of explants from juvenile shoots and in 6.7% of explants from rejuvenated shoots. The age of donor plant and type of explant affect the organogenic potential of C. montana. This study contributes to the understanding of this species’ response under in vitro conditions.

  10. Induction of bone formation in biphasic calcium phosphate scaffolds by bone morphogenetic protein-2 and primary osteoblasts.

    Science.gov (United States)

    Strobel, L A; Rath, S N; Maier, A K; Beier, J P; Arkudas, A; Greil, P; Horch, R E; Kneser, U

    2014-03-01

    Bone tissue engineering strategies mainly depend on porous scaffold materials. In this study, novel biphasic calcium phosphate (BCP) matrices were generated by 3D-printing. High porosity was achieved by starch consolidation. This study aimed to characterise the porous BCP-scaffold properties and interactions of osteogenic cells and growth factors under in vivo conditions. Five differently treated constructs were implanted subcutaneously in syngeneic rats: plain BCP constructs (group A), constructs pre-treated with BMP-2 (group B; 1.6 µg BMP-2 per scaffold), seeded with primary osteoblasts (OB) (group C), seeded with OB and BMP-2 (group D) and constructs seeded with OB and pre-cultivated in a flow bioreactor for 6 weeks (group E). After 2, 4 and 6 weeks, specimens were explanted and subjected to histological and molecular biological analyses. Explanted scaffolds were invaded by fibrovascular tissue without significant foreign body reactions. Morphometric analysis demonstrated significantly increased bone formation in samples from group D (OB + BMP-2) compared to all other groups. Samples from groups B-E displayed significant mRNA expression of bone-specific genes after 6 weeks. Pre-cultivation in the flow bioreactor (group E) induced bone formation comparable with group B. In this study, differences in bone distribution between samples with BMP-2 or osteoblasts could be observed. In conclusion, combination of osteoblasts and BMP-2 synergistically enhanced bone formation in novel ceramic scaffolds. These results provide the basis for further experiments in orthotopic defect models with a focus on future applications in orthopaedic and reconstructive surgery.

  11. Bone x-ray

    Science.gov (United States)

    ... or broken bone Bone tumors Degenerative bone conditions Osteomyelitis (inflammation of the bone caused by an infection) ... Multiple myeloma Osgood-Schlatter disease Osteogenesis imperfecta Osteomalacia Osteomyelitis Paget disease of the bone Rickets X-ray ...

  12. Application of Bladder Acellular Matrix in Urinary Bladder Regeneration: The State of the Art and Future Directions

    Directory of Open Access Journals (Sweden)

    Marta Pokrywczynska

    2015-01-01

    Full Text Available Construction of the urinary bladder de novo using tissue engineering technologies is the “holy grail” of reconstructive urology. The search for the ideal biomaterial for urinary bladder reconstruction has been ongoing for decades. One of the most promising biomaterials for this purpose seems to be bladder acellular matrix (BAM. In this review we determine the most important factors, which may affect biological and physical properties of BAM and its regeneration potential in tissue engineered urinary bladder. We also point out the directions in modification of BAM, which include incorporation of exogenous growth factors into the BAM structure. Finally, we discuss the results of the urinary bladder regeneration with cell seeded BAM.

  13. Universal tetanus, diphtheria, acellular pertussis (Tdap) vaccination of adults: What the Canadian public knows and wants to know.

    Science.gov (United States)

    Halperin, B A; MacDougall, D; MacKinnon-Cameron, D; Li, L; McNeil, S A; Langley, J M; Halperin, S A

    2015-11-27

    Tetanus, diphtheria, and acellular pertussis vaccine (Tdap) is recommended for all adults in Canada but uptake is low. This study measured the knowledge, attitudes, beliefs, and behaviors of Canadian adults to identify potential barriers and facilitators to Tdap uptake. A survey was undertaken on a geographically representative sample of Canadian adults (n=4023) and 8 focus groups (62 participants) were conducted nationwide. The survey revealed that knowledge about pertussis and Tdap was low (38.3% correct answers). Only 36.0% of respondents reported being aware that all adults were recommended to receive Tdap and only 10.7% reported being immunized; 36.7% did not know whether they had received Tdap. Respondents who were aware of the immunization recommendations were twice as likely to be immunized (16.6% vs. 8.3%; pvaccination with Tdap are not reaching the general public in Canada and an alternative strategy will be required to improve Tdap vaccine uptake.

  14. Differential effect of 17-beta-estradiol on smooth muscle cell and aortic explant MMP2.

    Science.gov (United States)

    Woodrum, Derek T; Ford, John W; Cho, Brenda S; Hannawa, Kevin K; Stanley, James C; Henke, Peter K; Upchurch, Gilbert R

    2009-07-01

    The present investigation tested the hypothesis that intrinsic gender-related differences exist in rat aortic smooth muscle cell matrix metalloproteinase 2 (MMP2). This investigation comprised 3 sets of experiments. Experiment I: Adult male and female rat aortic smooth muscle cells (RASMCs) at passages 4-8 were stimulated in serum-free media for 48 h with interleukin(IL)1beta at doses encountered in human abdominal aortic aneurysms (2 ng/mL). Messenger RNA was extracted from the RASMCs, and gene expression of MMP2 and tissue inhibitor of metalloproteinase 2 (TIMP2), a major MMP2 inhibitor, was measured by real-time polymerase chain reaction. MMP2 protein levels in conditioned media were measured by Western blotting, and MMP2 and TIMP2 activity quantified by standard and reverse gelatin zymography. Experiment II: Male and female RASMCs were incubated for 48 h in Dulbecco's modified Eagler's medium containing IL-1beta and 17-beta-estradiol at doses from 1x10(-10) to 1x10(-6) molar. MMP2 activity in the conditioned media was then determined. Experiment III: Male rats underwent sustained 17-beta-estradiol exposure for 21 d using extended-release, subcutaneously implanted pellets prior to sacrifice and aortic explantation. Aortas from males, females, and estradiol-treated males were stimulated with IL-1beta for 48-h, and MMP2 activity in the conditioned media was determined. Experiment I: MMP2 gene expression was 3-fold higher in male compared with female IL-1beta stimulated RASMCs (P<0.0001). MMP2:TIMP2 gene expression ratio was 7.5-fold greater in male versus female RASMCs. MMP2 protein levels were 3-fold higher (2.68 versus 0.96 o.d./mg total protein, P=0.003) in male versus female RASMCs. Gelatinolytic activity was more than 6-fold higher (15,010 versus 2,472 o.d./mg total protein, P=0.002) in male versus female RASMCs. Experiment II: MMP2 activity in male and female RASMCs was not altered by a wide range of 17-beta-estradiol concentrations. Experiment III: When

  15. Differential Effect of 17-β-Estradiol on Smooth Muscle Cell and Aortic Explant MMP2

    Science.gov (United States)

    Woodrum, Derek T.; Ford, John W; Cho, Brenda S.; Hannawa, Kevin K.; Stanley, James C.; Henke, Peter K.; Upchurch, Gilbert R.

    2010-01-01

    Objective The present investigation tested the hypothesis that intrinsic gender-related differences exist in rat aortic smooth muscle cell MMP2. Methods This investigation comprised three sets of experiments. Experiment I: Adult male and female rat aortic smooth muscle cells (RASMCs) at passages 4–8 were stimulated in serum-free media for 48 hours with IL1β at doses encountered in human AAAs (2ng/mL). Messenger RNA was extracted from the RASMCs, and gene expression of MMP2 and tissue inhibitor of metalloproteinase2 (TIMP2),a major MMP2 inhibitor, was measured by real-time polymerase chain reaction. MMP2 protein levels in conditioned media were measured by Western Blotting, and MMP2 and TIMP2 activity quantified by standard and reverse gelatin zymography. Experiment II: Male and female RASMCs were incubated for 48 hrs in DMEM containing IL-1β and 17-β-estradiol at doses from 1×10−10 to 1×10−6 molar. MMP2 activity in the conditioned media was then determined. Experiment III: Male rats underwent sustained 17-β-estradiol exposure for 21 days using extended-release, subcutaneously implanted pellets prior to sacrifice and aortic explantation. Aortas from males, females, and estradiol-treated males were stimulated with IL1β for 48 hrs, and MMP2 activity in the conditioned media was determined. Results Experiment I: MMP2 gene expression was 3-fold higher in male compared to female IL1β stimulated RASMCs (P<0.0001). MMP2: TIMP2 gene expression ratio was 7.5 fold greater in male vs. female RASMCs. MMP2 protein levels were 3-fold higher (2.68 vs. 0.96 O.D./mg total protein, P=0.003) in male vs. female RASMCs. Gelatinolytic activity was more than 6-fold higher (15,010 vs. 2,472 O.D./mg total protein, P=0.002) in male vs. female RASMCs. Experiment II: MMP2 activity in male and female RASMCs was not altered by a wide range of 17-β-estradiol concentrations. Experiment III: When pre-treated with 17-β-estradiol, MMP2 activity in the media of male rat whole

  16. ORGANOGÊNESE INDIRETA A PARTIR DE EXPLANTES FOLIARES E MULTIPLICAÇÃO IN VITRO DE BROTAÇÕES DE Eucalyptus benthamii X Eucalyptus dunnii

    Directory of Open Access Journals (Sweden)

    Yohana de Oliveira-Cauduro

    2014-01-01

    Full Text Available The aims of this research were to evaluate different culture media for indirect organogenesis and shoot multiplication of Eucalyptus benthamii x Eucalyptus dunnii . For organogenesis, leaf explants were used to test the following treatments: two culture media (MS N/2 and JADS supplemented with 0.1 μM 1-naphthaleneacetic acid (NAA and thidiazuron (TDZ (0.1 or 0.5 μ M, with or without PVP- 40 (250 mg L -1 . The percentage of oxidized explants, callus forming explants, explants with anthocyanin,buds, shoots and the shoot number per explant were evaluated. In the multiplication experiment, isolated shoots were cultivated in MS, JADS and WPM media, all supplemented with 1.11 μ M BAP. Four subcultures were carried out every 28 days. In every subculture the explant oxidation, partial or total leaf chlorosis, fresh mass and mean number of shoot per explant were evaluated. The MS N/2 medium supplemented with 0.1 μM NAA and 0.5 μM TDZ promoted the highest rate of organogenesis (8.3% and the culture media MS supplemented with 1.11 μ M BAP the multiplication rate was higher than in the other media, in the first and the second subcultures (9.28 and 9.24, respectively, without differences between the three media in the following subcultures.

  17. Bone pain

    DEFF Research Database (Denmark)

    Frost, Charlotte Ørsted; Hansen, Rikke Rie; Heegaard, Anne-Marie

    2016-01-01

    Skeletal conditions are common causes of chronic pain and there is an unmet medical need for improved treatment options. Bone pain is currently managed with disease modifying agents and/or analgesics depending on the condition. Disease modifying agents affect the underlying pathophysiology...... of the disease and reduce as a secondary effect bone pain. Antiresorptive and anabolic agents, such as bisphosphonates and intermittent parathyroid hormone (1-34), respectively, have proven effective as pain relieving agents. Cathepsin K inhibitors and anti-sclerostin antibodies hold, due to their disease...... modifying effects, promise of a pain relieving effect. NSAIDs and opioids are widely employed in the treatment of bone pain. However, recent preclinical findings demonstrating a unique neuronal innervation of bone tissue and sprouting of sensory nerve fibers open for new treatment possibilities....

  18. Bone graft

    Science.gov (United States)

    ... around the area. The bone graft can be held in place with pins, plates, or screws. Why ... Orthopaedic Surgery, San Francosco, CA. Also reviewed by David Zieve, MD, MHA, Isla Ogilvie, PhD, and the ...

  19. In vitro plant regeneration of Albizia lebbeck (L. from seed explants

    Directory of Open Access Journals (Sweden)

    S. Perveen

    2013-07-01

    Full Text Available Objectives: An efficient and reproducible regeneration protocol for rapid multiplication of Albizia lebbeck (L. was developed by using intact seed explants.Methods: Murashige and Skoog's (MS medium supplemented with different hormones (BA, Kn, GA3 and TDZ was used for the induction of multiple shoots from the seed explants. Ex-vitro rooting was performed by using pulse treatment method in auxins (IBA and NAA and the complete plantlets were transferred to the field.Results: High frequency direct shoot induction was found in aseptic seed cultures of A. lebbeck on Murashige and Skoog medium supplemented with 5.0 µM TDZ (Thiadiazuron. Seeds were germinated after 7 days of culture and induced maximum 8 shoots from the region adjacent to the apex of the primary shoot of the seedling upto 25 days of incubation. Proliferating shoot cultures with increased shoot length was established by sub-culture of excised sprouting epicotyls on MS medium supplied with reduced concentrations of TDZ. Maximum shoot regeneration frequency (76 % with  highest number of shoots (21 and shoot length (5.1 cm per sprouting epicotyl was observed in the MS medium supplemented with 0.5 µM TDZ after 8 weeks of culture. Different concentrations of Indole-3-butyric acid (IBA and α-naphthalene acetic acid (NAA were tested to determine the optimal conditions for ex-vitro rooting of the microshoots. The best treatment for maximum ex-vitro root induction frequency (81 % was accomplished with IBA (250 µM pulse treatment given to the basal end of the microshoots for 30 min followed by their transfer in plastic cups containing soilrite and eventually established in normal garden soil + soilrite (1:1 with 78 % survival rate. In addition, histological study was undertaken to gain a better understanding of the regenerated shoots from the epicotyl region.Conclusion: The findings will be fruitful in getting a time saving and cost effective protocol for the in vitro propagation of Albizia

  20. Construction and evaluation of acellular matrix for ureter tissue engineering%输尿管无细胞基质移植物的制备和评价

    Institute of Scientific and Technical Information of China (English)

    沈海波; 潘俊; 陈方

    2009-01-01

    的种子细胞具备一定的生长能力.%BACKGROUND:Compared to small-intestine submucosa,acellular vascular grafts have natural tubic structure,which is similar to ureter.When it is used as replacement for ureter,end-to-end anastomosis is used.It is characterized by simple operation,smooth vessel wall,collection and preparation method.OBJECTIVE:To prepare acellular vascular matrix as scaffold in tissue-engineered ureter in vitro.DESIGN,TIME AND SETTING:The observational experiment was performed at the Animal Experimental Center of Xinhua Hospital Affiliated to School of Medicine,Shanghai Jiao Tong University from September 2006 to June 2008.MATERIALS:Swines were supplied by Shanghai Song Lian experimental animals company.Eight healthy adult rats were supplied by Animal Experimental Center Affiliated to School of Medicine,Shanghai Jiao Tong University for the animal toxicity study of acellular vascular matrix.METHODS:Swine carotid artery membrana was removed and placed in phosphate-buffed saline(PBS,pH 7.1).The tissue was stirred at 4 ℃ with 0.5%sodium dodecylsulfate for 24 hours.Then the tissue was treated by double distilled water for 1 week at 4 ℃.Double distilled water was changed twice every day.Vessel with muscle were digested in mixed digestive juice at 37 ℃ for 2 hours before washing.The acellular matrix was stored in penicillin and streptomycin solution at 4 ℃.MAIN OUTCOME MEASURES:The components of acellular scaffold was investigated by optical and electron microscopes.Allogenic endothelial progenitor cells following proliferation were transplanted into acellular vascular matrix to observe cell growth.Animal toxicity study of acellular vascular matrix was performed.Tensile force study was employed to understand the contractility of acellular vascular matrix.RESULTS:Acellular vascular matrix was without cell component.Acellular vascular matrix was mainly composed of collagen.Under scanning electron microscope,cells and cell debris were not found

  1. Low Bone Density

    Science.gov (United States)

    ... Information › Bone Density Exam/Testing › Low Bone Density Low Bone Density Low bone density is when your ... compared to people with normal bone density. Detecting Low Bone Density A bone density test will determine ...

  2. Plant regeneration of Rhabdadenia Ragonesei (Apocynaceae by in vitro culturing of leaf explants REGENERACIÓN DE PLANTAS DE RHABDADENIA RAGONESEI (APOCYNACEAE POR CULTIVO IN VITRO DE EXPLANTES FOLIARES

    Directory of Open Access Journals (Sweden)

    Eduardo Flachsland

    2010-08-01

    Full Text Available Plants of Rhabdadenia Ragonesei Woodson (Apocynaceae were regenerated in vitro from leaves explants. The procedure employed includes: 1 Surface sterilization of leaves by immersion in 70% ethanol (10 s followed by 1,1%NaOCl (15 min and three wash with sterile distilled water. 2 Callus and buds induction by culture on Murashige and Skoog medium (MS + 3 mg/L benzyladenine (BAP. 3 Subculture of callus and buds on MS + 1 mg/L BAP, and 4 Rooting on MS + 0.5 mg/L naftalenacetic acid Se regeneraron plantas de Rhabdadenia Ragonesei Woodson (Apocynaceae mediante el cultivo in vitro de explantes foliares en condiciones ambientales controladas. El procedimiento consistió en: 1 Desinfección de las hojas por inmersión en etanol al 70% (10 s seguida de Inmersión en NaOCl al 1,1% (15 min y lavado tres veces con agua destilada estéril. 2 Inducción de callos y yemas mediante el cultivo de explantes foliares en el medio de Murashige y Skoog (MS + 3 mg/L de benciladenina (BAP. 3 Subcultivo de callos y yemas en MS + 1 mg/L de BAP y 4 Enraizamiento de los vastagos obtenidos en MS + 0,5 mg/L de ácido naftalenacético

  3. ORGANOGÊNESE IN VITRO DE Citrus EM FUNÇÃO DE CONCENTRAÇÕES DE BAP E SECCIONAMENTO DO EXPLANTE CITRUS IN VITRO ORGANOGENESIS RELATED TO BAP CONCENTRATIONS AND EXPLANT SECTION

    Directory of Open Access Journals (Sweden)

    THAÍS LACAVA DE MOURA

    2001-08-01

    Full Text Available O sucesso de técnicas biotecnológicas no melhoramento in vitro de Citrus depende diretamente do desenvolvimento de protocolos eficientes para regeneração de plantas. Objetivou-se avaliar o efeito de concentrações de 6-benzilaminopuria (BAP na organogênese in vitro de limão-'Cravo' e laranja-'Pêra', bem como o efeito do seccionamento do explante em laranja-'Valência'. Para o limão-'Cravo', foram utilizados como explante, segmentos internodais de plântulas germinadas in vitro, cultivados em meio MT e variando-se as concentrações de BAP em 0; 2,5; 5; 7,5 e 10 mg.L-1. Nas laranjas-'Pêra' e 'Valência' os explantes foram segmentos do epicótilo de plântulas germinadas in vitro. Os explantes de laranja-'Pêra' foram cultivados em meio MT variando-se as concentrações de BAP em 0; 1; 2; 3 e 4 mg.L-1. Para a laranja-'Valência', metade dos explantes foram seccionados e cultivados em meio MT acrescido de 1,0 mg.L-1 de BAP. Todas as brotações obtidas foram alongadas no meio de cultura MT + 25 g.L-1 de sacarose + 1 mg.L-1 de ácido giberélico (GA3 e enraizadas no meio MT + 25 g.L-1 de sacarose + 0,5 g.L-1 de carvão ativado + 1 mg.L-1 de ácido naftaleno acético (ANA. O melhor resultado para o número de brotações adventícias foi obtido na concentração 2,5 mg.L-1 de BAP para limão-'Cravo', e nas concentrações 1,0 e 2,0 mg.L-1 de BAP para laranja-'Pêra'. O seccionamento dos explantes favoreceu a organogênese in vitro da laranja-'Valência', porém as brotações apresentaram menor índice de enraizamento.The establishment of efficient plant regeneration protocols is essential for the success and application of in vitro breeding biotechnologies in Citrus. The objective of this work was to verify the effect of 6-benzilaminopurine (BAP on the in vitro organogenesis of Rangpur lime (Citrus limonia (L. Osbeck and 'Pera' sweet orange (Citrus sinensis (L. Osbeck, and the effect of cutting the explant on the in vitro organogenesis of

  4. Cells that emerge from embryonic explants produce fibers of type IV collagen.

    Science.gov (United States)

    Chen, J M; Little, C D

    1985-10-01

    Double immunofluorescence staining experiments designed to examine the synthesis and deposition of collagen types I and IV in cultured explants of embryonic mouse lung revealed the presence of connective tissue-like fibers that were immunoreactive with anti-type IV collagen antibodies. This observation is contrary to the widely accepted belief that type IV collagen is found only in sheet-like arrangements beneath epithelia or as a sheath-like layer enveloping bundles of nerve or muscle cells. The extracellular matrix produced by cells that migrate from embryonic mouse lung rudiments in vitro was examined by double indirect immunofluorescence microscopy. Affinity-purified monospecific polyclonal antibodies were used to examine cells after growth on glass or native collagen substrata. The data show that embryonic mesenchymal cells can produce organized fibers of type IV collagen that are not contained within a basement membrane, and that embryonic epithelial cells deposit fibers and strands of type IV collagen beneath their basal surface when grown on glass; however, when grown on a rat tail collagen substratum the epithelial cells produce a fine meshwork. To our knowledge this work represents the first report that type IV collagen can be organized by cells into a fibrous extracellular matrix that is not a basement membrane.

  5. Visible effects of rapamycin (sirolimus) on human skin explants in vitro.

    Science.gov (United States)

    Peramo, Antonio; Marcelo, Cynthia L

    2013-03-01

    In this manuscript, we report observations of the effects of rapamycin in an organotypic culture of human skin explants. The tissues were cultured for 5 days at the air-liquid interface or in submersed conditions with media with and without rapamycin at 2 nM concentration. Histological analysis of tissue sections indicated that rapamycin-treated samples maintained a better epidermal structure in the upper layers of the tissue than untreated samples, mostly evident when skin was cultured in submersed conditions. A significant decrease in the number of positive proliferative cells using the Ki67 antigen was observed when specimens were treated with rapamycin, in both air-liquid and submersed conditions but apoptosis differences between treated and untreated specimens, as seen by cleaved caspase-3 positive cells, were only observed in submersed specimens. Finally, a decrease and variability in the location in the expression of the differentiation marker involucrin and in E-cadherin were also evident in submersed samples. These results suggest that the development of topical applications containing rapamycin, instead of systemic delivery, may be a useful tool in the treatment of skin diseases that require reduction of proliferation and modulation or control of keratinocyte differentiation.

  6. Flask sealing on in vitro seed germination and morphogenesis of two types of ornamental pepper explants

    Directory of Open Access Journals (Sweden)

    Diego Silva Batista

    Full Text Available ABSTRACT: The influence of flask sealing and explant source on the in vitro morphogenesis of pepper, were evaluated in Capsicum annuum. Seeds were sterilized and inoculated in Murashige and Skoog media supplemented with vitamins, myo-inositol, sucrose, and agar. Gas exchange was evaluated in the germination stage by comparing 3 flask-sealing systems: rigid polypropylene lids (PLs without vents, PLs with 1 vent, and PLs with 2 vents covered with membranes. In the regeneration stage, cotyledon and hypocotyl segments were transferred to the organogenesis-inducing media, being the different sealing types also tested in a factorial scheme. Photosynthetic pigments, morphological and hystological analyses were conducted for each treatment. Plants maintained in glass flasks capped with vented lids showed more vigorous growth and differentiated anatomical structures. These treatments resulted in taller plants, higher numbers and more expanded leaves, higher fresh and dry weights, and an increase in photosynthetic pigments. Cultivation of C. annuum in flasks with reduced gas exchange was more effective on callus induction. During the regeneration stage, hypocotyls were more effective than cotyledons. Sealing type influenced the morphogenic responses of pepper, demonstrating that an increase in gas exchange has a positive effect on biomass production and acclimatization of the plantlets.

  7. Morphogenic responses of three explants of Lupinus montanus (H.B.K. cultured in vitro

    Directory of Open Access Journals (Sweden)

    Gabriel Ramírez-González

    2015-01-01

    Full Text Available Resumen El necrosamiento y la respuesta morfogénica de explantes de cotiledón, epicótilo e hipocótilo de plántulas de Lupinus montanus (H.B.K. germinadas in vitro se evaluaron para establecer las condiciones básicas de micropropagación. El necrosamiento se evaluó en medio MS con 0.40 mg·litro-1 de tiamina, 100 mg·litro-1 de mioinositol, 3 % de sacarosa y 7 g·litro-1 de agar-agar y distintas dosis de carbón activado (CA y ácido cítrico. Los tratamientos con CA (50 mg·litro-1 y 100 mg·litro-1 fueron los mejores en el control del necrosamiento (P = 0.001; el cotiledón tuvo mayor nivel de necrosamiento ( 70 % a diferencia del cotiledón, cuya respuesta fue primordialmente callogénica (50 %. El epicótilo cultivado en medio MS con 3.0 μM de AIA y 1.0 μM de BA tuvo el mayor número de brotes (10 y altura (11.4 ± 2.6 cm.

  8. In Vitro Regeneration of Shoots From Nodal Explants of Dendrobium Chrysotoxum Lindl

    Directory of Open Access Journals (Sweden)

    Kaur Saranjeet

    2017-06-01

    Full Text Available Transverse sections (2 mm thickness of stem-nodes from in vitro raised seedlings had morphogenic potential on semisolid and liquid Murashige and Skoog medium supplemented with cytokinins N6-benzyladenine (BA 4.44 μM, furfurylaminopurine (KIN 4.65 μM and auxin α-naphthalene acetic acid (NAA 5.37 μM individually and in combinations. The regeneration response was influenced by both the type of growth regulator and physical state of the medium. The explants produced either shoot buds on cytokinincontaining media or protocorm-like bodies (PLBs on NAA containing media both solid and liquid. More neo-formations were produced on liquid media, especially those containing only NAA. They were formed at nodal and inter-nodal regions. The secondary buds were produced on the surface of primary PLBs. The plantlets were developed on MS medium containing banana homogenate 50 g·dm-3. The current study is the first ever report on successful regeneration of Dendrobium chrysotoxum from stem-node segments.

  9. Epidermal DNA synthesis in organ culture explants. A study of hairless mouse ear epidermis.

    Science.gov (United States)

    Hansteen, I L; Iversen, O H; Refsum, S B

    1979-10-01

    Explants of split mouse ear were incubated in organ culture for up to 48 h, and the cell proliferation was studied by the addition of Thymidine-methyl-3-H (3HTdR) to the medium during different time periods, mainly for the first 14 h of incubation. Cultures were started at 0900, 2130 and 2300. In all cases the labelling index remained stable for 6-8 h, and then increased. The mean grain count, however, was falling and so was the epidermal DNA-specific uptake of 3HTdR. Based on the experimental results, calculations can be made of the flux of cells through S. It is concluded that the increasing LI is not due to inherent diurnal variation in cell proliferation, and is not a sign of real growth but caused instead by a complete block of the cell exit from S, probably combined with periods of an increased entrance rate into S. Other methodological factors, however, may also contribute to the increasing LI. Hence, this system is not suited for the measurement of factors that influence epidermal DNA synthesis.

  10. Activin A induces Langerhans cell differentiation in vitro and in human skin explants.

    Directory of Open Access Journals (Sweden)

    Tiziana Musso

    Full Text Available Langerhans cells (LC represent a well characterized subset of dendritic cells located in the epidermis of skin and mucosae. In vivo, they originate from resident and blood-borne precursors in the presence of keratinocyte-derived TGFbeta. In vitro, LC can be generated from monocytes in the presence of GM-CSF, IL-4 and TGFbeta. However, the signals that induce LC during an inflammatory reaction are not fully investigated. Here we report that Activin A, a TGFbeta family member induced by pro-inflammatory cytokines and involved in skin morphogenesis and wound healing, induces the differentiation of human monocytes into LC in the absence of TGFbeta. Activin A-induced LC are Langerin+, Birbeck granules+, E-cadherin+, CLA+ and CCR6+ and possess typical APC functions. In human skin explants, intradermal injection of Activin A increased the number of CD1a+ and Langerin+ cells in both the epidermis and dermis by promoting the differentiation of resident precursor cells. High levels of Activin A were present in the upper epidermal layers and in the dermis of Lichen Planus biopsies in association with a marked infiltration of CD1a+ and Langerin+ cells. This study reports that Activin A induces the differentiation of circulating CD14+ cells into LC. Since Activin A is abundantly produced during inflammatory conditions which are also characterized by increased numbers of LC, we propose that this cytokine represents a new pathway, alternative to TGFbeta, responsible for LC differentiation during inflammatory/autoimmune conditions.

  11. In vitro neurotoxic effects of 1 GeV/n iron particles assessed in retinal explants.

    Science.gov (United States)

    Vazquez, M E; Kirk, E

    2000-01-01

    The heavy ion component of the cosmic radiation remains problematic to the assessment of risk in manned space flight. The biological effectiveness of HZE particles has yet to be established, particularly with regard to nervous tissue. Using heavy ions accelerated at the AGS of Brookhaven National Laboratory, we study the neurotoxic effects of iron particles. We exposed retinal explants, taken from chick embryos, to determine the dose response relationships for neurite outgrowth. Morphometric techniques were used to evaluate the in vitro effects of 1 GeV/a iron particles (LET 148 keV/micrometer). Iron particles produced a dose-dependent reduction of neurite outgrowth with a maximal effect achieved with a dose of 100 cGy. Doses as low as 10-50 cGy were able to induce reductions of the neurite outgrowth as compared to the control group. Neurite generation is a more sensitive parameter than neurite elongation, suggesting different mechanism of radiation damage in our model. These results showed that low doses/fluences of iron particles could impair the retinal ganglion cells' capacity to generate neurites indicating the highly neurotoxic capability of this heavy charged particle.

  12. In Vitro Callus Induction and Embryogenesis of Oil Palm (Elaeis guineensis Jacq. from Leaf Explants

    Directory of Open Access Journals (Sweden)

    DWI HAPSORO

    2011-06-01

    Full Text Available This research was to study in vitro callus induction and somatic embryogenesis in oil palm from leaf explants. Young leaf segments from mature oil palm were cultured on MS medium supplemented with different concentrations of 2,4-D with or without addition of 2 g/l activated charcoal (AC or 2,4-D and picloram. Embryogenesis induction was done using MS medium containing 2,4-D 450 M and benziladenine 4.4 M with 3g/l activated charcoal. The treatment of 2,4-D 15 M resulted in the highest percentage of callus induction. The treatment of 2,4-D and AC showed that 2,4-D 450 M and AC led to higher percentage of callus induction than that of 2,4-D 400 M and 2 g/l AC. Embryogenesis occured in 27 out of 250 clumps of primary callus was occurred after 2-3 times subcultures. Somatic embryo development occurred when the embryogenic callus was transferred on the same basal medium supplemented with casein hydrolysate with 1 M BA or growth regulator free basal medium with 2 g/l activated charcoal.

  13. In Vitro Shoot Bud Differentiation from Hypocotyl Explants of Chili Peppers (Capsicum annuum L.

    Directory of Open Access Journals (Sweden)

    Owk ANIEL KUMAR

    2010-03-01

    Full Text Available Chili pepper (Capsicum annuum L. is an economically important spice crop in tropical and subtropical countries. In vitro plant regeneration was obtained from 15th day old hypocotyl explants of three chili pepper cultivars (Capsicum annuum L., var. �X-235�, var. �PC-1� and var. �Pusa Jwala�. Among the genotypes of Capsicum L. var. �X-235� responded better than the var. �PC-1� and var. �Pusa Jwala�. MS medium containing BAP (4.0 mg/l and IAA (0.5 mg/l was found to be the best medium for the production of maximum number of shoot buds in all the genotypes of chili pepper i.e., 6.80�0.16 (var. �X-235�, 5.00�0.19 (var. �PC-1� and 4.80�0.12 (var. �Pusa Jwala�. The shoots were rooted on MS medium fortified with IBA (0.5 mg/l. Rooted plants were hardened and transplanted to the soil. The plants showed 80-90% survival during transplantation.

  14. In Vitro Shoot Bud Differentiation from Hypocotyl Explants of Chili Peppers (Capsicum annuum L.

    Directory of Open Access Journals (Sweden)

    Owk ANIEL KUMAR

    2010-03-01

    Full Text Available Chili pepper (Capsicum annuum L. is an economically important spice crop in tropical and subtropical countries. In vitro plant regeneration was obtained from 15th day old hypocotyl explants of three chili pepper cultivars (Capsicum annuum L., var. X-235, var. PC-1 and var. Pusa Jwala. Among the genotypes of Capsicum L. var. X-235 responded better than the var. PC-1 and var. Pusa Jwala. MS medium containing BAP (4.0 mg/l and IAA (0.5 mg/l was found to be the best medium for the production of maximum number of shoot buds in all the genotypes of chili pepper i.e., 6.800.16 (var. X-235, 5.000.19 (var. PC-1 and 4.800.12 (var. Pusa Jwala. The shoots were rooted on MS medium fortified with IBA (0.5 mg/l. Rooted plants were hardened and transplanted to the soil. The plants showed 80-90% survival during transplantation.

  15. Efficient soybean regeneration and Agrobacterium-mediated transformation using a whole cotyledonary node as an explant.

    Science.gov (United States)

    Zhang, Fuli; Chen, Can; Ge, Honglian; Liu, Jinmei; Luo, Yunling; Liu, Kun; Chen, Long; Xu, Kedong; Zhang, Yi; Tan, Guangxuan; Li, Chengwei

    2014-01-01

    An optimized regeneration and Agrobacterium-mediated transformation protocol based on whole cotyledonary node explants was developed in soybean (Glycine max) cultivar Zhong Huang 13. Adding 6-benzylaminopurine (BAP) in a germinating medium could significantly increase regeneration efficiency; the optimal BAP concentration for shoot formation was 0.5 mg/L. The concentrations of plant growth regulators in a shoot induction medium were optimized by the orthogonal test [L9 (3(3))]. The best combination for shoot regeneration was a medium of Murashige & Skoog salts with B5 vitamins (MSB) supplemented with 3.5 mg/L BAP, 0.2 mg/L indole-3-butyric acid (IBA), and 0.2 mg/L kinetin (KT). Under this favorable condition, one node could regenerate 28-30 shoots. Soybean whole cotyledonary nodes were transformed by inoculation with A. tumefaciens strain EHA105 harboring a vector pBI121 containing a β-glucuronidase gene (gus). GUS assay, polymerase chain reaction, and Southern blot analysis indicated that the gus gene was transformed into soybean plants with 23.1% transformation efficiency. Transgenic plants could be obtained within 5-6 weeks, which was about 4 weeks less than that of a traditional single cotyledonary node method.

  16. In vitro plant regeneration of two cucumber (Cucumis sativum L. genotypes: Effects of explant types and culture medium

    Directory of Open Access Journals (Sweden)

    Grozeva Stanislava

    2014-01-01

    Full Text Available The effect of different phytohormone concentrations on callusogenesis and organogenesis in two cucumber genotypes were studied. It was established that the rate of plant regeneration depends on genotype, explant type and culture medium. Hypocotyls were found to be more responsive than cotyledons in morphogenesis. In vitro planlet-regenerants have been obtained in hypocotyls explants on culture medium with 1.0 and 2.0 mgL-1 BA for cultivar Gergana and in 1.0 and 3.0 mgL-1K-line 15B. Induction of regeneration in cotyledons were established only in cultivar Gergana on culture medium supplemented with 3.0 mgL-1 BA and in combination of 0.5 mgL-1IAA.

  17. Changes of free, soluble conjugated and bound polyamine titers of jojoba explants under sodium chloride salinity in vitro.

    Science.gov (United States)

    Roussos, Peter A; Pontikis, Constantine A

    2007-07-01

    Jojoba (Simmondsia chinensis L.) single node explants were cultured in a basal medium supplemented with 17.8 microM 6-benzyladenine and four levels of sodium chloride concentration (0, 56.41, 112.82 and 169.23 mM). The free, the soluble conjugated and the insoluble bound forms of polyamines (PAs) (putrescine (Put), spermidine (Spd) and spermine (Spm)) were determined monthly during a 3-month proliferation stage. Free Put and Spd were found in higher levels in the control treatment, while Spm content was higher in the salt treatments. All soluble conjugated PAs were found to be in lower concentrations in explants growing on medium supplemented with salt, while the opposite was true for the insoluble bound PAs. It appeared that certain PAs and PAs forms could play a significant role in the adaptation mechanism of jojoba under saline conditions.

  18. Effect of growth regulators and explant origin on in vitro propagation of Ceratonia siliqua L. via cuttings

    Directory of Open Access Journals (Sweden)

    Boussaid M.

    2008-01-01

    Full Text Available The present work was undertaken to develop a basic and simple protocol for micropropagation of Ceratonia siliqua. Axillaries bud sprouting and shoot development were stimulated on MS supplemented with BAP (0.5 mg.l-1, IBA (0.1 mg.l-1 and GA3 (0.5 mg.l-1, shoot multiplication was obtained on MS supplemented with BAP (2 mg.l-1 and rooting of microshoots was achieved on MS supplemented with IBA (2 mg.l-1 and charcoal (2 mg.l-1. The best results were obtained with herbaceous explants taken from juvenile trees. Significant differences in proliferation, multiplication and rooting due to the type and origin of explant and to the concentration of growth regulators were found.

  19. The effect of protease inhibitors on the induction of osteoarthritis-related biomarkers in bovine full-depth cartilage explants

    DEFF Research Database (Denmark)

    He, Yi; Zheng, Qinlong; Jiang, Mengmeng

    2015-01-01

    Objective The specific degradation of type II collagen and aggrecan by matrix metalloproteinase (MMP)-9, -13 and ADAMTS-4 and -5 (aggrecanase-1 and -2) in the cartilage matrix is a critical step in pathology of osteoarthritis (OA). The aims of this study were: i) To investigate the relative...... contribution of ADAMTS-4 and ADAMTS-5 to cartilage degradation upon catabolic stimulation; ii) To investigate the effect of regulating the activities of key enzymes by mean of broad-spectrum inhibitors. Methods Bovine full-depth cartilage explants stimulated with tumor necrosis factor alpha (TNF......I (ADAMTS-degraded aggrecan), AGNxII (MMP-degraded aggrecan), and CTX-II (MMP-derived type II collagen) were quantified in the explants-conditioned media. Results We found that: i) Active ADAMTS-4, MMP-9, -13 were released in the late stage of TNF-α/ OSM stimulation, whereas no significant active ADAMTS-5...

  20. Microvesicle-mediated release of soluble LH/hCG receptor (LHCGR from transfected cells and placenta explants

    Directory of Open Access Journals (Sweden)

    Randeva Harpal

    2011-05-01

    Full Text Available Abstract Placental hCG and pitutary LH transduce signals in target tissues through a common receptor (LHCGR. We demonstrate that recombinant LHCGR proteins which include the hormone-binding domain are secreted from transfected cells and that natural LHCGR is also secreted from human placental explants. LHCGR recombinant proteins representing varying lengths of the N-terminal extracellular domain were expressed in Chinese Hamster Ovary cells in suspension culture. Secretion was minimal up to 72h but by 96h 24-37% of the LHCGR had been released into the culture medium. The secreted proteins were folded and sensitive to glycosidases suggesting N-linked glycosylation. Secretion was independent of recombinant size and was mediated via structurally defined membrane vesicles (50-150nm. Similarly cultured human early pregnancy placental explants also released LHCGR via microvesicles. These studies provide the first experimental evidence of the possible mechanistic basis of the secretion of LHCGR.

  1. Hypotonic challenge modulates cell volumes differently in the superficial zone of intact articular cartilage and cartilage explant.

    Science.gov (United States)

    Turunen, Siru M; Lammi, Mikko J; Saarakkala, Simo; Koistinen, Arto; Korhonen, Rami K

    2012-05-01

    The objective of this study was to evaluate the effect of sample preparation on the biomechanical behaviour of chondrocytes. We compared the volumetric and dimensional changes of chondrocytes in the superficial zone (SZ) of intact articular cartilage and cartilage explant before and after a hypotonic challenge. Calcein-AM labelled SZ chondrocytes were imaged with confocal laser scanning microscopy through intact cartilage surfaces and through cut surfaces of cartilage explants. In order to clarify the effect of tissue composition on cell volume changes, Fourier Transform Infrared microspectroscopy was used for estimating the proteoglycan and collagen contents of the samples. In the isotonic medium (300 mOsm), there was a significant difference (p integrity of the mechanical environment of chondrocytes.

  2. Bone marrow transplant

    Science.gov (United States)

    Transplant - bone marrow; Stem cell transplant; Hematopoietic stem cell transplant; Reduced intensity nonmyeloablative transplant; Mini transplant; Allogenic bone marrow transplant; Autologous bone marrow transplant; Umbilical ...

  3. Vacina acelular contra pertússis para adolescentes Acellular pertussis vaccine for adolescents

    Directory of Open Access Journals (Sweden)

    Aroldo P. de Carvalho

    2006-07-01

    vacina é em torno de 6 a 12 anos. As avaliações sobre o impacto econômico do uso rotineiro da vacina em adolescentes evidenciam uma relação custo-benefício positiva. Os resultados do impacto epidemiológico dependem da qualidade do diagnóstico para que os dados reflitam a realidade da doença. CONCLUSÕES: Embora existam algumas questões a serem esclarecidas, a literatura disponível sinaliza a possibilidade para a solução do .ressurgimento. da coqueluche com o uso da vacina dTpa. Talvez a estratégia da utilização de uma dose de reforço na adolescência, substituindo a vacina dupla contra difteria e tétano, seja uma medida a ser prontamente indicada.BACKGROUND: The use of whole-cell pertussis vaccine has led to a significant decline in incidence of the disease among children. This change in the epidemiological profile led to an increased number of cases among teenagers and adults, as a result of loss of immunity to the disease or vaccine after approximately 10 years. An increased number of cases was also observed among non-immunized or partially immunized infants. Licensure of the DTP vaccine against diphtheria, tetanus, and acellular pertussis formulated specifically for patients over 10 years of age (Tdap suggests the possibility of controlling pertussis in the most affected age groups over the past few years. SOURCES OF DATA: Data were collected from MEDLINE. The research was limited to the period between January 1995 and January 2006. SUMMARY OF THE FINDINGS: In some countries there are two Tdap vaccines licensed for patients over 10 years of age. One of them contains five immunogenic components of Bordetella pertussis (pertussis toxin, filamentous hemagglutinin, fimbriae 2 and 3, and pertactin, and the other contains three components (pertactin, filamentous hemagglutinin, and inactivated pertussis toxin, the latter being the only one licensed in Brazil up to now. Although the composition of the two vaccines differs, studies show that they have

  4. Organotypic explant culture of adult rat retina for in vitro investigations of neurodegeneration, neuroprotection and cell transplantation

    OpenAIRE

    sprotocols

    2015-01-01

    This protocol details a method for isolating retinal tissue from adult rats as an organotypic culture to study neurobiological processes in mature tissue. It combines the efficiency and control common to in vitro techniques with close imitation of the in vivo environment. Eyes from adult rats are enucleated and the neural retina is isolated. Tissue is cut into quarters, yielding eight retinal explants per animal, and cultured at a fluid/air interface on organotypic culture membranes. Explanta...

  5. An efficient and reproducible indirect shoot regeneration from female leaf explants of Simmondsia chinensis, a liquid-wax producing shrub.

    Science.gov (United States)

    Bala, Raman; Beniwal, Vijay Singh; Laura, Jitender Singh

    2015-04-01

    Simmondsia chinensis (Link) Schneider is a perennial, dioecious, drought resistant and multipurpose seed oil crop grown in arid and semi-arid conditions throughout the world. A reproducible and more efficient method for indirect shoot organogenesis from female leaf explants has been standardized. The leaf explants cultured on Murashige and Skoog (MS) medium with 1.0 mg l(-1) 2,4-dichlorophenoxyacetic acid (2,4-D) alone produced the highest frequency of callus compared with 1.5 mg l(-1) IBA. Maximum proliferation of callus was observed on MS medium containing a combination of 1.0 mg l(-1) 2,4-D with 0.5 mg l(-1) BAP. For shoot differentiation, the proliferated callus was subcultured on MS medium supplemented with 6-benzylaminopurine (BAP) (1.0-4.0 mg l(-1)) along with 40 mg l(-1) adenine sulphate as additive or in combination with α-naphthalene acetic acid (NAA) or Indole-3-butyric acid (IBA). Optimum shoots differentiated from callus was obtained on MS medium supplemented with 2.0 mg l(-1) BAP and 0.2 mg l(-1) NAA. On this medium, 100 % cultures were responded with an average number of 14.44 shoots per explant with their mean length of 4.78 cm. In vitro rooting (6.22 roots per explant) was achieved on half strength MS medium containing 2 % sucrose with 3.0 mg l(-1) IBA and 300 mg l(-1) activated charcoal (AC). Rooted plantlets were successfully hardened under control conditions and acclimatized under field conditions with 90 % success rate. The present protocol is highly efficient, reproducible and economically viable for large scale production of female plants.

  6. An investigation of donor and culture parameters which influence epithelial outgrowths from cultured human cadaveric limbal explants.

    Science.gov (United States)

    Baylis, Oliver; Rooney, Paul; Figueiredo, Francisco; Lako, Majlinda; Ahmad, Sajjad

    2013-05-01

    Limbal stem cell deficiency is a blinding disease which affects the cornea at the front of the eye. The definitive cure involves replacing the corneal epithelial (limbal) stem cells, for example by transplanting cultured limbal epithelial cells. One method of performing cultures is to grow a sheet of epithelial cells from a limbal explant on human amniotic membrane. The growth of limbal tissue can be variable. The aim of this study is to investigate how different donor and culture factors influence the ex vivo growth of cadaveric limbal explants. Limbal explant cultures were established from 10 different cadaveric organ cultured corneo-scleral discs. The growth rate and the time taken for growth to be established were determined. Statistical analysis was performed to assess correlation between these factors and donor variables including donor age, sex, time from donor death to enucleation, time from enucleation to organ culture storage and duration in organ culture. Growth curves consistently showed a lag phase followed by a steeper linear growth phase. Donor age, time between death and enucleation, and time between enucleation and organ culture were not correlated to the lag time or the growth rate. Time in organ culture had a significant correlation with the duration of lag time (P = 0.003), but no relationship with the linear growth rate. This study shows that an important factor correlating with growth variation is the duration of corneo-scleral tissue in organ culture. Interestingly, donor age was not correlated with limbal explant growth. Copyright © 2012 Wiley Periodicals, Inc.

  7. Plant regeneration of non-toxic Jatropha curcas—impacts of plant growth regulators, source and type of explants

    KAUST Repository

    Kumar, Nitish

    2011-01-28

    Jatropha curcas is an oil bearing species with multiple uses and considerable economic potential as a biofuel plant, however, oil and deoiled cake are toxic. A non-toxic variety of J. curcas is reported from Mexico. The present investigation explores the effects of different plant growth regulators (PGRs) viz. 6-benzyl aminopurine (BAP) or thidiazuron (TDZ) individually and in combination with indole-3-butyric acid (IBA), on regeneration from in vitro and field-grown mature leaf explants, in vitro and glasshouse-grown seedlings cotyledonary leaf explants of non-toxic J. curcas. In all the tested parameters maximum regeneration efficiency (81.07%) and the number of shoot buds per explants (20.17) was observed on 9.08 μM TDZ containing Murashige and Skoog’s (MS) medium from in vitro cotyledonary leaf explants. The regenerated shoot buds were transferred to MS medium containing 10 μM kinetin (Kn), 4.5 μM BAP and 5.5 μM α-naphthaleneacetic acid (NAA) for shoot proliferation. The proliferated shoots could be elongated on MS medium supplemented with 2.25 μM BAP and 8.5 μM IAA. Rooting was achieved when the basal cut end of elongated shoots were dipped in half strength MS liquid medium containing different concentrations and combinations of IBA, IAA and NAA for four days followed by transfer to growth regulators free half strength MS medium supplemented 0.25 mg/l activated charcoal. The rooted plants could be established in soil with more than 90% survival rate.

  8. A comparative evaluation of the effectiveness of subpedicle acellular dermal matrix allograft with subepithelial connective tissue graft in the treatment of isolated marginal tissue recession: A clinical study

    OpenAIRE

    Shori, Tony; Kolte, Abhay; Kher, Vishal; Dharamthok, Swarup; Shrirao, Tushar

    2013-01-01

    Introduction: The most common problem encountered in our day to day practice is exposed root surface or a tooth getting long. The main indication for root coverage procedures are esthetics and/or cosmetic demands followed by the management of root hypersensitivity, root caries or when it hampers proper plaque removal. Over the years, various techniques have been used to achieve root coverage. Aim and Objectives: The aim of this study was to compare the effectiveness of subpedicle acellular de...

  9. Effect of gamma rays on different explants of callus treatment of multiple shoots in Cucumis melo cv. Bathasa.

    Science.gov (United States)

    Venkateshwarlu, M

    2008-09-01

    A mutagenesis programme was carried out using physical mutagens (gamma rays) on Cucumis melo cv. Bathasa. In irradiated seeds the number of shoots formed in the lower doses was significantly higher than the controls. Decrease in the number of shoots and shoot bud formation was observed with higher doses in all the explants studies. There was complete lethality in the 10, 15 and 20 kR doses. In irradiated stem cultures the maximum number of shoots were observed in 2 kR. The number of shoots decreased with increasing doses of irradiation. At higher doses of 4 and 5kR, light green compact callus was formed in almost all the explants. The effect of lower doses of gamma irradiation on shoot bud formation and rooting efficiency from nodal explants cultured on MS +2.0 mgl(-1) L-glutamic acid + 0.5 mgl(-1) BAP, stimulation of shoot and root induction were studied. The number of shoots and root lengths decreased with the increasing dosage of irradiation. The irradiated callus was grown on solidified MS medium with containing 2.0 mgl(-1) BAP + 1.0 mgl(-1) IAA. There was a significant stimulation of growth in the callus at lower doses. At higher doses like 15 and 20 kRs growth was drastically reduced.

  10. Thidiazuron: A potent cytokinin for efficient plant regeneration in Himalayan poplar (Populus ciliata Wall. using leaf explants

    Directory of Open Access Journals (Sweden)

    Gaurav Aggarwal

    2012-11-01

    Full Text Available Populus species are important resource for certain branches of industry and have special roles for scientific study on biological and agricultural systems. The present investigation was undertaken with an objective of enhancing the frequency of plant regeneration in Himalayan poplar (Populus ciliata Wall.. The effect of Thiadizuron (TDZ alone and in combination with adenine and α-Naphthalene acetic acid (NAA were studied on the regeneration potential of leaf explants. A high efficiency of shoot regeneration was observed in leaf (80.00% explants on MS basal medium supplemented with 0.024 mg/l TDZ and 79.7 mg/l adenine. Elongation and multiplication of shoots were obtained on Murashige and Skoog (MS basal medium, containing 0.5 mg/l 6. Benzyl aminopurine (BAP + 0.2mg/l Indole 3-acetic acid (IAA + 0.3 mg/l Gibberellic acid (GA3. High frequency root regeneration from in vitro developed shoots was observed on MS basal medium supplemented with 0.10 mg/l Indole 3-butyric acid(IBA. Maximum of the in vitro rooted plantlets were well accomplished to the mixture of sand: soil (1:1 and exhibited similar morphology with the field plants. A high efficiency plant regeneration protocol has been developedfrom leaf explants in Himalayan poplar (Populus ciliata Wall..

  11. Effect of ZnO nanoparticles on Brassica nigra seedlings and stem explants: growth dynamics and antioxidative response

    Directory of Open Access Journals (Sweden)

    Hira eZafar

    2016-04-01

    Full Text Available Nanoparticles (NPs have diverse properties in comparison to respective chemicals due to structure, surface area ratio, morphology, and reactivity. Toxicological effects of metallic NPs to organisms including plants have been reported. However, to the authors’ knowledge there is no report on the effect of NPs on in vitro culture of plant explants. In this study, ZnO NPs at 500-1500 mg/L badly affected Brassica nigra seed germination and seedling growth and raised antioxidative activities and antioxidants concentrations. On the other hand, culturing the stem explants of B. nigra on Murashige and Skoog (MS medium in presence of low concentration of ZnO NPs (1-20 mg/L produced white thin roots with thick root hairs. At 10 mg/L ZnO NPs shoots emergence was also observed. The developed calli/roots showed 79% DPPH (2,2-diphenyl-1-picryl hydrazyl radical scavenging activity at 10 mg/L. While total antioxidant and reducing power potential were also significantly different in presence of ZnO NPs. Non enzymatic antioxidative molecules, phenolics (up to 0.15 µg GAE/mg FW and flavonoids (up to 0.22 µg QE/mg FW, also raised and found NPs concentration dependent. We state that ZnO NPs may induce roots from explants cultured on appropriate medium and can be cultured for production of valuable secondary metabolites.

  12. Cholangiocarcinoma Secondary to Primary Sclerosing Cholangitis in Explanted Livers: A Single-Center Study in the South of Iran

    Directory of Open Access Journals (Sweden)

    Geramizadeh

    2015-12-01

    Full Text Available Background Primary sclerosing cholangitis (PSC is a chronic disease, characterized by chronic inflammation and fibrosis of bile duct epithelial cells. This is a significant contributory factor to the development of malignancy, most commonly cholangiocarcinoma (CCA, which is the second most common malignant liver tumor. Objectives For the first time in Iran, we intend to describe our experience with cases of PSC, with and without CCA, in explanted livers, and compare our results with those found in other areas of the world. Patients and Methods The study population comprised 181 individuals with a diagnosis of PSC who had undergone liver transplantation in the main liver transplant center of Iran, the largest center of hepatobiliary surgery in the south of that country, over a 3-year period between 2012 and 2014. All explanted livers, with and without CCA, were evaluated. Results Of the 181 patients, 16 were found to have CCA, two of whom had been diagnosed after pathologic study of the explanted livers. Therefore it appeared that 8.8% of the patients with PSC in our center had developed CCA before liver transplantation. Conclusions A comparison of our results with those obtained from other centers in both Western and Asian countries (which reported CCA in 3.6% - 36.5% of patients with PSC, shows that the incidence of CCA in the patients we studied is intermediate.

  13. An efficient protocol devised for rapid callus induction from leaf explants of Biophytum sensitivum (lDC.

    Directory of Open Access Journals (Sweden)

    Sirigiri Chandra Kala

    2014-03-01

    Full Text Available The Cell cultures are used extensively for in vitro secondary metabolite productions were obtained from callus tissue through cell suspension culture.  The establishment of callus cultures has considerable potential for the production of known and novel secondary metabolites. The objective of the study was to scientifically assess callus culture of Biophytum sensitivum (L DC. was established from leaf explants with different growth regulators greatly influenced the growth of callus cultures. The callus from leaf explants is induced by inoculating the young leaf bits on MS medium supplemented with various auxins (2, 4- Dichlorophenoxyacetic acid (2, 4-D, α-Naphthalene Acetic Acid (NAA and Indole Buteric Acid (IBA, cytokinins (6-Benzyladenine (BA and Kinetin (KN and cytokinin-auxin combination (BA+NAA in different concentrations were (0.5 to 5.0 mg/l used. BA 1mg/l, in combination with NAA (1.0 mg/l also produced maximum amount of callus.  So, this research is concluded that the plant leaf explants cultured on MS medium with 1 mg/ l  BA with 0.5 mg/l NAA was found most efficient for callus induction, provided calli with quite good in texture and friable in nature.

  14. Evaluation of taper joints with combined fatigue and crevice corrosion testing: Comparison to human explanted modular prostheses

    Energy Technology Data Exchange (ETDEWEB)

    Reclaru, L., E-mail: lucien.reclaru@pxgroup.com [PX Group S.A., Dep R and D Corrosion and Biocompatibility Group, Bd. des Eplatures 42, CH-2304 La Chaux-de-Fonds (Switzerland); Brooks, R.A. [Orthopaedic Research, Addenbrooke' s Hospital, University of Cambridge, Box 180 Hills Road, CB2 0QQ Cambridge (United Kingdom); Zuberbühler, M. [Smith and Nephew Orthopaedics AG, Schachenalle 29, 5001 Aarau (Switzerland); Eschler, P.-Y.; Constantin, F. [PX Group S.A., Dep R and D Corrosion and Biocompatibility Group, Bd. des Eplatures 42, CH-2304 La Chaux-de-Fonds (Switzerland); Tomoaia, G. [University of Medicine and Pharmacy Iuliu Hateganu of Cluj-Napoca, Dept. of Orthopaedics and Traumatology, Cluj-Napoca (Romania)

    2014-01-01

    The requirement for revision surgery of total joint replacements is increasing and modular joint replacement implants have been developed to provide adjustable prosthetic revision systems with improved intra-operative flexibility. An electrochemical study of the corrosion resistance of the interface between the distal and proximal modules of a modular prosthesis was performed in combination with a cyclic fatigue test. The complexity resides in the existence of interfaces between the distal part, the proximal part, and the dynamometric screw. A new technique for evaluating the resistance to cyclic dynamic corrosion with crevice stimulation was used and the method is presented. In addition, two components of the proximal module of explanted Ti6Al4V and Ti6Al7Nb prostheses were investigated by optical and electron microscopy. Our results reveal that: The electrolyte penetrates into the interface between the distal and proximal modules during cyclic dynamic fatigue tests, the distal module undergoes cracking and corrosion was generated at the interface between the two models; The comparison of the explanted proximal parts with the similar prostheses evaluated following cyclic dynamic crevice corrosion testing showed that there were significant similarities indicating that this method is suitable for evaluating materials used in the fabrication of modular prostheses. - Highlights: • Electrochemical crevice corrosion testing combined with fatigue test conducted on Ti6Al7Nb and Ti6Al4V modular prostheses • Cations released from integral prostheses • Comparison of human explanted modular prostheses with the similar prostheses evaluated in cyclic dynamic crevice corrosion.

  15. Effects of variations in culture media and hormonal treatments upon callus induction potential in endosperm explant of Barringtonia racemosa L.

    Institute of Scientific and Technical Information of China (English)

    Nurul Izzati Osman; Norrizah Jaafar Sidik; Asmah Awal

    2016-01-01

    Objective: To induce callus from the medicinally valuable species, Barringtonia racemosa L.(B. racemosa) whereby the formation of callus is essential for micropropagation studies and in vitro plant secondary metabolites production.Methods: The callus induction potential in B. racemosa was assessed from endosperm explant cultured on different culture media and plant hormonal treatments. Lloyd and Mc Cown’s woody plant medium and Murashige and Skoog’s medium were used in the study as culture media. On the other hand, various concentrations and combinations of2,4-dichlorophenoxyacetic acid(1.0–2.0 mg/L) and kinetin(0.5–2.5 mg/L) had been incorporated in the culture media to exert the effects of auxin and cytokinin on callus induction.Results: From the present study, it was found that the profuse [(1.681 ± 0.770) g fresh weight,(0.239 ± 0.239) g dry weight] and friable callus formation was optimally produced with desirable morphology and considerable percentage of callus induction(56.70%) in endosperm explants cultured on 1.0 mg/L 2,4-dichlorophenoxyacetic acid and 1.5 mg/L kinetin in Murashige and Skoog’s medium.Conclusions: A reliable protocol for inducing callus formation of profuse and friable morphology in endosperm explant of B. racemosa had therefore been successfully established.

  16. In vitro effect of biogenic silver nanoparticles on sterilisation of tobacco leaf explants and for higher yield of protoplasts.

    Science.gov (United States)

    Bansod, Sunita; Bawskar, Manisha; Rai, Mahendra

    2015-08-01

    Isolation of protoplasts from leaves is useful in plant research. The standard reference methods for isolation of protoplasts are tedious, cause cell damage, are low-yield, time consuming and prone to microbial contamination. To overcome this problem, the authors used silver nanoparticles (AgNPs) for the control of microbial contamination and with low concentration of enzyme mixture for rapid release of protoplasts. The leaf explants were sterilised with 95% ethanol for 30 s followed by biologically synthesised AgNPs (1, 5, 10 and 15 mg/l) for 10 to 20 min. The authors found that 10 mg/l concentration of AgNPs treatment on explants showed remarkable inhibitory effect on microbial contamination with high level of tolerance. Moreover, during protoplasts isolation, the addition of 10 mg/l AgNPs in leaf incubation buffer yielded 34% viable protoplasts in 3 h. This is the first report of AgNPs synthesis from waste plant medium, which was applied for the sterilisation of explants and rapid isolation of protoplasts.

  17. Direct regeneration of Periwinkle (Catharanthus roseus via node explants culture and different combinations of plant growth regulators

    Directory of Open Access Journals (Sweden)

    M. Talebi

    2012-09-01

    Full Text Available Periwinkle (Catharanthus roseus L., Apocynaceae contains more than 130 different terpenoid indole alkaloids (TIAs, of which two dimeric alkaloids, Vinblastine and Vincristine, have antineoplastic activity and are useful in treatment of various cancers. Specific production of some alkaloids in differentiated tissues such as leaf and stem led to use direct regeneration of explants in order to increase the production of these important alkaloids in the plant. In this research, 30 combinations of plant growth regulators and activated charcoal were used in MS media for direct regeneration of node explants. Application of BAP in media containing 1 g/l activated charcoal showed the best direct regeneration of node explants and shoot proliferation. Although application of activated charcoal is necessary for periwinkle growth in media due to many phenolic compounds, but it has negative effects on adsorption of plant growth regulators and consequently reduce shoot proliferation. Therefore, it seems that 1 g/l activated charcoal is an appropriate concentration for preparing shoot proliferation media. In addition, transporting regenerated shoots to culture media containing NAA resulted in increasing shoot length. Proliferated shoots rooted in media without PGR and with 2 g/l activated charcoal and acclimated with environmental conditions after transferring to the soil.

  18. DIRECT IN VITRO SHOOTS PROLIFERATION OF CHICK PEA (CICER ARIETINUM L. FROM SHOOT TIP EXPLANTS INDUCED BY THIDIAZURON

    Directory of Open Access Journals (Sweden)

    Shaheena Parveen

    2012-01-01

    Full Text Available A rapid, simple and efficient protocol for direct in vitro multiple shoot induction and plantlet regeneration was achieved from shoot tip explants of Cicer arietinum. The shoot tips were cultured on MS medium fortified with Thidiazuron (TDZ (1.0-7.0 mg/L for multiple shoot induction. Multiple shoots proliferation was best observed at 3.0 mg/L TDZ from the shoot tip explants within three weeks of culture. Shoot number per explant ranged between 2 and 10. Individual shoots were aseptically excised and sub cultured in the same media for shoot elongation. The elongated shoots were transferred to Indole Butyric Acid (IBA (1.0mg/L–5.0mg/L for root induction. Rooting was observed within two weeks of culture. Rooted plantlets were successfully hardened under culture conditions and subsequently established in the field conditions. The recorded survival rate of the plants was 86%. Plants looked healthy with no visually detectable phenotypic variations.

  19. Plantlet regeneration from mature zygotic embryos and embryonic explants of masson pine (Pinus massoniana Lamb.)

    Institute of Scientific and Technical Information of China (English)

    HUANGJIANQIU; ZHIMINGWEI; 等

    1994-01-01

    Excised zygotic embryos,cotyledons and hypocotyls of juvenile seedlings of masson pine were grown on DCR medium supplemented with several concentrations of various plant phytohormones.BA(1.0mg/L) in combination with NAA(0.05mg/L) in DCR medium was found to increase the formation of adventitious buds from mature zygotic embryos,but most of them were formed at the tips of embryonic cotyledons.Adventitious buds were obtained from cotyledons and hypocotyls from juvenile seedlings when they were cultured on DCR medium containing BA 3-5 mg/L and NAA 0.1-0.2 mg/L.Elongation of buds were observed on hormone-free DCR medium with or without activated charcoal(0.5%).Root initiation was achieved with full or half strength DCR medium supplemented with IBA 1.0 mg/L and NAA 0.25-0.5 mg/L.Approximately 11-20 axillary buds formed on each explant when juvenile seedling explants were treated(3-20h) with BA 50-100 mg/L,followed by transfer to hormone-free DCR medium.The maximum number of shoots obtained per explant within six months was 33.

  20. Investigating the Potential of Amnion-Based Scaffolds as a Barrier Membrane for Guided Bone Regeneration.

    Science.gov (United States)

    Li, Wuwei; Ma, Guowu; Brazile, Bryn; Li, Nan; Dai, Wei; Butler, J Ryan; Claude, Andrew A; Wertheim, Jason A; Liao, Jun; Wang, Bo

    2015-08-11

    Guided bone regeneration is a new concept of large bone defect therapy, which employs a barrier membrane to afford a protected room for osteogenesis and prevent the invasion of fibroblasts. In this study, we developed a novel barrier membrane made from lyophilized multilayered acellular human amnion membranes (AHAM). After decellularization, the AHAM preserved the structural and biomechanical integrity of the amnion extracellular matrix (ECM). The AHAM also showed minimal toxic effects when cocultured with mesenchymal stem cells (MSCs), as evidenced by high cell density, good cell viability, and efficient osteogenic differentiation after 21-day culturing. The effectiveness of the multilayered AHAM in guiding bone regeneration was evaluated using an in vivo rat tibia defect model. After 6 weeks of surgery, the multilayered AHAM showed great efficiency in acting as a shield to avoid the invasion of the fibrous tissues, stabilizing the bone grafts and inducing the massive bone growth. We hence concluded that the advantages of the lyophilized multilayered AHAM barrier membrane are as follows: preservation of the structural and mechanical properties of the amnion ECM, easiness for preparation and handling, flexibility in adjusting the thickness and mechanical properties to suit the application, and efficiency in inducing bone growth and avoiding fibrous tissues invasion.

  1. Indirect organogenesis from various explants of Hildegardia populifolia (Roxb. Schott & Endl. – A threatened tree species from Eastern Ghats of Tamil Nadu, India

    Directory of Open Access Journals (Sweden)

    A.R. Lavanya

    2014-12-01

    Full Text Available Hildegardia species are an important resource for fiber industry. This investigation was conducted to develop a plant regeneration protocol for Hildegardia populifolia (Roxb. Schott & Endl. via indirect organogenesis Callus was obtained from leaf, internode and petiole explants, among these explants internode explant gave best result on MS medium supplemented with different concentrations of 2,4-Dichlorophenoxy acetic acid (2,4-D. The highest percentage (100% of regeneration was obtained with benzyladenine (BA (2.0 mg/l + indole-3-acetic acid (IAA (0.1 mg/l + glutamine (25 mg/l + thidiazuron (TDZ (0.5 mg/l from internode explants. Shootlets were highly rooted on MS medium supplemented with 3.0 mg/l indole-3-butyric acid (IBA. In vitro rooted seedlings were successfully acclimatized. This in vitro regeneration system will facilitate further development of reliable procedures for this genus.

  2. Sensitivity of Allium and Nicotiana in cellular and acellular comet assays to assess differential genotoxicity of direct and indirect acting mutagens.

    Science.gov (United States)

    Bandyopadhyay, Atrayee; Mukherjee, Anita

    2011-05-01

    We have evaluated the extent of DNA damage induced by direct and indirect mutagens by cellular and acellular comet assays in two plant systems, Nicotiana tabacum (wild type tobacco) and Allium cepa (common onion). The objectives of this study were: (1) to generate dose-response curves for DNA migration values from root and shoot nuclei of A. cepa and N. tabacum treated with the direct acting mutagens, ethyl methanesulphonate (EMS), hydrogen peroxide (H(2)O(2)) and the indirect acting mutagen, cadmium chloride (CdCl(2)), (2) to assess the differential response between isolated nuclei and nuclei of root and shoot and of both plants and (3) to examine the differences of sensitivity between direct and indirect acting mutagens by cellular and acellular comet assays. Similar sensitivities were evident in both plant systems to direct and indirect acting mutagens. The combination of cellular and acellular comet assays provided valuable insight to the mode of action of the genotoxicants used. The data obtained demonstrated the estimable capacity of the two plant systems to evaluate genotoxicity under different stress conditions and suggests Allium is a more desirable test system for rapid monitoring of genotoxicity.

  3. Bone development

    DEFF Research Database (Denmark)

    Tatara, M.R.; Tygesen, Malin Plumhoff; Sawa-Wojtanowicz, B.

    2007-01-01

    The objective of this study was to determine the long-term effect of alpha-ketoglutarate (AKG) administration during early neonatal life on skeletal development and function, with emphasis on bone exposed to regular stress and used to serve for systemic changes monitoring, the rib. Shropshire ram...... the groups were recorded int erms of: (1) growth rate, (2) body weight at days 14, 28 and 130 of age or (3) final body weight. The weight and length of ribs were, however, significantly increased in the lambs given AKG for the first 14 days of neonatal life by 8.2% and 3.2%, respectively (P....01). Furthermore, AKG administration induced significantly higher bone mineral density of the cortical bone by 7.1% (P

  4. [Bone transplant].

    Science.gov (United States)

    San Julián, M; Valentí, A

    2006-01-01

    We describe the methodology of the Bone and Soft Tissue Bank, from extraction and storage until use. Since the year 1986, with the creation of the Bone Bank in the University Clinic of Navarra, more than 3,000 grafts have been used for very different types of surgery. Bone grafts can be classified into cortical and spongy; the former are principally used in surgery to save tumour patients, in large post-traumatic reconstructions and in replacement surgery where there are massive bone defects and a structural support is required. The spongy grafts are the most used due to their numerous indications; they are especially useful in filling cavities that require a significant quantity of graft when the autograft is insufficient, or as a complement. They are also of special help in treating fractures when there is bone loss and in the treatment of delays in consolidation and pseudoarthrosis in little vascularized and atrophic zones. They are also used in prosthetic surgery against the presence of cavity type defects. Allografts of soft tissues are specially recognised in multiple ligament injuries that require reconstructions. Nowadays, the most utilised are those employed in surgery of the anterior cruciate ligament although they can be used for filling any ligament or tendon defect. The principal difficulties of the cortical allografts are in the consolidation of the ends with the bone itself and in tumour surgery, given that these are patients immunodepressed by the treatment, the incidence of infection is increased with respect to spongy grafts and soft tissues, which is irrelevant. In short, the increasingly widespread use of allografts is an essential therapeutic weapon in orthopaedic surgery and traumatology. It must be used by expert hands.

  5. Bone mineral content and bone metabolism in young adults with severe periodontitis

    DEFF Research Database (Denmark)

    Wowern von, N.; Westergaard, J.; Kollerup, G.

    2001-01-01

    Bone loss, bone markers, bone metabolism, bone mineral content, osteoporosis, severe periodontitis......Bone loss, bone markers, bone metabolism, bone mineral content, osteoporosis, severe periodontitis...

  6. Effect Of Explant Source And Different Medium Culture On Friable Embryogenic Callus Induction Of Four Cultivars Of Cassava Manihot Esculenta Crantz

    Directory of Open Access Journals (Sweden)

    Simplice Prosper Yandia

    2015-08-01

    Full Text Available In order to obtain Friable Embryogenic Callus FEC for protoplast isolation we have evaluated in this research the competance for Friable Embryogenic Callus FEC of four cassava cultivars M61033 Rendre Yalipe and Six-mois in media containing MS supplemented with 8mgl 24-D MS supplemented with 10 mgl BAP and GD supplemented with 12mgl picloram using apical bud AB and immature leaves lobes ILL as explants. In general in the medium GD12mgl picloram the highest efficiencies of FEC ranged from 58 to 87 and the highest score of FEC ranged from 4.2 to 5.4 with explants AB however we have observed with explants ILL the efficiencies of somatic embryos ranged form 41 to 75 and the score ranged from 4.1 to 4.4. The mediums MS28 mgl 24-D have induced with explants AB the efficiencies of FEC ranged from 43 to 57 and the score ranged from 3.1 to 3.8 however with ILL explants the efficiencies of FEC ranged from 39 to 49 and the score ranged from 2.9 to 3.7. The least FEC were observed in the medium MS210 mgl BAP with BA explants however the efficiencies ranged from 6 to 11 and the score ranged from 1.1 to 1.8. Whereas the efficiencies of FEC with ILL explants ranged from 4 to 7 and the score ranged from 0.5 to 0.8. All of four cultivars showed capability of producing FEC although their efficiency varied according to gonotype donors explants and medium taking into acount. Abbreviations GD Gressoff and Doy MS Murashige and Skoog 24-dichloro phenoxyacetic acid BAP Benzylamino-purin-Acid AB Apical Bud ILL Immature Leaves lobes

  7. Replication of avian, human and swine influenza viruses in porcine respiratory explants and association with sialic acid distribution

    Directory of Open Access Journals (Sweden)

    Nauwynck Hans J

    2010-02-01

    Full Text Available Abstract Background Throughout the history of human influenza pandemics, pigs have been considered the most likely "mixing vessel" for reassortment between human and avian influenza viruses (AIVs. However, the replication efficiencies of influenza viruses from various hosts, as well as the expression of sialic acid (Sia receptor variants in the entire porcine respiratory tract have never been studied in detail. Therefore, we established porcine nasal, tracheal, bronchial and lung explants, which cover the entire porcine respiratory tract with maximal similarity to the in vivo situation. Subsequently, we assessed virus yields of three porcine, two human and six AIVs in these explants. Since our results on virus replication were in disagreement with the previously reported presence of putative avian virus receptors in the trachea, we additionally studied the distribution of sialic acid receptors by means of lectin histochemistry. Human (Siaα2-6Gal and avian virus receptors (Siaα2-3Gal were identified with Sambucus Nigra and Maackia amurensis lectins respectively. Results Compared to swine and human influenza viruses, replication of the AIVs was limited in all cultures but most strikingly in nasal and tracheal explants. Results of virus titrations were confirmed by quantification of infected cells using immunohistochemistry. By lectin histochemistry we found moderate to abundant expression of the human-like virus receptors in all explant systems but minimal binding of the lectins that identify avian-like receptors, especially in the nasal, tracheal and bronchial epithelium. Conclusions The species barrier that restricts the transmission of influenza viruses from one host to another remains preserved in our porcine respiratory explants. Therefore this system offers a valuable alternative to study virus and/or host properties required for adaptation or reassortment of influenza viruses. Our results indicate that, based on the expression of Sia

  8. The effects of Brazilian propolis on etiological agents of mastitis and the viability of bovine mammary gland explants.

    Science.gov (United States)

    Fiordalisi, Samira A L; Honorato, Luciana A; Loiko, Márcia R; Avancini, César A M; Veleirinho, Maria B R; Machado Filho, Luiz C P; Kuhnen, Shirley

    2016-03-01

    The objective of this study was to evaluate in vitro the antimicrobial activity of Brazilian propolis from Urupema, São Joaquim, and Agua Doce (Santa Catarina State) and green propolis from Minas Gerais State, and the effects of propolis on bovine mammary gland explant viability. The propolis samples differed in flavonoid content and antioxidant activity. Green propolis showed the highest content of flavonoids, followed by the sample from São Joaquim. The propolis from Urupema showed the lowest flavonoid content along with the lowest antioxidant activity. The total phenolics were similar across all studied samples. Despite phytochemical differences, the propolis samples from Minas Gerais, São Joaquim, and Urupema presented the same level of antimicrobial activity against Staphylococcus aureus strains. The reduction in S. aureus growth was, on average, 1.5 and 4 log10 times at 200 and 500 μg/mL, respectively. At concentrations of 1,000 μg/mL, all propolis reduced bacterial growth to zero. On the other hand, when the propolis were tested against strains of Escherichia coli, the samples presented weak antimicrobial activity. Mammary explants were maintained in culture for 96h without a loss in viability, demonstrating the applicability of the model in evaluating the toxicity of propolis. The origin and chemical composition of the propolis had an effect on mammary explant viability. We encountered inhibitory concentrations of 272.4, 171.8, 63.85, and 13.26 μg/mL for the propolis from Água Doce, Urupema, São Joaquim, and Mina Gerais, respectively. A clear association between greater antimicrobial activity and toxicity for mammary explants was observed. Of all propolis tested, the Urupema sample was noteworthy, as it showed antimicrobial activity at less toxic concentrations than the other samples, reducing bacterial growth to an average of 9.3 × 10(2) cfu/mL after 6h of contact using 200 μg/mL of extract. The results demonstrate the potential for Brazilian

  9. Growth of vegetative explant Moringa oleifera on different composition of auxin and cytokinin and its synthetic seed germination

    Science.gov (United States)

    Muslihatin, Wirdhatul; Jadid, Nurul; Puspitasari, Ika D.; Safitri, Chusnul E.

    2017-06-01

    The spread of Moringa oleifera is also rare for seed germination and viability or survival are low, and the lack of vegetative propagation method. The purpose of this study are to determine the effect of auxin and cytokinin on growth vegetative explants Moringa oleifera and its synthetic seed germination. The explants grown on MS medium with sucrose content of 30% and a range of additional hormone. Addition concentration and different types of hormone made in order to know the sensitivity and response explant growth on a variety of media to get a good callus and embryosomatic. The composition of the hormone given is MS + 2.4 D 3 ppm; MS + 2,4D 2 ppm + BAP 2 ppm; MS + NAA + 0.5 ppm kinetin 1 ppm; MS + NAA 1 ppm + kinetin 1 ppm; MS + NAA 1 ppm + 0.5 ppm kinetin. The explants were incubated at a temperature of 18-20 ° C with a photoperiod 16/8. Explants and MS medium is incubated to form embryonic callus. Seeds synthetic made from embryonic callus growing on medium 1 ppm kinetin + NAA 1 ppm with encapsulation method with sodium alginate 2%. Seed synthetic germinated in some kind of medium that medium ms0 solid (M1), ms0 liquid (M2), MS0 semi-solid (M3), MS solid NAA 1ppm + Kinetin 1 ppm (M4), MS liquid NAA 1 ppm + kinetin (M5), and semi-solid MS + NAA 1 ppm kinetin 1 ppm (M6). Synthetic seed viability was observed with the parameters of the fresh weight of synthetic seed, germination percentage and seedling. Chlorophyll content was measured by spectrophotometric method with solvent asseton. Best callus generated in this study are embryonic callus that grew on media NAA 1 ppm + kinetin 1 ppm. Embryonic callus on M6 + NAA 1 ppm kinetin 1 ppm capable of germination with an average weight of callus and sprouts of 40.38 mg. Of the entire amount of a synthetic seed on M6, just 5 seed germinate, so the percentage of germination of seeds is equal to 41.67%. with an average length of sprouts 1 cm with an average total chlorophyll content of 8.66 mg / g.

  10. Bone lesion biopsy

    Science.gov (United States)

    Bone biopsy; Biopsy - bone ... needle is gently pushed and twisted into the bone. Once the sample is obtained, the needle is ... sample is sent to a lab for examination. Bone biopsy may also be done under general anesthesia ...

  11. Facts about Broken Bones

    Science.gov (United States)

    ... Room? What Happens in the Operating Room? Broken Bones KidsHealth > For Kids > Broken Bones Print A A ... sticking through the skin . What Happens When a Bone Breaks? It hurts to break a bone! It's ...

  12. Calcium and bones

    Science.gov (United States)

    Bone strength and calcium ... calcium (as well as phosphorus) to make healthy bones. Bones are the main storage site of calcium in ... your body does not absorb enough calcium, your bones can get weak or will not grow properly. ...

  13. Broken Bones (For Parents)

    Science.gov (United States)

    ... Feeding Your 1- to 2-Year-Old Broken Bones KidsHealth > For Parents > Broken Bones Print A A ... bone fragments in place. When Will a Broken Bone Heal? Fractures heal at different rates, depending upon ...

  14. Bone biopsy (image)

    Science.gov (United States)

    A bone biopsy is performed by making a small incision into the skin. A biopsy needle retrieves a sample of bone and it ... examination. The most common reasons for bone lesion biopsy are to distinguish between benign and malignant bone ...

  15. Early somatic embryogenesis in Heliconia chartacea Lane ex Barreiros cv. Sexy Pink ovary section explants

    Directory of Open Access Journals (Sweden)

    Cláudia Ulisses

    2010-02-01

    Full Text Available The present work evaluated the development of embryogenic callus from transversal ovary sections. The experiments were carried out under two experimental regimes using combinations of IAA (0; 5.71; 8.56; 11.42; 14.27μM and 2,4-D (0; 13.57; 18.10; 22.62μM or combinations of 2,4-D with BA (0; 4.43; 6.65; 8.87; 11.09μM. Assessments were made of anatomical aspects of the callus and for the presence of embryogenic structures using cytochemical and histological analyses and stereomicroscopic and scanning electronic microscopic observations. Treatments with 2,4-D and IAA produced friable calluses demonstrating cellular acquisition of morphogenetic competence as well as the formation of pro-embryogenic sectors. The expression of embryogenic program could be observed, with proembryogenic cell clusters developing into globular embryos. These results offer the possibility of using new types of explants for culturing helicons that avoid the growth of endophytic bacteria.Este trabalho teve como objetivo avaliar a resposta de secções transversais de ovários e o desenvolvimento de calos embriogênicos. O experimento constou de dois ensaios. No primeiro avaliou-se combinações entre AIA (0; 5.71; 8.56; 11.42; 14.27μM e 2,4-D (0; 13.57; 18.10; 22.62μM e no segundo avaliou-se as concentrações de 2,4-D supracitadas, combinadas com concentrações de BA (0; 4.43; 6.65; 8.87; 11.09μM. Os calos formados foram avaliados quanto à presença de estruturas embriogênicas utilizando-se estereomicroscópio, microscópio eletrônico de varredura, além de análises citoquímicas e histológicas. Combinações entre 2,4-D e AIA induziram a formação de calos friáveis com setores pró-embriogênicos, refletindo a aquisição de competência morfogenética. Posteriormente foi observada a expressão do programa embriogênico quando massas pró-embriogências desenvolveram-se formando embriões somáticos. Esses resultados apresentam uma alternativa para a utiliza

  16. Induction of shoot buds, multiplication and plantlet formation in seedling explants of bell pepper (Capsicum annuum L. cv. Bryza in vitro

    Directory of Open Access Journals (Sweden)

    Andrzej Gatz

    2014-01-01

    Full Text Available In vitro shoot bud induction and multiplication as well as plantlets formation from different parts of 21-d old seedlings (shoot tip, cotyledonary node, distal part of cotyledon, acropetal section of hypocotyl of Capsicum annuum L., cv. Bryza were compared. During 4 weeks of primary explant culture on initiation media, first shoot bud primordia appeared; they reminded leaf primordia and subsequently some of them underwent enlargement, some developed into leaves and leaf-like structures (mainly on cotyledon explants. The highest number of shoot bud primordia was noted on cotyledonary node explants, but they were smaller than those on the remaining types of the explants. The best response of shoot regeneration showed cotyledon explants on which most of shoot buds were formed in each from four treated passages. From shoot buds on elongation media after 4 weeks of culture rooted rosettes of leaves were achieved, and the extension of the culture time to eight weeks without subculture caused that the rosettes developed into plantlets. Throughout four successive passages plantlets were obtained from cotyledon and shoot tip explants.

  17. Efficient plant regeneration protocol through callus for Saussurea obvallata (DC.) Edgew. (Asteraceae): effect of explant type, age and plant growth regulators.

    Science.gov (United States)

    Dhar, Uppeandra; Joshi, Mitali

    2005-06-01

    A callus induction and in vitro plantlet regeneration system for the endangered state flower of Uttaranchal (Saussurea obvallata) was optimized by studying the influence of explant type (root, hypocotyl, cotyledon and leaf), age and different concentrations of plant growth regulators. Explants from 10 to 15-day-old seedlings showed maximum callus induction. Callus formation and shoot differentiation was initiated on Murashige-Skoog (MS) medium containing 6-benzyladenine (BA) and alpha-naphthalene acetic acid (NAA) in all explant types. The best results were obtained using leaf explants: 100% callusing was achieved in MS medium supplemented with 2.5 microM BA and 1.0 microM NAA, and 100% differentiation along with a multiplication rate of 12 shoots per explant with a combination of 5.0 microM BA and 1.0 microM NAA. However, the results reflected the existence of high inter-explant variability in response to growth regulators. In vitro rooting of shoots was achieved at an efficiency of 100% in one-half strength MS medium supplemented with 2.5 microM indole-3-butyric acid. Application of this protocol has potential for mass multiplication of the target species in a limited time period.

  18. Safety and immunogenicity of a combined Tetanus, Diphtheria, recombinant acellular Pertussis vaccine (TdaP) in healthy Thai adults

    Science.gov (United States)

    Sirivichayakul, Chukiat; Chanthavanich, Pornthep; Limkittikul, Kriengsak; Siegrist, Claire-Anne; Wijagkanalan, Wassana; Chinwangso, Pailinrut; Petre, Jean; Hong Thai, Pham; Chauhan, Mukesh; Viviani, Simonetta

    2017-01-01

    ABSTRACT Background: An acellular Pertussis (aP) vaccine containing recombinant genetically detoxified Pertussis Toxin (PTgen), Filamentous Hemagglutinin (FHA) and Pertactin (PRN) has been developed by BioNet-Asia (BioNet). We present here the results of the first clinical study of this recombinant aP vaccine formulated alone or in combination with tetanus and diphtheria toxoids (TdaP). Methods: A phase I/II, observer-blind, randomized controlled trial was conducted at Mahidol University in Bangkok, Thailand in healthy adult volunteers aged 18–35 y. The eligible volunteers were randomized to receive one dose of either BioNet's aP or Tetanus toxoid-reduced Diphtheria toxoid-acellular Pertussis (TdaP) vaccine, or the Tdap Adacel® vaccine in a 1:1:1 ratio. Safety follow-up was performed for one month. Immunogenicity was assessed at baseline, at 7 and 28 d after vaccination. Anti-PT, anti-FHA, anti-PRN, anti-tetanus and anti-diphtheria IgG antibodies were assessed by ELISA. Anti-PT neutralizing antibodies were assessed also by CHO cell assay. Results: A total of 60 subjects (20 per each vaccine group) were enrolled and included in the safety analysis. Safety laboratory parameters, incidence of local and systemic post-immunization reactions during 7 d after vaccination and incidence of adverse events during one month after vaccination were similar in the 3 vaccine groups. One month after vaccination, seroresponse rates of anti-PT, anti-FHA and anti-PRN IgG antibodies exceeded 78% in all vaccine groups. The anti-PT IgG, anti-FHA IgG, and anti-PT neutralizing antibody geometric mean titers (GMTs) were significantly higher following immunization with BioNet's aP and BioNet's TdaP than Adacel® (Pdiphtheria GMTs at one month after immunization were comparable in all vaccine groups. All subjects had seroprotective titers of anti-tetanus and anti-diphtheria antibodies at baseline. Conclusion: In this first clinical study, PTgen-based BioNet's aP and TdaP vaccines showed

  19. Bladder tissue regeneration using acellular bi-layer silk scaffolds in a large animal model of augmentation cystoplasty.

    Science.gov (United States)

    Tu, Duong D; Chung, Yeun Goo; Gil, Eun Seok; Seth, Abhishek; Franck, Debra; Cristofaro, Vivian; Sullivan, Maryrose P; Di Vizio, Dolores; Gomez, Pablo; Adam, Rosalyn M; Kaplan, David L; Estrada, Carlos R; Mauney, Joshua R

    2013-11-01

    Acellular scaffolds derived from Bombyx mori silk fibroin were investigated for their ability to support functional tissue regeneration in a porcine model of augmentation cystoplasty. Two bi-layer matrix configurations were fabricated by solvent-casting/salt leaching either alone (Group 1) or in combination with silk film casting (Group 2) to yield porous foams buttressed by heterogeneous surface pore occlusions or homogenous silk films, respectively. Bladder augmentation was performed with each scaffold group (6 × 6 cm(2)) in juvenile Yorkshire swine for 3 m of implantation. Augmented animals exhibited high rates of survival (Group 1: 5/6, 83%; Group 2: 4/4, 100%) and voluntary voiding over the course of the study period. Urodynamic evaluations demonstrated mean increases in bladder capacity over pre-operative levels (Group 1: 277%; Group 2: 153%) which exceeded nonsurgical control gains (144%) encountered due to animal growth.In addition, animals augmented with both matrix configurations displayed increases in bladder compliance over pre-operative levels(Group 1: 357%; Group 2: 338%) similar to growth-related elevations observed in non-surgical controls (354%) [corrected]. Gross tissue evaluations revealed that both matrix configurations supported extensive de novo tissue formation throughout the entire original implantation site which exhibited ultimate tensile strength similar to nonsurgical counterparts. Histological and immunohistochemical analyses showed that both implant groups promoted comparable extents of smooth muscle regeneration and contractile protein (α-smooth muscle actin and SM22α) expression within defect sites similar to controls. Parallel evaluations demonstrated the formation of a transitional, multi-layered urothelium with prominent cytokeratin, uroplakin, and p63 protein expression in both matrix groups. De novo innervation and vascularization processes were evident in all regenerated tissues indicated by synaptophysin-positive neuronal

  20. Desmosomal cadherins are decreased in explanted arrhythmogenic right ventricular dysplasia/cardiomyopathy patient hearts.

    Directory of Open Access Journals (Sweden)

    Alexia Vite

    Full Text Available AIMS: Arrhythmogenic right ventricular Dysplasia/cardiomyopathy (ARVD/C is an autosomal dominant inherited cardiomyopathy associated with ventricular arrhythmia, heart failure and sudden death. Genetic studies have demonstrated the central role of desmosomal proteins in this disease, where 50% of patients harbor a mutation in a desmosmal gene. However, clinical diagnosis of the disease remains difficult and molecular mechanisms appears heterogeneous and poorly understood. The aim of this study was to characterize the expression profile of desmosomal proteins in explanted ARVD/C heart samples, in order to identify common features of the disease. METHODS AND RESULTS: We examined plakophilin-2, desmoglein-2, desmocollin-2, plakoglobin and β-catenin protein expression levels from seven independent ARVD/C heart samples compared to two ischemic, five dilated cardiomyopathy and one healthy heart sample as controls. Ventricular and septum sections were examined by immunoblot analysis of total heart protein extracts and by immunostaining. Immunoblots indicated significant decreases in desmoglein-2 and desmocollin-2, independent of any known underlying mutations, whereas immune-histochemical analysis showed normal localization of all desmosomal proteins. Quantitative RT-PCR revealed normal DSG2 and DSC2 mRNA transcript levels, suggesting increased protein turn-over rather than transcriptional down regulation. CONCLUSION: Reduced cardiac desmoglein-2 and desmocollin-2 levels appear to be specifically associated with ARVD/C, independent of underlying mutations. These findings highlight a key role of desmosomal cadherins in the pathophysiology of ARVD/C. Whether these reductions could be considered as specific markers for ARVD/C requires replication analysis.

  1. Effects of compression on the loss of newly synthesized proteoglycans and proteins from cartilage explants

    Energy Technology Data Exchange (ETDEWEB)

    Sah, R.L.; Doong, J.Y.; Grodzinsky, A.J.; Plaas, A.H.; Sandy, J.D. (Department of Electrical Engineering and Computer Science, Massachusetts Institute of Technology Harvard-M.I.T., Cambridge (United States))

    1991-04-01

    The effects of mechanical compression of calf cartilage explants on the catabolism and loss into the medium of proteoglycans and proteins radiolabeled with (35S)sulfate and (3H)proline were examined. A single 2- or 12-h compression of 3-mm diameter cartilage disks from a thickness of 1.25 to 0.50 mm, or slow cyclic compression (2 h on/2 h off) from 1.25 mm to 1.00, 0.75, or 0.50 mm for 24 h led to transient alterations and/or sustained increases in loss of radiolabeled macromolecules. The effects of imposing or removing loads were consistent with several compression-induced physical mediators including fluid flow, diffusion, and matrix disruption. Cyclic compression induced convective fluid flow and enhanced the loss of 35S- and 3H-labeled macromolecules from tissue into medium. In contrast, prolonged static compression induced matrix consolidation and appeared to hinder the diffusional transport and loss of 35S- and 3H-labeled macromolecules. Since high amplitude cyclic compression led to a sustained increase in the rate of loss of 3H- and 35S-labeled macromolecules that was accompanied by an increase in the rate of loss of (3H)hydroxyproline residues and an increase in tissue hydration, such compression may have caused disruption of the collagen meshwork. The 35S-labeled proteoglycans lost during such cyclic compression were of smaller average size than those from controls, but contained a similarly low proportion (approximately 15%) that could form aggregates with excess hyaluronate and link protein. The size distribution and aggregability of the remaining tissue proteoglycans and 35S-labeled proteoglycans were not markedly affected. The loss of tissue proteoglycan paralleled the loss of 35S-labeled macromolecules.

  2. Specific inhibition of TRPV4 enhances retinal ganglion cell survival in adult porcine retinal explants.

    Science.gov (United States)

    Taylor, Linnéa; Arnér, Karin; Ghosh, Fredrik

    2017-01-01

    Signaling through the polymodal cation channel Transient Receptor Potential Vanilloid 4 (TRPV4) has been implicated in retinal neuronal degeneration. To further outline the involvement of this channel in this process, we here explore modulation of Transient Receptor Potential Vanilloid 4 (TRPV4) activity on neuronal health and glial activation in an in vitro model of retinal degeneration. For this purpose, adult porcine retinal explants were cultured using a previously established standard protocol for up to 5 days with specific TRPV4 agonist GSK1016790A (GSK), or specific antagonist RN-1734, or culture medium only. Glial and neuronal cell health were evaluated by a battery of immunohistochemical markers, as well as morphological staining. Specific inhibition of TRPV4 by RN-1734 significantly enhanced ganglion cell survival, improved the maintenance of the retinal laminar architecture, reduced apoptotic cell death and attenuated the gliotic response as well as preserved the expression of TRPV4 in the plexiform layers and ganglion cells. In contrast, culture controls, as well as specimens treated with GSK, displayed rapid remodeling and neurodegeneration as well as a downregulation of TRPV4 and the Müller cell homeostatic mediator glutamine synthetase. Our results indicate that TRPV4 signaling is an important contributor to the retinal degeneration in this model, affecting neuronal cell health and glial homeostasis. The finding that pharmacological inhibition of the receptor significantly attenuates neuronal degeneration and gliosis in vitro, suggests that TRPV4 signaling may be an interesting pharmaceutical target to explore for treatment of retinal degenerative disease.

  3. In vitro cell cultures obtained from different explants of Corylus avellana produce Taxol and taxanes

    Directory of Open Access Journals (Sweden)

    Cavalli Francesca

    2006-12-01

    Full Text Available Abstract Background Taxol is an effective antineoplastic agent, originally extracted from the bark of Taxus brevifolia with a low yield. Many attempts have been made to produce Taxol by chemical synthesis, semi-synthesis and plant tissue cultures. However, to date, the availability of this compound is not sufficient to satisfy the commercial requirements. The aim of the present work was to produce suspension cell cultures from plants not belonging to Taxus genus and to verify whether they produced Taxol and taxanes. For this purpose different explants of hazel (Corylus avellana species were used to optimize the protocol for inducing in vitro callus, an undifferentiated tissue from which suspension cell cultures were established. Results Calli were successfully induced from stems, leaves and seeds grown in various hormone concentrations and combinations. The most suitable callus to establish suspension cell cultures was obtained from seeds. Media recovered from suspension cell cultures contained taxanes, and showed antiproliferative activity on human tumour cells. Taxol, 10-deacetyltaxol and 10-deacetylbaccatin III were the main taxanes identified. The level of Taxol recovered from the media of hazel cultures was similar to that found in yew cultures. Moreover, the production of taxanes in hazel cell cultures increased when elicitors were used. Conclusion Here we show that hazel cell cultures produce Taxol and taxanes under controlled conditions. This result suggests that hazel possesses the enzymes for Taxol production, which until now was considered to be a pathway particular to Taxus genus. The main benefit of producing taxanes through hazel cell cultures is that hazel is widely available, grows at a much faster rate in vivo, and is easier to cultivate in vitro than yew. In addition, the production of callus directly from hazel seeds shortens the culture time and minimizes the probability of contamination. Therefore, hazel could become a