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Sample records for acellular bone explants

  1. 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.

  2. 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.

  3. 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.

  4. 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.

  5. 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.

  6. 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。

  7. 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.

  8. 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

  9. 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.

  10. 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.

  11. 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

  12. 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...

  13. 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

  14. 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

  15. 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.

  16. 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

  17. 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

  18. 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

  19. 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.

  20. 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.

  1. 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.

  2. 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

  3. 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.

  4. 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.

  5. 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

  6. 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

  7. 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.

  8. 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.

  9. 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.

  10. 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

  11. 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.

  12. 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

  13. CALLUS INDUCTION FROM FLORAL EXPLANTS OF CUPUASSU

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    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.

  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. 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

  16. 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...

  17. 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.

  18. 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.

  19. 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.

  20. 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.

  1. 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.

  2. 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.

  3. 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...

  4. 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.

  5. 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.

  6. 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.

  7. 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

  8. 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.

  9. 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.

  10. 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.

  11. 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.

  12. 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.

  13. 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.

  14. 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...

  15. 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

  16. 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

  17. 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.

  18. 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.

  19. 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.

  20. 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.

  1. 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.

  2. 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.

  3. 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.

  4. 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.

  5. 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.

  6. 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.

  7. 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.

  8. 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.

  9. 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.

  10. 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...... opacification, the findings were calcifications in 2 of the 3 lenses (66.6%) and proteins in 2 lenses (66.6%). Conclusions A marked variation in surface changes was observed by SEM. Findings did not correlate with cause for explantation. Scanning electron microscopy is a useful tool that provides exclusive...

  11. 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.

  12. 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.

  13. 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.

  14. 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.

  15. 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 ...

  16. 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

  17. 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...

  18. 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.

  19. [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.

  20. 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.

  1. 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.

  2. 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.

  3. 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

  4. 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

  5. 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...

  6. 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 ...

  7. 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...

  8. 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.

  9. 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

  10. 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.

  11. 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.

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

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    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.

  13. 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.

  14. 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.

  15. 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.

  16. 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.

  17. 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...

  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. 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.

  20. 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.

  1. 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.

  2. 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.

  3. 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.

  4. 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

  5. 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.

  6. 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.

  7. 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 ...

  8. 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.

  9. 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 ...

  10. 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

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

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    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

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

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    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.

  13. 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.

  14. 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.

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

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    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.

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

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    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.

  17. 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.

  18. 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

  19. 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.

  20. 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

  1. 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

  2. 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.

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

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    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.

  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. 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.

  6. 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.

  7. 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.

  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. 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.

  11. 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.

  12. 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.

  13. 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.

  14. 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.

  15. 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

  16. 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

  17. 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.

  18. 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.

  19. 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

  20. 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

  1. 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.

  2. 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

  3. 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.

  4. 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.

  5. 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. ...

  6. 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.

  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. 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)

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

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    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.

  10. 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...

  11. 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.

  12. 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...

  13. 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.

  14. 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.

  15. 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.

  16. 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.

  17. 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.

  18. 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.

  19. 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.

  20. 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

  1. 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.

  2. 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...

  3. 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.

  4. Evaluation of respiratory model employing conventional NIH mice to access the immunity induced by cellular and acellular pertussis vaccines

    Directory of Open Access Journals (Sweden)

    Alexandre Alves de Souza de Oliveira Dias

    2006-11-01

    Full Text Available The increasing number of pertussis cases reported on the last twenty years and the existence of new acellular vaccines reinforce the need of research for experimental models to assure the quality of available pertussis vaccines. In this study, allotments of whole-cell and acellular pertussis vaccines were tested through the Intranasal Challenge Model (INM using conventional NIH mice. The results have been compared to those achieved by the "Gold standard" Intracerebral Challenge Model (ICM. In contrast to ICM, INM results did not show intralaboratorial variations. Statistical analysis by Anova and Ancova tests revealed that the INM presented reproducibility and allowed identification and separation of different products, including three-component and four-component accellular pertussis vaccines. INM revealed differences between pertussis vaccines. INM provides lower distress to the mice allowing the reduction of mice number including the possibility of using conventional mice (less expensive under non-aseptic environment. Thus, INM may be used as an alternative method of verifying the consistence of allotment production, including acellular pertussis vaccines.

  5. 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, ...

  6. 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 ...

  7. 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 ...

  8. 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 ...

  9. 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

  10. 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.

  11. 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.

  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. 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.

  14. 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.

  15. 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.

  16. 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.

  17. 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.

  18. 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

  19. 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.

  20. 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.

  1. 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.

  2. 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

  3. 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.

  4. 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.

  5. 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.

  6. 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.

  7. 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.

  8. 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...

  9. 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.

  10. 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...

  11. 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...

  12. 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.

  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. 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.

  15. 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.

  16. 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.

  17. 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.

  18. 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.

  19. 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.

  20. 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.

  1. 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.

  2. 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.

  3. 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

  4. 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.

  5. 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....

  6. 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.

  7. 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

  8. 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 ...

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

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    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.

  10. 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

  11. 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.

  12. 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

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    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

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

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    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

  14. 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.

  15. 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.

  16. Using porcine acellular collagen matrix (Pelvicol® in bladder augmentation: experimental study

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    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.

  17. 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.

  18. 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.

  19. 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.

  20. 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

  1. 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.

  2. 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.

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

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    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.

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

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    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.

  5. 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.

  6. 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.

  7. 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

  8. 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

  9. 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.

  10. 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.

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

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    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.

  12. 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.

  13. Sustained release of VEGF from PLGA nanoparticles embedded thermo-sensitive hydrogel in full-thickness porcine bladder acellular matrix

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    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.

  14. 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.

  15. 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.

  16. 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.

  17. EFEITO DE REGULADORES DE CRESCIMENTO E TIPO DE EXPLANTES NA MORFOGÊNESE in vitro DE Capsicum annuum L.

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    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.

  18. 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.

  19. Organotypic Culture of Breast Tumor Explants as a Multicellular System for the Screening of Natural Compounds with Antineoplastic Potential

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    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.

  20. 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

  1. 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.

  2. 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

  3. 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

  4. 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

  5. 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.

  6. 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.

  7. 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.

  8. 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.

  9. 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.

  10. 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.

  11. 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.

  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. 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.

  14. 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.

  15. 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.

  16. 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.

  17. 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.

  18. 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.

  19. 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.

  20. 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.

  1. 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.

  2. 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

  3. 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...

  4. 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.

  5. 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.

  6. 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.

  7. 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

  8. 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.

  9. 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.

  10. 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.

  11. 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.

  12. 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.

  13. 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.

  14. 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.

  15. 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.

  16. 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...

  17. 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.

  18. 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

    a stronger cytotoxic response and higher level of insulin resistance in the co-cultured hepatocytes. In conclusion, our results reveal depot-dependent effects of ATEs on co-cultured primary hepatocytes, which in part may be related to a more pronounced infiltration of stromal vascular cells (SVCs......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...

  19. 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.

  20. 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.

  1. 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.

  2. 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.

  3. 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...

  4. 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.

  5. 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.

  6. 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.

  7. 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.

  8. 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.

  9. 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 ...

  10. 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

  11. 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

  12. 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...

  13. 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....

  14. 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 ...

  15. 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.

  16. 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.

  17. In vitro plant regeneration of Albizia lebbeck (L. from seed explants

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    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

  18. 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 ...

  19. 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.

  20. 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

  1. 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

  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

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    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

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    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. 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 ...

  5. Flask sealing on in vitro seed germination and morphogenesis of two types of ornamental pepper explants

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    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.

  6. 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.

  7. Activin A induces Langerhans cell differentiation in vitro and in human skin explants.

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    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.

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

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    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.

  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. 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.

  11. 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.

  12. 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.

  13. 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...

  14. 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.

  15. 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.

  16. 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.

  17. 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.

  18. 化学去细胞尿道在大鼠皮下的埋置%Subcutaneous embedment of the acellular urethra in rats

    Institute of Scientific and Technical Information of China (English)

    刘小燕; 王远征; 李莹; 李琳; 毛丹; 李航旭

    2014-01-01

    BACKGROUND:The nature of graft rejective reaction is the antigen immunological reaction of recipient’s immune system in response to donor’s graft. OBJECTIVE:To observe the effect of chemical extraction on the immunogenicity of acellular peripheral urethral isografts. METHODS:Twenty urethras of Sprague-Dawley rats, 1 cm long, were included in this study. Ten urethras were treated with chemical acellular method to remove immunogenic component, thus prepared into acellular urethras;another ten urethras were fresh, without any acellular treatment. Acellular urethras were implanted into the back of ten Wistar rats, while fresh urethras into another ten rats. 6 weeks after transplantation, the specimens were harvested for gross and histological observations. RESULTS AND CONCLUSION:After acellular or fresh urethras were implanted into the rats, no autotomy was observed, the incision healed wel , the diet and activities of the rat were normal, and no ulcer formation was found. At 6 weeks, tissue membrane formed around the acellular urethra, while no formation was visible around fresh urethra by gross observation. Hematoxylin-eosin staining showed that, acellular urethras were gradual y absorbed and replaced by smooth muscular tissue. And unequal blood vessels grew in the acellular urethra, even formed blood sinusoid. In the control group, the urethras were not absorbed and no blood vessels were grown. The acellular urethras can gradual y form normal urethra in rats, indicating that the acellular urethral autograft has a lower antigenicity and can replace the cavernous body of normal urethra.%背景:有研究提出了移植排斥反应的实质是受者免疫系统对供者移植物抗原的免疫应答反应。  目的:验证化学萃取去细胞处理对同种异体尿道移植抗原性的影响。  方法:取SD大鼠尿道共20根,每段长1 cm。其中10根采用化学去细胞法处理大鼠尿道的免疫原性成分,使其

  19. 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.

  20. 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.

  1. 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...

  2. 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...

  3. 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.

  4. 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.

  5. 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.

  6. 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.

  7. 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.

  8. 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..

  9. 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.

  10. 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.

  11. 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.

  12. 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.

  13. 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.

  14. 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.

  15. [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.

  16. Decreased Laminin Expression by Human Lung Epithelial Cells and Fibroblasts Cultured in Acellular Lung Scaffolds from Aged Mice.

    Science.gov (United States)

    Godin, Lindsay M; Sandri, Brian J; Wagner, Darcy E; Meyer, Carolyn M; Price, Andrew P; Akinnola, Ifeolu; Weiss, Daniel J; Panoskaltsis-Mortari, Angela

    2016-01-01

    The lung changes functionally and structurally with aging. However, age-related effects on the extracellular matrix (ECM) and corresponding effects on lung cell behavior are not well understood. We hypothesized that ECM from aged animals would induce aging-related phenotypic changes in healthy inoculated cells. Decellularized whole organ scaffolds provide a powerful model for examining how ECM cues affect cell phenotype. The effects of age on ECM composition in both native and decellularized mouse lungs were assessed as was the effect of young vs old acellular ECM on human bronchial epithelial cells (hBECs) and lung fibroblasts (hLFs). Native aged (1 year) lungs demonstrated decreased expression of laminins α3 and α4, elastin and fibronectin, and elevated collagen, compared to young (3 week) lungs. Proteomic analyses of decellularized ECM demonstrated similar findings, and decellularized aged lung ECM contained less diversity in structural proteins compared to young ECM. When seeded in old ECM, hBECs and hLFs demonstrated lower gene expression of laminins α3 and α4, respectively, as compared to young ECM, paralleling the laminin deficiency of aged ECM. ECM changes appear to be important factors in potentiating aging-related phenotypes and may provide clues to mechanisms that allow for aging-related lung diseases.

  17. Decreased Laminin Expression by Human Lung Epithelial Cells and Fibroblasts Cultured in Acellular Lung Scaffolds from Aged Mice.

    Directory of Open Access Journals (Sweden)

    Lindsay M Godin

    Full Text Available The lung changes functionally and structurally with aging. However, age-related effects on the extracellular matrix (ECM and corresponding effects on lung cell behavior are not well understood. We hypothesized that ECM from aged animals would induce aging-related phenotypic changes in healthy inoculated cells. Decellularized whole organ scaffolds provide a powerful model for examining how ECM cues affect cell phenotype. The effects of age on ECM composition in both native and decellularized mouse lungs were assessed as was the effect of young vs old acellular ECM on human bronchial epithelial cells (hBECs and lung fibroblasts (hLFs. Native aged (1 year lungs demonstrated decreased expression of laminins α3 and α4, elastin and fibronectin, and elevated collagen, compared to young (3 week lungs. Proteomic analyses of decellularized ECM demonstrated similar findings, and decellularized aged lung ECM contained less diversity in structural proteins compared to young ECM. When seeded in old ECM, hBECs and hLFs demonstrated lower gene expression of laminins α3 and α4, respectively, as compared to young ECM, paralleling the laminin deficiency of aged ECM. ECM changes appear to be important factors in potentiating aging-related phenotypes and may provide clues to mechanisms that allow for aging-related lung diseases.

  18. Persistence of antibodies 3 years after booster vaccination of adults with combined acellular pertussis, diphtheria and tetanus toxoids vaccine.

    Science.gov (United States)

    Weston, Wayde; Messier, Marc; Friedland, Leonard R; Wu, Xiangfeng; Howe, Barbara

    2011-11-01

    The duration of protection after vaccination with reduced antigen content diphtheria, tetanus and acellular pertussis vaccines (Tdap) is not known. Long-term post-vaccination serological data will help to improve understanding of the duration of humoral immunity and guide vaccination policy for the timing of repeat dose administration. The persistence of antibodies to Tdap antigens was measured 3 years after vaccination of adults 19-64 years of age with one of 2 Tdap vaccines (Boostrix(®), GlaxoSmithKline Biologicals; Tdap-B: or Adacel(®), Sanofi Pasteur; Tdap-A). In both groups, geometric mean concentrations for antibodies to diphtheria, tetanus, and pertussis vaccine antigens were decreased at year 3 relative to levels observed 1 month and 1 year following vaccination, but remained higher than pre-vaccination levels. Seroprotection rates for diphtheria and tetanus remained high for both Tdap vaccines (for diphtheria, 96.9% and 97.8% for the Tdap-B and Tdap-A groups, respectively; for tetanus, 98.1% and 99.6%, respectively).

  19. Universal tetanus, diphtheria, acellular pertussis (Tdap) vaccination of adults: What Canadian health care providers know and need to know.

    Science.gov (United States)

    MacDougall, D; Halperin, B A; MacKinnon-Cameron, D; Li, L; McNeil, S A; Langley, J M; Halperin, S A

    2015-01-01

    The tetanus, diphtheria, and acellular pertussis vaccine (Tdap) is recommended for all adults in both Canada and the United States. There are few data on the proportion of Canadian adults vaccinated with Tdap; however, anecdotal reports indicate that uptake is low. This study aimed to explore the knowledge, attitudes, beliefs, and behaviors of Canadian health care providers (HCPs) in an attempt to identify potential barriers and facilitators to Tdap uptake. HCPs were surveyed and a geographic and practice representative sample was obtained (N =1,167). In addition, 8 focus groups and 4 interviews were conducted nationwide. Results from the survey indicate that less than half (47.5%) of all respondents reported being immunized with Tdap themselves, while 58.5% routinely offer Tdap to their adult patients. Knowledge scores were relatively low (63.2% correct answers). The best predictor of following the adult Tdap immunization guidelines was awareness of and agreement with those recommendations. Respondents who were aware of the recommendations were more likely to think that Tdap is safe and effective, that their patients are at significant risk of getting pertussis, and to feel that they have sufficient information (p vaccine, and vaccine hesitancy were identified as barriers to compliance with the national recommendations for universal adult immunization, and suggestions were provided to better translate recommendations to front-line practitioners.

  20. Persistence of T-cell immune response induced by two acellular pertussis vaccines in children five years after primary vaccination.

    Science.gov (United States)

    Palazzo, Raffaella; Carollo, Maria; Bianco, Manuela; Fedele, Giorgio; Schiavoni, Ilaria; Pandolfi, Elisabetta; Villani, Alberto; Tozzi, Alberto E; Mascart, Françoise; Ausiello, Clara M

    2016-01-01

    The resurgence of pertussis suggests the need for greater efforts to understand the long-lasting protective responses induced by vaccination. In this paper we dissect the persistence of T memory responses induced by primary vaccination with two different acellular pertussis (aP) vaccines, hexavalent Hexavac® vaccine (Hexavac) (Sanofi Pasteur MSD) and Infanrix hexa® (Infanrix) (Glaxo-SmithKline Biologicals). We evaluated magnitude and duration of T-cell responses to pertussis toxin (PT) by measuring T-cell proliferation, cytokines (IL-2 and IFNγ) production and memory subsets in two groups of children 5 years after primary vaccination. Some of the enrolled children received only primary vaccination, while others had the pre-school boost dose. Positive T-cell responses to PT were detected in 36% of children. Percentage of responsive children, T-cell proliferation and CD4IL-2+ cells were significantly higher in the children primed with Hexavac than in those who received Infanrix vaccine. No major effects of the boost on PT-specific proliferation were observed. Overall, our data documented a persistence of T-cell memory against PT in a minor fraction of children 5 years after primary vaccination. The different responses induced by Hexavac and Infanrix vaccine could rely on differences in PT inactivation process or excipients/adjuvants formulations.

  1. High contrast microstructural visualization of natural acellular matrices by means of phase-based x-ray tomography

    Science.gov (United States)

    Hagen, Charlotte K.; Maghsoudlou, Panagiotis; Totonelli, Giorgia; Diemoz, Paul C.; Endrizzi, Marco; Rigon, Luigi; Menk, Ralf-Hendrik; Arfelli, Fulvia; Dreossi, Diego; Brun, Emmanuel; Coan, Paola; Bravin, Alberto; de Coppi, Paolo; Olivo, Alessandro

    2015-12-01

    Acellular scaffolds obtained via decellularization are a key instrument in regenerative medicine both per se and to drive the development of future-generation synthetic scaffolds that could become available off-the-shelf. In this framework, imaging is key to the understanding of the scaffolds’ internal structure as well as their interaction with cells and other organs, including ideally post-implantation. Scaffolds of a wide range of intricate organs (esophagus, lung, liver and small intestine) were imaged with x-ray phase contrast computed tomography (PC-CT). Image quality was sufficiently high to visualize scaffold microarchitecture and to detect major anatomical features, such as the esophageal mucosal-submucosal separation, pulmonary alveoli and intestinal villi. These results are a long-sought step for the field of regenerative medicine; until now, histology and scanning electron microscopy have been the gold standard to study the scaffold structure. However, they are both destructive: hence, they are not suitable for imaging scaffolds prior to transplantation, and have no prospect for post-transplantation use. PC-CT, on the other hand, is non-destructive, 3D and fully quantitative. Importantly, not only do we demonstrate achievement of high image quality at two different synchrotron facilities, but also with commercial x-ray equipment, which makes the method available to any research laboratory.

  2. 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 ...

  3. 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 ...

  4. 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 ...

  5. 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. ...

  6. 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 ...

  7. 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

  8. 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

  9. 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.

  10. 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.

  11. 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

  12. Bone densitometry

    DEFF Research Database (Denmark)

    Ravn, Pernille; Alexandersen, P; Møllgaard, A

    1999-01-01

    The bisphosphonates have been introduced as alternatives to hormone replacement therapy (HRT) for the treatment and prevention of postmenopausal osteoporosis. The expected increasing application in at clinical practice demands cost-effective and easily handled methods to monitor the effect on bone...

  13. Shoot regeneration from cotyledonary leaf explants of jatropha curcas: A biodiesel plant

    KAUST Repository

    Kumar, Nitish Chandramohana

    2010-03-07

    A simple, high frequency, and reproducible method for plant regeneration through direct organogenesis from cotyledonary leaf explants of Jatropha curcas was developed using Murashige and Skoog (MS) medium supplemented with different concentrations of thidiazuron (TDZ) or 6-benzyl aminopurine (BAP). Medium containing TDZ has greater influence on regeneration as compared to BAP. The induced shoot buds were transferred to MS medium containing 10 lM kinetin (Kn), 4.5 lM BAP, and 5.5 lM a-naphthaleneacetic acid (NAA) for shoot proliferation. The proliferated shoots could be elongated on MS medium supplemented with different concentrations and combinations of BAP, indole-3-acetic acid (IAA), NAA, and indole-3-butyric acid (IBA). MS medium with 2.25 lM BAP and 8.5 lM IAA was found to be the best combination for shoot elongation. However, significant differences in plant regeneration and shoot elongation were observed among the genotypes studied. Rooting was achieved when the basal cut end of elongated shoots were dipped in half strength MS liquid medium containing dif- ferent concentrations and combinations of IBA, IAA, and NAA for 4 days, followed by transfer to growth regulators free half strength MS medium supplemented 0.25 mg l-1 activated charcoal. Elongated shoot treated with 15 lM IBA, 5.7 lM IAA, and 11 lM NAA resulted in highest percent rooting. The rooted plants could be established in soil with more than 90% survival rate. The method developed may be useful in improvement of J. curcas through genetic modification. © Franciszek Górski Institute of Plant Physiology, Polish Academy of Sciences, Kraków 2010.

  14. Mechanical spectroscopy of retina explants at the protein level employing nanostructured scaffolds.

    Science.gov (United States)

    Mayazur Rahman, S; Reichenbach, Andreas; Zink, Mareike; Mayr, Stefan G

    2016-04-14

    Development of neuronal tissue, such as folding of the brain, and formation of the fovea centralis in the human retina are intimately connected with the mechanical properties of the underlying cells and the extracellular matrix. In particular for neuronal tissue as complex as the vertebrate retina, mechanical properties are still a matter of debate due to their relation to numerous diseases as well as surgery, where the tension of the retina can result in tissue detachment during cutting. However, measuring the elasticity of adult retina wholemounts is difficult and until now only the mechanical properties at the surface have been characterized with micrometer resolution. Many processes, however, such as pathological changes prone to cause tissue rupture and detachment, respectively, are reflected in variations of retina elasticity at smaller length scales at the protein level. In the present work we demonstrate that freely oscillating cantilevers composed of nanostructured TiO2 scaffolds can be employed to study the frequency-dependent mechanical response of adult mammalian retina explants at the nanoscale. Constituting highly versatile scaffolds with strong tissue attachment for long-term organotypic culture atop, these scaffolds perform damped vibrations as fingerprints of the mechanical tissue properties that are derived using finite element calculations. Since the tissue adheres to the nanostructures via constitutive proteins on the photoreceptor side of the retina, the latter are stretched and compressed during vibration of the underlying scaffold. Probing mechanical response of individual proteins within the tissue, the proposed mechanical spectroscopy approach opens the way for studying tissue mechanics, diseases and the effect of drugs at the protein level.

  15. Depot-dependent effects of adipose tissue explants on co-cultured hepatocytes.

    Directory of Open Access Journals (Sweden)

    Zhen-Yu Du

    Full Text Available 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 adipose tissues. Expressions of inflammation related genes (IL-6, TNF-α, COX-2 were higher in the inguinal than the epididymal ATE. Similarly, expressions of marker genes of macrophage and monocyte (MPEG-1, CD68, F4/80, CD64 were higher in the stromal vascular fraction (SVF isolated from inguinal ATE than that from epididymal ATE. However, expressions of lipolysis related genes (ATGL, HSL, perilipin-1 were higher in the epididymal adipocytes than inguinal adipocytes. Moreover, secretion of IL-6 and PGE(2 was higher from inguinal ATEs than from epididymal ATEs. There was a trend that the total levels of IL-6, TNF-α 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 a stronger cytotoxic response and higher level of insulin resistance in the co-cultured hepatocytes. In conclusion, our results reveal depot-dependent effects of ATEs on co-cultured primary hepatocytes, which in part may be related to a more pronounced infiltration of stromal vascular cells (SVCs, particularly macrophages, in inguinal adipose tissue resulting in stronger responses in terms of hepatotoxicity and insulin-resistance.

  16. 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.

  17. 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

  18. 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.

  19. Culture of Adult Transgenic Zebrafish Retinal Explants for Live-cell Imaging by Multiphoton Microscopy.

    Science.gov (United States)

    Lahne, Manuela; Gorsuch, Ryne A; Nelson, Craig M; Hyde, David R

    2017-02-24

    An endogenous regeneration program is initiated by Müller glia in the adult zebrafish (Danio rerio) retina following neuronal damage and death. The Müller glia re-enter the cell cycle and produce neuronal progenitor cells that undergo subsequent rounds of cell divisions and differentiate into the lost neuronal cell types. Both Müller glia and neuronal progenitor cell nuclei replicate their DNA and undergo mitosis in distinct locations of the retina, i.e. they migrate between the basal Inner Nuclear Layer (INL) and the Outer Nuclear Layer (ONL), respectively, in a process described as Interkinetic Nuclear Migration (INM). INM has predominantly been studied in the developing retina. To examine the dynamics of INM in the adult regenerating zebrafish retina in detail, live-cell imaging of fluorescently-labeled Müller glia/neuronal progenitor cells is required. Here, we provide the conditions to isolate and culture dorsal retinas from Tg[gfap:nGFP](mi2004) zebrafish that were exposed to constant intense light for 35 h. We also show that these retinal cultures are viable to perform live-cell imaging experiments, continuously acquiring z-stack images throughout the thickness of the retinal explant for up to 8 h using multiphoton microscopy to monitor the migratory behavior of gfap:nGFP-positive cells. In addition, we describe the details to perform post-imaging analysis to determine the velocity of apical and basal INM. To summarize, we established conditions to study the dynamics of INM in an adult model of neuronal regeneration. This will advance our understanding of this crucial cellular process and allow us to determine the mechanisms that control INM.

  20. Adenoviral Mediated Expression of BMP2 by Bone Marrow Stromal Cells Cultured in 3D Copolymer Scaffolds Enhances Bone Formation.

    Directory of Open Access Journals (Sweden)

    Sunita Sharma

    Full Text Available Selection of appropriate osteoinductive growth factors, suitable delivery method and proper supportive scaffold are critical for a successful outcome in bone tissue engineering using bone marrow stromal cells (BMSC. This study examined the molecular and functional effect of a combination of adenoviral mediated expression of bone morphogenetic protein-2 (BMP2 in BMSC and recently developed and characterized, biodegradable Poly(L-lactide-co-є-caprolactone{poly(LLA-co-CL}scaffolds in osteogenic molecular changes and ectopic bone formation by using in vitro and in vivo approaches. Pathway-focused custom PCR array, validation using TaqMan based quantitative RT-PCR (qRT-PCR and ALP staining showed significant up-regulation of several osteogenic and angiogenic molecules, including ALPL and RUNX2 in ad-BMP2 BMSC group grown in poly(LLA-co-CL scaffolds both at 3 and 14 days. Micro CT and histological analyses of the subcutaneously implanted scaffolds in NOD/SCID mice revealed significantly increased radiopaque areas, percentage bone volume and formation of vital bone in ad-BMP2 scaffolds as compared to the control groups both at 2 and 8 weeks. The increased bone formation in the ad-BMP2 group in vivo was paralleled at the molecular level with concomitant over-expression of a number of osteogenic and angiogenic genes including ALPL, RUNX2, SPP1, ANGPT1. The increased bone formation in ad-BMP2 explants was not found to be associated with enhanced endochondral activity as evidenced by qRT-PCR (SOX9 and FGF2 and Safranin O staining. Taken together, combination of adenoviral mediated BMP-2 expression in BMSC grown in the newly developed poly(LLA-co-CL scaffolds induced expression of osteogenic markers and enhanced bone formation in vivo.

  1. Regeneración de Solanum tuberosum L. variedad pastusa suprema a partir de explantes internodales

    Directory of Open Access Journals (Sweden)

    Chaparro Giraldo Alejandro

    2006-06-01

    Full Text Available Una de las vías posibles para aumentar la eficacia de los métodos de transformación genética mediante Agrobacterium tumefaciens es la optimización de la capacidad de regeneración de las plantas de interés. En la Universidad Nacional de Colombia, Sede Bogotá, el grupo de Ingeniería Genética de Plantas (IGP, ha venido trabajando en la obtención de plantas transgénicas a partir de la variedad de papa Diacol Capiro. Es de interés llevar a cabo este tipo de procedimientos en otras variedades de papa colombiana. La variedad Pastusa Suprema
    es un cultivar obtenido mediante mejoramiento genético convencional, que presenta cualidades superiores a otros cultivares. Es importante desarrollar un sistema eficiente de regeneración en esta variedad, como soporte para ulteriores estudios de transferencia de genes que permitan optimizar aún más su rendimiento. En este trabajo se evaluó el efecto de diferentes combinaciones hormonales adicionadas a un medio básico compuesto de sales Murashige y Skogg (1962, sobre el comportamiento callogénico y regenerativo de la variedad
    de Pastusa Suprema. Se realizó un diseño experimental factorial, en arreglo completamente al azar. Los factores corresponden a las hormonas ácido naftalénacetico (ANA, ácido giberélico (AG3 y ZR (Zeatina Ribósido, y los niveles son las concentraciones por factor ZR: 2,0 mg/L y 3,0 mg/L; ANA 0,0 mg/L, 0,02 mg/L, y 0,2 mg/L; y AG3: 0,02 mg/L, 1,0 mg/L. Los tratamientos probados corresponden a todas las combinaciones posibles entre los niveles de hormona evaluados, dando lugar a un total de doce tratamientos. La eficiencia del medio de cultivo se evaluó con base en las variables: porcentaje de inducción de callos, porcentaje de regeneración, y número de regenerantes por explante. El análisis de los datos se realizó con el software estadístico
    SAS versión 8.0. La presencia de ANA demostró ser esencial en la respuesta callogénica y regenerativa de

  2. The Stimulatory Effect of Notochordal Cell-Conditioned Medium in a Nucleus Pulposus Explant Culture

    NARCIS (Netherlands)

    de Vries, Stefan A H; van Doeselaar, Marina; Meij, Björn P; Tryfonidou, Marianna A; Ito, K

    2016-01-01

    Objectives: Notochordal cell-conditioned medium (NCCM) has previously shown to have a stimulatory effect on nucleus pulposus cells (NPCs) and bone marrow stromal cells (BMSCs) in alginate and pellet cultures. These culture methods provide a different environment than the nucleus pulposus (NP) tissue

  3. The Stimulatory Effect of Notochordal-Cell Conditioned Medium in a Nucleus Pulposus Explant Culture

    NARCIS (Netherlands)

    de Vries, Stefan; Doeselaar, Marina van; Meij, Björn; Tryfonidou, M; Ito, Keita

    2015-01-01

    OBJECTIVES: Notochordal cell-conditioned medium (NCCM) has previously shown to have a stimulatory effect on nucleus pulposus cells (NPCs) and bone marrow stromal cells (BMSCs) in alginate and pellet cultures. These culture methods provide a different environment than the nucleus pulposus (NP) tissue

  4. Effect of 6-BA on nodal explant bud sproutings of Coffea arabica cv. Mundo Novo Efeito de 6-BA na brotação de gemas de explantes nodais de Coffea arabica cv. Mundo Novo

    Directory of Open Access Journals (Sweden)

    Luis Carlos da Silva Ramos

    2005-01-01

    Full Text Available Coffee plants can be micropropagated by nodal bud sprouting using the 6-benzylaminopurine (6-BA hormone. However, literature reports the use of a wide range of 6-BA, from 0.5 to 88.8 µM L-1. So, this study was performed to narrow that range. Nodal explants of Coffea arabica cv Mundo Novo obtained from in vitro plantlets were inoculated on gelled-MS medium supplemented with different concentrations of 6-BA. Two assays were carried out: in the first one, 6-BA was used at concentrations of 0, 5, 25, 50, and 100 µM L-1, being evaluated at 43 and 123 days. In the second experiment, dosis of 10, 20 and 30 µM L-1, have evaluated at 65 and 100 days. Treatments with 6-BA induced multiple sprouting from the nodal explants, which were best characterized around 100 days after inoculation. The nodal explants grew taller and showed multiple shoots, whereas the effect of 6-BA at 5 to 25 µM L-1 was similar to that with higher concentrations (50 and 100 µM L-1. Nodal explants yielded from 2.9 to 6.0 buds per node, achieving height of 1.3 to 1.5 cm at 5 to 25 µM L-1 of 6-BA, whereas they yielded from 4.3 to 4.9 buds per node but the sprouting grew about 0.8 cm at 50 and 100 µM L-1 of 6-BA. This study indicated that multiple sprouting of lateral buds can be induced by lower concentrations of 6-BA, for example, from 10 to 30 µM L-1, diminishing possible risks of somaclonal variation due to high levels of hormone concentration.O cafeeiro pode ser micropropagado via brotação de gemas laterais, aplicando o regulador de crescimento 6-benzilaminopurina (6-BA. Entretanto, a literatura apresenta ampla variação da dose empregada, desde 0.5 a 88.8 µM L-1. Assim, este estudo visou otimizar doses para explantes nodais do cafeeiro C. arabica cv Mundo Novo. Explantes nodais, obtidos de plântulas cultivadas in vitro, foram inoculados em meio MS geleificado, com adição de diferentes concentrações de 6-BA. Foram feitos dois experimentos: no primeiro, 6-BA foi

  5. A randomised, double-blind, non-inferiority clinical trial on the safety and immunogenicity of a tetanus, diphtheria and monocomponent acellular pertussis (TdaP) vaccine in comparison to a tetanus and diphtheria (Td) vaccine when given as booster vaccinations to healthy adults

    DEFF Research Database (Denmark)

    Thierry-Carstensen, Birgit; Jordan, Karina; Uhlving, Hilde Hylland;

    2012-01-01

    Increasing incidence of pertussis in adolescents and adults has stimulated the development of safe and immunogenic acellular pertussis vaccines for booster vaccination of adolescents and adults.......Increasing incidence of pertussis in adolescents and adults has stimulated the development of safe and immunogenic acellular pertussis vaccines for booster vaccination of adolescents and adults....

  6. Regeneration of human epidermis on acellular dermis is impeded by small-molecule inhibitors of EGF receptor tyrosine kinase.

    Science.gov (United States)

    Forsberg, Sofi; Ostman, Arne; Rollman, Ola

    2008-10-01

    The family of human epidermal growth factor receptors (EGFR, HER2-4) exerts key functions in normal and malignant epithelial cells. Both EGFR and HER2 are valuable targets for anti-cancer drugs by interfering with ligand binding, receptor dimerization, or tyrosine kinase activity. A similar therapeutic strategy has been advocated for chronic psoriasis since plaque lesions overexpress EGFR and its ligands. Our aim was to characterize EGFR/HER2 protein expression in skin cultures and to evaluate the effects of tyrosine kinase inhibitors on epidermal outgrowth, morphology, and EGFR activation. Human skin explants were established on cell-free dermis and cultured at the air-liquid interface. The impact of small-molecule HER inhibitors on outgrowth was assayed by fluorescence-based image analysis and histometry. Effects of a dual EGFR/HER2 kinase inhibitor, PKI166, on neoepidermis were studied by immunohistochemistry and Western blot. Receptor immunostaining showed in vivo-like distributions with highest EGFR intensity in the proliferative layers whereas HER2 was mainly expressed by suprabasal keratinocytes. Reepithelialization was associated with EGFR autophosphorylation irrespective of exogenous ligand stimulation. PKI166 inhibited neoepidermal EGFR activation, keratinocyte proliferation, and outgrowth from normal and psoriatic skin explants. The rate of epidermalization in presence of other HER inhibitors varied suggesting that drug specificity, potency, and reversibility determine the dynamic outcome. Overall, agents predominantly targeting EGFR kinase were more efficient inhibitors of epidermal regeneration than an HER2-selective drug. The study illustrates the usefulness of a dynamic skin model and emphasizes the potential of HER-directed approaches to control epidermal growth in hyperproliferative skin disorders.

  7. Clinical Evaluation of Heterogeneous Acellular Dermal Matrix for Guided Tissue Regeneration in Periodontal Disease%异种脱细胞真皮基质膜在引导牙周组织再生中应用的临床效果评价

    Institute of Scientific and Technical Information of China (English)

    章立群; 邓碧霞; 谢安琪; 孙辉

    2012-01-01

    Objective: To explore the clinical effectiveness of heterogeneous acellular dermal matrix( AMD) membrane for guided tissue regeneration in treatment of periodontal defects. Methods: 30 periodontal defects were randomly assigned into two treatments; 20 for testing group using a combination of AMD and coralline hydroxyapatite (HA) grafting and 10 for control group using HA grafting alone. After six months of surgery, the change of clinical index and radiographic alveolar bone level were compared statistically. Results: The reduction of pocket probing depth and clinical attachment loss and increasing level of alveolar bone of testing group were all higher than those in control group while the recession quantity was lower than that in control group(P<0. 05). Conclusion: Heterogeneous acellular dermal matrix membrane is effective for the treatment of periodontal defects.%目的:评价异种脱细胞真皮基质膜在引导牙周组织再生中应用的临床效果.方法:30例牙周缺损区患牙随机分两组:实验组20例,异种脱细胞真皮基质膜和羟基磷灰石修复;对照组10例仅羟基磷灰石修复.统计学比较6个月时两组各项临床指标和牙槽骨水平改变.结果:实验组牙周袋深度、临床附着丧失的减少量和牙槽骨水平的增加量均较对照组高(P<0.05),牙龈退缩量低于对照组(P<0.05).结论:异种脱细胞真皮基质膜应用于引导牙周组织再生有较好的临床效果,值得推广.

  8. Indução de calos friáveis em explantes foliares de Salix (Salyx humboldtiana Willd Induction of friable callus in leaf explants of Salix (Salyx humboldtiana Willd

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    Breno Régis Santos

    2005-06-01

    Full Text Available O salix é uma árvore ornamental adaptada a terrenos úmidos, sendo considerada uma espécie importante para a recomposição de áreas ciliares degradadas. A madeira pode ser empregada na indústria em geral. Apesar de produzir grande quantidade de sementes, estas não possuem alta percentagem de germinação. O presente trabalho teve como objetivo realizar o estabelecimento in vitro de salix através da indução e formação de calos friáveis em explantes foliares, visando a uma posterior regeneração de plantas para a propagação massal desta espécie. Explantes foliares foram inoculados em meio de cultura MS (MURASHIGE & SKOOG, 1962, acrescido de diferentes concentrações de ácido 2,4-diclorofenoxiacético (0; 1,0; 2,0; 4,0; 6,0; 8,0; 10,0 e 12,0mg L-1 e combinações entre ácido naftalenoacético e benzilaminopurina, ambos em concentrações de 0,0; 1,0; 2,0; 4,0; 6,0 e 8,0mg L-1. Os resultados demonstram que explantes inoculados na ausência de reguladores de crescimento não apresentam a formação de calos friáveis. Significativa produção de calos friáveis (90% é obtida utilizando-se 6,0mg L-1 de ácido 2,4-diclorofenoxiacético. A utilização de concentração individual de ácido naftalenoacético ou benzilaminopurina, além de induzir a calogênese também foi capaz de promover rizogênese.Salix is an ornamental tree adapted to humid soils being considered an important species used in depleted areas. Its wood may be used in all kinds of industries. Although the production of seeds is large, the germination is reduced. The objective of this work was to establish salix in vitro through the induction and formation of friable callus of leaf explants to provide future regeneration of plants for mass propagation of the species. Leaf explants were inoculated in MS medium (MURASHIGE & SKOOG, 1962 supplemented with different concentrations of 2-4-diclorofenoxiacetic acid (0; 1.0; 2.0; 4.0; 6.0; 8.0 and 12.0mg L-1 and combinations

  9. Indução de brotações em explantes de segmentos de folhas de plântulas de urucueiro em diferentes citocininas Induction of shoot buds in leaf explants of annatto seedlings in differents cytokinins

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    Jacqueline Leite Almeida

    1996-04-01

    Full Text Available Segmentos de folhas de plântulas de urucum (Bixa orellana L, foram cultivados in vitro, em um meio básico idealizado por Murashige & Skoog (1962, suplementado com 0,5mg/l dos reguladores do crescimento, benzilaminopurina (BAP, cinetina (KN e isopentenil-adenina (2iP, isolados ou combinados entre si, com o objetivo de avaliar qual a melhor citocinina ou combinação destas para indução de brotações de boa qualidade visando à micropropagação. Após 60 dias de cultivo, as gemas obtidas foram transferidas para um meio MS modificado, contendo a metade da formulação de sais minerais, sem a suplementação de reguladores do crescimento. Gemas de melhor qualidade foram obtidas nos tratamentos onde a KN e o BAP estavam em combinação. A KN sozinha não foi eficiente na indução de brotações. O maior número de gemas por explante foi obtido com KN, BAP e 2iP e no tratamento com apenas BAP.Excised leaf explants of annatto (Bixa orellana L. seedlings were cultivated on a Murashige & Skoog basal medium containing benzylaminopurine (BAP, kinetin (KN and isopentenyl-adenine (2iP individually or in combination, each at concentration of 0.5mg/l, with the purpose of evaluated the best cytokinins or cytokinins combination for shoot bud formation. After 60 days, regenerated shoots were transferred to half strenght MS medium without growth regulators. Best shoots were observed where KN and BAP were added in combination. Isolated KN was ineffective for shoot bud formation. The maximum average number of shoot buds per leaf explant was obtained on medium with BAP, KN and 2iP in combination and on medium supplied of isolated BAP.

  10. The biomechanical behavior and host response to porcine-derived small intestine submucosa, pericardium and dermal matrix acellular grafts in a rat abdominal defect model.

    Science.gov (United States)

    Zhang, Jian; Wang, Guan Yu; Xiao, Yi Pin; Fan, Lie Ying; Wang, Qiang

    2011-10-01

    Several porcine-derived acellular biologic grafts are increasingly used in abdominal wall reconstruction due to the limitations of synthetic meshes in many clinical situations. However, relatively little is known so far about their comparative mechanical characteristics and performance after defect repair. We therefore investigated three most commonly used porcine-derived acellular biomaterials, small intestine submucosa (P-SIS), pericardium (P-PC) and acellular dermal matrix (P-ADM) immediately after prepared, and their effectiveness, biomechanical and histological characteristics in repairing full-thickness abdominal defect in a rat model. P-PC had the best native performance in the burst strength, tensile strength and ball burst among the three porcine-derived scaffolds. P-SIS showed a significantly higher water vapor transmission in comparison with P-PC or P-ADM. Abdominal wall defects in rats were all satisfied repaired with P-SIS, P-PC or P-ADM. No laxity or fistula was observed in the repaired abdominal wall in the P-SIS group up to 8 weeks after surgery. However, there was a tendency for high postoperative abdominal eventration in the P-ADM and P-PC groups as compared with the P-SIS group. With regard to overall aspects of the postoperative laxity, intra-abdominal adhesion formation, tensile stress, stretchability, and degree of tissue ingrowth in terms of collagen deposition and neovascularization, P-SIS exhibits clear advantages over P-PC as well as P-ADM after large abdominal wall defect reconstruction.

  11. Cellular versus acellular matrix devices in treatment of diabetic foot ulcers: study protocol for a comparative efficacy randomized controlled trial

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    Lev-Tov Hadar

    2013-01-01

    Full Text Available Abstract Background Diabetic foot ulcers (DFUs represent a significant source of morbidity and an enormous financial burden. Standard care for DFUs involves systemic glucose control, ensuring adequate perfusion, debridement of nonviable tissue, off-loading, control of infection, local wound care and patient education, all administered by a multidisciplinary team. Unfortunately, even with the best standard of care (SOC available, only 24% or 30% of DFUs will heal at weeks 12 or 20, respectively. The extracellular matrix (ECM in DFUs is abnormal and its impairment has been proposed as a key target for new therapeutic devices. These devices intend to replace the aberrant ECM by implanting a matrix, either devoid of cells or enhanced with fibroblasts, keratinocytes or both as well as various growth factors. These new bioengineered skin substitutes are proposed to encourage angiogenesis and in-growth of new tissue, and to utilize living cells to generate cytokines needed for wound repair. To date, the efficacy of bioengineered ECM containing live cellular elements for improving healing above that of a SOC control group has not been compared with the efficacy of an ECM devoid of cells relative to the same SOC. Our hypothesis is that there is no difference in the improved healing effected by either of these two product types relative to SOC. Methods/Design To test this hypothesis we propose a randomized, single-blind, clinical trial with three arms: SOC, SOC plus Dermagraft® (bioengineered ECM containing living fibroblasts and SOC plus Oasis® (ECM devoid of living cells in patients with nonhealing DFUs. The primary outcome is the percentage of subjects that achieved complete wound closure by week 12. Discussion If our hypothesis is correct, then immense cost savings could be realized by using the orders-of-magnitude less expensive acellular ECM device without compromising patient health outcomes. The article describes the protocol proposed to test

  12. Clinical application of cultured epithelial autografts on acellular dermal matrices in the treatment of extended burn injuries.

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    Fang, Taolin; Lineaweaver, William C; Sailes, Frederick C; Kisner, Carson; Zhang, Feng

    2014-11-01

    Achieving permanent replacement of skin in extensive full-thickness and deep partial-thickness burn injuries and chronic wounds remains one of the fundamental surgical problems. Presently, split-thickness skin grafts are still considered the best material for surgical repair of an excised burn wound. However, in burns that affect greater than 50% of total body surface area, the patient has insufficient areas of unaffected skin from which split-thickness skin grafts can be harvested. The use of cultured epithelial (or epidermal) autografts (CEAs) has achieved satisfactory results. But the take rate of CEAs is poor in full-thickness bed or in chronically infected area. Providing temporary cover with allograft skin, or a more permanent allodermis, may increase clinical take. This review aims to (1) describe the use of CEAs in the regeneration of the epidermis, (2) introduce the application of the acellular dermal matrices (ADMs) in the clinics, and (3) enhance understanding of the CEAs applied with ADM as an appropriate strategy to treat the extended burn injuries. The current evidence regarding the cultured epithelial cell or keratinocyte autograft and dermal grafts applied in the treatment of burn injuries was investigated with an extensive electronic and manual search (MEDLINE and EMBASE). The included literature (N=136 publications) was critically evaluated focusing on the efficacy and safety of this technique in improving the healing of the deep dermal and full-thickness burn injuries. This review concluded that the use of ADM with CEAs is becoming increasingly routine, particularly as a life-saving tool after acute thermal trauma.

  13. Vaccination with tetanus, diphtheria, and acellular pertussis vaccine of pregnant women enrolled in Medicaid--Michigan, 2011-2013.

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    Housey, Michelle; Zhang, Fan; Miller, Corinne; Lyon-Callo, Sarah; McFadden, Jevon; Garcia, Erika; Potter, Rachel

    2014-09-26

    In October 2011, the Advisory Committee on Immunization Practices (ACIP) first recommended the routine administration of a tetanus, diphtheria, and acellular pertussis vaccine (Tdap) during pregnancy as a strategy to protect infants from pertussis (also known as whooping cough). This recommendation applied to women previously unvaccinated with Tdap and specified the optimal vaccination time as late second or third trimester (after 20 weeks' gestation). By vaccinating pregnant women, infants, who are at highest risk for mortality and morbidity from pertussis, gain passive immunity from maternal antibodies transferred to them in utero. Since this recommendation was made, little has been published on the percentage of women receiving Tdap during pregnancy. In Michigan, Medicaid pays for costs of pregnancy for approximately 40% of births. Infants enrolled in Medicaid are a particularly vulnerable population; in Michigan, their all-cause mortality is higher than that of privately insured infants. To assess vaccination coverage among pregnant women enrolled in a publicly funded insurance program in Michigan, Medicaid administrative claims data and statewide immunization information system data for mothers of infants born during November 2011-February 2013 were analyzed. This report describes the results of that analysis, which indicated that only 14.3% of these women received Tdap during pregnancy, with rates highest (17.6%) among non-Hispanic, non-Arab whites and lowest (6.8%) among Arab women. Vaccination was related to maternal age and gestational age at birth, but not to adequacy of prenatal care. In 2013, recognizing the importance of Tdap for every pregnancy, ACIP revised its guidelines to include a Tdap dose during every pregnancy. Ensuring that all infants receive the protection against pertussis afforded by maternal vaccination will require enhanced efforts to vaccinate pregnant women.

  14. Acellular bi-layer silk fibroin scaffolds support tissue regeneration in a rabbit model of onlay urethroplasty.

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    Yeun Goo Chung

    Full Text Available Acellular scaffolds derived from Bombyx mori silk fibroin were investigated for their ability to support functional tissue regeneration in a rabbit model of urethra repair. A bi-layer silk fibroin matrix was fabricated by a solvent-casting/salt leaching process in combination with silk fibroin film casting to generate porous foams buttressed by homogeneous silk fibroin films. Ventral onlay urethroplasty was performed with silk fibroin grafts (Group 1, N = 4 (Width × Length, 1 × 2 cm(2 in adult male rabbits for 3 m of implantation. Parallel control groups consisted of animals receiving small intestinal submucosa (SIS implants (Group 2, N = 4 or urethrotomy alone (Group 3, N = 3. Animals in all groups exhibited 100% survival prior to scheduled euthanasia and achieved voluntary voiding following 7 d of initial catheterization. Retrograde urethrography of each implant group at 3 m post-op revealed wide urethral calibers and preservation of organ continuity similar to pre-operative and urethrotomy controls with no evidence of contrast extravasation, strictures, fistulas, or stone formation. Histological (hematoxylin and eosin and Masson's trichrome, immunohistochemical, and histomorphometric analyses demonstrated that both silk fibroin and SIS scaffolds promoted similar extents of smooth muscle and epithelial tissue regeneration throughout the original defect sites with prominent contractile protein (α-smooth muscle actin and SM22α and cytokeratin expression, respectively. De novo innervation and vascularization were also evident in all regenerated tissues indicated by synaptophysin-positive neuronal cells and vessels lined with CD31 expressing endothelial cells. Following 3 m post-op, minimal acute inflammatory reactions were elicited by silk fibroin scaffolds characterized by the presence of eosinophil granulocytes while SIS matrices promoted chronic inflammatory responses indicated by mobilization of mononuclear cell infiltrates. The results

  15. Tissue-engineered conduit using bladder acellular matrix and bladder epithelial cells for urinary diversion in rabbits

    Institute of Scientific and Technical Information of China (English)

    LIAO Wen-biao; SONG Chao; LI Yong-wei; YANG Si-xing; MENG Lin-chao; LI Xin-hui

    2013-01-01

    Background For muscle invasive bladder cancer,radical cystectomy is the most effective treatment now and urinary diversion is often necessary.The use of intestinal tissue for urinary diversion is frequently associated with complications.In this study,we aimed to make a tissue-engineered conduit (TEC) using bladder epithelial cells and bladder acellular matrix (BAM) for urinary diversion in rabbits.Methods Bladder epithelial cells of rabbit were cultivated and expanded in vitro,then seeded on BAM,and cultured for 7 days.Then cell-seeded graft was used to make TEC.In the experimental group,most of bladder of the rabbit was removed while bladder trigone was retained.The proximal end of TEC was anastomosed with bladder trigone and the distal end was anastomosed with the abdominal stoma.In the control group,TEC was made using unseeded BAM.Haematoxylin and eosin staining was conducted,respectively,at 1,2,4,and 8 weeks postoperatively.Immunohistochemistry was performed 8 weeks postoperatively.Intravenous urography,retrograde pyelography,and cystoscopy of TEC were made at 12 weeks postoperatively.Results All animals were alive in the experimental group.Haematoxylin and eosin staining showed epithelial coverage in TEC.Immunohistochemistry showed anti-cytokeratin AE1/AE3 antibody and anti-ZO1 antibody positive,confirming there were mature and functional epithelial cells on the lumen of TEC.Retrograde pyelography and intravenous urography showed that TEC developed well and that there was no obstruction.In the control group,four rabbits were dead within 2 weeks and scar formation,atresia,and severe hydronephrosis were found.Conclusions We successfully made TEC using BAM and bladder epithelial cells for urinary diversion in rabbits.The lumen of this new TEC covered mature epithelial cells and could prevent urinary extravasation.

  16. Degradation of polysaccharide hydrogels seeded with bone marrow stromal cells.

    Science.gov (United States)

    Jahromi, Shiva H; Grover, Liam M; Paxton, Jennifer Z; Smith, Alan M

    2011-10-01

    In order to produce hydrogel cell culture substrates that are fit for the purpose, it is important that the mechanical properties are well understood not only at the point of cell seeding but throughout the culture period. In this study the change in the mechanical properties of three biopolymer hydrogels alginate, low methoxy pectin and gellan gum have been assessed in cell culture conditions. Samples of the gels were prepared encapsulating rat bone marrow stromal cells which were then cultured in osteogenic media. Acellular samples were also prepared and incubated in standard cell culture media. The rheological properties of the gels were measured over a culture period of 28 days and it was found that the gels degraded at very different rates. The degradation occurred most rapidly in the order alginate > Low methoxy pectin > gellan gum. The ability of each hydrogel to support differentiation of bone marrow stromal cells to osteoblasts was also verified by evidence of mineral deposits in all three of the materials. These results highlight that the mechanical properties of biopolymer hydrogels can vary greatly during in vitro culture, and provide the potential of selecting hydrogel cell culture substrates with mechanical properties that are tissue specific.

  17. Concentration Dependent Influence of Lipopolysaccharides on Separation of Hoof Explants and Supernatant Lactic Acid Concentration in an Ex Vivo/In Vitro Laminitis Model.

    Science.gov (United States)

    Reisinger, Nicole; Schaumberger, Simone; Nagl, Veronika; Hessenberger, Sabine; Schatzmayr, Gerd

    2015-01-01

    Laminitis is one of the most common diseases in horses. It is not only painful for the animal, but also has a significant financial impact on the equine industry. This multifactorial disease affects the connective tissue of the hoof. However, the pathogenesis of laminitis is still not fully understood. Endotoxins, also known as lipopolysaccharides (LPS), and bacterial exotoxins seem to play an important role during the development of laminitis. The aim of our study was to investigate the effect of increasing LPS concentrations (0, 2.5, 5, 10, and 100 μg/mL) on cell viability of isolated epidermal and dermal hoof cells as well as on the tissue integrity of hoof explants. Furthermore, glucose, acetic acid, lactic acid, and propionic acid concentrations in explant supernatants were measured to evaluate the energy metabolism in the hoof tissue. LPS did not exhibit cytotoxic effects on epidermal or dermal cells. Force required to separate LPS treated hoof explants decreased in a concentration dependent manner. Specifically, explants incubated with 10 and 100 μg/mL needed significantly less force to separate compared to control explants. Lactic acid concentrations were significantly decreased in explants incubated with 5, 10, or 100 μg/mL LPS, while glucose, acetic acid and propionic acid concentrations were unaffected by LPS treatment. Our study indicates that LPS has no cytotoxic effect on epidermal and dermal cells isolated from hoof tissue, but impairs integrity of hoof explants. In addition, LPS led to an alteration of the lactic acid production in the lamellar tissue. Since our data highlight that LPS can affect the integrity of the equine hoof tissue in vitro, endotoxins should be further explored for their contribution to facilitate the development of laminitis.

  18. Concentration Dependent Influence of Lipopolysaccharides on Separation of Hoof Explants and Supernatant Lactic Acid Concentration in an Ex Vivo/In Vitro Laminitis Model.

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    Nicole Reisinger

    Full Text Available Laminitis is one of the most common diseases in horses. It is not only painful for the animal, but also has a significant financial impact on the equine industry. This multifactorial disease affects the connective tissue of the hoof. However, the pathogenesis of laminitis is still not fully understood. Endotoxins, also known as lipopolysaccharides (LPS, and bacterial exotoxins seem to play an important role during the development of laminitis. The aim of our study was to investigate the effect of increasing LPS concentrations (0, 2.5, 5, 10, and 100 μg/mL on cell viability of isolated epidermal and dermal hoof cells as well as on the tissue integrity of hoof explants. Furthermore, glucose, acetic acid, lactic acid, and propionic acid concentrations in explant supernatants were measured to evaluate the energy metabolism in the hoof tissue. LPS did not exhibit cytotoxic effects on epidermal or dermal cells. Force required to separate LPS treated hoof explants decreased in a concentration dependent manner. Specifically, explants incubated with 10 and 100 μg/mL needed significantly less force to separate compared to control explants. Lactic acid concentrations were significantly decreased in explants incubated with 5, 10, or 100 μg/mL LPS, while glucose, acetic acid and propionic acid concentrations were unaffected by LPS treatment. Our study indicates that LPS has no cytotoxic effect on epidermal and dermal cells isolated from hoof tissue, but impairs integrity of hoof explants. In addition, LPS led to an alteration of the lactic acid production in the lamellar tissue. Since our data highlight that LPS can affect the integrity of the equine hoof tissue in vitro, endotoxins should be further explored for their contribution to facilitate the development of laminitis.

  19. Development of efficient catharanthus roseus regeneration and transformation system using agrobacterium tumefaciens and hypocotyls as explants

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    Wang Quan

    2012-06-01

    Full Text Available Abstract Background As a valuable medicinal plant, Madagascar periwinkle (Catharanthus roseus produces many terpenoid indole alkaloids (TIAs, such as vindoline, ajamlicine, serpentine, catharanthine, vinblastine and vincristine et al. Some of them are important components of drugs treating cancer and hypertension. However, the yields of these TIAs are low in wild-type plants, and the total chemical synthesis is impractical in large scale due to high-cost and their complicated structures. The recent development of metabolic engineering strategy offers a promising solution. In order to improve the production of TIAs in C. roseus, the establishment of an efficient genetic transformation method is required. Results To develop a genetic transformation method for C. roseus, Agrobacterium tumefaciens strain EHA105 was employed which harbors a binary vector pCAMBIA2301 containing a report β-glucuronidase (GUS gene and a selectable marker neomycin phosphotransferase II gene (NTPII. The influential factors were investigated systematically and the optimal transformation condition was achieved using hypocotyls as explants, including the sonication treatment of 10 min with 80 W, A. tumefaciens infection of 30 min and co-cultivation of 2 d in 1/2 MS medium containing 100 μM acetosyringone. With a series of selection in callus, shoot and root inducing kanamycin-containing resistance media, we successfully obtained stable transgenic regeneration plants. The expression of GUS gene was confirmed by histochemistry, polymerase chain reaction, and genomic southern blot analysis. To prove the efficiency of the established genetic transformation system, the rate-limiting gene in TIAs biosynthetic pathway, DAT, which encodes deacetylvindoline-4-O-acetyltransferase, was transferred into C. roseus using this established system and 9 independent transgenic plants were obtained. The results of metabolite analysis using high performance liquid chromatography (HPLC

  20. In Vitro propagation of enterolobium cyclocarpum (guanacaste from nodal explants of axenic seedlings

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    Araceli Rodríguez Sahagún

    2007-01-01

    Full Text Available Enterolobium cyclocarpum (Jacq. Griseb. es un árbol leguminoso de uso múltiple, el cual es considerado una especie amenazada, resultado de la sobreexplotación y las bajas tasas de propagación natural debidas a las características intrínsecas del árbol. Una alternativa para superar este problema es el establecimiento de sistemas para su propagación masiva en tiempos cortos. En este trabajo, se investigó un protocolo para la propagación in vitro de E. cyclocarpum utilizando los segmentos nodales axénicos obtenidos de plántulas germinadas in vitro. Las semillas colectadas en dos comunidades mexicanas fueron germinadas tanto ex vitro como in vitro, y se evaluó el efecto de un pre-tratamiento de escarificación térmica. Para los experimentos de propagación se seleccionaron semillas provenientes de sólo una de las comunidades, debido a que presentaban una menor variabilidad genética de acuerdo con marcadores RAPD y a que existía una gran variación en las respuestas observadas en lotes de semillas mezclados. Esta variación fisiológica presente en semillas mezcladas, probablemente refleja un efecto del genotipo.Los segmentos nodales obtenidos de las plántulas fueron cultivados en medio basal MS suplementado con 30 g/L de sacarosa en presencia de distintas concentraciones de ácido 1-naftalenacético (ANA en combinación con benziladenina (BA o kinetina (KIN. La mayor tasa de multiplicación (de 4.75 brotes por explante en promedio se obtuvo cuando el medio MS fue suplementado con 2.2 µM BA y 10.7 µM ANA. Los brotes obtenidos fueron enraizados en medio MS con la mitad de concentración de sales y sin reguladores de crecimiento. Las plántulas micropropagadas fueron aclimatadas y transferidas exitosamente a suelo con una tasa de sobrevivencia del 90%. Estas plantas eran morfológicamente similares a la planta madre y no se detectó variación entre ellas por el uso de marcadores RAPD, lo cual hace posible el uso de este

  1. Plant regeneration from cotyledonary explants of Eucalyptus camaldulensis dehn and histological study of organogenesis in vitro

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    Roberson Dibax

    2010-04-01

    Full Text Available The present work aimed at regenerating plants of Eucalyptus camaldulensis from the cotyledonary explants and describing the anatomy of the tissues during callogenesis and organogenesis processes, in order to determine the origin of the buds. The cotyledonary leaves of E. camaldulensis were cultured in Murashige and Skoog (MS, WPM and JADS media supplemented with 2.7 µM NAA and 4.44 µM BAP. The best results for bud regeneration were obtained on MS and WPM media (57.5 and 55% of calluses formed buds, respectively. Shoot elongation and rooting (80% were obtained on MS/2 medium (with half-strength salt concentration with 0.2% activated charcoal. Acclimatization was performed in the growth chamber for 48 h and then the plants were transferred to a soil:vermiculite mixture and cultured in a greenhouse. Histological studies revealed that the callogenesis initiated in palisade parenchyma cells and that the adventitious buds were formed from the calluses, indicating indirect organogenesis.Este trabalho teve como objetivo a obtenção de plantas de Eucalyptus camaldulensis a partir de folhas cotiledonares e o estudo da anatomia dos tecidos durante a calogênese e organogênese para determinar a origem das gemas. Folhas cotiledonares foram cultivadas em meios de cultura MS, WPM e JADS suplementados com 2,7 µM de ANA e 4,44 µM de BAP. Os melhores resultados para a regeneração de gemas foram obtidos com os meios MS e WPM. Para o alongamento e enraizamento, o meio de cultura MS/2 contendo 0,2% de carvão ativado apresentou-se eficiente para ambas as etapas. A aclimatização foi realizada mediante a abertura dos frascos na sala de crescimento por 48 horas, seguido da transferência para casa-de-vegetação com nebulização intermitente. Estudos histológicos foram conduzidos e revelaram que a calogênese teve início nas células do parênquima paliçádico e que as gemas adventícias formaram-se a partir dos calos, indicando a organogênese indireta.

  2. A novel method for the culture and polarized stimulation of human intestinal mucosa explants.

    Science.gov (United States)

    Tsilingiri, Katerina; Sonzogni, Angelica; Caprioli, Flavio; Rescigno, Maria

    2013-05-01

    Few models currently exist to realistically simulate the complex human intestine's micro-environment, where a variety of interactions take place. Proper homeostasis directly depends on these interactions, as they shape an entire immunological response inducing tolerance against food antigens while at the same time mounting effective immune responses against pathogenic microbes accidentally ingested with food. Intestinal homeostasis is preserved also through various complex interactions between the microbiota (including food-associated beneficial bacterial strains) and the host, that regulate the attachment/degradation of mucus, the production of antimicrobial peptides by the epithelial barrier, and the "education" of epithelial cells' that controls the tolerogenic or immunogenic phenotype of unique, gut-resident lymphoid cells' populations. These interactions have been so far very difficult to reproduce with in vitro assays using either cultured cell lines or peripheral blood mononuclear cells. In addition, mouse models differ substantially in components of the intestinal mucosa (mucus layer organization, commensal bacteria community) with respect to the human gut. Thus, studies of a variety of treatments to be brought in the clinics for important stress-related or pathological conditions such as irritable bowel syndrome, inflammatory bowel disease or colorectal cancer have been difficult to carry out. To address these issues, we developed a novel system that enables us to stimulate explants of human intestinal mucosa that retain their in situ conditioning by the host microbiota and immune response, in a polarized fashion. Polarized apical stimulation is of great importance for the outcome of the elicited immune response. It has been repeatedly shown that the same stimuli can produce completely different responses when they bypass the apical face of the intestinal epithelium, stimulating epithelial cells basolaterally or coming into direct contact with lamina

  3. Evaluation of bone response to synthetic bone grafting material treated with argon-based atmospheric pressure plasma

    Energy Technology Data Exchange (ETDEWEB)

    Beutel, Bryan G., E-mail: bryanbeutel@gmail.com; Danna, Natalie R.; Gangolli, Riddhi; Granato, Rodrigo; Manne, Lakshmiprada; Tovar, Nick; Coelho, Paulo G.

    2014-12-01

    Bone graft materials are utilized to stimulate healing of bone defects or enhance osseointegration of implants. In order to augment these capabilities, various surface modification techniques, including atmospheric pressure plasma (APP) surface treatment, have been developed. This in vivo study sought to assess the effect of APP surface treatment on degradation and osseointegration of Synthograft™, a beta-tricalcium phosphate (β-TCP) synthetic bone graft. The experimental (APP-treated) grafts were subjected to APP treatment with argon for a period of 60 s. Physicochemical characterization was performed by environmental scanning electron microscopy, surface energy (SE), and x-ray photoelectron spectroscopy analyses both before and after APP treatment. Two APP-treated and two untreated grafts were surgically implanted into four critical-size calvarial defects in each of ten New Zealand white rabbits. The defect samples were explanted after four weeks, underwent histological analysis, and the percentages of bone, soft tissue, and remaining graft material were quantified by image thresholding. Material characterization showed no differences in particle surface morphology and that the APP-treated group presented significantly higher SE along with higher amounts of the base material chemical elements on it surface. Review of defect composition showed that APP treatment did not increase bone formation or reduce the amount of soft tissue filling the defect when compared to untreated material. Histologic cross-sections demonstrated osteoblastic cell lines, osteoid deposition, and neovascularization in both groups. Ultimately, argon-based APP treatment did not enhance the osseointegration or degradation of the β-TCP graft. Future investigations should evaluate the utility of gases other than argon to enhance osseointegration through APP treatment. - Highlights: • Degradation/osseointegration of bone graft treated with argon-based APP is studied. • APP treatment did

  4. Enhanced functional integration of human photoreceptor precursors into human and rodent retina in an ex vivo retinal explant model system.

    Science.gov (United States)

    Yanai, Anat; Laver, Christopher R J; Gregory-Evans, Cheryl Y; Liu, Ran R; Gregory-Evans, Kevin

    2015-06-01

    Retinal disease is the major cause of irreversible blindness in developed countries. Transplantation of photoreceptor precursor cells (PPCs) derived from human embryonic stem cells (hESCs) is a promising and widely applicable approach for the treatment of these blinding conditions. Previously, it has been shown that after transplantation into the degenerating retina, the percentage of PPCs that undergo functional integration is low. The factors that inhibit PPC engraftment remain largely unknown, in part, because so many adverse factors could be at play during in vivo experiments. To advance our knowledge in overcoming potential adverse effects and optimize PPC transplantation, we have developed a novel ex vivo system. Harvested neural retina was placed directly on top of cultured retinal pigment epithelial (RPE) cells from a number of different sources. To mimic PPC transplantation into the subretinal space, hESC-derived PPCs were inserted between the retinal explant and underlying RPE. Explants cocultured with hESC-derived RPE maintained normal gross morphology and viability for up to 2 weeks, whereas the explants cultured on ARPE19 and RPE-J failed by 7 days. Furthermore, the proportion of PPCs expressing ribbon synapse-specific proteins BASSOON and RIBEYE was significantly higher when cocultured with hESC-derived RPE (20% and 10%, respectively), than when cocultured with ARPE19 (only 6% and 2%, respectively). In the presence of the synaptogenic factor thrombospondin-1 (TSP-1), the proportion of BASSOON-positive and RIBEYE-positive PPCs cocultured with hESC-derived RPE increased to ∼30% and 15%, respectively. These data demonstrate the utility of an ex vivo model system to define factors, such as TSP-1, which could influence integration efficiency in future in vivo experiments in models of retinal degeneration.

  5. High-frequency in vitro plantlet regeneration from apical bud as a novel explant of Carum copticum L.

    Institute of Scientific and Technical Information of China (English)

    Mansoureh Salehi; Bahman Hosseini; Zohreh Jabbarzadeh

    2014-01-01

    Objective: To develop an in vitro regeneration system to increase the recovery of Carum copticum L. plantlets as a part of developing a metabolic engineering program.Methods:3-acetic acid and indole butyric acid on direct shoot regeneration and rooting of ajowan from apical bud explants were assessed. All explants were cultured on Murashige and Skoog (MS) medium supplemented with different combinations of 6-benzyl amino purine (BAP) (0, 2.2, 4.4, 8.8µ The efficacy of different concentrations and combinations of 6-benzyladenine, indole-Results: The maximum shoot regeneration frequency (97.5%) and the highest number of shoots produced from apical buds (34 shoots per explant) were obtained on MS medium fortified with BAP (4.4 µmol/L) and IAA (0.5 µmol/L). Low shoot regeneration frequency was observed in BAP free treatments. The effects of different strengths of MS medium and various concentrations of IAA and indole-3- butyric acid on rooting rate, length and average number of roots were also investigated. Application of indole-3- butyric acid (6 µmol/L) in full-strength MS medium, was more effective than IAA and resulted in highest shoot regeneration frequency with the rooting rate of 100% and highest mean number of roots per shoot (41.8). The rooted plantlets were acclimatized successfully in greenhouse conditions with a survival rate of 90%. mol/L) and indole-3-acetic acid (IAA) (0, 0.5, 1.1, 2.2 µmol/L). Conclusion: In this study, a simple and reliable regeneration and acclimatization protocol for Carum copticum has been presented. This protocol can be found very advantageous for a variety of purposes, including mass multiplication of Carum species, medicinal plant breeding studies and transgenic plant production.

  6. Indução de calos friáveis em explantes foliares de Salix (Salyx humboldtiana Willd)

    OpenAIRE

    Santos,Breno Régis; Paiva,Renato; Martinotto,Cristiano; Nogueira,Raírys Cravo; Paiva,Patrícia Duarte de Oliveira

    2005-01-01

    O salix é uma árvore ornamental adaptada a terrenos úmidos, sendo considerada uma espécie importante para a recomposição de áreas ciliares degradadas. A madeira pode ser empregada na indústria em geral. Apesar de produzir grande quantidade de sementes, estas não possuem alta percentagem de germinação. O presente trabalho teve como objetivo realizar o estabelecimento in vitro de salix através da indução e formação de calos friáveis em explantes foliares, visando a uma posterior regeneração de ...

  7. Usefulness of a bioengineered oral mucosa model for preventing palate bone alterations in rabbits with a mucoperiostial defect.

    Science.gov (United States)

    Fernández-Valadés-Gámez, Ricardo; Garzón, Ingrid; Liceras-Liceras, Esther; España-López, Antonio; Carriel, Víctor; Martin-Piedra, Miguel-Ángel; Muñoz-Miguelsanz, María-Ángeles; Sánchez-Quevedo, Maria-Carmen; Alaminos, Miguel; Fernández-Valadés, Ricardo

    2016-02-19

    The use of mucoperiostial flaps during cleft palate surgery is associated with altered palatal bone growth and development. We analyzed the potential usefulness of a bioengineered oral mucosa in an in vivo model of cleft palate. First, a 4 mm palate defect was created in one side of the palate oral mucosa of 3 week-old New Zealand rabbits, and a complete autologous bioengineered oral mucosa (BOM) or acellular fibrin-agarose scaffold (AS) was implanted. No material was implanted in the negative controls (NC), and positive controls were not subjected to palatal defect (PC). Animals were allowed to grow for 6 months and the results were analyzed morphologically (palate mucosa and bone size) and histologically. Results show that palatal mucosa and bone growth and development were significantly altered in NC and AS animals, whereas BOM animals had similar results to PC and the bioengineered oral mucosa was properly integrated in the host palate. The amount and compaction of collagen fibers was similar between BOM and PC, and both groups of animals had comparable contents of proteoglycans and glycoproteins at the palate bone. No differences were found for decorin, osteocalcin and BMP2. The use of bioengineered oral mucosa substitutes is able to improve palate growth and maturation by preventing the alterations found in animals with denuded palate bone. These results support the potential clinical usefulness of BOM substitutes for the treatment of patients with cleft palate and other conditions in which palate mucosa grafts are necessary with consequent bone denudation.

  8. Dating of cremated bones

    NARCIS (Netherlands)

    Lanting, JN; Aerts-Bijma, AT; van der Plicht, J; Boaretto, E.; Carmi, I.

    2001-01-01

    When dating unburnt bone, bone collagen, the organic fraction of the bone, is used. Collagen does not survive the heat of the cremation pyre, so dating of cremated bone has been considered impossible. Structural carbonate in the mineral fraction of the bone, however, survives the cremation process.

  9. ANATOMICAL STUDIES OF IN VITRO ORGANOGENESIS INDUCED IN LEAF-DERIVED EXPLANTS OF PASSIONFRUIT ESTUDOS ANATÔMICOS DA ORGANOGÊNESE IN VITRO INDUZIDA EM EXPLANTES DE FOLHA DE MARACUJÁ

    Directory of Open Access Journals (Sweden)

    BEATRIZ APPEZZATO DA GLORIA

    1999-11-01

    Full Text Available With the aim of studying the organogenesis in vitro in Passiflora edulis Sims f. flavicarpa Deg., the passionfruit, leaf-derived explants were cultured on media containing NAA or BAP and incubated either in continuous darkness or in light. The histological events leading to de novo organ formation were evaluated. Darkness induces rhizogenesis in the presence of NAA, whereas direct shoot regeneration is stimulated by light and BAP. This latter condition is recommended for passionfruit micropropagation as several adventitious shoot buds were formed from meristemoids of parenchymal origin.Com o objetivo de estudar a organogênese in vitro em Passiflora edulis Sims f. flavicarpa Deg., o maracujá-amarelo, explantes derivados de folha foram cultivados em meio contendo NAA ou BAP, no escuro e na presença de luz. Foram descritos os eventos histológicos que levam à formação de novo de órgãos. Concluiu-se que o escuro induz a rizogênese, na presença de NAA, enquanto a regeneração de brotos é estimulada pela luz e BAP. Esta condição é recomendada para micropropagar o maracujá uma vez que vários brotos adventícios são formados a partir de meristemóides de origem parenquimática.

  10. Clinical use of acellular allogenic dermis or acellular porcine dermis with split- thickaess autologous skin graftin 119 cases%去细胞异体真皮、去细胞猪真皮和自体刃厚皮移植在临床中的应用

    Institute of Scientific and Technical Information of China (English)

    柴家科; 杨红明; 李利根; 郭振荣; 盛志勇; 许明火; 陈宝驹; 贾晓明; 京萨; 陆江阳; 李国红

    2000-01-01

    Objective To observe the effect of acellular allogenic dermis or acelludar porcine dermis together with split-thickness autologous skin graft on coverage of deep burn wound and the wound of scar excision. Methods Acellular allogenic dermis or acellular porcine dermis produced by our unit, with split-thickness autologous skin graft, were used in repairing various wounds in 119 cases. The take rate of various wounds was compared, and the take rate of wounds in which autologous skin fr-om different sites was used to cover either acellular allogenic dermis or acellular porcine dermis was also compared. Histological examination and follow-up were made in some cases. Results After tangential excision, eschar excision and scar excision, the wounds were covered with either allogenic acellular dernis or porcine acellular dermis and autologous split-thickness skin. The take rate was found to be (93.4 ± 3.4) %, ( 92.1 ± 4.6) %, (94.5 ± 3.5) %, respectively. There was no a significant defference in take rate( P > 0.05). No significant difference in take rate between the transplantation of allogenic acellular dermis with autologous split-thickness skin and transplantation of porcine acellular demis with autologous split-thickness skin was found (P>0.05). When autologous split-thickness skin harvested from the trunk or extremites was used, the take rate was(93.1 ±4.8)%, (89.0±6.2)%, respectively, whereas the take rate was only about 70% ~ 80% when skin harvested from the scalp was used. The take rate was only about 40% when ace llalar allogenic dermis and autologous micro-skin graft were used to cover the wound. Histological examination 19 months after the transplantation revealed normal structure. Collagenous fibers were orderly arranged. There was no apparent hyperplasia of collagenous fibers. Hair follicles, sweat glands and other skin appendages were not found in the healed area. Grossly, no obvious pigmentation was

  11. Explant culture: a simple, reproducible, efficient and economic technique for isolation of mesenchymal stromal cells from human adipose tissue and lipoaspirate.

    Science.gov (United States)

    Priya, Nancy; Sarcar, Shilpita; Majumdar, Anish Sen; SundarRaj, Swathi

    2014-09-01

    Adipose tissue has emerged as a preferred source of mesenchymal stem/stromal cells (MSC), due to its easy accessibility and high MSC content. The conventional method of isolation of adipose tissue-derived stromal cells (ASC) involves enzymatic digestion and centrifugation, which is a costly and time-consuming process. Mechanical stress during isolation, use of bacterial-derived products and potential contamination with endotoxins and xenoantigens are other disadvantages of this method. In this study, we propose explant culture as a simple and efficient process to isolate ASC from human adipose tissue. This technique can be used to reproducibly isolate ASC from fat tissue obtained by liposuction as well as surgical resection, and yields an enriched ASC population free from contaminating haematopoietic cells. We show that explanting adipose tissue results in a substantially higher yield of ASC at P0 per gram of initial fat tissue processed, as compared to that obtained by enzymatic digestion. We demonstrate that ASC isolated by explant culture are phenotypically and functionally equivalent to those obtained by enzymatic digestion. Further, the explant-derived ASC share the immune privileged status and immunosuppressive properties implicit to MSC, suggesting that they are competent to be tested and applied in allogeneic clinical settings. As explant culture is a simple, inexpensive and gentle method, it may be preferred over the enzymatic technique for obtaining adipose tissue-derived stem/stromal cells for tissue engineering and regenerative medicine, especially in cases of limited starting material.

  12. A protocol for sonication-assisted Agrobacterium rhizogenes-mediated transformation of haploid and diploid sugar beet (Beta vulgaris L.) explants.

    Science.gov (United States)

    Klimek-Chodacka, Magdalena; Baranski, Rafal

    2014-01-01

    Hairy root cultures obtained after Agrobacterium rhizogenes-mediated genetic transformation can serve as a model system for studying plant metabolism and physiology, or can be utilized for the production of secondary metabolites. So far no efficient protocol of hairy root development in sugar beet has been publically released. In this work, two A. rhizogenes strains (A4T and LBA1334) carrying a binary vector pBIN-m-gfp5-ER or pCAMBIA1301 possessing gfp and uidA reporter genes were used to transform petiole explants of haploid and diploid sugar beet genotypes. Five treatment combinations of sonicated-assisted Agrobacterium-mediated transformation were compared. Hairy roots appeared on 0% to 54% of explants depending on the treatment combination used. The highest frequency was achieved when explants of a diploid genotype were sonicated for 15 s in the inoculum containing A. rhizogenes of OD600=0.5 and then co-cultured for three days. Using the same treatment combinations the explants of haploid genotypes developed hairy roots with the frequency ranging from 10% to 36%. Transformation efficiency was independent on the bacterial strain used. The results indicate that haploid sugar beet explants are amenable to transformation using A. rhizogenes, and that the efficiency of that process can be increased by applying short ultrasound treatment.

  13. Effect on cochlea function of guinea pig after controlled release recombinant human bone morphogenetic protein 2 transplanted into the middle ear

    Institute of Scientific and Technical Information of China (English)

    LI Xue-sheng; SUN Jian-jun; JIANG Wei; LIU Xiao

    2010-01-01

    Background The recombinant human bone morphogenetic protein 2 (rhBMP-2) has been used to induce osteogenesis in animals' middle ear and this technique is possible to be used to reconstruct the defects of ossicles. The side effects of the rhBMP-2 in middle ear should be observed before using in clinic. Thus we prepared the controlled release rhBMP-2 and implanted it into the acoustic bulla of guinea pigs. The effect on the cochlea was observed. Methods We prepared the acellular cancellous bone, accompanied with rhBMP-2. The material accompanied with rhBMP-2 was implanted into one acoustic bulla of the animal and the opposite side of the acoustic bulla was implanted with acellular cancellous bone without rhBMP-2. Totally 20 guinea pigs were undergone this procedure. After the operation, the auditory brainstem response (ABR) of the animals was tested according to the time sequence. Three months after the operation, the animals were sacrificed. The osteogenesis induced by rhBMP-2, the acoustic bulla and cochlea affected by rhBMP-2 were observed. The structures of hair cells were observed after silver nitrate staining. Results The animals were recovered soon after surgery. The hearing thresholds of the animals were declined slightly just after the surgery and come back completely after 3 months. Also, the bulla and cochlea were normal in shape. The osteogenesis occurred in the pore of the acellular cancellous bone with rhBMP-2. There was not any abnormal hyperplasia of bone in the bulla and cochlea. The articulation between the stapes and oval window was not merged. The shapes of the hair cells were normal and there was no obvious deletion of the hair cells compared with control group. Conclusions The controlled release rhBMP-2 transplanted into the middle ear could induce osteogenesis in the bulla of the animals. It did not affect the shape of the bulla and the hearing threshold of the animal, and did not induce the abnormal hyperplasia of bone in the bulla and might

  14. Synthetic scaffold coating with adeno-associated virus encoding BMP2 to promote endogenous bone repair.

    Science.gov (United States)

    Dupont, Kenneth M; Boerckel, Joel D; Stevens, Hazel Y; Diab, Tamim; Kolambkar, Yash M; Takahata, Masahiko; Schwarz, Edward M; Guldberg, Robert E

    2012-03-01

    Biomaterial scaffolds functionalized to stimulate endogenous repair mechanisms via the incorporation of osteogenic cues offer a potential alternative to bone grafting for the treatment of large bone defects. We first quantified the ability of a self-complementary adeno-associated viral vector encoding bone morphogenetic protein 2 (scAAV2.5-BMP2) to enhance human stem cell osteogenic differentiation in vitro. In two-dimensional culture, scAAV2.5-BMP2-transduced human mesenchymal stem cells (hMSCs) displayed significant increases in BMP2 production and alkaline phosphatase activity compared with controls. hMSCs and human amniotic-fluid-derived stem cells (hAFS cells) seeded on scAAV2.5-BMP2-coated three-dimensional porous polymer Poly(ε-caprolactone) (PCL) scaffolds also displayed significant increases in BMP2 production compared with controls during 12 weeks of culture, although only hMSC-seeded scaffolds displayed significantly increased mineral formation. PCL scaffolds coated with scAAV2.5-BMP2 were implanted into critically sized immunocompromised rat femoral defects, both with or without pre-seeding of hMSCs, representing ex vivo and in vivo gene therapy treatments, respectively. After 12 weeks, defects treated with acellular scAAV2.5-BMP2-coated scaffolds displayed increased bony bridging and had significantly higher bone ingrowth and mechanical properties compared with controls, whereas defects treated with scAAV2.5-BMP2 scaffolds pre-seeded with hMSCs failed to display significant differences relative to controls. When pooled, defect treatment with scAAV2.5-BMP2-coated scaffolds, both with or without inclusion of pre-seeded hMSCs, led to significant increases in defect mineral formation at all time points and increased mechanical properties compared with controls. This study thus presents a novel acellular bone-graft-free endogenous repair therapy for orthotopic tissue-engineered bone regeneration.

  15. Healing rate and autoimmune safety of full-thickness wounds treated with fish skin acellular dermal matrix versus porcine small-intestine submucosa: a noninferiority study.

    Science.gov (United States)

    Baldursson, Baldur Tumi; Kjartansson, Hilmar; Konrádsdóttir, Fífa; Gudnason, Palmar; Sigurjonsson, Gudmundur F; Lund, Sigrún Helga

    2015-03-01

    A novel product, the fish skin acellular dermal matrix (ADM) has recently been introduced into the family of biological materials for the treatment of wounds. Hitherto, these products have been produced from the organs of livestock. A noninferiority test was used to compare the effect of fish skin ADM against porcine small-intestine submucosa extracellular matrix in the healing of 162 full-thickness 4-mm wounds on the forearm of 81 volunteers. The fish skin product was noninferior at the primary end point, healing at 28 days. Furthermore, the wounds treated with fish skin acellular matrix healed significantly faster. These results might give the fish skin ADM an advantage because of its environmental neutrality when compared with livestock-derived products. The study results on these acute full-thickness wounds might apply for diabetic foot ulcers and other chronic full-thickness wounds, and the shorter healing time for the fish skin-treated group could influence treatment decisions. To test the autoimmune reactivity of the fish skin, the participants were tested with the following ELISA (enzyme-linked immunosorbent assay) tests: RF, ANA, ENA, anti ds-DNA, ANCA, anti-CCP, and anticollagen I and II. These showed no reactivity. The results demonstrate the claims of safety and efficacy of fish skin ADM for wound care.

  16. Time-dependent bladder tissue regeneration using bilayer bladder acellular matrix graft-silk fibroin scaffolds in a rat bladder augmentation model.

    Science.gov (United States)

    Zhao, Yang; He, Yi; Zhou, Zhe; Guo, Jian-hua; Wu, Jia-sheng; Zhang, Ming; Li, Wei; Zhou, Juan; Xiao, Dong-dong; Wang, Zhong; Sun, Kang; Zhu, Ying-jian; Lu, Mu-jun

    2015-09-01

    With advances in tissue engineering, various synthetic and natural biomaterials have been widely used in tissue regeneration of the urinary bladder in rat models. However, reconstructive procedures remain insufficient due to the lack of appropriate scaffolding, which should provide a waterproof barrier function and support the needs of various cell types. To address these problems, we have developed a bilayer scaffold comprising a porous network (silk fibroin [SF]) and an underlying natural acellular matrix (bladder acellular matrix graft [BAMG]) and evaluated its feasibility and potential for bladder regeneration in a rat bladder augmentation model. Histological (hematoxylin and eosin and Masson's trichrome staining) and immunohistochemical analyses demonstrated that the bilayer BAMG-SF scaffold promoted smooth muscle, blood vessel, and nerve regeneration in a time-dependent manner. At 12weeks after implantation, bladders reconstructed with the BAMG-SF matrix displayed superior structural and functional properties without significant local tissue responses or systemic toxicity. These results demonstrated that the bilayer BAMG-SF scaffold may be a promising scaffold with good biocompatibility for bladder regeneration in the rat bladder augmentation model.

  17. Accelerated craniofacial bone regeneration through dense collagen gel scaffolds seeded with dental pulp stem cells

    Science.gov (United States)

    Chamieh, Frédéric; Collignon, Anne-Margaux; Coyac, Benjamin R.; Lesieur, Julie; Ribes, Sandy; Sadoine, Jérémy; Llorens, Annie; Nicoletti, Antonino; Letourneur, Didier; Colombier, Marie-Laure; Nazhat, Showan N.; Bouchard, Philippe; Chaussain, Catherine; Rochefort, Gael Y.

    2016-12-01

    Therapies using mesenchymal stem cell (MSC) seeded scaffolds may be applicable to various fields of regenerative medicine, including craniomaxillofacial surgery. Plastic compression of collagen scaffolds seeded with MSC has been shown to enhance the osteogenic differentiation of MSC as it increases the collagen fibrillary density. The aim of the present study was to evaluate the osteogenic effects of dense collagen gel scaffolds seeded with mesenchymal dental pulp stem cells (DPSC) on bone regeneration in a rat critical-size calvarial defect model. Two symmetrical full-thickness defects were created (5 mm diameter) and filled with either a rat DPSC-containing dense collagen gel scaffold (n = 15), or an acellular scaffold (n = 15). Animals were imaged in vivo by microcomputer tomography (Micro-CT) once a week during 5 weeks, whereas some animals were sacrificed each week for histology and histomorphometry analysis. Bone mineral density and bone micro-architectural parameters were significantly increased when DPSC-seeded scaffolds were used. Histological and histomorphometrical data also revealed significant increases in fibrous connective and mineralized tissue volume when DPSC-seeded scaffolds were used, associated with expression of type I collagen, osteoblast-associated alkaline phosphatase and osteoclastic-related tartrate-resistant acid phosphatase. Results demonstrate the potential of DPSC-loaded-dense collagen gel scaffolds to benefit of bone healing process.

  18. Outgrowth of fibroblast cells from goat skin explants in three different culture media and the establishment of cell lines.

    Science.gov (United States)

    Singh, Mahipal; Sharma, Anil K

    2011-02-01

    Three different commercially available media, known to support human and porcine-specific fibroblast cultures, were tested for their growth potential on goat skin explants. Although outgrowth of fibroblasts was observed in all media tested, irrespective of breed, porcine-specific media exhibited higher rate of growth. Using this media, three fibroblast cell lines (GSF289, GSF737, and GSF2010) from ear skin explants of normal healthy dairy goats of Kiko and Saanen breed were successfully established in culture. Liquid nitrogen stocks of these frozen cells had a viability rate of 96.2% in in vitro cultures. These cells were morphologically indistinguishable from the cell stocks prior to freezing. Analysis of the growth of a fifth passage culture revealed an 'S' shaped growth curve with a population doubling time of 25 h. The cell lines were found negative for microbial, fungal, and mycoplasma contaminations. These goat skin fibroblast lines and the simple method of their isolation and freezing with high rate of viability will provide additional tools to study molecular mechanisms that regulate fibroblast function and for genetic manipulation of small ruminants.

  19. How to explant a diseased liver for living donor liver transplantation after previous gastrectomy with severe adhesion (with video).

    Science.gov (United States)

    Eguchi, Susumu; Soyama, Akihiko; Takatsuki, Mitsuhisa; Hidaka, Masaaki; Adachi, Tomohiko; Kitasato, Amane; Baimakhanov, Zhassulan; Kuroki, Tamotsu

    2014-08-01

    We performed living donor liver transplantation (LDLT) in a patient who had undergone distal gastrectomy for gastric ulcer disease with Billroth I reconstruction 30 years before the LDLT. The adhesion was very severe between remnant stomach and hepatic hilum as well as left liver lobe with shortening of hepatoduodenal structures. After dissection of the infrahepatic inferior vena cava, the Spiegel lobe was identified from the dorsal side. The Spiegel lobe was then penetrated with a right angle dissector so that a plastic tape could be placed around the whole adhesion, including important structures in the hepatoduodenal ligament. Next, the right hepatic vein was transected with a vascular stapler using Pringle's maneuver using the plastic tape to fasten the entire adhesional structure. Subsequently, the trunk of the middle and left hepatic vein was transected after clamping. The remaining short hepatic veins in the left side were divided completely from the cranial to the caudal direction to dissect Spiegel's lobe. Finally, the hepatoduodenal ligament was identified from the attached remnant stomach and the duodenum and a vascular clamp was placed on the entire hepatoduaodenal ligament. Finally, the diseased liver was explanted for graft implantation. Thus, retrograde explantation of the liver was effective in decreasing the risk of damaging vital elements in the hepatoduodenal ligament, the remnant stomach, and the duodenum.

  20. Programming jammed Codman Hakim programmable valves: study of an explanted valve and successful programming in a patient.

    Science.gov (United States)

    Wong, Sui-To; Wen, Eleanor; Fong, Dawson

    2013-08-01

    Malfunction of a Codman Hakim programmable valve due to jamming of its programmable component may necessitate shunt revision. The authors report a method for programming jammed Codman Hakim programmable valves by using a Strata II magnet and additional neodymium magnets. The programming method was derived after studying a jammed valve in the laboratory that was explanted from an 10-year-old boy with a history of fourth ventricle ependymoma. Programming the explanted valve with a Codman programmer failed, but rotating a Strata II magnet above the valve resulted in rotation of the spiral cam in the valve. It was found that the Strata II magnet could be used to program the jammed valve by rotating the magnet 90° or multiples of 90° above the valve. The strength of the magnetic field of the Strata II magnet was able to be increased by putting neodymium magnets on it. The programming method was then successfully used in a patient with a jammed Codman Hakim programmable valve. After successful programming using this method, clinical and radiological follow-up of the patient was advised.

  1. MRI screening for silicone breast implant rupture: accuracy, inter- and intraobserver variability using explantation results as reference standard

    Energy Technology Data Exchange (ETDEWEB)

    Maijers, M.C.; Ritt, M.J.P.F. [VU University Medical Centre, Department of Plastic, Reconstructive and Hand Surgery, De Boelelaan 1117, PO Box 7057, Amsterdam (Netherlands); Niessen, F.B. [VU University Medical Centre, Department of Plastic, Reconstructive and Hand Surgery, De Boelelaan 1117, PO Box 7057, Amsterdam (Netherlands); Jan van Goyen Clinic, Department of Plastic Surgery, Amsterdam (Netherlands); Veldhuizen, J.F.H. [MRI Centre, Amsterdam (Netherlands); Manoliu, R.A. [MRI Centre, Amsterdam (Netherlands); VU University Medical Centre, Department of Radiology, Amsterdam (Netherlands)

    2014-06-15

    The recall of Poly Implant Prothese (PIP) silicone breast implants in 2010 resulted in large numbers of asymptomatic women with implants who underwent magnetic resonance imaging (MRI) screening. This study's aim was to assess the accuracy and interobserver variability of MRI screening in the detection of rupture and extracapsular silicone leakage. A prospective study included 107 women with 214 PIP implants who underwent explantation preceded by MRI. In 2013, two radiologists blinded for previous MRI findings or outcome at surgery, independently re-evaluated all MRI examinations. A structured protocol described the MRI findings. The ex vivo findings served as reference standard. In 208 of the 214 explanted prostheses, radiologists agreed independently about the condition of the implants. In five of the six cases they disagreed (2.6 %), but subsequently reached consensus. A sensitivity of 93 %, specificity of 93 %, positive predictive value of 77 % and negative predictive value of 98 % was found. The interobserver agreement was excellent (kappa value of 0.92). MRI has a high accuracy in diagnosing rupture in silicone breast implants. Considering the high kappa value of interobserver agreement, MRI appears to be a consistent diagnostic test. A simple, uniform classification, may improve communication between radiologist and plastic surgeon. (orig.)

  2. Pre-culturing of nodal explants in thidiazuron supplemented liquid medium improves in vitro shoot multiplication of Cassia angustifolia.

    Science.gov (United States)

    Siddique, I; Abdullwahab Bukhari, N; Perveen, K; Siddiqui, I; Anis, M

    2013-09-01

    An in vitro propagation system for Cassia angustifolia Vahl. has been developed. Due to the presence of sennosides, the demand of this plant has increased manyfold in global market. Multiple shoots were induced by culturing nodal explants excised from mature plants on a liquid Murashige and Skoog [8] medium supplemented with 5-100 μM of thidiazuron (TDZ) for different treatment duration (4, 8, 12 and 16 d). The optimal level of TDZ supplemented to the culture medium was 75 μM for 12 d induction period followed by subculturing in MS medium devoid of TDZ as it produced maximum regeneration frequency (87%), mean number of shoots (9.6 ± 0.33) and shoot length (4.4 ± 0.46 cm) per explant. A culture period longer than 12 d with TDZ resulted in the formation of fasciated or distorted shoots. Ex vitro rooting was achieved when the basal cut end of regenerated shoots was dipped in 200 μM indole-3-butyric acid (IBA) for half an hour followed by their transplantation in plastic pots filled with sterile soilrite where 85% plantlets grew well and all exhibited normal development. The present findings describe an efficient and rapid plant regeneration protocol that can further be used for genetic transformation studies.

  3. The Effects of Gamma Irradiation on the Growth and Propagation of In-Vitro Chrysanthemum Shoot Explants (cv. Yellow Puma

    Directory of Open Access Journals (Sweden)

    I. Dwimahyani

    2010-08-01

    Full Text Available The study on the effect of gamma irradiation on in-vitro shoot growth of chrysanthemum cv. Yellow Puma has been carried out. The aim of the study was to observe genetic variability of shoot growth caused by gamma irradiation. Shoot explants with four leaves were irradiated by gamma with dose of 10, 15 and 20 Gy with 3 replications at each of dose. The irradiated shoot explants were then transferred into fresh MS solid medium and placed in a growth room. Observation was performed on number of leaves and branches on M1V0 generation, while plantlets height and number of branches were observed a M1V1 generation. Number of survival plantlets and multiplication rate on three subsequent subcultures were observed as well. Results showed that gamma rays with dose of 20 Gy inhibited growth of leaves as much as 50% compared to control (shoots without irradiation, and branches 73.7% in three weeks. Observation on multiplication rate at M1V1 generation showed that gamma irradiation with dose of 10 Gy promoted multiplication rate as much as 10% higher than control. It can be concluded that in vitro mutagenesis using gamma iradiation with dose of 10 to 15 Gy can be used for inducing genetic variability of chrysanthemum cv. Yellow Puma.

  4. THE PHENOLS ACCUMULATION IN TRANSFORMED ROOT CULTURES OF DIFFERENT EXPLANTS SOURCES OF COMMON BUCKWHEAT (Fagopyrum esculentum Moench

    Directory of Open Access Journals (Sweden)

    O. V. Sytar

    2013-06-01

    Full Text Available The growth parameters of transformed root cultures, total phenolic content and phenolic acids composition has been studied in root cultures, which were obtained from various explants of buckwheat by Agrobacterium rhizogenes strains A4. The methods of obtaining of the transformed root cultures, total phenol estimation, gas-liquid chromatography and polymerase chain reaction has been used. Elevated levels of total phenols in transformed roots of buckwheat from different sources of explants have been found. The high content of chlorogenic, p-hydroxybenzoic, p-anisic and caffeic acids has been discovered in the root cultures, which can be used for their industrial production. Maximal root growth was equal 21.2 g/l of dry weight in the roots as source for root culture, 17.7 g/l with leaves and 14.6 g/l with stems at 3 week after placement. Molecular analysis by polymerase chain reaction amplification was confirmed that the rol B gene (652 bp which transferred info hairy roots from Ri-plasmid in Agrobacterium rhizogenes is responsible for induction of root from plant species.

  5. Effect of radiation on the growth of normal and malignant human oesophageal explant cultures pre-treated with bleomycin

    Energy Technology Data Exchange (ETDEWEB)

    Seymour, C.B.; Cusack, A.; Mothersill, C.; Hennessy, T.P.

    1988-05-01

    A method has been developed for testing the response of oesophageal explants from tumour and surrounding normal tissue in the same patient to chemotherapy and ..gamma..-radiation, singly and in combination. The test allows treatment combinations, time and order of administration of agents to the tissue to be accurately controlled. Cytotoxicity, determined by measuring the area of outgrowth from an explant 2 weeks after plating, is the most useful short-term end-point, although many other are possible. Results showing differential cytotoxicity of belomycin with and without radiation in squamous and adenocarcinoma of the oesophagus and surrounding normal tissue from the same patient indicate tumour cells are relatively resistant to radiation alone, low levels of belomycin with or without radiation preferentially spare tumour cells and high levels, in combination with any radiation dose tested, but not without radiation, spare normal cells and give a significantly high amount of relative tumour cell kill. Belomycin must be added to the cells just before or just after irradiation to obtain the normal-tissue sparing effect.

  6. Smoking and Bone Health

    Science.gov (United States)

    ... supported by your browser. Home Bone Basics Lifestyle Smoking and Bone Health Publication available in: PDF (85 ... late to adopt new habits for healthy bones. Smoking and Osteoporosis Cigarette smoking was first identified as ...

  7. [Influence of genotype, explant type and component of culture medium on in vitro callus induction and shoot organogenesis of tomato (Solanum lycopersicum L.)].

    Science.gov (United States)

    Khaliluev, M R; Bogoutdinova, L R; Baranova, G B; Baranova, E N; Kharchenko, P N; Dolgov, S V

    2014-01-01

    The influence of explant type as well as of the type of growth regulators and concentration on callus induction processes and somatic organogenesis of shoots was studied in vitro on four tomato genotypes of Russian breeding. Cytological study of callus tissue was conducted. It was established that tomato varieties possess a substantially greater ability to indirect shoot organogenesis compared with the F1 hybrid. The highest frequency of somatic organogenesis of shoots, as well as their number per explant, was observed for most of the genotypes studied during the cultivation of cotyledons on Murashige-Skoog culture medium containing 2 mg/l of zeatin in combination with 0.1 mg/l of 3-indoleacetic acid. An effective protocol of indirect somatic organogenesis of shoots from different explants of tomato varieties with a frequency of more than 80% was developed.

  8. Suppression of MMP activity in bovine cartilage explants cultures has little if any effect on the release of aggrecanase-derived aggrecan fragments

    DEFF Research Database (Denmark)

    Wang, Bijue; Chen, Pingping; Jensen, Anne-Christine Bay

    2009-01-01

    - and aggrecanase-derived fragments of aggrecan and type II collagen into the supernatant of bovine cartilage explants cultures using neo-epitope specific immunoassays, and to associate the release of these fragments with the activity of proteolytic enzymes using inhibitors. FINDINGS: Bovine cartilage explants were...... cultured in the presence or absence of the catabolic cytokines oncostatin M (OSM) and tumor necrosis factor alpha (TNFalpha). In parallel, explants were co-cultured with protease inhibitors such as GM6001, TIMP1, TIMP2 and TIMP3. Fragments released into the supernatant were determined using a range of neo......-epitope specific immunoassays; (1) sandwich (342)FFGVG-G2 ELISA, (2) competition NITEGE(373)ELISA (3) sandwich G1-NITEGE(373 )ELISA (4) competition (374)ARGSV ELISA, and (5) sandwich (374)ARGSV-G2 ELISA all detecting aggrecan fragments, and (6) sandwich CTX-II ELISA, detecting C-telopeptides of type II collagen...

  9. Comparison of cardiac power output and exercise performance in patients with left ventricular assist devices, explanted (recovered) patients, and those with moderate to severe heart failure.

    Science.gov (United States)

    Jakovljevic, Djordje G; George, Robert S; Donovan, Gay; Nunan, David; Henderson, Keiran; Bougard, Robert S; Yacoub, Magdi H; Birks, Emma J; Brodie, David A

    2010-06-15

    Peak cardiac power output (CPO), as a direct measurement of overall cardiac function, has been shown to be a most powerful predictor of prognosis for patients with chronic heart failure. The present study assessed CPO and exercise performance in patients implanted with a left ventricular assist device (LVAD), those explanted due to myocardial recovery, and those with moderate to severe heart failure. Hemodynamic and respiratory gas exchange measurements were undertaken at rest and at peak graded exercise. These were performed in 54 patients-20 with moderate to severe heart failure, 18 with implanted LVADs, and 16 with explanted LVADs. At rest there was a nonsignificant difference in CPO among groups (p >0.05). Peak CPO was significantly higher in the explanted LVAD than in the heart failure and implanted LVAD groups (heart failure 1.90 +/- 0.45 W, implanted LVAD 2.37 +/- 0.55 W, explanted LVAD 3.39 +/- 0.61 W, p <0.01) as was peak cardiac output (heart failure 9.1 +/- 2.1 L/min, implanted LVAD 12.4 +/- 2.2 L/min, explanted LVD 14.6 +/- 2.9 L/min, p <0.01). Peak oxygen consumption was higher in the explanted LVAD than in the heart failure and implanted LVAD groups (heart failure 15.8 +/- 4.1 ml/kg/min, implanted LVAD 19.8 +/- 5.8 ml/kg/min, explanted LVAD 28.2 +/- 5.0 ml/kg/min, p <0.05) as was anaerobic threshold (heart failure 11.2 +/- 1.9 ml/kg/min, implanted LVAD 14.7 +/- 4.9 ml/kg/min, explanted LVAD 21.4 +/- 5.0 ml/kg/min, p <0.05). In conclusion, peak CPO differentiates well during cardiac restoration using LVADs and emphasizes the benefits of this therapy. CPO has the potential to be a key physiologic marker of heart failure severity and can guide management of patients with LVAD.

  10. Biocompatibility and superiority of lyophilized acellular ligament scaffolds%冻干韧带脱细胞支架材料的生物相容性及优势

    Institute of Scientific and Technical Information of China (English)

    王辉; 陈雄生; 周盛源; 黄俊俊; 蔡弢艺

    2011-01-01

    BACKGROUND: Acellular matrix ligament removes the cellular components within the ligament tissue and reduce the immunogenicity through a variety of acellular ways. Simultaneously, the damage to scaffold structure is mild in the process of decellularization, and it retains the mechanical properties of the extracellular matrix.OBJECTIVE: To verify the biocompatibility and superiority of rabbit patellar tendon acellular scaffold after frozen and lyophilized processing.METHODS: Patellar ligaments were treated with 1% sodium deoxycholate for preparation of acellular ligaments with or without lyophilization. RESULTS AND CONCLUSION: No residual nuclear component was detected in all ligaments. Collagen structure was maintained.No significant differences in elastic modulus and ultimate tensile stress were found between non-lyophilized acellular scaffolds and lyophilized ones. The in vitro cytotoxicity test showed the cells grew well in all groups with or without extracts from lyophilized acellular scaffold. No significant difference was found between the control group and the experiment group. Toxicity symptoms were not obvious.Pyrogen detection experiment showed that no pyrogen was found in the lyophilized acellular scaffold extracts. Percutaneous stimulation test was negative as primary stimulation index was 0. In vivo implantation experiment showed that lyophilized acellular ligament scaffold showed the characteristics of little immunogenicity and light inflammation. Lyophilized acellular ligament scaffold treated with 1% DCA method not only maintains the mechanical characteristics of the non-lyophilized ones, but also has good biological compatibility. Because of its preparation, disinfection, packaging and preservation was easy and convenient, the lyophilized acellular ligament scaffold will be an ideal scaffold for tissue engineering ligament.%背景:韧带脱细胞基质是通过各种脱细胞方法将韧带组织内的细胞成分清除,降低免疫

  11. 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...... lambs were randomly assigned to two weight-matched groups at birth. During the first 14 days of life AKG was administered orally to the experimental group (n=12) at the dosage of 0.1 g/kg body weight per day, while the control group (n=11) received an equal dose of the vehicle. Lambs were slaughtered...... has a long-term effect on skeletal development when given early in neonatal life, and that changes in rib properties serve to improve chest mechanics and functioning in young animals. Moreover, neonatal administration of AKG may be considered as an effective factor enhancing proper development...

  12. Synovial explant inflammatory mediator production corresponds to rheumatoid arthritis imaging hallmarks

    DEFF Research Database (Denmark)

    Andersen, Martin; Boesen, Mikael; Ellegaard, Karen

    2014-01-01

    (BME), synovitis and erosion scores were estimated on the basis of the rheumatoid arthritis magnetic resonance imaging score (RAMRIS). Mixed models were used for the statistical analyses. Parsimony was achieved by omitting covariates with P > 0.1 from the statistical model. RESULTS: Tissue samples from......INTRODUCTION: Despite the widespread use of magnetic resonance imaging (MRI) and Doppler ultrasound for the detection of rheumatoid arthritis (RA) disease activity, little is known regarding the association of imaging-detected activity and synovial pathology. The purpose of this study...... procedure of the hand joints. The synovial tissue specimens were incubated for 72 hours, and spontaneous release of monocyte chemoattractant protein 1 (MCP-1), interleukin 6 (IL-6), macrophage inflammatory protein 1β (MIP-1β) and IL-8 was measured by performing multiplex immunoassays. Bone marrow oedema...

  13. Engineering tubular bone using mesenchymal stem cell sheets and coral particles

    Energy Technology Data Exchange (ETDEWEB)

    Geng, Wenxin [Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, College of Life Science, Northwest University, No.229 North Taibai Road, Xi’an 710069 (China); Ma, Dongyang [Department of Oral and Maxillofacial Surgery, Lanzhou General Hospital, Lanzhou Command of PLA, BinHe 333 South Road, Lanzhou 730052 (China); Yan, Xingrong; Liu, Liangqi; Cui, Jihong; Xie, Xin; Li, Hongmin [Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, College of Life Science, Northwest University, No.229 North Taibai Road, Xi’an 710069 (China); Chen, Fulin, E-mail: chenfl@nwu.edu.cn [Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, College of Life Science, Northwest University, No.229 North Taibai Road, Xi’an 710069 (China)

    2013-04-19

    Highlights: • We developed a novel engineering strategy to solve the limitations of bone grafts. • We fabricated tubular constructs using cell sheets and coral particles. • The composite constructs showed high radiological density and compressive strength. • These characteristics were similar to those of native bone. -- Abstract: The development of bone tissue engineering has provided new solutions for bone defects. However, the cell-scaffold-based approaches currently in use have several limitations, including low cell seeding rates and poor bone formation capacity. In the present study, we developed a novel strategy to engineer bone grafts using mesenchymal stem cell sheets and coral particles. Rabbit bone marrow mesenchymal stem cells were continuously cultured to form a cell sheet with osteogenic potential and coral particles were integrated into the sheet. The composite sheet was then wrapped around a cylindrical mandrel to fabricate a tubular construct. The resultant tubular construct was cultured in a spinner-flask bioreactor and subsequently implanted into a subcutaneous pocket in a nude mouse for assessment of its histological characteristics, radiological density and mechanical property. A similar construct assembled from a cell sheet alone acted as a control. In vitro observations demonstrated that the composite construct maintained its tubular shape, and exhibited higher radiological density, compressive strength and greater extracellular matrix deposition than did the control construct. In vivo experiments further revealed that new bone formed ectopically on the composite constructs, so that the 8-week explants of the composite sheets displayed radiological density similar to that of native bone. These results indicate that the strategy of using a combination of a cell sheet and coral particles has great potential for bone tissue engineering and repairing bone defects.

  14. Immunogenicity of a low-dose diphtheria, tetanus and acellular pertussis combination vaccine with either inactivated or oral polio vaccine compared to standard-dose diphtheria, tetanus, acellular pertussis when used as a pre-school booster in UK children: A 5-year follow-up of a randomised controlled study.

    Science.gov (United States)

    John, T; Voysey, M; Yu, L M; McCarthy, N; Baudin, M; Richard, P; Fiquet, A; Kitchin, N; Pollard, A J

    2015-08-26

    This serological follow up study assessed the kinetics of antibody response in children who previously participated in a single centre, open-label, randomised controlled trial of low-dose compared to standard-dose diphtheria booster preschool vaccinations in the United Kingdom (UK). Children had previously been randomised to receive one of three combination vaccines: either a combined adsorbed tetanus, low-dose diphtheria, 5-component acellular pertussis and inactivated polio vaccine (IPV) (Tdap-IPV, Repevax(®); Sanofi Pasteur MSD); a combined adsorbed tetanus, low-dose diphtheria and 5-component acellular pertussis vaccine (Tdap, Covaxis(®); Sanofi Pasteur MSD) given concomitantly with oral polio vaccine (OPV); or a combined adsorbed standard-dose diphtheria, tetanus, 2-component acellular pertussis and IPV (DTap-IPV, Tetravac(®); Sanofi Pasteur MSD). Blood samples for the follow-up study were taken at 1, 3 and 5 years after participation in the original trial (median, 5.07 years of age at year 1), and antibody persistence to each vaccine antigen measured against defined serological thresholds of protection. All participants had evidence of immunity to diphtheria with antitoxin concentrations greater than 0.01IU/mL five years after booster vaccination and 75%, 67% and 79% of children who received Tdap-IPV, Tdap+OPV and DTap-IPV, respectively, had protective antitoxin levels greater than 0.1IU/mL. Long lasting protective immune responses to tetanus and polio antigens were also observed in all groups, though polio responses were lower in the sera of those who received OPV. Low-dose diphtheria vaccines provided comparable protection to the standard-dose vaccine and are suitable for use for pre-school booster vaccination.

  15. Bone grafting: An overview

    Directory of Open Access Journals (Sweden)

    D. O. Joshi

    2010-08-01

    Full Text Available Bone grafting is the process by which bone is transferred from a source (donor to site (recipient. Due to trauma from accidents by speedy vehicles, falling down from height or gunshot injury particularly in human being, acquired or developmental diseases like rickets, congenital defects like abnormal bone development, wearing out because of age and overuse; lead to bone loss and to replace the loss we need the bone grafting. Osteogenesis, osteoinduction, osteoconduction, mechanical supports are the four basic mechanisms of bone graft. Bone graft can be harvested from the iliac crest, proximal tibia, proximal humerus, proximal femur, ribs and sternum. An ideal bone graft material is biologically inert, source of osteogenic, act as a mechanical support, readily available, easily adaptable in terms of size, shape, length and replaced by the host bone. Except blood, bone is grafted with greater frequency. Bone graft indicated for variety of orthopedic abnormalities, comminuted fractures, delayed unions, non-unions, arthrodesis and osteomyelitis. Bone graft can be harvested from the iliac crest, proximal tibia, proximal humerus, proximal femur, ribs and sternum. By adopting different procedure of graft preservation its antigenicity can be minimized. The concept of bone banking for obtaining bone grafts and implants is very useful for clinical application. Absolute stability require for successful incorporation. Ideal bone graft must possess osteogenic, osteoinductive and osteocon-ductive properties. Cancellous bone graft is superior to cortical bone graft. Usually autologous cancellous bone graft are used as fresh grafts where as allografts are employed as an alloimplant. None of the available type of bone grafts possesses all these properties therefore, a single type of graft cannot be recomm-ended for all types of orthopedic abnormalities. Bone grafts and implants can be selected as per clinical problems, the equipments available and preference of

  16. Orchestration of bone remodeling

    NARCIS (Netherlands)

    Moester, Martiene Johanna Catharina

    2014-01-01

    In healthy individuals, a balance exists between bone formation and resorption. Disruption of this balance can lead to higher or lower bone mass, and disease such as osteoporosis. Treatment for osteoporosis generally inhibits bone resorption, but does not rebuild bone to a healthy strength. More kno

  17. Bone grafts in dentistry

    Directory of Open Access Journals (Sweden)

    Prasanna Kumar

    2013-01-01

    Full Text Available Bone grafts are used as a filler and scaffold to facilitate bone formation and promote wound healing. These grafts are bioresorbable and have no antigen-antibody reaction. These bone grafts act as a mineral reservoir which induces new bone formation.

  18. Bone Marrow Diseases

    Science.gov (United States)

    Bone marrow is the spongy tissue inside some of your bones, such as your hip and thigh bones. It contains stem cells. The stem cells can ... the platelets that help with blood clotting. With bone marrow disease, there are problems with the stem ...

  19. Foam-like scaffolds for bone tissue engineering based on a novel couple of silicate-phosphate specular glasses: synthesis and properties.

    Science.gov (United States)

    Vitale-Brovarone, Chiara; Baino, Francesco; Bretcanu, Oana; Verne, Enrica

    2009-11-01

    Glass-ceramic scaffolds mimicking the structure of cancellous bone were produced via sponge replication technique by using a polyurethane foam as template and glass powder below 30 lm as inorganic phase. Specifically, a SiO₂-based glass of complex composition and its corresponding P₂O₅-based "specular" glass were used as materials for scaffolding. The polymeric sponge was thermally removed and the glass powders were sintered to obtain a replica of the template structure. The scaffolds were investigated and compared from a structural, morphological and mechanical viewpoint by assessing their crystalline phases, volumetric shrinkage, pores content and interconnection, mechanical strength. In addition, the scaffolds were soaked in acellular simulated body fluid to investigate their in vitro behaviour. The produced scaffolds have a great potential for bone reconstructive surgery because their features, such as shape, strength, bioactivity and bioresorption, can be easily tailored according to the end use.

  20. Bone Health and Osteoporosis.

    Science.gov (United States)

    Lupsa, Beatrice C; Insogna, Karl

    2015-09-01

    Osteoporosis is characterized by low bone mass and microarchitectural deterioration of bone tissue leading to decreased bone strength and an increased risk of low-energy fractures. Central dual-energy X-ray absorptiometry measurements are the gold standard for determining bone mineral density. Bone loss is an inevitable consequence of the decrease in estrogen levels during and following menopause, but additional risk factors for bone loss can also contribute to osteoporosis in older women. A well-balanced diet, exercise, and smoking cessation are key to maintaining bone health as women age. Pharmacologic agents should be recommended in patients at high risk for fracture.

  1. Nicotine promotes rooting in leaf explants of in vitro raised seedlings of tomato, Lycopersicon esculentum Miller var. Pusa Ruby.

    Science.gov (United States)

    Bamel, Kiran; Gupta, Rajendra; Gupta, Shrish C

    2015-11-01

    Nicotine promotes rooting in leaf explants of tomato (Lycopersicon esculentum Miller var. Pusa Ruby). Nicotine at 10(-9) to 10(-3) M concentrations was added to the MS basal medium. The optimum response (three-fold increase in rooting) was obtained at 10(-7) M nicotine-enriched MS medium. At the same level i.e. 10(-7) M Nicotine induced dramatic increase (11-fold) in the number of secondary roots per root. We have shown earlier that exogenous acetylcholine induces a similar response in tomato leaves. Since nicotine is an agonist of one of the two acetylcholine receptors in animals, its ability to simulate ACh action in a plant system suggests the presence of the same molecular mechanism operative in both, animal and plant cells.

  2. In vitro establishment of a highly effective method of castor bean (Ricinus communisL.) regeneration using shoot explants

    Institute of Scientific and Technical Information of China (English)

    ZHANG Ji-xing; CHI Yue; CHEN Yong-sheng; WANG Xiao-yu; FENG Zi-zhou; GENG Xue-jun; MU Sha-moli; HUO Hong-yan; TONG Huan; LI Meng-zhu; LI Yi

    2016-01-01

    An efifcient plant regeneration protocol was established for castor bean (Ricinus communisL.), in which 0.3 mg L–1 thidiazuron (TDZ) induced shoot clusters and increased the number of adventitious shoots from hypocotyl tissue. Our results showed that treatment under dark conditions signiifcantly promoted the average number of shoots per explant to 37.36±4.54 (with a 6-d treatment). Modiifed 1/2 Murashige and Skoog (MS) basal medium supplemented with 440 mg L–1 Ca2+, 0.2 mg L–1 gibberelic acid and 0.1 mg L–1 TDZ signiifcantly increased shoot elongation rates and lowered vitriifcation rates. Further-more, 1/2 MS media supplemented with 0.2 mg L–1 1-naphthaleneacetic acid induced a higher rooting rate compared with other culture conditions.

  3. BONE IN OSTEOPETROSIS

    Directory of Open Access Journals (Sweden)

    Ramkumar

    2014-04-01

    Full Text Available Osteopetrosis, a generalized developmental bone disease due to genetic disturbances, characterized by failure of bone re sorption and continuous bone formation making the bone hard, dense and brittle. Bones of intramembranous ossification and enchondrial ossification are affected genetically and symmetrically. During the process of disease the excess bone formation obliterates the cranial foramina and presses the optic, auditory and facial nerves resulting in defective vision, impaired hearing and facial paralysis. The bone formation in osteopetrosis affects bone marrow function leading to severe anemia and deficient of blood cells. The bone devoid of blood supply due to compression of blood vessels by excess formation of bone are prone to osteomyelitic changes with suppuration and pathological fracture if exposed to infection. Though the condition is chronic progressive, it produces changes leading to fatal condition, it should be studied thoroughly by everyone and hence this article presents a classical case of osteopetrosis with detailed description and discussion for the benefit of readers

  4. Replication kinetics of neurovirulent versus non-neurovirulent equine herpesvirus type 1 strains in equine nasal mucosal explants.

    Science.gov (United States)

    Vandekerckhove, Annelies P; Glorieux, S; Gryspeerdt, A C; Steukers, L; Duchateau, L; Osterrieder, N; Van de Walle, G R; Nauwynck, H J

    2010-08-01

    Equine herpesvirus type 1 (EHV-1) is the causative agent of equine herpes myeloencephalopathy, of which outbreaks are reported with increasing frequency throughout North America and Europe. This has resulted in its classification as a potentially emerging disease by the US Department of Agriculture. Recently, it was found that a single nucleotide polymorphism (SNP) in the viral DNA polymerase gene (ORF30) at aa 752 (N-->D) is associated with the neurovirulent potential of EHV-1. In the present study, equine respiratory mucosal explants were inoculated with several Belgian isolates typed in their ORF30 as D(752) or N(752), to evaluate a possible difference in replication in the upper respiratory tract. In addition, to evaluate whether any observed differences could be attributed to the SNP associated with neurovirulence, the experiments were repeated with parental Ab4 (reference neurovirulent strain), parental NY03 (reference non-neurovirulent strain) and their N/D revertant recombinant viruses. The salient findings were that EHV-1 spreads plaquewise in the epithelium, but plaques never cross the basement membrane (BM). However, single EHV-1-infected cells could be observed below the BM at 36 h post-inoculation (p.i.) for all N(752) isolates and at 24 h p.i. for all D(752) isolates, and were identified as monocytic cells and T lymphocytes. Interestingly, the number of infected cells was two to five times higher for D(752) isolates compared with N(752) isolates at every time point analysed. Finally, this study showed that equine respiratory explants are a valuable and reproducible model to study EHV-1 neurovirulence in vitro, thereby reducing the need for horses as experimental animals.

  5. A valued Indian medicinal plant – Begonia malabarica Lam. : Successful plant regeneration through various explants and field performance

    Directory of Open Access Journals (Sweden)

    Sevanan Rajeshkumar

    2009-05-01

    Full Text Available A cost-effective and efficient protocol has been described in the present work for large-scale and rapid in vitro propagation of a valuable medicinal herb Begonia malabarica Lam. (Begoniaceae by shoot auxillary-bud proliferation and organogenesis on MS medium supplemented with 6-benzylaminopurine (BA; 0.0-8.8 mg/l and indole-3-acetic acid (IAA; 0.0-2.88 mg/l at different concentrations, either alone or in combinations. Initiation of callus formation from the base of the leaf lamina was observed on MS supplemented with BA, IAA and adenine sulphate. Root induction on shoots was achieved on full strength MS with IAA/ indole-3-butyric acid (IBA at different concentrations. MS medium with 4.4 mg/l BA and 1.4 mg/l IAA elicited the maximum number of shoots (10 multiple shoots from nodal explants. Leaf-based callus differentiated into more than 28 shoots on MS with 150 mg/l adenine sulphate. The regenerated shoots were rooted on MS with 1.2 mg/l IBA within ten days. Almost 95% of the rooted shoots survived hardening when transferred to the field. The regenerated plants did not show any morphological change and variation in levels of secondary metabolites when compared with the mother stock. Thus, a reproduction of B. malabarica was established through nodal and leaf explants. This protocol can be exploited for conservation and commercial propagation of this medical plant in the Indian subcontinent and might be useful for genetic improvement programs.

  6. Multiscale Modeling of Bone

    Science.gov (United States)

    2014-12-01

    DISEASE Both age and disease can affect the structure of bone, the effects of which are often similar. The most common bone disease is osteoporosis ... Osteoporosis is a disease that results in reduced bone mass and density. This reduction of bone mass and density has a greater impact on trabecular...Bone loss in females is linked to a decrease in estrogen ; the decrease of estrogen associated with menopause increases osteoclast activity [89]. This

  7. TRANSFORMING GROWTH FACTOR-β AND FIBROBLAST GROWTH FACTOR INDUCE LENS EPITHELIAL EXPLANT METAPLASIA: IMPLICATIONS FOR THE FORMATION OF SUBCAPSULAR OPACIFICATION

    Institute of Scientific and Technical Information of China (English)

    刘颉; 叶俊杰

    1998-01-01

    Objective. This study was to investigate the effects of transforming growth factor-β(TGFβ) and fibroblast growth factor (FGF) in the subcapsular opaeification formation of the lens. Methods. Lens epithelial explants from 10-day-old rats were cultured with TGFβ1 or TGFβ2 in the presence of FGF for 5 days, then were examined by light and electron microscopy, and by immunolocalization of α-smooth muscle(α-sm) actin and type Ⅰ collagen. Resets. In TGFβ/FGF-treated explants,extensive proliferation oeeured, with formation of spindle and star-shaped cells. These cells showed ultrastructure and biochemical features of fibroblast or myofibroblast.Prominent Golgi apparatus and rough endoplaie reticulum were observed in scene cells, Intracellular microfilaments with cytoplasmic dense bodies and membrane associated dense bodies, features of smooth muscle cells, were also observed. Some cells showed reactivity to α-sin actin antibody. TGFβ/FGF-treated explants were strongly stained with type I collagen antibody. Conclusion. In the presence of FGF, TGFβ1 and TGFβ2 induced lens epithelial cell(LEC)proliferation and transformation into fibroblast or myofibroblast like ceils, with producing of abundant collagen matrix in the explants. The changes are similar to the metaplasia that oeeurrs in subeapsular opacification of the lens. The findings suggest that TGFβ and FGF plays a role in the pathogenesis of subcapsular opacification of the lens.

  8. Induction of Hairy roots of Pegagan (Centella Asiatica (l. Urban using Several Explant Sources with Several Agrobacterium Rhizogenes Strains in Vitro

    Directory of Open Access Journals (Sweden)

    Zahanis Zahanis

    2014-01-01

    Full Text Available Induction of hairy roots of pegagan using A. rhizogenes strains is important to obtain high production of triterpenoid as secondary metabolite.  The objectives of this research were to obtain the best A. rhizogenes strains and the best explant sources for C. asiatica hairy root induction. The research consisted of two stages. Stage I was A. rhizogenes arranged in  Completely Randomized Design  (CRD with 3 treatments and 6 replications. As a treatment was inoculation with A. rhizogenes strains LBA 9457, A4,  R1000 and control.  Stage II  was explant sources trial arranged in CRD with 3 treatments and 6 replications. The treatments were: internodus, apical bud and leaves. The results showed that all three strains of A. rhizogenes were able to induce C. asiatica plant to produce hairy roots, with R1000 as the best strain.The best explant source was explant leave with  15 day after inoculation and the percentage of  C. asiatica  hairyroot formation was 100 %. The β-glucoronidase (  GUS test result has proven that Ri plasmid T-DNA of A. rhizogenes  was integrated into the genome of C. asiatica plant.

  9. Establishment of high frequency shoot regeneration system in Himalayan poplar (Populus ciliata Wall. ex Royle) from petiole explants using Thidiazuron cytokinin as plant growth regulator

    Institute of Scientific and Technical Information of China (English)

    G Aggarwal; A Gaur; D K Srivastava

    2015-01-01

    Populus species are important resources for industry and in scientific study on biological and agricul-tural systems. Our objective was to enhance the frequency of plant regeneration in Himalayan poplar (Populus ciliata wall. ex Royle). The effect of TDZ alone and in combi-nation with adenine and NAA was studied on the regen-eration potential of petiole explants. The explants were excised from Himalayan poplar plants grown in glass-houses. After surface sterilization the explants were cul-tured on shoot induction medium. High percentage shoot regeneration (86%) was recorded on MS medium sup-plemented with 0.004 mg L-1 TDZ and 79.7 mg L-1 adenine. The regenerated shoots for elongation and multi-plication were transferred to MS ? 0.5 mg L-1 BAP ? 0.2 mg L-1 IAA ? 0.3 mg L-1 GA3. Root re-generation from shoots developed in vitro was observed on MS medium supplemented with 0.10 mg L-1 IBA. Hi-malayan poplar plantlets could be produced within 2 months after acclimatization in a sterile mixture of sand and soil. We developed a high efficiency plant regeneration protocol from petiole explants of P. ciliata.

  10. Protocols for Callus and Somatic Embryo Initiation for Hibiscus sabdariffa L. (Malvaceae): Influence of Explant Type, Sugar, and Plant Growth Regulators

    Science.gov (United States)

    A significant work on callus induction and somatic embryogenesis was realized for Hibiscus sabdariffa. Two genotypes (Hibiscus sabdariffa and Hibiscus sabdariffa var. altissima) two sugars (sucrose and glucose) and three concentrations (1 %, 2%, 3%) of each sugar, 3 explant types (root, hypocotyl, c...

  11. Combined periodontal and restorative approach to the treatment of gingival recessions with noncarious cervical lesions: a case treated with acellular dermal matrix allograft and compomer restorations.

    Science.gov (United States)

    Efeoğlu, Ahmet; Hanzade, Mete; Sari, Esra; Alpay, Hande; Karakaş, Ozan; Koray, Fatma

    2012-08-01

    Treatment of gingival recessions has become one of the most challenging procedures in periodontal plastic surgery. Various surgical options with predictable outcomes are available, but in cases with cervical lesions or restorations, optimal functional and esthetic results may require the combination of periodontal and restorative procedures. In this case report, one patient treated with acellular dermal matrix allograft and a coronally positioned flap in combination with compomer cervical restorations is presented. Clinical parameters were recorded immediately prior to surgery and after 12 months. Postoperatively, significant root coverage, reductions in probing depths, and gains in clinical attachment were observed. The final clinical results, esthetics, color match, and tissue contours were acceptable to both the patient and clinicians.

  12. Reduced-antigen, combined diphtheria, tetanus and acellular pertussis vaccine, adsorbed (Boostrix®): a review of its properties and use as a single-dose booster immunization.

    Science.gov (United States)

    McCormack, Paul L

    2012-09-10

    Reduced-antigen, combined diphtheria, tetanus and three-component acellular pertussis vaccine (Tdap; Boostrix®) is indicated for booster vaccination against diphtheria, tetanus and pertussis in individuals from age four years onwards in Europe and from age 10 years onwards in the US. Compared with infant formulations used for primary vaccination, Tdap contains reduced quantities (10-50%) of all toxoids and antigens, which are adsorbed to either ≤0.39 mg/dose (US licensed formulation) or 0.5 mg/dose (rest-of-world formulation) of aluminium adjuvant. The reduced antigen content is designed to avoid the increasing reactogenicity historically seen with the fourth and fifth doses of infant vaccine. This article reviews the immunogenicity, protective efficacy and reactogenicity of Tdap booster administered to children, adolescents and adults, including those aged ≥65 years. In clinical trials, a single booster dose of Tdap induced seroprotective levels of antibodies to diphtheria and tetanus toxoids in virtually all children and adolescents, and in a high proportion of adults and elderly individuals at approximately 1 month post-vaccination irrespective of their vaccination history. In all age groups, seropositivity rates for antibodies against pertussis antigens were ≥90% (including in unvaccinated adolescents), and booster response rates were high. Tdap was safely co-administered with other common vaccines without significantly affecting the immune responses. The immunogenicity and reactogenicity profiles of booster doses of Tdap were generally similar to those of infant diphtheria-tetanus-whole-cell pertussis vaccine and infant diphtheria-tetanus-acellular pertussis vaccine in children aged 4-6 years, and infant diphtheria-tetanus vaccine in older children. In adolescents and adults, the immunogenicity and reactogenicity of Tdap were generally similar to those of reduced-antigen diphtheria-tetanus vaccine, reduced-antigen diphtheria

  13. Licensure of a Diphtheria and Tetanus Toxoids and Acellular Pertussis Adsorbed and Inactivated Poliovirus Vaccine and Guidance for Use as a Booster Dose.

    Science.gov (United States)

    Liang, Jennifer; Wallace, Greg; Mootrey, Gina

    2015-09-04

    On March 24, 2015, the Food and Drug Administration licensed an additional combined diphtheria and tetanus toxoids and acellular pertussis adsorbed (DTaP) and inactivated poliovirus (IPV) vaccine (DTaP-IPV) (Quadracel, Sanofi Pasteur Inc.). Quadracel is the second DTaP-IPV vaccine to be licensed for use among children aged 4 through 6 years in the United States (1). Quadracel is approved for administration as a fifth dose in the DTaP series and as a fourth or fifth dose in the IPV series in children aged 4 through 6 years who have received 4 doses of DTaP-IPV-Hib (Pentacel, Sanofi Pasteur) and/or DTaP (Daptacel, Sanofi Pasteur) vaccine (2,3). This report summarizes the indications for Quadracel vaccine and provides guidance from the Advisory Committee on Immunization Practices (ACIP) for its use.

  14. Bone cysts: unicameral and aneurysmal bone cyst.

    Science.gov (United States)

    Mascard, E; Gomez-Brouchet, A; Lambot, K

    2015-02-01

    Simple and aneurysmal bone cysts are benign lytic bone lesions, usually encountered in children and adolescents. Simple bone cyst is a cystic, fluid-filled lesion, which may be unicameral (UBC) or partially separated. UBC can involve all bones, but usually the long bone metaphysis and otherwise primarily the proximal humerus and proximal femur. The classic aneurysmal bone cyst (ABC) is an expansive and hemorrhagic tumor, usually showing characteristic translocation. About 30% of ABCs are secondary, without translocation; they occur in reaction to another, usually benign, bone lesion. ABCs are metaphyseal, excentric, bulging, fluid-filled and multicameral, and may develop in all bones of the skeleton. On MRI, the fluid level is evocative. It is mandatory to distinguish ABC from UBC, as prognosis and treatment are different. UBCs resolve spontaneously between adolescence and adulthood; the main concern is the risk of pathologic fracture. Treatment in non-threatening forms consists in intracystic injection of methylprednisolone. When there is a risk of fracture, especially of the femoral neck, surgery with curettage, filling with bone substitute or graft and osteosynthesis may be required. ABCs are potentially more aggressive, with a risk of bone destruction. Diagnosis must systematically be confirmed by biopsy, identifying soft-tissue parts, as telangiectatic sarcoma can mimic ABC. Intra-lesional sclerotherapy with alcohol is an effective treatment. In spinal ABC and in aggressive lesions with a risk of fracture, surgical treatment should be preferred, possibly after preoperative embolization. The risk of malignant transformation is very low, except in case of radiation therapy.

  15. Fibrin gel-immobilized primary osteoblasts in calcium phosphate bone cement: in vivo evaluation with regard to application as injectable biological bone substitute.

    Science.gov (United States)

    Kneser, U; Voogd, A; Ohnolz, J; Buettner, O; Stangenberg, L; Zhang, Y H; Stark, G B; Schaefer, D J

    2005-01-01

    Osteogenic injectable bone substitutes may be useful for many applications. We developed a novel injectable bone substitute based on osteoblast-fibrin glue suspension and calcium phosphate bone cement (BC). Human osteoblasts were isolated from trabecular bone samples and cultured under standard conditions. Osteoblasts were suspended in fibrinogen solution (FS). BC was cured with thrombin solution. 8 x 4 mm injectable bone discs were prepared using silicon molds and a custom-made applicator device. Discs containing BC, BC/FS, or BC/FS/osteoblasts were implanted subcutaneously into athymic nude mice. After 3, 9 and 24 weeks, specimens were explanted and subjected to morphologic and biomechanical evaluation. In vitro fibrin gel-embedded osteoblasts displayed a differentiated phenotype as evidenced by alkaline phosphatase, collagen type 1 and von Kossa stains. A proportion of osteoblasts appeared morphologically intact over a 3-day in vitro period following application into the BC. BC/FS and BC/FS/osteoblast discs were sparsely infiltrated with vascularized connective tissue. There was no bone formation in implants from all groups. However, positive von Kossa staining only in BC/FS/osteoblast groups suggests engraftment of at least some of the transplanted cells. Biomechanical evaluation demonstrated initial stability of the composites. Young's modulus and maximal load did not differ significantly in the BC/FS and BC/FS/osteoblast groups. The practicability of osteoblast-containing injectable bone could be demonstrated. The dense microstructure and the suboptimal initial vascularization of the composites may explain the lack of bone formation. Modifications with regard to enhanced osteoblast survival are mandatory for a possible application as injectable osteogenic bone replacement system.

  16. Antibody responses to individual Bordetella pertussis fimbrial antigen Fim2 or Fim3 following immunization with the five-component acellular pertussis vaccine or to pertussis disease.

    Science.gov (United States)

    Alexander, Frances; Matheson, Mary; Fry, Norman K; Labram, Briony; Gorringe, Andrew R

    2012-11-01

    Bordetella pertussis expresses two serologically distinct fimbriae (Fim2 and Fim3) which are included in the Sanofi Pasteur 5-component acellular pertussis vaccine, and antibody responses to these antigens have been shown to be associated with protection. Studies to date have assessed the IgG response to this vaccine using a copurified mixture of Fim2 and Fim3, and the response to the individual antigens has not been characterized. We have purified separate Fim2 and Fim3 from strains that express either Fim2 or Fim3 and have used these antigens in an enzyme-linked immunosorbent assay (ELISA) to quantify IgG responses following immunization with 5-component acellular pertussis vaccine in 15-month-old, 4- to 6-year-old, and 11- to 18-year-old subjects. All individuals showed increases in Fim2 and Fim3 IgG concentrations following immunization, with 3-fold-greater Fim2 than Fim3 IgG concentrations seen in the younger two age groups. Fim2 IgG concentrations were 1.5-fold greater than Fim3 IgG concentrations in the 11- to 18-year-olds. We have also compared Fim2 and Fim3 IgG concentrations in individuals with prolonged cough who were diagnosed as having recent pertussis using a pertussis toxin (Ptx) IgG ELISA with individuals with prolonged cough but without elevated Ptx IgG concentrations. Individuals with evidence of recent pertussis had greater Fim3 IgG concentrations, consistent with the predominant serotype of isolates obtained in the United Kingdom. However, a surprising number of individuals had moderate Fim2 IgG concentrations despite very few isolates of that serotype obtained in the sampling period.

  17. Quality of the Haemophilus influenzae type b (Hib) antibody response induced by diphtheria-tetanus-acellular pertussis/Hib combination vaccines.

    Science.gov (United States)

    Denoël, Philippe A; Goldblatt, David; de Vleeschauwer, Isabel; Jacquet, Jeanne-Marie; Pichichero, Michael E; Poolman, Jan T

    2007-10-01

    It has been repeatedly observed that mixing Haemophilus influenzae type b (Hib) conjugate vaccines with acellular pertussis-containing vaccines (diphtheria-tetanus-acellular pertussis [DTPa]) resulted in a reduced magnitude of the anti-polyriboseribitolphosphate antibody response compared to that obtained when Hib vaccines were administered separately and not mixed. Nevertheless, the quality and functionality of the immune responses have been shown to be the same. With the purpose of investigating the quality of the anti-Hib immune responses that are elicited under different vaccination regimens, we report here four primary and booster-based pediatric clinical trials in which Hib vaccine was either mixed with DTPa or diphtheria-tetanus-whole-cell pertussis (DTPw)-based vaccines or was coadministered. Our results show that avidity maturation of the antibodies was lower when primary vaccination involved DTPa mixed with Hib compared to when DTPa and Hib were coadministered. No such difference was observed between mixed and separately administered Hib when associated with DTPa-hepatitis B virus-inactivated poliovirus or DTPw-based vaccines. All different combinations and regimens elicited the same opsonophagocytic and bactericidal activity as well as the same ability to protect in a passive infant rat protection assay. The functional activity of mixed DTPa-based and Hib vaccines was similar to that of mixed DTPw-based/Hib combinations. In conclusion, in vitro and in vivo data as well as postmarketing vaccine effectiveness data attest to the ability of DTPa-based/Hib combination vaccines to effectively prevent Hib-induced disease in children.

  18. Immunogenicity and safety after booster vaccination of diphtheria, tetanus, and acellular pertussis in young adults: an open randomized controlled trial in Japan.

    Science.gov (United States)

    Hara, Megumi; Okada, Kenji; Yamaguchi, Yuko; Uno, Shingo; Otsuka, Yasuko; Shimanoe, Chisato; Nanri, Hinako; Horita, Mikako; Ozaki, Iwata; Nishida, Yuichiro; Tanaka, Keitaro

    2013-12-01

    The recent increase of pertussis in young adults in Japan is hypothesized to be due in part to waning protection from the acellular pertussis vaccine. While a booster immunization may prevent an epidemic of pertussis among these young adults, little is known about the safety and immunogenicity of such a booster with the diphtheria, tetanus, and acellular pertussis vaccine (DTaP), which is currently available in Japan. One hundred and eleven medical students with a mean age of 19.4 years were randomly divided into 2 groups of 55 and 56 subjects and received, respectively, 0.2 or 0.5 ml of DTaP. Immunogenicity was assessed by performing the immunoassay using serum, and the geometric mean concentration (GMC), GMC ratio (GMCR), seropositive rate, and booster response rate were calculated. Adverse reactions and adverse events were monitored for 7 days after vaccination. After booster vaccination in the two groups, significant increases were found in the antibodies against pertussis toxin, filamentous hemagglutinin, diphtheria toxoid, and tetanus toxoid, and the booster response rates for all subjects reached 100%. The GMCs and GMCRs against all antigens were significantly higher in the 0.5-ml group than in the 0.2-ml group. No serious adverse events were observed. Frequencies of local reactions were similar in the 2 groups, although the frequency of severe local swelling was significantly higher in the 0.5-ml group. These data support the acceptability of booster immunization using both 0.2 and 0.5 ml of DTaP for young adults for controlling pertussis. (This study was registered at UMIN-CTR under registration number UMIN000010672.).

  19. Preservation of micro-architecture and angiogenic potential in a pulmonary acellular matrix obtained using intermittent intra-tracheal flow of detergent enzymatic treatment

    Science.gov (United States)

    Maghsoudlou, Panagiotis; Georgiades, Fanourios; Tyraskis, Athanasios; Totonelli, Giorgia; Loukogeorgakis, Stavros P.; Orlando, Giuseppe; Shangaris, Panicos; Lange, Peggy; Delalande, Jean-Marie; Burns, Alan J.; Cenedese, Angelo; Sebire, Neil J.; Turmaine, Mark; Guest, Brogan N.; Alcorn, John F.; Atala, Anthony; Birchall, Martin A.; Elliott, Martin J.; Eaton, Simon; Pierro, Agostino; Gilbert, Thomas W.; De Coppi, Paolo

    2013-01-01

    Tissue engineering of autologous lung tissue aims to become a therapeutic alternative to transplantation. Efforts published so far in creating scaffolds have used harsh decellularization techniques that damage the extracellular matrix (ECM), deplete its components and take up to 5 weeks to perform. The aim of this study was to create a lung natural acellular scaffold using a method that will reduce the time of production and better preserve scaffold architecture and ECM components. Decellularization of rat lungs via the intratracheal route removed most of the nuclear material when compared to the other entry points. An intermittent inflation approach that mimics lung respiration yielded an acellular scaffold in a shorter time with an improved preservation of pulmonary micro-architecture. Electron microscopy demonstrated the maintenance of an intact alveolar network, with no evidence of collapse or tearing. Pulsatile dye injection via the vasculature indicated an intact capillary network in the scaffold. Morphometry analysis demonstrated a significant increase in alveolar fractional volume, with alveolar size analysis confirming that alveolar dimensions were maintained. Biomechanical testing of the scaffolds indicated an increase in resistance and elastance when compared to fresh lungs. Staining and quantification for ECM components showed a presence of collagen, elastin, GAG and laminin. The intratracheal intermittent decellularization methodology could be translated to sheep lungs, demonstrating a preservation of ECM components, alveolar and vascular architecture. Decellularization treatment and methodology preserves lung architecture and ECM whilst reducing the production time to 3 h. Cell seeding and in vivo experiments are necessary to proceed towards clinical translation. PMID:23727263

  20. UV-independent induction of beta defensin 3 in neonatal human skin explants [v2; ref status: indexed, http://f1000r.es/53b

    Directory of Open Access Journals (Sweden)

    Erin Wolf Horrell

    2015-02-01

    Full Text Available In order to determine the effect of UV radiation on β-defensin 3 (BD3 expression in human skin, freshly-isolated UV-naïve skin was obtained from newborn male infants undergoing planned circumcision.  Skin explants sustained ex vivo dermis side down on RPMI media were exposed to 0.5 kJ/m2 UVB, and biopsies were taken from the explant through 72 hours after radiation.  mRNA expression was measured by qRTPCR and normalized to TATA-binding protein.  BD3 expression at each time point was compared with an untreated control taken at time 0 within each skin sample.  Extensive variability in both the timing and magnitude of BD3 induction across individuals was noted and was not predicted by skin pigment phenotype, suggesting that BD3 induction was not influenced by epidermal melanization.  However, a mock-irradiated time course demonstrated UV-independent BD3 mRNA increases across multiple donors which was not further augmented by treatment with UV radiation, suggesting that factors other than UV damage promoted increased BD3 expression in the skin explants.  We conclude that BD3 expression is induced in a UV-independent manner in human skin explants processed and maintained in standard culture conditions, and that neonatal skin explants are an inappropriate model with which to study the effects of UV on BD3 induction in whole human skin.

  1. UV-independent induction of beta defensin 3 in neonatal human skin explants [v1; ref status: indexed, http://f1000r.es/4s2

    Directory of Open Access Journals (Sweden)

    Erin Wolf Horrell

    2014-11-01

    Full Text Available In order to determine the effect of UV radiation on β-defensin 3 (BD3 expression in human skin, freshly-isolated UV-naïve skin was obtained from newborn male infants undergoing planned circumcision.  Skin explants sustained ex vivo dermis side down on RPMI media were exposed to 0.5 kJ/m2 UVB, and biopsies were taken from the explant through 72 hours after radiation.  mRNA expression was measured by qRTPCR and normalized to TATA-binding protein.  BD3 expression at each time point was compared with an untreated control taken at time 0 within each skin sample.  Extensive variability in both the timing and magnitude of BD3 induction across individuals was noted and was not predicted by skin pigment phenotype, suggesting that BD3 induction was not influenced by epidermal melanization.  However, a mock-irradiated time course demonstrated UV-independent BD3 mRNA increases across multiple donors which was not further augmented by treatment with UV radiation, suggesting that factors other than UV damage promoted increased BD3 expression in the skin explants.  We conclude that BD3 expression is induced in a UV-independent manner in human skin explants processed and maintained in standard culture conditions, and that neonatal skin explants are an inappropriate model with which to study the effects of UV on BD3 induction in whole human skin.

  2. Changes in the metabolic footprint of placental explant-conditioned medium cultured in different oxygen tensions from placentas of small for gestational age and normal pregnancies.

    LENUS (Irish Health Repository)

    Horgan, R P

    2012-01-31

    Being born small for gestational age (SGA) confers significantly increased risks of perinatal morbidity and mortality. Accumulating evidence suggests that an SGA fetus results from a poorly perfused and abnormally developed placenta. Some of the placental features seen in SGA, such as abnormal cell turnover and impaired nutrient transport, can be reproduced by culture of placental explants in hypoxic conditions. Metabolic footprinting offers a hypothesis-generating strategy to investigate factors absorbed by and released from this tissue in vitro. Previously, metabolic footprinting of the conditioned culture media has identified differences in placental explants cultured under normoxic and hypoxic conditions and between normal pregnancies and those complicated by pre-eclampsia. In this study we aimed to examine the differences in the metabolic footprint of placental villous explants cultured at different oxygen (O(2)) tensions between women who deliver an SGA baby (n = 9) and those from normal controls (n = 8). Placental villous explants from cases and controls were cultured for 96 h in 1% (hypoxic), 6% (normoxic) and 20% (hyperoxic) O(2). Metabolic footprints were analysed by Ultra Performance Liquid Chromatography coupled to an electrospray hybrid LTQ-Orbitrap Mass Spectrometry (UPLC-MS). 574 metabolite features showed significant difference between SGA and normal at one or more of the oxygen tensions. SGA explant media cultured under hypoxic conditions was observed, on a univariate level, to exhibit the same metabolic signature as controls cultured under normoxic conditions in 49% of the metabolites of interest, suggesting that SGA tissue is acclimatised to hypoxic conditions in vivo. No such behaviour was observed under hyperoxic culture conditions. Glycerophospholipid and tryptophan metabolism were highlighted as areas of particular interest.

  3. Effect of explant density and volume of cultivation medium on in-vitro multiplication of blueberry (Vaccinium corymbosum L. varieties "Brigitta" and "Legacy"

    Directory of Open Access Journals (Sweden)

    Mario Rodríguez Beraud

    2015-03-01

    Full Text Available The objective of the investigation was to evaluate the in-vitro multiplication of two varieties of blueberry (Vaccinium corymbosum L., “Brigitta” and “Legacy” in response to five explants densities (5, 10, 15, 20 and 25 and four flask volumes (10, 20, 30 and 40 mL for cultivation. For both varieties the cultivation medium WPM (Woody Plant Medium was used. The experiment was completely randomized with 20 treatments and 12 repetitions per treatment. After 45 days of cultivation we evaluated the height of shoots, number of shoots/explant, number of nodes/shoot and number of shoots/flask. Variety “Brigitta” had highest shoots at higher densities and flask volumes, while variety “Legacy” had the highest average shoot height with intermediate densities and high volumes. Regarding the number of shoots/explant, the volume of the medium had no influence on “Brigitta”, however, higher plant densities affected this parameter. With variety “Legacy” the maximum number of shoots was achieved with lower plant densities and intermediate culture volumes per flask. In relation to the number of nodes per explant "Brigitta had lower numbers as compared to “Legacy”, but with both varieties the number of nodes decresed with smaller volumes of medium in the flasks. For the number of shoots per flask, “Brigitta” responsed best at higher densities exceeding 40 shoots per flask. In contrast, “Legacy” produced maximum results at density of 25 explants in 30 mL of medium. It is concluded that for the optimum multiplication of both varieties the correct selection of both, the planting density and the volume of multiplication medium are important.

  4. Effect of phytic acid from rice and corn on morphology, cell proliferation, apoptosis and cyclooxygenase-2 expression in swine jejunal explants

    Directory of Open Access Journals (Sweden)

    Elisângela Olegário da Silva

    2014-06-01

    Full Text Available Phytic acid (IP6 is a potent antioxidant present in several natural foods. Beneficial effects on colon cancer and inflammation have been associated to IP6 in several studies, however, scarce data about the effect on small intestine are available. The aim of the present study was to evaluate the effect of different doses of IP6 from rice and corn on intestinal morphology, cellular proliferation, apoptosis and cyclooxygenase-2 (Cox-2 expression using swine jejunal explants as experimental model. This report demonstrated that explants treated with 0.5 mM, 2.5 mM and 5 mM of IP6 from rice and 2.5 mM and 5 mM from corn showed higher villi height compared to control. Explants treated with 2.5 mM and 5 mM IP6 from rice exhibited a significant reduction on intestinal histological changes (villi atrophy and fusion, edema, lymphatic vessel dilation, loss of apical enterocytes, cell vacuolation, necrotic debris, morphology of enterocytes and microvilli and number of villi. The cellular proliferation decreased in the explants treated with the dosages of 2.5 mM and 5 mM from rice and a significant decrease in cell apoptosis was observed in the treatments with 2.5 mM IP6 from rice and 5 mM IP6 from corn compared to the control. The explants treated with 2.5 mM and 5 mM IP6 from rice and corn showed a significant reduction of the Cox-2 expression. Higher dosages of IP6 from rice and corn used in this experiment increased the viability and preservation of intestinal tissue as evidenced by morphological and immunohistochemical assays.

  5. Flower-bud formation in explants of photoperiodic and day-neutral Nicotiana biotypes and its bearing on the regulation of flower formation

    Energy Technology Data Exchange (ETDEWEB)

    Rajeevan, M.S.; Lang, A. (Michigan State Univ., East Lansing (United States))

    1993-05-15

    The capacity to form flower buds in thin-layer explants was studied in Nicotiana of several species, cultivars, and lines of differing in their response to photoperiod. This capacity was found in all biotypes examined and could extend into sepals and corolla. It varied depending on genotype, source tissue and its developmental state, and composition of the culture medium, particularly the levels of glucose, auxin, and cytokinin. It was greatest in the two day-neutral plants examined, Samsun tobacco and Nicotiana rustica, where it extended from the inflorescence region down the vegetative stem, in a basipetally decreasing gradient; it was least in the two qualitative photoperiodic plants studied, the long-day plant Nicotiana silvestris and the short-day plant Maryland Mammoth tobacco, the quantitative long-day plant Nicotiana alata and the quantitative short-day plant Nicotiana otophora line 38-G-81, where it was limited to the pedicels (and, in some cases, the sepals). Regardless of the photoperiodic response of the source plants, the response was the same in explants cultured under long and short days. The capacity to form flow buds in explants is present in all Nicotiana biotypes studied supports the idea that it is regulated by the same mechanism(s), regardless of the plant's photoperiodic character. However, flower formation in the explants is not identical with de novo flower formation in a hitherto vegetative plant: it is rather the expression of a floral state already established in the plant, although it can vary widely in extent and spatial distribution. Culture conditions that permit flower-bud formation in an explant are conditions that maintain the floral state and encourage its expression; conditions under which no flower buds are formed reduce this state and/or prevent its expression. 14 refs., 5 figs., 3 tabs.

  6. Endochondral ossification for enhancing bone regeneration: converging native extracellular matrix biomaterials and developmental engineering in vivo.

    Science.gov (United States)

    Dennis, S Connor; Berkland, Cory J; Bonewald, Lynda F; Detamore, Michael S

    2015-06-01

    provide necessary osteo- and chondro-conductive and inductive features for enhancing EC ossification. In addition, critical perspectives on existing stem cell-based therapeutic strategies will be discussed with a focus on their use as an extension of the acellular ECM-based designs for specific clinical indications. Within this framework, a novel realm of unexplored design strategies for bone tissue engineering will be introduced into the collective consciousness of the regenerative medicine field.

  7. Regulation of Bone Metabolism.

    Science.gov (United States)

    Shahi, Maryam; Peymani, Amir; Sahmani, Mehdi

    2017-04-01

    Bone is formed through the processes of endochondral and intramembranous ossification. In endochondral ossification primary mesenchymal cells differentiate to chondrocytes and then are progressively substituted by bone, while in intramembranous ossification mesenchymal stem cells (MSCs) differentiate directly into osteoblasts to form bone. The steps of osteogenic proliferation, differentiation, and bone homeostasis are controlled by various markers and signaling pathways. Bone needs to be remodeled to maintain integrity with osteoblasts, which are bone-forming cells, and osteoclasts, which are bone-degrading cells.In this review we considered the major factors and signaling pathways in bone formation; these include fibroblast growth factors (FGFs), bone morphogenetic proteins (BMPs), wingless-type (Wnt) genes, runt-related transcription factor 2 (RUNX2) and osteoblast-specific transcription factor (osterix or OSX).

  8. Regulation of Bone Metabolism

    Science.gov (United States)

    Shahi, Maryam; Peymani, Amir; Sahmani, Mehdi

    2017-01-01

    Bone is formed through the processes of endochondral and intramembranous ossification. In endochondral ossification primary mesenchymal cells differentiate to chondrocytes and then are progressively substituted by bone, while in intramembranous ossification mesenchymal stem cells (MSCs) differentiate directly into osteoblasts to form bone. The steps of osteogenic proliferation, differentiation, and bone homeostasis are controlled by various markers and signaling pathways. Bone needs to be remodeled to maintain integrity with osteoblasts, which are bone-forming cells, and osteoclasts, which are bone-degrading cells.In this review we considered the major factors and signaling pathways in bone formation; these include fibroblast growth factors (FGFs), bone morphogenetic proteins (BMPs), wingless-type (Wnt) genes, runt-related transcription factor 2 (RUNX2) and osteoblast-specific transcription factor (osterix or OSX). PMID:28367467

  9. In vitro culture and characterization of alveolar bone osteoblasts isolated from type 2 diabetics

    Energy Technology Data Exchange (ETDEWEB)

    Sun, Dao-Cai [Department of Implant Dentistry, School of Stomatology, Fourth Military Medical University, Xi' an (China); Department of Stomatology, The 291st Hospital of P.L.A, Baotou (China); Li, De-Hua [Department of Implant Dentistry, School of Stomatology, Fourth Military Medical University, Xi' an (China); Ji, Hui-Cang [Military Sanatorium of Retired Cadres, Baotou (China); Rao, Guo-Zhou [Center of Laboratory, School of Stomatology, Xi' an Jiaotong University, Xi' an (China); Liang, Li-Hua [Department of Implant Dentistry, School of Stomatology, Fourth Military Medical University, Xi' an (China); Ma, Ai-Jie [Xi' an Technology University, Xi' an (China); Xie, Chao; Zou, Gui-Ke; Song, Ying-Liang [Department of Implant Dentistry, School of Stomatology, Fourth Military Medical University, Xi' an (China)

    2012-04-05

    In order to understand the mechanisms of poor osseointegration following dental implants in type 2 diabetics, it is important to study the biological properties of alveolar bone osteoblasts isolated from these patients. We collected alveolar bone chips under aseptic conditions and cultured them in vitro using the tissue explants adherent method. The biological properties of these cells were characterized using the following methods: alkaline phosphatase (ALP) chemical staining for cell viability, Alizarin red staining for osteogenic characteristics, MTT test for cell proliferation, enzyme dynamics for ALP contents, radio-immunoassay for bone gla protein (BGP) concentration, and ELISA for the concentration of type I collagen (COL-I) in the supernatant. Furthermore, we detected the adhesion ability of two types of cells from titanium slices using non-specific immunofluorescence staining and cell count. The two cell forms showed no significant difference in morphology under the same culture conditions. However, the alveolar bone osteoblasts received from type 2 diabetic patients had slower growth, lower cell activity and calcium nodule formation than the normal ones. The concentration of ALP, BGP and COL-I was lower in the supernatant of alveolar bone osteoblasts received from type 2 diabetic patients than in that received from normal subjects (P < 0.05). The alveolar bone osteoblasts obtained from type 2 diabetic patients can be successfully cultured in vitro with the same morphology and biological characteristics as those from normal patients, but with slower growth and lower concentration of specific secretion and lower combining ability with titanium than normal ones.

  10. Influência do número de gemas, presença ou ausência de folhas e posição do explante na multiplicação in vitro da batata Influence of bud numbers, presence or absence of leaves and explant position on the in vitro multiplication of potato

    Directory of Open Access Journals (Sweden)

    Jonny Everson S. Pereira

    2005-03-01

    Full Text Available Avaliou-se a posição, presença ou ausência de folhas e número de gemas iniciais do explante na multiplicação in vitro da batata. O meio de cultura foi formado pelos sais e vitaminas de MS, acrescido de 100 mg L-1 de mio-inositol, 30 g L-1 de sacarose e 6 g L-1 de ágar. Utilizaram-se diferentes tipos de segmentos nodais (basais e apicais, com e sem folhas, contendo uma, duas e três gemas axilares. Após a inoculação o material foi mantido em sala de crescimento com temperatura de 25±2ºC, fotoperíodo de 16 horas e 19 µE m-2 s-1 de irradiância por 32 dias. Para altura e número médio de brotações regeneradas, os melhores resultados foram obtidos com explantes oriundos da posição basal e com três gemas axilares. A taxa de multiplicação do material em cultivo foi maior nos explantes inoculados inicialmente com uma única gema, independentemente da posição do explante ser basal ou apical e, somente nos explantes basais a presença de folhas proporcionou taxa de multiplicação significativamente superior ao apical. Conclui-se que quando se trabalha com material vegetal heterogêneo, sob condições in vitro, as características iniciais dos explantes podem provocar variações na resposta final, causando erros na estimativa da multiplicação.The position, presence or absence of leaves and bud numbers of the explant were evaluated on the in vitro multiplication of potatoes. The culture medium was constituted by salts and vitamins of MS, added with 100 mg L-1 myo-inositol, 30 g L-1 sucrose and 6 g L-1 agar. Different types of nodal segments were used (basal and apical, with and without leaves, having, one, two and three axillary buds. The material was maintained in growth room at 25±2ºC, 16 hours photoperiod and 19 µE m-2 s-1 irradiance during 32 days. For height and number of regenerate sprouts, the best results were obtained with explants originating from the basal position and with three axillary buds. The multiplication

  11. Regeneração in vitro de urucum (Bixa orellana L. a partir de diferentes tipos de explantes In vitro regeneration of annatto (Bixa orellana L. from various explants

    Directory of Open Access Journals (Sweden)

    Jane Fiuza Rodrigues Portela de Carvalho

    2005-12-01

    Full Text Available Este trabalho teve como objetivo avaliar a regeneração in vitro de plantas de urucum (Bixa orellana L. a partir de diferentes tipos de explantes. Para definir o meio de cultura adequado para indução de brotações, diferentes concentrações e, ou, combinações da auxina AIA e das citocininas BAP e ZEA foram testadas. As melhores respostas de regeneração para segmentos de hipocótilo, nós cotiledonares e hipocótilos invertidos foram observadas em meios suplementados de ZEA (2,28 µM e AIA (0,30 µM, ZEA (4,56 µM e ZEA (4,56 µM, respectivamente. O meio de enraizamento mais eficaz foi o MS, com a metade de sua concentração salina e 5 µM de AIB. Análises citológicas, realizadas antes da aclimatação, confirmaram a estabilidade cromossômica das plantas cultivadas in vitro, não sendo detectado variação com relação ao número de cromossomos metafásicos (2n = 14.The present work aimed the establishment of a regeneration protocol for annatto (Bixa orellana L. from different juvenile explants. In order to promote shoot induction, different concentrations and/or combinations of IAA and the cytokinins BAP and ZEA were assessed. Better regeneration responses were achieved when segmented hypocotyl, cotiledonary nodes and inverted hypocotyl were cultured onto MS-based medium supplemented with ZEA (2.28 µM and IAA (0.30 µM, ZEA (4.56 µM or ZEA (4.56 µM, respectively. Rooting of elongated shoots displayed higher frequencies when half-strength MS medium with IBA (5 µM was used. No genetic variation was detected among regenerants as revealed by cytological analysis based on metaphasic chromosome countings (2n = 14.

  12. Clinical analysis of intraocular lens explantation in 68 eyes%人工晶状体取出术68只眼临床分析

    Institute of Scientific and Technical Information of China (English)

    楚美芳; 柴芳; 张倩; 马波

    2016-01-01

    目的 通过对人工晶状体取出病例的原因分析,探讨人工晶状体植人手术的适应证及并发症的防治.方法 回顾在西安市第四医院眼科自2013年1月至2015年1月行人工晶状体取出术共65例68只眼进行临床分析.结果 导致人工晶状体取出的常见原因依次为人工晶状体脱位或半脱位、视网膜脱离、人工晶状体度数误差、眼内炎等.人工晶状体植入至取出间隔时间最短1d,最长为15年,人工晶状体植入多在术后1月内和2~5年发生严重并发症导致人工晶状体取出.取出的人工晶状体:后房型64只眼,ICL3只眼,前房型1只眼.术后视力:53只眼(77.9%)有不同程度的提高,14只眼(20.6%)无明显变化,1只眼因并发眼内炎,术后视力由光感变为无光感.结论 人工晶状体植入术后的严重并发症是导致人工晶状体取出的直接原因.术前严格掌握人工晶状体植入的适应证,有效控制术中、术后的并发症,是避免术后人工晶状体取出的重要保证,及时准确地取出人工晶状体,同时酌情植入适当的人工晶状体,可有效提高患者视力和生活质量.%Objective To analyze the causes of explantation of intraocular lens (IOL) postoperation,and explore surgery accommodation and prevent surgery complications.Methods The causes for 65 cases (68 eyes) of lens explantation from January 2013 to January 2015 were retrospectively analyzed.Results The most common causes of IOL explantation were IOL decentration and dislocation,retinal detachment,postoperative refractive error and endophthalmitis.The shortest interval of IOL implantation to explantation was one day,while the longest interval was 15 years.Intraocular lens implantation surgery in one month and 2 ~ 5 years was more prone to occur serious complications leading to IOL explantation.IOL types of explantation including:posterior chamber IOL in 64 eyes,ICL (implantable collamer lens) in 3 eyes,and anterior chamber IOL

  13. What Is Bone Cancer?

    Science.gov (United States)

    ... start in bone, muscle, fibrous tissue, blood vessels, fat tissue, as well as some other tissues. They can develop anywhere in the body. There are several different types of bone tumors. Their names are based on ...

  14. Osteochondroma (Bone Tumor)

    Science.gov (United States)

    ... to be the most common benign bone tumor, accounting for 35% to 40% of all benign bone ... imaging scans. Doctors may also request computed tomography (CT) scans or magnetic resonance imaging (MRI) scans to ...

  15. What causes bone loss?

    Science.gov (United States)

    ... bone biology. In: Melmed S, Polonsky KS, Larsen PR, Kronenberg HM, eds. Williams Textbook of Endocrinology . 13th ed. Philadelphia, PA: Elsevier; 2016:chap 29. Maes C, Kronenberg HM. Bone development and remodeling. In: Jameson JL, ...

  16. Bone mineral density test

    Science.gov (United States)

    BMD test; Bone density test; Bone densitometry; DEXA scan; DXA; Dual-energy x-ray absorptiometry; p-DEXA; Osteoporosis-BMD ... need to undress. This scan is the best test to predict your risk of fractures. Peripheral DEXA ( ...

  17. Application of acellular dermal matrix embedded in socket after wisdom tooth extraction%脱细胞异体真皮组织补片在智齿拔除中的应用

    Institute of Scientific and Technical Information of China (English)

    白忠诚; 施生根; 李莉莉; 牛忠英; 张艳茹

    2011-01-01

    BACKGROUND: Few reports are found with J-1 acellular dermal matrix to prevent postoperative complications after impactedmandibular third molar extraction.OBJECTIVE: To evaluate the effects of acellular dermal matrix embedded in socket after wisdom tooth extraction.METHODS: 400 patients with impacted mandibular third molar were divided into two groups at random with 200 in each group. Ingroup A, the acellular dermal matrix was embedded in the sockets after wisdom tooth extraction; group B was the blank control.Postoperative complicati ons of the two groups were observed after treatment.RESULTS AND CONCLUSION: No acellular dermal matrix lost from wisdom tooth extraction sokets. Blooding after toothextraction decreased when acellular dermal matrix was embedded in the sokets, hemorrhage incidence rate was lower in group Athan in group B. Acellular dermal matrix could prevent blood clot lose from sockets, and also can prevent food residual enteringinto the sockets. Dry socket incidence rate was lower in group A than group B. Acellular embedded in the sockets had no effectson swelling incidence rate. Acellular dermal matrix embedded in socket after tooth extraction can prevent dry socket andhemorrhage, but can not prevent swelling.%背景:使用J-1型脱细胞异体真皮组织补片覆盖拔牙创口的报告较少.目的:探讨异体脱细胞组织补片置入拔牙窝对预防拔牙后并发症的影响.方法:将400例阻生智齿拔除患者随机分为2组,实验组智齿拔除后拔牙窝内放置医用组织补片;对照组智齿拔除后不放置医用组织补片.分别观察拔牙后组织补片脱落率、肿胀发生率、拔牙窝内血凝块存留和食物残渣残留情况、牙龈是否红肿、对拔牙后出血的影响以及干槽症的发生率.结果与结论:拔牙后出血的百分比,血凝块留存率,拔牙窝内食物残渣残留百分比,干槽症发生率实验组均明显低于对照组.放置组织补片对术后

  18. Rod and cone photoreceptor cells produce ROS in response to stress in a live retinal explant system.

    LENUS (Irish Health Repository)

    Bhatt, Lavinia

    2010-01-01

    PURPOSE: The production of reactive oxygen species (ROS) can lead to oxidative stress, which is a strong contributory factor to many ocular diseases. In this study, the removal of trophic factors is used as a model system to investigate the effects of stress in the retina. The aims were to determine if both rod and cone photoreceptor cells produce ROS when they are deprived of trophic factor support and to demonstrate if the nicotinamide adenine dinucleotide phosphate (NADPH) oxidase (Nox) enzymes are responsible for this ROS production. METHODS: Retinas were explanted from mice aged between postnatal days 8-10 and cultured overnight. The following morning, confocal microscopy combined with various fluorescent probes was used to detect the production of ROS. Each time peanut agglutinin (PNA), a cone photoreceptor marker, was used to facilitate orientation of the retina. Dihydroethidium and dihydrorhodamine 123 (DHR123) were used to determine which cells produce ROS. Subsequently, western blots of retinal serial sections were used to detect the presence of Noxs in the different retinal layers. The Nox inhibitor apocynin was then tested to determine if it altered the production of ROS within these cells. RESULTS: Live retinal explants, viewed at high magnifications using confocal microscopy, displayed an increase in the fluorescent products of dihydroethidium and DHR123 upon serum removal when compared to controls. DHR123 fluorescence, once oxidized, localized to mitochondria and was found in the same focal plane as the PNA staining. This showed that cones and rods produced ROS when stressed. Retinal serial sectioning established that the photoreceptor layer expressed Nox4, dual oxidase (Duox) 1, and Duox2 at varying levels. Finally, the Nox inhibitor apocynin decreased the burst stimulated by the stress of serum removal. CONCLUSIONS: Confocal microscopy and PNA staining allowed differentiation of cell types within the outermost layers of the retina, demonstrating

  19. Identification of small molecule lead compounds for visceral leishmaniasis using a novel ex vivo splenic explant model system.

    Directory of Open Access Journals (Sweden)

    Yaneth Osorio

    Full Text Available BACKGROUND: New drugs are needed to treat visceral leishmaniasis (VL because the current therapies are toxic, expensive, and parasite resistance may weaken drug efficacy. We established a novel ex vivo splenic explant culture system from hamsters infected with luciferase-transfected Leishmania donovani to screen chemical compounds for anti-leishmanial activity. METHODOLOGY/PRINCIPAL FINDINGS: THIS MODEL HAS ADVANTAGES OVER IN VITRO SYSTEMS IN THAT IT: 1 includes the whole cellular population involved in the host-parasite interaction; 2 is initiated at a stage of infection when the immunosuppressive mechanisms that lead to progressive VL are evident; 3 involves the intracellular form of Leishmania; 4 supports parasite replication that can be easily quantified by detection of parasite-expressed luciferase; 5 is adaptable to a high-throughput screening format; and 6 can be used to identify compounds that have both direct and indirect anti-parasitic activity. The assay showed excellent discrimination between positive (amphotericin B and negative (vehicle controls with a Z' Factor >0.8. A duplicate screen of 4 chemical libraries containing 4,035 compounds identified 202 hits (5.0% with a Z score of <-1.96 (p<0.05. Eighty-four (2.1% of the hits were classified as lead compounds based on the in vitro therapeutic index (ratio of the compound concentration causing 50% cytotoxicity in the HepG(2 cell line to the concentration that caused 50% reduction in the parasite load. Sixty-nine (82% of the lead compounds were previously unknown to have anti-leishmanial activity. The most frequently identified lead compounds were classified as quinoline-containing compounds (14%, alkaloids (10%, aromatics (11%, terpenes (8%, phenothiazines (7% and furans (5%. CONCLUSIONS/SIGNIFICANCE: The ex vivo splenic explant model provides a powerful approach to identify new compounds active against L. donovani within the pathophysiologic environment of the infected spleen

  20. Estabelecimento e desenvolvimento in vitro de Eugenia involucrata DC.: influência do tipo de explante e do meio nutritivo

    Directory of Open Access Journals (Sweden)

    Diego Pascoal Golle

    2012-03-01

    Full Text Available http://dx.doi.org/10.5902/198050985092Apresentando diversas características de interesse nos setores da silvicultura, fruticultura, meio ambiente e medicinal, Eugenia involucrata DC. (Myrtaceae é uma espécie florestal nativa de vários Estados brasileiros. Considerada a dificuldade da propagação por sementes, que são recalcitrantes, perdendo rapidamente sua viabilidade após a colheita, este trabalho objetivou avaliar a influência do uso de segmentos apicais e nodais cultivados em diferentes meios nutritivos no estabelecimento e no desenvolvimento in vitro desta espécie. Segmentos apicais e nodais coletados em plantas de três anos de idade, mantidas em casa de vegetação, foram inoculados nos meios nutritivos MS, ½ MS e WPM. Em um primeiro momento, adicionaram-se aos meios de cultura 1 μM de ANA e 5 μM de TDZ e, após, os explantes foram transferidos para frascos contendo os respectivos meios nutritivos frescos, na ausência de reguladores de crescimento, mas com o acréscimo de 1 g L-1 de carvão ativado, sendo cultivados por 30 dias adicionais. Os meios ½ MS e WPM são adequados para o estabelecimento e o desenvolvimento in vitro a partir do cultivo de segmentos nodais, enquanto que, com segmentos apicais, é apropriado o meio ½ MS. O meio ½ MS possibilitou os melhores índices de enraizamento nos explantes, juntamente com o meio WPM. O estabelecimento e o enraizamento in vitro foram promovidos pelo cultivo de segmentos apicais e nodais de Eugenia involucrata DC. Os meios nutritivos ½ MS e WPM são apropriados para o cultivo in vitro de segmentos nodais de E. involucrata, recomendando-se o meio ½ MS pela economia e praticidade. Para segmentos apicais, o uso do meio de cultura ½ MS é mais adequado que o dos meios MS e WPM.

  1. Androgens and bone.

    Science.gov (United States)

    Vanderschueren, Dirk; Vandenput, Liesbeth; Boonen, Steven; Lindberg, Marie K; Bouillon, Roger; Ohlsson, Claes

    2004-06-01

    Loss of estrogens or androgens increases the rate of bone remodeling by removing restraining effects on osteoblastogenesis and osteoclastogenesis, and also causes a focal imbalance between resorption and formation by prolonging the lifespan of osteoclasts and shortening the lifespan of osteoblasts. Conversely, androgens, as well as estrogens, maintain cancellous bone mass and integrity, regardless of age or sex. Although androgens, via the androgen receptor (AR), and estrogens, via the estrogen receptors (ERs), can exert these effects, their relative contribution remains uncertain. Recent studies suggest that androgen action on cancellous bone depends on (local) aromatization of androgens into estrogens. However, at least in rodents, androgen action on cancellous bone can be directly mediated via AR activation, even in the absence of ERs. Androgens also increase cortical bone size via stimulation of both longitudinal and radial growth. First, androgens, like estrogens, have a biphasic effect on endochondral bone formation: at the start of puberty, sex steroids stimulate endochondral bone formation, whereas they induce epiphyseal closure at the end of puberty. Androgen action on the growth plate is, however, clearly mediated via aromatization in estrogens and interaction with ERalpha. Androgens increase radial growth, whereas estrogens decrease periosteal bone formation. This effect of androgens may be important because bone strength in males seems to be determined by relatively higher periosteal bone formation and, therefore, greater bone dimensions, relative to muscle mass at older age. Experiments in mice again suggest that both the AR and ERalpha pathways are involved in androgen action on radial bone growth. ERbeta may mediate growth-limiting effects of estrogens in the female but does not seem to be involved in the regulation of bone size in males. In conclusion, androgens may protect men against osteoporosis via maintenance of cancellous bone mass and

  2. Gracile bone dysplasias

    Energy Technology Data Exchange (ETDEWEB)

    Kozlowski, Kazimierz [Department of Medical Imaging, The Children' s Hospital at Westmead, Locked Bag 4001, Westmead 2145, NSW (Australia); Masel, John [Department of Radiology, Royal Children' s Hospital, Brisbane (Australia); Sillence, David O. [Department of Paediatrics and Child Health, The University of Sydney (Australia); Arbuckle, Susan [Department of Anatomical Pathology, The Children' s Hospital at Westmead, NSW (Australia); Juttnerova, Vera [Oddeleni Lekarske Genetiky, Hradec Kralove (Czech Republic)

    2002-09-01

    Gracile bone dysplasias constitute a group of disorders characterised by extremely slender bones with or without fractures. We report four newborns, two of whom showed multiple fractures. Two babies had osteocraniostenosis and one had features of oligohydramnios sequence. The diagnosis in the fourth newborn, which showed thin long bones and clavicles and extremely thin, poorly ossified ribs, is uncertain. Exact diagnosis of a gracile bone dysplasia is important for genetic counselling and medico-legal reasons. (orig.)

  3. Enzymatic maceration of bone

    DEFF Research Database (Denmark)

    Uhre, Marie-Louise; Eriksen, Anne Marie; Simonsen, Kim Pilkjær;

    2015-01-01

    the bones. The DNA analysis showed that DNA was preserved on all the pieces of bones which were examined. Finally, the investigation suggests that enzyme maceration could be gentler on the bones, as the edges appeared less frayed. The enzyme maceration was also a quicker method; it took three hours compared...

  4. Precision extruding deposition (PED) fabrication of polycaprolactone (PCL) scaffolds for bone tissue engineering

    Energy Technology Data Exchange (ETDEWEB)

    Shor, Lauren; Gueceri, Selcuk; Chang, Robert; Sun Wei [Department of Mechanical Engineering and Mechanics, Drexel University, Philadelphia, PA (United States); Gordon, Jennifer; Kang Qian; Hartsock, Langdon; An Yuehuei [Department of Orthopedic Surgery, Medical University of South Carolina, Charleston, SC (United States)], E-mail: st963bya@drexel.edu, E-mail: guceri@drexel.edu, E-mail: rcc34@drexel.edu, E-mail: sunwei@drexel.edu, E-mail: kangqk@musc.edu, E-mail: hartsock@musc.edu, E-mail: any@musc.edu

    2009-03-01

    Bone tissue engineering is an emerging field providing viable substitutes for bone regeneration. Recent advances have allowed scientists and engineers to develop scaffolds for guided bone growth. However, success requires scaffolds to have specific macroscopic geometries and internal architectures conducive to biological and biophysical functions. Freeform fabrication provides an effective process tool to manufacture three-dimensional porous scaffolds with complex shapes and designed properties. A novel precision extruding deposition (PED) technique was developed to fabricate polycaprolactone (PCL) scaffolds. It was possible to manufacture scaffolds with a controlled pore size of 350 {mu}m with designed structural orientations using this method. The scaffold morphology, internal micro-architecture and mechanical properties were evaluated using scanning electron microscopy (SEM), micro-computed tomography (micro-CT) and mechanical testing, respectively. An in vitro cell-scaffold interaction study was carried out using primary fetal bovine osteoblasts. Specifically, the cell proliferation and differentiation was evaluated by Alamar Blue assay for cell metabolic activity, alkaline phosphatase activity and osteoblast production of calcium. An in vivo study was performed on nude mice to determine the capability of osteoblast-seeded PCL to induce osteogenesis. Each scaffold was implanted subcutaneously in nude mice and, following sacrifice, was explanted at one of a series of time intervals. The explants were then evaluated histologically for possible areas of osseointegration. Microscopy and radiological examination showed multiple areas of osseous ingrowth suggesting that the osteoblast-seeded PCL scaffolds evoke osteogenesis in vivo. These studies demonstrated the viability of the PED process to fabricate PCL scaffolds having the necessary mechanical properties, structural integrity, and controlled pore size and interconnectivity desired for bone tissue engineering.

  5. Multiplicação in vitro do porta-enxerto de macieira cv. Marubakaido: efeito da orientação do explante no meio de cultura In vitro multiplication of the apple rootstock cv. Marubakaido: effect of the orientation of explant in the medium of culture

    Directory of Open Access Journals (Sweden)

    ALAN CRISTIANO ERIG

    2002-08-01

    Full Text Available Objetivou-se avaliar o efeito da orientação do explante, vertical ou horizontal, no meio de cultura, na multiplicação in vitro, do porta-enxerto de macieira cv. Marubakaido. O meio de cultura utilizado foi o MS com N (nitrogênio reduzido a ¾ da concentração original, 100mg.L-1 de mio-inositol, 40g.L-1 de sacarose e 6g.L-1 de ágar, suplementado com 4,44mM de BAP (6-benzilaminopurina e 0,2ml.L-1 de PPM TM ("Plant Preservative Mixture". Segmentos caulinares com duas gemas e o ápice excisado foram utilizados como explantes. Após a inoculação, os frascos com os explantes foram incubados a 16 horas de fotoperíodo, à temperatura de 25±2ºC, com radiação de 25µmoles.m-2.s-1. O número de brotações, o número de gemas por explante, a taxa de multiplicação e a altura da brotação maior foram avaliados aos quarenta dias de cultivo. O maior número de brotações, o maior número de gemas e a maior taxa de multiplicação foram obtidos com o explante na orientação horizontal no meio de cultura. Não houve diferença significativa quanto à orientação vertical e horizontal do explante no meio de cultura para a altura da brotação maior.The aim of this study was evaluate the effect of the vertical and the horizontal orientation of the explant in the culture medium, in the in vitro multiplication, for the apple rootstock cv. Marubakaido. The culture medium used was the MS with N reduced to ¾ of the original concentration, myo-inositol (100mg.L-1, sucrose (40g.L-1 and agar (6g.L-1, suplemented with BAP (4.44mM and PPM TM (0.2ml.L-1. Stem segments with two buds and the apex excised were used as explants. After the inoculation, the flasks with the explants were incubated at 16 hour of photoperiod, 25±2ºC temperature, with irradiation of 25µmoles.m-2.s-1. The number of shoots and buds, the rate of multiplication and the height of the larger shoot were evaluated after 40 days of cultivation. The highests shoot number, number of buds

  6. Oxytocin and bone

    Science.gov (United States)

    Sun, Li; Zaidi, Mone; Zallone, Alberta

    2014-01-01

    One of the most meaningful results recently achieved in bone research has been to reveal that the pituitary hormones have profound effect on bone, so that the pituitary-bone axis has become one of the major topics in skeletal physiology. Here, we discuss the relevant evidence about the posterior pituitary hormone oxytocin (OT), previously thought to exclusively regulate parturition and breastfeeding, which has recently been established to directly regulate bone mass. Both osteoblasts and osteoclasts express OT receptors (OTR), whose stimulation enhances bone mass. Consistent with this, mice deficient in OT or OTR display profoundly impaired bone formation. In contrast, bone resorption remains unaffected in OT deficiency because, even while OT stimulates the genesis of osteoclasts, it inhibits their resorptive function. Furthermore, in addition to its origin from the pituitary, OT is also produced by bone marrow osteoblasts acting as paracrine-autocrine regulator of bone formation modulated by estrogens. In turn, the power of estrogen to increase bone mass is OTR-dependent. Therefore, OTR−/− mice injected with 17β-estradiol do not show any effects on bone formation parameters, while the same treatment increases bone mass in wild-type mice. These findings together provide evidence for an anabolic action of OT in regulating bone mass and suggest that bone marrow OT may enhance the bone-forming action of estrogen through an autocrine circuit. This established new physiological role for OT in the maintenance of skeletal integrity further suggests the potential use of this hormone for the treatment of osteoporosis. PMID:25209411

  7. A Single Center Study Comparing the Stainable Iron Depositions in 1000 Explanted Cirrhotic Livers of Different Causes

    Directory of Open Access Journals (Sweden)

    Geramizadeh

    2015-12-01

    Full Text Available Background There have been very few studies evaluating the close association between excess iron and cirrhosis; however, cirrhosis could be regarded as an iron-loading disorder. Objectives In this study, the goal was to show the levels of the iron content in the liver tissue in certain types of cirrhosis. Patients and Methods In this 7 year study (2008 - 2014, in 1000 explanted livers, the amount of iron was scored and compared according to the cause of the cirrhosis. The amount of iron in the liver was determined via the histochemical staining of the liver tissue, using Prussian-blue staining. Additionally, in each patient, the serum iron was determined and compared according to the cause of cirrhosis. Results The highest content of iron has been found in cirrhosis caused by chronic hepatitis (i.e. hepatitis B, C, and autoimmune hepatitis, as well as in alcoholic cirrhosis. The least amount of stainable iron has been shown in biliary cirrhosis. Conclusions The presence of high stainable iron in patients with cirrhosis, secondary to chronic viral hepatitis, autoimmune hepatitis, and alcoholic hepatitis, should not be considered indicative of the presence of hereditary hemochromatosis; however, in those patients with biliary cirrhosis, a high iron content is rare, and can be a sign of the presence of the high iron Fe (HFE gene mutation, or another type of hereditary hemochromatosis.

  8. Micropropagation ofTigridia pavonia (L.f) DC-a potential floricultural plant from twin scale explants

    Institute of Scientific and Technical Information of China (English)

    Lekha Kumar; Sincy Joseph; Narmatha Bai

    2012-01-01

    Objective:The present study was performed to standardize an effective protocol for micropropagation ofTigridia pavonia using tissue culture.Methods: The explants were cultured on Murashige and Skoog (MS) medium supplemented with cytokinins like thidiazuron (TDZ), zeatin, kinetin and auxins such as indole-3-acetic acid (IAA), 1-naphthalene acetic acid (NAA) and indole-3-butyric acid (IBA), individually at different concentrations.Results:Multiple shoots were obtained on MS medium containing either 2.0 mg/L TDZ or 2.0 mg/L IAA or 0.5 mg/L IBA and in the same medium for a long period (120 d) produced tiny bulbs at the base of the senescent leaves. TDZ favored only multiple shoots without roots, whereas IAA or IBA individually or in combination with TDZ produced rooted shoots. Shoots developed on MS medium supplemented with TDZ were rooted on MS medium containing either IBA or NAA at 0.5 mg/L. The plantlets were acclimatized in pots containing garden soil. Regenerated plantlets developed into normal plants. The plants showed 99% survival.Conclusions:The highest number of bulblets obtained in the present study represents an effective alternative to the conventional method.

  9. Accuracy of hepatocellular carcinoma detection on multidetector CT in a transplant liver population with explant liver correlation

    Energy Technology Data Exchange (ETDEWEB)

    Addley, H.C., E-mail: helenclareaddley@hotmail.co.u [Department of Radiology, Addenbrooke' s Hospital, Cambridge University Hospitals NHS Foundation Trust, Cambridge (United Kingdom); Griffin, N.; Shaw, A.S.; Mannelli, L. [Department of Radiology, Addenbrooke' s Hospital, Cambridge University Hospitals NHS Foundation Trust, Cambridge (United Kingdom); Parker, R.A. [Department of Medical Statistics, Addenbrooke' s Hospital, Cambridge University Hospitals NHS Foundation Trust, Cambridge (United Kingdom); Aitken, S.; Wood, H. [Department of Radiology, Addenbrooke' s Hospital, Cambridge University Hospitals NHS Foundation Trust, Cambridge (United Kingdom); Davies, S. [Department of Histopathology, Addenbrooke' s Hospital, Cambridge University Hospitals NHS Foundation Trust, Cambridge (United Kingdom); Alexander, G.J. [Department of Hepatology, Addenbrooke' s Hospital, Cambridge University Hospitals NHS Foundation Trust, Cambridge (United Kingdom); Lomas, D.J. [Department of Radiology, Addenbrooke' s Hospital, Cambridge University Hospitals NHS Foundation Trust, Cambridge (United Kingdom)

    2011-04-15

    Aim: To evaluate the diagnostic accuracy of multidetector computed tomography (MDCT) for hepatocellular carcinoma (HCC) in cirrhotic patients undergoing liver transplantation. Secondary aims were to examine the effect of radiologist experience and lesion size on diagnostic accuracy. Materials and methods: Thirty-nine patients (72% male with a mean age of 56.5 years) underwent liver transplantation following preoperative triple-phase MDCT examination of the liver. MDCT examinations were retrospectively independently reviewed by three radiologists for the presence and location of suspected HCCs, with the diagnostic confidence recorded using a five-point confidence scale. MDCT examinations were compared with explant specimens for histopathological correlation. Results: Histopathological results demonstrated 46 HCCs in 29 of the 39 patients. Analysis demonstrated a sensitivity of 65-75% and specificity of 47-88% for detection of HCC lesions. The sensitivity dropped to 48-57% for lesions of size {<=}20 mm. As the diagnostic confidence increased, there was a further decrease in the sensitivity (4-26%). The radiologist with the greatest number of years experience was found to have a significantly higher accuracy of detection of HCC lesions compared with the least experienced radiologist. Conclusion: Larger lesion size of HCC and greater number of years experience of the radiologist resulted in significantly higher accuracy of HCC lesion detection. The overall sensitivity and specificity results for MDCT detection of HCC are comparable to previous helical CT imaging.

  10. An analysis of flexible anterior chamber lenses with special reference to the normalized rate of lens explantation.

    Science.gov (United States)

    Lim, E S; Apple, D J; Tsai, J C; Morgan, R C; Wasserman, D; Assia, E I

    1991-02-01

    A survey of 1204 closed-loop anterior chamber intraocular lenses (AC-IOLs) and 310 open-loop AC-IOLs accessioned between November 1982 and January 1990 was conducted at the Center for Intraocular Lens Research. An analysis of complication rates was done after normalization of data with respect to market share totals. The results establish that an unacceptable complication rate is associated with the closed-loop design when compared with either the tripod or quadripod lens styles. Furthermore, the closed-loop designs, while comprising an estimated 45% of the total number of AC-IOLs estimated to be implanted in the United States (n = 674,000), were responsible for 80% of the AC-IOLs explanted after complications and accessioned at the authors' center. A rethinking of the extreme condemnation of all anterior chamber IOLs that has surfaced in recent years is warranted. This is particularly true with respect to indications for use of sutured posterior chamber (PC) IOLs as well as with regard to possible use of open-loop AC-IOLs in less-industrialized nations.

  11. Increased Agrobacterium-mediated transformation and rooting efficiencies in canola (Brassica napus L.) from hypocotyl segment explants

    Science.gov (United States)

    Cardoza, V.; Stewart, C. N.

    2003-01-01

    An efficient protocol for the production of transgenic Brassica napus cv. Westar plants was developed by optimizing two important parameters: preconditioning time and co-cultivation time. Agrobacterium tumefaciens-mediated transformation was performed using hypocotyls as explant tissue. Two variants of a green fluorescent protein (GFP)-encoding gene--mGFP5-ER and eGFP--both under the constitutive expression of the cauliflower mosaic virus 35S promoter, were used for the experiments. Optimizing the preconditioning time to 72 h and co-cultivation time with Agrobacterium to 48 h provided the increase in the transformation efficiency from a baseline of 4% to 25%. With mGFP5-ER, the transformation rate was 17% and with eGFP it was 25%. Transgenic shoots were selected on 200 mg/l kanamycin. Rooting efficiency was 100% on half-strength Murashige and Skoog medium with 10 g/l sucrose and 0.5 mg/l indole butyric acid in the presence of kanamycin.

  12. Acetylcholine suppresses shoot formation and callusing in leaf explants of in vitro raised seedlings of tomato, Lycopersicon esculentum Miller var. Pusa Ruby.

    Science.gov (United States)

    Bamel, Kiran; Gupta, Rajendra; Gupta, Shirish C

    2016-06-02

    We present experimental evidence to show that acetylcholine (ACh) causes decrease in shoot formation in leaf explants of tomato (Lycopersicon esculentum Miller var Pusa Ruby) when cultured on shoot regeneration medium. The optimum response was obtained at 10(-4) M ACh-enriched medium. ACh also causes decrease in percentage of cultures forming callus and reduces the callus mass. Inhibitors of enzymatic hydrolysis of ACh, neostigmine and physostigmine, also suppresses callogenesis and caulogenesis. On the other hand, the breakdown products of Ach, choline and acetate, do not alter the morphogenic response induced on the shoot regeneration medium. Neostigmine showed optimal reduction in shoot formation at 10(-5) M. The explants cultured on neostigmine augmented medium showed decline in the activity of ACh hydrolyzing enzyme acetylcholinesterase. ACh and neostigmine added together showed marked reduction in callus mass. These results strongly support the role of ACh as a natural regulator of morphogenesis in tomato plants.

  13. Evaluation for Plant Regeneration Potential of Root Explants in Echinacea purpurea%松果菊根外植体植株再生能力的评价

    Institute of Scientific and Technical Information of China (English)

    Dahanayake NILANTHI; 赵福成; 杨跃生; 吴鸿

    2009-01-01

    为了评价松果菊Echinacea purpurea L.根外植体的再生能力,将从松果菊无菌小苗得到的根外植体和叶片以及叶柄外植体接种到含有不同种类和浓度的细胞分裂素和生长素的培养基上,诱导不定芽的再生.结果表明,在多数情况下,根外植体的再生能力显著高于叶片,和叶柄类似.0.3 mg/L的苄基腺嘌呤和0.01 mg/L的萘乙酸是诱导根外植体不定芽再生最合适的激素种类和质量浓度组合.根外植体培养的不定芽再生频率为100%,每个根外植体得到再生芽1.75个.当把这些由根再生的不定芽从母体组织切开并转移培养到含有0.01 mg/L萘乙酸的培养基后,很容易生根并成为完整的植株.可见根是组培快繁松果菊理想的外植体材料.%For evaluation of the plant regeneration potential of root explants, explants of root, leaf and petiole were taken from in vitro grown purple coneflower, Echinacea purpurea L. Plantlets and cultured on adventitious bud inducing media with different cytokinins and auxins at various concentrations. In most of the cases, the regeneration potential of root explants was much higher than that of leaf ones and similar to that of petiole explants, and a combination of 0.3 mg/L benzyladeine with 0.01 mg/L naphthaleneacetic acid was the most effective combination and concentrations for inducing adventitious bud regen-eration. Although the best result of bud regeneration was obtained from culture of petiole explants, a good result in regenera-tion rate of 100% and a high number of 1.75 buds per exphnt were obtained from culture of root explants. Buds regenerated from root explants initiated roots and became intact plants readily upon transfer to a medium containing 0.01 mg/L naphthale-neacetic acid. Results of the experiments indicated that root was an ideal explant source for rapid propagation by means of tis-sue culture in this plant species.

  14. Effect of polyamines and silver nitrate on the high frequency regeneration from cotyledon explants of bottle gourd (Lagenaria siceraria; sp. asiatica).

    Science.gov (United States)

    Shyamali, Saha; Hattori, Kazumi

    2007-04-15

    In this study, we have investigated the effect ofpolyamines (PA) and silver nitrate (AgNO3) on the high frequency regeneration from cotyledon explants of bottle gourd containing Murashige and Skoog (MS) media supplemented with different kind of Cytokinin alone or in the combination. Synergistic effect of kinetin (1 mg L(-1)) and benzyl adenine (BA) (2 mg L(-1)) itself showed highest shoot regeneration (80.6%) efficiency than BA or Kinetin alone in cotyledon explants of bottle gourd without adding AgNO3 or PAs. We have also observed that PAs and AgNO3 show their sensitivity on the regeneration, which is hormonal dependent. Regenerated shoots were rooted in half strength MS media containing 0.1 mg L(-1) IAA.

  15. Bone regeneration with cultured human bone grafts

    Energy Technology Data Exchange (ETDEWEB)

    Yoshikawa, T.; Nakajima, H. [Nara Medical Univ., Kashihara City (Japan). Dept. of Pathology; Nara Medical Univ., Kashihara City (Japan). Dept. of Orthopedic Surgery; Ohgushi, H.; Ueda, Y.; Takakura, Y. [Nara Medical Univ., Kashihara City (Japan). Dept. of Orthopedic Surgery; Uemura, T.; Tateishi, T. [National Inst. for Advanced Interdisciplinary Research (NAIR), Ibaraki (Japan). Tsukuba Research Center; Enomoto, Y.; Ichijima, K. [Nara Medical Univ., Kashihara City (Japan). Dept. of Pathology

    2001-07-01

    From 73 year old female patient, 3 ml of bone marrow was collected from the ilium. The marrow was cultured to concentrate and expand the marrow mesenchymal cells on a culture dish. The cultured cells were then subculturedeither on another culture dish or in porous areas of hydroxyapatite ceramics in the presence of dexamethasone and beta-glycerophosphate (osteo genic medium). The subculturedtissues on the dishes were analyzed by scanning electron microscopy (SEM), and subculturedtissues in the ceramics were implanted intraperitoneally into athymic nude mice. Vigorous growth of spindle-shaped cells and a marked formation of bone matrix beneath the cell layers was observed on the subculture dishes by SEM. The intraperitoneally implanted ceramics with cultured tissues revealed thick layer of lamellar bone together with active osteoblasts lining in many pore areas of the ceramics after 8 weeks. The in vitro bone formations on the culture dishes and in vivo bone formation in porous ceramics were detected. These results indicate that we can assemble an in vitro bone/ceramic construct, and due to the porous framework of the ceramic, the construct has osteogenic potential similar to that of autologous cancellous bone. A significant benefit of this method is that the construct can be made with only a small amount of aspirated marrow cells from aged patients with little host morbidity. (orig.)

  16. An Injectable Hydrogel as Bone Graft Material with Added Antimicrobial Properties

    Science.gov (United States)

    Tommasi, Giacomo; Perni, Stefano

    2016-01-01

    Currently, the technique which provides the best chances for a successful bone graft, is the use of bone tissue from the same patient receiving it (autograft); the main limitations are the limited availability and the risks involved in removing living bone tissue, for example, explant site pain and morbidity. Allografts and xenografts may overcome these limitations; however, they increase the risk of rejection. For all these reasons the development of an artificial bone graft material is particularly important and hydrogels are a promising alternative for bone regeneration. Gels were prepared using 1,4-butanediol diacrylate as crosslinker and alpha tricalciumphosphate; ZnCl2 and SrCl2 were added to the aqueous phase. MTT results demonstrated that the addition of strontium had a beneficial effect on the osteoblast cells density on hydrogels, and zinc instead did not increase osteoblast proliferation. The amount of calcium produced by the osteoblast cells quantified through the Alizarin Red protocol revealed that both strontium and zinc positively influenced the formation of calcium; furthermore, their effect was synergistic. Rheology properties were used to mechanically characterize the hydrogels and especially the influence of crosslinker's concentration on them, showing the hydrogels presented had extremely good mechanical properties. Furthermore, the antimicrobial activity of strontium and zinc in the hydrogels against methicillin-resistant Staphylococcus aureus and Staphylococcus epidermidis was determined. PMID:27174392

  17. CRESCIMENTO E OXIDAÇÃO DE EXPLANTES DE BANANEIRA-PRATA (Musa AAB IN VITRO: I. CONCENTRAÇÕES DE SAIS DE FERRO, COBRE E ZINCO

    Directory of Open Access Journals (Sweden)

    UTINO SERGIO

    2001-01-01

    Full Text Available Este experimento teve como objetivo avaliar a influência de diferentes concentrações de ferro, cobre e zinco do meio MS (MURASHIGE & SKOOG, 1962 no controle da oxidação de explantes iniciais de bananeira-Prata (Musa AAB. Foram utilizadas três concentrações (100, 50 e 0 miM de FeEDTA, duas concentrações (0,1 e 0miM de (CuSO4.5H2O e duas concentrações (30 e 0miM de (ZnSO4.7H2O, num delineamento inteiramente casualizado, arranjado em um fatorial completo 3 x 2 x 2, utilizando-se de 15 repetições. Ápices caulinares foram inoculados em meio MS modificado e, decorridos 28 dias após a inoculação, avaliaram-se a massa de matéria fresca, altura e grau de oxidação. Observou-se que esses micronutrientes são essenciais para o crescimento dos explantes e que a concentração de ferro influencia na oxidação de explantes, sendo que maiores graus de escurecimento foram observados nas concentrações mais elevadas. A redução ou retirada destes elementos do meio MS, isoladamente ou em combinações, não foi suficiente para eliminar a oxidação dos explantes.