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Sample records for acellular bone explants

  1. Combining acellular nerve allografts with brain-derived neurotrophic factor transfected bone marrow mesenchymal stem cells restores sciatic nerve injury better than either intervention alone

    Zhang, Yanru; Zhang, Hui; Zhang, Gechen; Ka, Ka; Huang, Wenhua

    2014-01-01

    In this study, we chemically extracted acellular nerve allografts from bilateral sciatic nerves, and repaired 10-mm sciatic nerve defects in rats using these grafts and brain-derived neurotrophic factor transfected bone marrow mesenchymal stem cells. Experiments were performed in three groups: the acellular nerve allograft bridging group, acellular nerve allograft + bone marrow mesenchymal stem cells group, and the acellular nerve allograft + brain-derived neurotrophic factor transfected bone...

  2. Socket Preservation Therapy with Acellular Dermal Matrix and Mineralized Bone Allograft After Tooth Extraction in Humans: A Clinical and Histomorphometric Study.

    Fernandes, Patricia Garani; Muglia, Valdir Antonio; Reino, Danilo Maeda; Maia, Luciana Prado; de Moraes Grisi, Marcio Fernando; de Souza, Sergio Luís; Taba, Mario; Palioto, Daniela Bazan; de Almeida, Adriana G; Novaes, Arthur Belém

    2016-01-01

    The aim of this study was to analyze through clinical and histomorphometric parameters the use of acellular dermal matrix (ADM) with or without mineralized bone allograft (AB) on bone formation in human alveoli after a 6- to 8-month healing period. A total of 19 patients in need of extraction of the maxillary anterior teeth were selected and randomly assigned to the test group (ADM plus AB) or to the control group (ADM only). Clinical and histomorphometric measurements and histologic analysis were recorded 6 to 8 months after ridge preservation procedures. Clinical parameters and amount of mineralized and nonmineralized tissue were measured and analyzed. In the clinical measurements, the test group showed reduced bone loss in the buccopalatal dimension after 6 to 8 months (intragroup analysis P acellular dermal matrix in association with mineralized bone allograft reduced alveolar bone loss in the anterior maxillae both in height and width after a follow-up period of 6 to 8 months. PMID:26901306

  3. 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

    Arthur B. Novaes Jr.; Patricia Garani Fernandes; Flávia Adelino Suaid; Marcio Fernando de Moraes Grisi; Sergio Luis Scombatti de Souza; Mario Taba Jr.; Daniela Bazan Palioto; Valdir Antonio Muglia

    2012-01-01

    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 a...

  4. Formation of acellular cementum-like layers, with and without extrinsic fiber insertion, along inert bone surfaces of aging c-Src gene knockout mice.

    Baba, Otto; Miyata, Atsushi; Abe, Tatsuhiko; Shibata, Shunichi; Nakano, Yukiko; Terashima, Tatsuo; Oda, Tsuyoshi; Kudo, Akira; Takano, Yoshiro

    2006-12-01

    To investigate the long-term effects of c-src deficiency on skeletal and dental tissues, we examined the lower jaws and long bones of c-src gene knockout (c-src KO) mice by histological and histochemical methods. Numerous multinucleated osteoclasts were distributed throughout the mandible in 5-wk-old c-src KO mice, but by 14 wk they had almost completely disappeared from the alveolar bone, leaving tartrate-resistant acid phosphatase (TRAP)-positive layers along the bone surface. Deposition of osteopontin-positive mineralized tissue, reminiscent of acellular afibrillar cementum (AAC), was confirmed along the TRAP-positive bone surface at 14 wk. The layer progressively thickened up to 21 months. A comparable mineralized layer was noted along the trabeculae of long bones as thickened cement lines. In the periostin-rich areas of jaw bones, but not in the long bones, portions of AAC-like mineralized layers were often replaced with and/or covered by acellular extrinsic fiber cementum (AEFC)-like tissue. These data suggest that the deposition of AAC-like mineralized tissue is a general phenomenon that may occur along inert or slowly remodeling bone surfaces under conditions characterized by reduced bone-resorbing activity, whereas the induction of AEFC-like tissue seems to be associated with the expression of certain molecules that are particularly abundant in the microenvironment of the periodontal ligament. PMID:17184236

  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

    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. Biocompatibility of acellular natural bone matrix with induced osteoblasts in vitro%脱细胞天然骨基质与诱导性成骨细胞的体外相容性

    李康杰; 孙抒

    2011-01-01

    背景:前期工作表明TritonX-100处理的脱细胞骨基质已满足组织学和免疫学方面的修复要求.如果细胞能在材料表面很好地生长,将利于进一步进行体内动物实验.目的:采用细胞培养法在体外评估脱细胞骨基质与诱导后成骨细胞的生物相容性.方法:第3代骨髓基质干细胞经成骨诱导分化培养液诱导分化为成骨细胞,接种于TritonX-100处理的脱细胞骨基质及羟基磷灰石表面,检测成骨细胞的碱性磷酸酶表达并用扫描电镜观察材料表面的细胞生长情况.结果与结论:碱性磷酸酶活性分析均表明,TritonX-100处理的脱细胞骨基质在培养48 h之后比羟基磷灰石更利于诱导成骨细胞生长;扫描电镜下可见,成骨细胞在脱细胞骨基质表面呈现立体生长方式,细胞呈球形,并且聚集成簇.体外实验结果显示成骨细胞与脱细胞天然骨基质有较好的生物相容性.%BACKGROUND:The repair requirements of histology and immunology can be met by acellular bone matrix treated by TritonX-100.The test cells will be able to grow nicely on the surface of test materials, which will benefit fu rther evaluating them in animals in vivo.OBJECTIVE:To evaluate the biocompatibility of acellular natural bone matrix prepared with TritonX-100 to osteoblasts by cell culture assay.METHODS:The third passage of bone marrow stromal stem cells were induced to differentiate into osteoblasts, then the identified osteoblasts were seeded on surface of acellular natural bone matrix prepared with TritonX-100 and calcium hydroxyapatite. Alkaline phosphatase vitality was measured by alkaline phosphatase detection kit and the growth of osteoblast-like cells on su rface of the material was observed with scanning electron microscopy.RESULTS AND CONCLUSION:Alkaline phosphatase detection kit analysis indicated that acellular natural bone matrix prepared with TritonX-100 were able to promote the growth of osteoblast-like cells compared

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

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

    2016-01-01

    Gel aspiration-ejection (GAE) has recently been introduced as an effective technique for the rapid production of injectable dense collagen (IDC) gel scaffolds with tunable collagen fibrillar densities (CFDs) and microstructures. Herein, a GAE system was applied for the advanced production and delivery of IDC and IDC-Bioglass® (IDC-BG) hybrid gel scaffolds for potential bone tissue engineering applications. The efficacy of GAE in generating mineralizable IDC-BG gels (from an initial 75-25 coll...

  8. Chemically modified RNA induces osteogenesis of stem cells and human tissue explants as well as accelerates bone healing in rats.

    Balmayor, Elizabeth R; Geiger, Johannes P; Aneja, Manish K; Berezhanskyy, Taras; Utzinger, Maximilian; Mykhaylyk, Olga; Rudolph, Carsten; Plank, Christian

    2016-05-01

    Limitations associated to the use of growth factors represent a major hurdle to musculoskeletal regeneration. On the one hand, they are needed to induce neo-tissue formation for the substitution of a necrotic or missing tissue. On the other hand, these factors are used in supraphysiological concentrations, are short lived and expensive and result in many side effects. Here we develop a gene transfer strategy based on the use of chemically modified mRNA (cmRNA) coding for human bone morphogenetic protein 2 (hBMP-2) that is non-immunogenic and highly stable when compared to unmodified mRNA. Transfected stem cells secrete hBMP-2, show elevated alkaline phosphatase levels and upregulated expression of RunX2, ALP, Osterix, Osteocalcin, Osteopontin and Collagen Type I genes. Mineralization was induced as seen by positive Alizarin red staining. hBMP-2 cmRNA transfected human fat tissue also yielded an osteogenic response in vitro as indicated by expression of hBMP-2, RunX2, ALP and Collagen Type I. Delivering hBMP-2 cmRNA to a femur defect in a rat model results in new bone tissue formation as early as 2 weeks after application of very low doses. Overall, our studies demonstrate the feasibility and therapeutic potential of a new cmRNA-based gene therapy strategy that is safe and efficient. When applied clinically, this approach could overcome BMP-2 growth factor associated limitations in bone regeneration. PMID:26923361

  9. Xenopus laevis Keller Explants

    sprotocols

    2014-01-01

    Authors: Hazel L. Sive, Robert M. Grainger and Richard M. Harland This protocol was adapted from “Microdissection,” Chapter 10, in [*Early Development of* *Xenopus laevis*](http://www.cshlpress.com/link/xenopus.htm) by Hazel L. Sive, Robert M. Grainger, and Richard M. Harland. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 2000. ### INTRODUCTION The basic Keller explant is a rectangle of dorsal mesendoderm and ectoderm from an early-gastrula-stage *Xenopus laevi...

  10. Acellular Dermal Matrix in Rotator Cuff Surgery.

    Cooper, Joseph; Mirzayan, Raffy

    2016-01-01

    The success of rotator cuff repair (RCR) surgery can be measured clinically (validated outcome scores, range of motion) as well as structurally (re-tear rates using imaging studies). Regardless of repair type or technique, most studies have shown that patients do well clinically. However, multiple studies have also shown that structurally, the failure rate can be very high. A variety of factors, including poor tendon quality, age over 63 years, smoking, advanced fatty infiltration into the muscle, and the inability of the tendon to heal to bone, have been implicated as the cause of the high re-tear rate in RCRs. The suture-tendon interface is felt to be the weakest link in the RCR construct, and suture pullout through the tendon is believed to be the most common method of failure. This review of the published literature seeks to determine if there is support for augmentation of RCR with acellular dermal matrices to strengthen the suture-tendon interface and reduce the re-tear rate. PMID:27552454

  11. Explant cultures of human colon

    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...

  12. Expression of vascular endothelial growth factor in repair of cranial bone defect with acellular bone matrix combined with platelet-rich plasma%VEGF在脱细胞骨基质复合富血小板血浆修复颅骨缺损中的表达

    袁道英; 杨佑成; 张彬; 牛怀恩; 李克义; 张巍峰

    2010-01-01

    目的:研究血管内皮生长因子(VEGF)在脱细胞骨基质(acellular bone matrix,ABM)复合富血小板血浆(platelet-rich plasma,PRP)修复兔颅骨缺损时的表达及分布,探讨富血小板血浆促进成骨的机制.方法:雄性新西兰大白兔24只,体质量1.5~2.0 kg.在兔颅骨两侧分别建立一个1cm×0.5cm全层骨缺损区,同时去除该区骨膜,注意勿伤及硬脑膜.随机选择一侧骨缺损作实验侧,植入复合PRP的ABM;另一侧为对照侧,仅植入ABM.术后第2、4、8、12周末分别处死6只兔取材.免疫组织化学法测定骨缺损修复区血管内皮细胞生长因子(Vascular endothelial growth factor,VEGF)的表达;应用Image-proplus 5.0图像分析软件测量VEGF表达强度的灰度值.采用SSPS 16.0软件包进行t检验.结果:术后2周,实验组VEGF呈强阳性表达,随后急剧下降,以后趋于平缓.对照组在术后2、4周呈阳性表达,以后平缓下降.两组相比,在第2、4周时差异均有显著性(P<0.05).第8、12周时,2组表达差异无显著性.结论:VEGF在实验组早期阶段的强阳性表达,说明血管形成活跃.PRP促进骨修复的作用发生在植入后早期,启动了早期活跃的成骨.

  13. Acellular Dermal Matrix in Postmastectomy Breast Reconstruction

    A.M.S. Ibrahim (Ahmed)

    2014-01-01

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

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

    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.

  15. Central role of pyrophosphate in acellular cementum formation.

    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

  16. Data from acellular human heart matrix.

    Sánchez, Pedro L; Fernández-Santos, M Eugenia; Espinosa, M Angeles; González-Nicolas, M Angeles; Acebes, Judith R; Costanza, Salvatore; Moscoso, Isabel; Rodríguez, Hugo; García, Julio; Romero, Jesús; Kren, Stefan M; Bermejo, Javier; Yotti, Raquel; Del Villar, Candelas Pérez; Sanz-Ruiz, Ricardo; Elizaga, Jaime; Taylor, Doris A; Fernández-Avilés, Francisco

    2016-09-01

    Perfusion decellularization of cadaveric hearts removes cells and generates a cell-free extracellular matrix scaffold containing acellular vascular conduits, which are theoretically sufficient to perfuse and support tissue-engineered heart constructs. This article contains additional data of our experience decellularizing and testing structural integrity and composition of a large series of human hearts, "Acellular human heart matrix: a critical step toward whole heat grafts" (Sanchez et al., 2015) [1]. Here we provide the information about the heart decellularization technique, the valve competence evaluation of the decellularized scaffolds, the integrity evaluation of epicardial and myocardial coronary circulation, the pressure volume measurements, the primers used to assess cardiac muscle gene expression and, the characteristics of donors, donor hearts, scaffolds and perfusion decellularization process. PMID:27331090

  17. Use of an Acellular Regenerative Tissue Matrix Over Chronic Wounds

    Stacey, D. Heath

    2013-01-01

    Objectives: Bioengineered skin grafts, including acellular dermal matrices, may be effective in treating lower extremity and trunk wounds that are not responsive to traditional wound management. Acellular dermal wound matrix is derived from human acellular dermal wound matrix (HADWM) tissue and provides a scaffold that supports cellular repopulation and revascularization. The major structural components of the dermis are retained during processing, and a single application has been shown to h...

  18. Acellularization-induced changes in tensile properties are organ specific

    Schleifenbaum, Stefan; Prietzel, Torsten; Aust, Gabriela; Boldt, Andreas; Fritsch, Sebastian; Keil, Isabel; Koch, Holger; Möbius, Robert; Scheidt, Holger A.; Wagner, Martin F. X.; Hammer, Niels

    2016-01-01

    Introduction: 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. Materials and methods: 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...

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

    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.

  20. Responsiveness of mouse calvaria to parathyroid hormone after explant cryopreservation: 45Ca release in vitro

    Wezeman, F.H.; Dungan, D.D.

    1986-08-01

    Newborn mouse calvaria prelabeled with /sup 45/Ca and cryopreserved at -196 degrees C in serum-free medium containing dimethylsulfoxide were compared to unpreserved explants for response to parathyroid hormone during subsequent culture. After short-term cryopreservation followed by rapid thawing, the viable explants continued to release /sup 45/Ca to the culture medium but additions of parathyroid hormone to the medium did not cause increased bone resorption. The data suggest that cryopreservation and thawing impairs mechanisms responsible for parathyroid hormone action on bone cells.

  1. Acellular organ scaffolds for tumor tissue engineering

    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.

  2. Pancreatic acellular matrix supports islet survival and function in a synthetic tubular device: in vitro and in vivo studies.

    De Carlo, E; Baiguera, S; Conconi, M T; Vigolo, S; Grandi, C; Lora, S; Martini, C; Maffei, P; Tamagno, G; Vettor, R; Sicolo, N; Parnigotto, P P

    2010-02-01

    Increasing pancreatic islet survival and function is a starting point for obtaining a valuable bioartificial pancreas for the treatment of type 1 diabetes. In this context, decellularized matrices, obtained after the removal of tissue cellular part, are known to support in vitro adhesion, growth, and function of several cell types. We demonstrate that a homologous acellular pancreatic matrix is a suitable scaffold for rat islet cultures maintaining their long-term viability and function. Islets adhered to the pancreatic matrix showed a constant glucose-induced insulin release during long-term in vitro incubation, while islets cultured without a matrix or on the liver matrix showed a progressive reduction. In order to obtain implantable devices, acellular matrix/islet cultures were entrapped into poly(vinyl alcohol) (PVA)/ poly(ethylene glycol) (PEG) tubes obtained by the freezing/thawing procedure. Under this condition, an in vitro constant insulin release was detected. The devices were then implanted into diabetic rats where reduced insulin requirement was noted suggesting insulin secretory activity of islets contained in the device. Indeed, immunofluorescence confirmed the presence of insulin- and glucagon-producing cells into the explanted devices. These data show that PVA/PEG semi-permeable membrane can obtain devices that restore, at least in part, insulin secretion. PMID:20043127

  3. 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

    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

  4. Bone

    Bone scanning provides information on the extent of primary bone tumors, on possible metastatic disease, on the presence of osteomyelitis prior to observation of roentgenographic changes so that earlier therapy is possible, on the presence of collagen diseases, on the presence of fractures not disclosed by x-ray films, and on the evaluation of aseptic necrosis. However, the total effect and contribution of bone scanning to the diagnosis, treatment, and ultimate prognosis of pediatric skeletal diseases is, as yet, unknown. (auth)

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

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

    2013-01-01

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

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

    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.

  7. Abdominal wall repair with human acellular dermal autograft

    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

  8. Failure analysis of explanted sternal wires.

    Shih, Chun-Ming; Su, Yea-Yang; Lin, Shing-Jong; Shih, Chun-Che

    2005-05-01

    To classify and understand the mechanisms of surface damages and fracture mechanisms of sternal wires, explanted stainless steel sternal wires were collected from patients with sternal dehiscence following open-heart surgery. Surface alterations and fractured ends of sternal wires were examined and analyzed. Eighty fractured wires extracted from 25 patients from January 1999 to December 2003, with mean implantation interval of 55+/-149 days (range 5-729 days) after cardiac surgery, were studied by various techniques. The extracted wires were cleaned and the fibrotic tissues were removed. Irregularities and fractured ends were assayed by a scanning electron microscopy. After stereomicroscopy and documentation, the explants were cleaned with 1% sodium hypochlorite to remove the blood and tissues and was followed by cleaned with deionized water and alcohol. The explants were examined by stereomicroscopy, and irregularities on surface and fracture surfaces of sternal wires were assayed by scanning electron microscopy, energy dispersive X-ray analysis (EDAX) and X-ray mapping. The explants with surrounding fibrotic tissue were stained and examined with stereomicroscopy and transmission electronic microscopy. Corrosion pits were found on the surface of explanted sternal wires. EDAX and X-ray mapping examinations revealed diminution of nickel concentration in the severely corroded pits on sternal wires. A feature of transgranular cracking was observed for stress corrosion cracking and striation character for typical corrosion fatigue was also identified. TEM examination of tissue showed the metallic particles in phagolysosomes of macrophages inside the surrounding sternal tissue. The synergic effect of hostile environment and the stress could be the precursors of failures for sternal wires. PMID:15576179

  9. Evaluation of lymphangiogenesis in acellular dermal matrix

    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

  10. Effects of microporous porcine acellular dermal matrix combined with bone marrow mesenchymal cells of rats on the regeneration of cutaneous appendages cells in nude mice%微孔化猪脱细胞真皮基质与大鼠骨髓间充质细胞对裸鼠皮肤附件细胞再生的作用

    罗旭; 辛国华; 曾逃方; 林才; 曾元临; 李郁葱; 邱泽亮

    2013-01-01

    色清亮渗液;A、D组均未见明显脓性分泌物.B、C组皮片外观与周围皮肤颜色接近.(5)移植术后5、7 d,A、B、C组真皮基质的微孔结构中已见血管化,其内可见有形红细胞;D组移植皮片部分干燥坏死.移植术后14 d,A、B、C组真皮基质的微孔结构中已完全血管化,其内可见大量的红细胞.纵切片中,A组微孔真皮基质成活,但与其上所覆盖的无孔猪ADM未紧密结合;B、C组皮片与真皮基质间连接紧密,皮片中均未见皮肤附属器,C组创面皮片与真皮基质交接处可见特殊的单层细胞.(6)D组移植皮片末能成活,故放弃电镜观察.移植术后7d,A、B、C组透射电镜图片未见明显差别.移植术后14 d,A、B组移植物中未见皮脂腺样及汗腺样细胞,也未见新生神经末梢,仅见Fb迁入.C组创面刃厚皮与真皮基质交接处可见大量新生毛细血管增生,Fb粗面内质网分裂增殖旺盛,可见新生的无髓神经末梢;在真皮基质浅层,出现单个游离的皮脂腺样及汗腺样细胞 结论 LPADM为骨髓间充质细胞群的迁移和分化提供了“干细胞龛”样微环境,联合刃厚皮片移植可在体诱导外源性BMSC分化,实现部分皮肤附件的重建.%Objective To observe the effects of microporous porcine acellular dermal matrix (ADM) combined with bone marrow mesenchymal cells (BMMCs) population containing bone mesenchymal stem cells (BMSCs) of rats on the regeneration of cutaneous appendages cells in nude mice.Methods Split-thickness dermal grafts,20 cm × 10 cm in size and 0.3 mm in thickness,were prepared from a healthy pig which was sacrificed under sanitary condition.Laser microporous porcine ADM (LPADM) was produced by laser punching,hypertonic saline solution acellular method,and crosslinking treatment,and nonporous porcine ADM (NPADM) was produced by the latter two procedures.Then the appearance observation,histological examination and scanning electron microscope observation were

  11. A comparative study of acellular nerve xenografts and allografts in repairing rat facial nerve defects.

    Huang, Haitao; Xiao, Hongxi; Liu, Huawei; Niu, Yu; Yan, Rongzeng; Hu, Min

    2015-10-01

    Acellular nerves are composed of a basal lamina tube, which retains sufficient bioactivity to promote axon regeneration, thereby repairing peripheral nerve gaps. However, the clinical application of acellular allografts has been restricted due to its limited availability. To investigate whether xenografts, a substitute to allograft acellular nerves in abundant supply, could efficiently promote nerve regeneration, rabbit and rat acellular nerve grafts were used to reconstruct 1 cm defects in Wistar rat facial nerves. Autologous peroneal nerve grafts served as a positive control group. A total of 12 weeks following the surgical procedure, the axon number, myelinated axon number, myelin sheath thickness, and nerve conduction velocity of the rabbit and rat‑derived acellular nerve grafts were similar, whereas the fiber diameter of the rabbit‑derived acellular xenografts decreased, as compared with those of rat‑derived acellular allografts. Autografts exerted superior effects on nerve regeneration; however, no significant difference was observed between the axon number in the autograft group, as compared with the two acellular groups. These results suggested that autografts perform better than acellular nerve grafts, and chemically extracted acellular allografts and xenografts have similar effects on the regeneration of short facial nerve defects. PMID:26239906

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

    Zhang, Yanru; Zhang, Hui; Katiella, Kaka; Huang, Wenhua

    2014-01-01

    A chemically extracted acellular allogeneic nerve graft can reduce postoperative immune rejection, 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 regeneration. This study investigated the repair of injured rat sciatic nerve using a chemically extracted acellular allogeneic nerve graft com...

  13. Biomechanical properties of peripheral nerve after acellular treatment

    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.

  14. Acellular dermal matrices: Use in reconstructive and aesthetic breast surgery

    Macadam, Sheina A; Lennox, Peter A

    2012-01-01

    Acellular dermal matrices (ADMs) were first described for use in breast surgery in 2001. Since this initial report, ADMs have become an increasingly common component of implant-based breast procedures. ADMs have shown promise for use in both aesthetic and reconstructive breast surgery; however, concerns about their use remain because of the significant costs associated with these products. The present article reviews the history of ADM use in breast surgery and the outcomes reported to date. ...

  15. Infection in the Nasal Tip Caused by Acellular Dermal Matrix.

    Lee, Kun Hee

    2015-12-01

    A 19-year-old female patient visited our clinic for rhinoplasty. She complained about her low take-off point, which was apparent in profile view, and wanted slight tip projection. She refused additional cartilage harvesting from ears or ribs but consented to the use of homologous tissue, including acellular dermal matrix, for her dorsum and tip. Septoturbinoplasty was performed, and only a very small amount of septal cartilage could be harvested. It was used as both the columellar strut and the alar rim graft. Nasal dorsum and tip were augmented with acellular dermal matrix. Three months postoperatively, she experienced a few episodes of edema and redness on her nasal tip, followed by pus exudation from the nasal skin. Six months postoperatively, she underwent revision rhinoplasty for removal of inflamed grafts, and onlay tip graft with homologous rib cartilage was performed. Nasal dorsum or tip grafts are an integral part of Asian rhinoplasty. Autogenous tissue is the gold standard for grafting materials. However, the limited availability of autogenous tissue and the preference of patients and surgeons for artificial surgical implants make Asian rhinoplasty challenging. Unavailability of autogenous cartilage and patient refusal of artificial implants led to the use of acellular dermal matrix (ADM) in the nasal dorsum and tip for this case. This is the first report of postoperative complication because of infection rather than absorption after ADM use. PMID:26894006

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

    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.

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

    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. Porcine acellular lung matrix for wound healing and abdominal wall reconstruction: A pilot study.

    Fernandez-Moure, Joseph S; Van Eps, Jeffrey L; Rhudy, Jessica R; Cabrera, Fernando J; Acharya, Ghanashyam S; Tasciotti, Ennio; Sakamoto, Jason; Nichols, Joan E

    2016-01-01

    Surgical wound healing applications require bioprosthetics that promote cellular infiltration and vessel formation, metrics associated with increased mechanical strength and resistance to infection. Porcine acellular lung matrix is a novel tissue scaffold known to promote cell adherence while minimizing inflammatory reactions. In this study, we evaluate the capacity of porcine acellular lung matrix to sustain cellularization and neovascularization in a rat model of subcutaneous implantation and chronic hernia repair. We hypothesize that, compared to human acellular dermal matrix, porcine acellular lung matrix would promote greater cell infiltration and vessel formation. Following pneumonectomy, porcine lungs were processed and characterized histologically and by scanning electron microscopy to demonstrate efficacy of the decellularization. Using a rat model of subcutaneou implantation, porcine acellular lung matrices (n = 8) and human acellular dermal matrices (n = 8) were incubated in vivo for 6 weeks. To evaluate performance under mechanically stressed conditions, porcine acellular lung matrices (n = 7) and human acellular dermal matrices (n = 7) were implanted in a rat model of chronic ventral incisional hernia repair for 6 weeks. After 6 weeks, tissues were evaluated using hematoxylin and eosin and Masson's trichrome staining to quantify cell infiltration and vessel formation. Porcine acellular lung matrices were shown to be successfully decellularized. Following subcutaneous implantation, macroscopic vessel formation was evident. Porcine acellular lung matrices demonstrated sufficient incorporation and showed no evidence of mechanical failure after ventral hernia repair. Porcine acellular lung matrices demonstrated significantly greater cellular density and vessel formation when compared to human acellular dermal matrix. Vessel sizes were similar across all groups. Cell infiltration and vessel formation are well-characterized metrics of incorporation

  19. Making post-mortem implantable cardioverter defibrillator explantation safe

    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...

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

    Zhang, Yanru; Zhang, Hui; Katiella, Kaka; Huang, Wenhua

    2014-07-15

    A chemically extracted acellular allogeneic nerve graft can reduce postoperative immune rejection, 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 regeneration. 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 anastomosis, but superior to chemically extracted acellular allogeneic nerve bridging alone. PMID:25221592

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

    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. Functional bone histology of zebrafish reveals two types of endochondral ossification, different types of osteoblast clusters and a new bone type.

    Weigele, Jochen; Franz-Odendaal, Tamara A

    2016-07-01

    The zebrafish is as an important vertebrate animal model system for studying developmental processes, gene functions and signalling pathways. It is also used as a model system for the understanding of human developmental diseases including those related to the skeleton. However, surprisingly little is known about normal zebrafish skeletogenesis and osteogenesis. As in most vertebrates, it is commonly known that the bones of adult zebrafish are cellular unlike that of some other teleosts. After careful histological analyses of each zebrafish adult bone, we identified several acellular bones, with no entrapped osteocytes in addition to several cellular bones. We show that both cellular and acellular bones can even occur within the same skeletal element and transitions between these two cell types can be found. Furthermore, we describe two types of osteoblast clusters during skeletogenesis and two different types of endochondral ossification. The epiphyseal plate, for example, lacks a zone of calcification and a degradation zone with osteoblasts. A new bone type that we term tubular bone was also identified. This bone is completely filled with adipose tissue, unlike spongy bones. This study provides important insight on how osteogenesis takes place in zebrafish, and especially on the transition from cellular to acellular bones. Overall, this study leads to a deeper understanding of the functional histological composition of adult zebrafish bones. PMID:27278890

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

    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

  4. Extended Eden model reproduces growth of an acellular slime mold

    Wagner, Geri; Halvorsrud, Ragnhild; Meakin, Paul

    1999-11-01

    A stochastic growth model was used to simulate the growth of the acellular slime mold Physarum polycephalum on substrates where the nutrients were confined in separate drops. Growth of Physarum on such substrates was previously studied experimentally and found to produce a range of different growth patterns [Phys. Rev. E 57, 941 (1998)]. The model represented the aging of cluster sites and differed from the original Eden model in that the occupation probability of perimeter sites depended on the time of occupation of adjacent cluster sites. This feature led to a bias in the selection of growth directions. A moderate degree of persistence was found to be crucial to reproduce the biological growth patterns under various conditions. Persistence in growth combined quick propagation in heterogeneous environments with a high probability of locating sources of nutrients.

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

    Brittany Busse; Hakan Orbay; Sahar, David E.

    2014-01-01

    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...

  6. Chest wall reconstruction with acellular dermal matrix (Strattice™) and a TRAM flap

    Brunbjerg, Mette Eline; Juhl, Alexander Andersen; Damsgaard, Tine Engberg

    2014-01-01

    Mette Eline Brunbjerg, Alexander Andersen Juhl, Tine E. Damsgaard. "Chest wall reconstruction with acellular dermal matrix (Strattice™) and a TRAM flap.” Acta Oncol. 2013 Jun;52(5):1052-4. Epub 2012 Oct 24. PMID: 23095144......Mette Eline Brunbjerg, Alexander Andersen Juhl, Tine E. Damsgaard. "Chest wall reconstruction with acellular dermal matrix (Strattice™) and a TRAM flap.” Acta Oncol. 2013 Jun;52(5):1052-4. Epub 2012 Oct 24. PMID: 23095144...

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

    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.

  8. DermACELL: a novel and biocompatible acellular dermal matrix in tissue expander and implant-based breast reconstruction

    Bullocks, Jamal M.

    2014-01-01

    Background Acellular dermal matrices present a new alternative to supporting expanders and implants for breast reconstruction in breast cancer patients following mastectomy. However, some studies have suggested that acellular dermal matrix may increase the complication rates in certain clinical settings. DermACELL acellular dermal matrix offers advanced processing in order to attempt to decrease bio-intolerance and complications. Methods Ten consecutive patients that presented for breast reco...

  9. Complications Following Expander/Implant Breast Reconstruction Utilizing Acellular Dermal Matrix: A Systematic Review and Meta-Analysis

    Hoppe, Ian C.; Yueh, Janet H.; Wei, Cindy H.; Ahuja, Naveen K.; Patel, Priti P.; Datiashvili, Ramazi O.

    2011-01-01

    Background: The recent increase in popularity of acellular dermal matrix assistance in immediate expander/implant breast reconstruction has led to variety of viewpoints. Many studies are published indicating an increase in complications with the use of acellular dermal matrix, while others indicate there is no increase in complications. Methods: This meta-analysis utilizes information from available studies that directly compare one specific type of acellular dermal matrix with traditional me...

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

    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. PMID:20700752

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

    Ken eYe

    2016-01-01

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

  12. Chondrogenesis of Human Infrapatellar Fat Pad Stem Cells on Acellular Dermal Matrix.

    Ye, Ken; Traianedes, Kathy; Choong, Peter F M; Myers, Damian E

    2016-01-01

    Acellular dermal matrix (ADM) has been in clinical use for decades in numerous surgical applications. The ability for ADM to promote cellular repopulation, revascularisation and tissue regeneration is well documented. Adipose stem cells have the ability to differentiate into mesenchymal tissue types, including bone and cartilage. The aim of this study was to investigate the potential interaction between ADM and adipose stem cells in vitro using TGFβ3 and BMP6. Human infrapatellar fat pad-derived adipose stem cells (IPFP-ASC) were cultured with ADM derived from rat dermis in chondrogenic (TGFβ3 and BMP6) medium in vitro for 2 and 4 weeks. Histology, qPCR, and immunohistochemistry were performed to assess for markers of chondrogenesis (collagen Type II, SOX9 and proteoglycans). At 4 weeks, cell-scaffold constructs displayed cellular changes consistent with chondrogenesis, with evidence of stratification of cell layers and development of a hyaline-like cartilage layer superficially, which stained positively for collagen Type II and proteoglycans. Significant cell-matrix interaction was seen between the cartilage layer and the ADM itself with seamless integration between each layer. Real time qPCR showed significantly increased COL2A1, SOX9, and ACAN gene expression over 4 weeks when compared to control. COL1A2 gene expression remained unchanged over 4 weeks. We believe that the principles that make ADM versatile and successful for tissue regeneration are applicable to cartilage regeneration. This study demonstrates in vitro the ability for IPFP-ASCs to undergo chondrogenesis, infiltrate, and interact with ADM. These outcomes serve as a platform for in vivo modelling of ADM for cartilage repair. PMID:26858950

  13. Blood Vessel-Derived Acellular Matrix for Vascular Graft Application

    Luigi Dall’Olmo

    2014-01-01

    Full Text Available To overcome the issues connected to the use of autologous vascular grafts and artificial materials for reconstruction of small diameter (<6 mm blood vessels, this study aimed to develop acellular matrix- (AM- based vascular grafts. Rat iliac arteries were decellularized by a detergent-enzymatic treatment, whereas endothelial cells (ECs were obtained through enzymatic digestion of rat skin followed by immunomagnetic separation of CD31-positive cells. Sixteen female Lewis rats (8 weeks old received only AM or previously in vitro reendothelialized AM as abdominal aorta interposition grafts (about 1 cm. The detergent-enzymatic treatment completely removed the cellular part of vessels and both MHC class I and class II antigens. One month after surgery, the luminal surface of implanted AMs was partially covered by ECs and several platelets adhered in the areas lacking cell coverage. Intimal hyperplasia, already detected after 1 month, increased at 3 months. On the contrary, all grafts composed by AM and ECs were completely covered at 1 month and their structure was similar to that of native vessels at 3 months. Taken together, our findings show that prostheses composed of AM preseeded with ECs could be a promising approach for the replacement of blood vessels.

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

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

    2007-09-15

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

  15. Explant exenisation for tissue culture in marine macroalgae

    Liu, Xuewu; Kloareg, Bernard

    1992-09-01

    Unialgal explants from Laminaria digitata, and from a variety of red algae, were obtained by hand removing the visible epiphytes, and stirring the tissue in the presence of glass beads. Two antibiotic mixtures were found to be efficient in removing the contaminating fungi and bacteria from the algae. The procedure proved suitable as a primary step in the tissue culture of the investigated species.

  16. Thidiazuron-induced high-frequency plant regeneration from leaf explants of Paulownia tomentosa mature trees

    CORREDOIRA, E.; Ballester, A.; Viéitez Martín, Ana María

    2008-01-01

    Attempts were made to study the effect of thidiazuron (TDZ) on adventitious shoot induction and plant development in Paulownia tomentosa explants derived from mature trees. Media with different concentrations of TDZ in combination with an auxin were used to induce adventitious shoot-buds in two explant types: basal leaf halves with the petiole attached (leaf explant) and intact petioles. Optimal shoot regeneration was obtained in leaf explants cultured on induction medium containing TDZ (22.7...

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

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

    2015-01-01

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

  18. Acellular comet assay: A tool for assessing variables influencing the alkaline comet assay

    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 60Co 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 60Co 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. (authors)

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

    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

  20. Bone Tissue Engineering with Multilayered Scaffolds-Part II: Combining Vascularization with Bone Formation in Critical-Sized Bone Defect.

    Sathy, Binulal Nelson; Watson, Brendan M; Kinard, Lucas A; Spicer, Patrick P; Dahlin, Rebecca L; Mikos, Antonios G; Nair, Shantikumar

    2015-10-01

    Our previous in vivo study showed that multilayered scaffolds made of an angiogenic layer embedded between an osteogenic layer and an osteoconductive layer, with layer thickness in the 100-400 μm range, resulted in through-the-thickness vascularization of the construct even in the absence of exogenous endothelial cells. The angiogenic layer was a collagen-fibronectin gel, and the osteogenic layer was made from nanofibrous polycaprolactone while the osteoconductive layer was made either from microporous hydroxyapatite or microfibrous polycaprolactone. In this follow-up study, we implanted these acellular and cellular multilayered constructs in critical-sized rat calvarial defects and evaluated their vascularization and bone formation potential. Vascularization and bone formation at the defect were evaluated and quantified using microcomputed tomography (microCT) followed by perfusion of the animals with the radio opaque contrast agent, MICROFIL. The extent of bony bridging and union within the critical-sized defect was evaluated using a previously established scoring system from the microCT data set. Similarly the new bone formation in the defect was quantified from the microCT data set as previously reported. Histological evaluation at 4 and 12 weeks validated the microCT findings. Our experimental results showed that acellular multilayered scaffolds with microscale-thick nanofibers and porous ceramic discs with angiogenic zone at their interface can regenerate functional vasculature and bone similar to that of cellular constructs in critical-sized calvarial defects. This result suggests that suitably bioengineered acellular multilayered constructs can be an improved and more translational approach in functional in vivo bone regeneration. PMID:26262560

  1. Method for detection of a suspect viral deoxyribonucleic acid in an acellular biological fluid

    A method for evaluating an acellular biological fluid for the presence of a suspect viral DNA, such as DNA of the Hepatitis-B virus, is described. The acellular biological fluid is treated to immobilize in denatured form the DNAs including the suspect viral DNA on a solid substrate. This substrate is contacted with a solution including radioisotopically-labelled suspect viral denatured DNA to renature the immobilized suspect viral native DNA. The solid substrate is then evaluated for radioisotopically-labelled suspect viral renatured DNA. (author)

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

    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.

  3. Laboratory analyses of two explanted hydrophobic acrylic intraocular lenses

    Yunhai Dai

    2014-01-01

    Full Text Available Two three-piece hydrophobic acrylic intraocular lenses (IOLs were explanted from two patients at 7 and 9 years, respectively, after implantation, because of poor fundus visualisation and/or a clinically significant decrease in visual acuity related to their opacified IOLs. In addition to light microscopy, scanning electron microscopy and energy dispersive X-ray spectroscopy, confocal laser scanning microscopy was used for the first time to observe the explanted IOLs. The clinical aspect seemed to correspond to the phenomenon of surface light scattering, while laboratory analyses showed dense glistenings in the central layer of the IOL optic, which had no change next to the surface. Further studies on these phenomena are needed.

  4. Bovine explant model of degeneration of the intervertebral disc

    Sivan Sarit; Menage Janis; Roberts Sally; Urban Jill PG

    2008-01-01

    Abstract Background Many new treatments for degeneration of the intervertebral disc are being developed which can be delivered through a needle. These require testing in model systems before being used in human patients. Unfortunately, because of differences in anatomy, there are no ideal animal models of disc degeneration. Bovine explant model systems have many advantages but it is not possible to inject any significant volume into an intact disc. Therefore we have attempted to mimic disc de...

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

    Uitterlinden EJ; Koevoet JLM; Verkoelen CF; Bierma-Zeinstra SMA; Jahr H; Weinans H; Verhaar JAN; GJVM van Osch

    2008-01-01

    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 s...

  6. Molecular weight characterization of virgin and explanted polyester arterial prostheses.

    Maarek, J M; Guidoin, R; Aubin, M; Prud'homme, R E

    1984-10-01

    The macromolecular properties of 17 virgin commercial arterial prostheses and a series of explanted prostheses, both manufactured from poly(ethylene terephthalate) (PET) yarns, have been studied by gel permeation chromatography (GPC) and by differential scanning calorimetry (DSC). Only small differences were found between the average molecular weights and the degree of crystallinity of the unused reference grafts. A broadening of the DSC curves was observed for the prostheses containing texturized yarns compared with those made solely from flat, untexturized yarns. This broadening may be due to greater heterogeneity of the crystal sizes caused by the texturizing process and to the use of two or more different yarns with dissimilar thermal histories in the same prosthesis. Average molecular weights of the explant series were significantly lower than those of the corresponding reference grafts but almost time independent. The polydispersity index and the degree of crystallinity of the explants remained constant as a function of time. These results are discussed in regard to others available in the literature. PMID:6242474

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

    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. Repair of a Gingival Fenestration Using an Acellular Dermal Matrix Allograft.

    Breault, Lawrence G; Brentson, Raquel C; Fowler, Edward B; Bisch, Frederick C

    2016-01-01

    A case report illustrating the successful treatment of a gingival fenestration with an acellular dermal matrix (ADM) allograft. After 2½ months of healing, the ADM was completely integrated into the soft tissues of the mandibular anterior gingiva with complete resolution of the gingival fenestration, resulting in excellent gingival esthetics. PMID:26874103

  9. Expanding the applications of Cadaveric skin - the properties and uses of an acellular dermal matrix

    The ability to transplant organs and tissues has been one of the most significant advances of modern medicine. The availability of cadaveric allograft skin has greatly facilitated the practice of aggressive, early excision of massive burn injuries. Due to its ultimate rejection however, the role of allograft skin has historically been limited to that of a temporary wound dressing. Development of an acellular dermal allograft has greatly expanded the applications for donated human skin. AlloDerm(r) preserved dermal graft (LifeCell, The Woodlands, TX) is prepared via ionic separation of allograft skin followed by detergent removal of antigenic cells. Acellular dermal grafts are then cryoprotected and freeze-dried. The process maintains the structural integrity of the extracellular matrix and preserves the biochemical composition of the basement membrane. The resultant immunologically inert allograft can be used in a variety of applications. In burn injuries, lack of an adequate dermal component at either the donor or wound site may result in complications including contraction, delayed healing, hypertrophic scarring and keloid formation. Utilizing allogenic dermis eliminates the need for autologous dermis at the wound site and minimizes donor site trauma by allowing procurement of ultra-thin (0.006) autografts. Expanding the scope of traditional uses for allograft skin, acellular dermal grafts have been successfully utilized in a variety of procedures including duraplasty, orbital reconstruction, and hemia repar. In periodontal surgery, allograft tissue eliminates the need for painful palatal autografts and has been used to increase attached gingiva and reduce gingival recession. Resorption of autologous grafts or extrusion of synthetic material often hampers repair or reconstruction of soft tissue deficits. Transplantation of acellular allograft dermis provides a biochemically and structurally intact matrix, which persists and is ultimately repopulated with

  10. PATTERN OF PLANT REGENERATION FROM SHOOT TIP EXPLANTS OF PIGEONPEA (CAJANUS CAJAN L MILLLSP) VAR LRG-41

    T. Raghavendra; P. Sudhakar

    2014-01-01

    An efficient direct shoot bud differentiation and multiple shoot induction from shoot tip explants of pigeon pea (Cajanus cajan L.) has been achieved. The frequency of shoot bud regeneration was influenced by the type of explants, genotype and concentrations of cytokinin. Explants viz. shoot tip isolated from 10 day old seedlings showed better explants response Explants were cultured on Murashige and skoog (MS) medium augmented with different concentrations of BAP and NAA. Among the various c...

  11. Mucus glycoprotein secretion by tracheal explants: effects of pollutants

    Tracheal slices incubated with radioactive precursors in tissue culture medium secrete labeled mucus glycoproteins into the culture medium. We have used an in vivtro approach, a combined method utilizing exposure to pneumotoxins in vivo coupled with quantitation of mucus secretion rates in vitro, to study the effects of inhaled pollutants on mucus biosynthesis by rat airways. In addition, we have purified the mucus glycoproteins secreted by rat tracheal explants in order to determine putative structural changes that might by the basis for the observed augmented secretion rates after exposure of rats to H2SO4 aerosols in combination with high ambient levels of ozone. After digestion with papain, mucus glycoproteins secreted by tracheal explants may be separated into five fractions by ion-exchange chromatography, with recovery in high yield, on columns of DEAE-cellulose. Each of these five fractions, one neutral and four acidic, migrates as a single unique spot upon cellulose acetate electrophoresis at pH values of 8.6 and 1.2. The neutral fraction, which is labeled with [3H] glucosamine, does not contain radioactivity when Na2 35SO4 is used as the precursor. Acidic fractions I to IV are all labeled with either 3H-glucosamine or Na2 35SO4 as precursor. Acidic fraction II contains sialic acid as the terminal sugar on its oligosaccharide side chains, based upon its chromatographic behavior on columns of wheat-germ agglutinin-Agarose. Treatment of this fraction with neuraminidase shifts its elution position in the gradient to a lower salt concentration, coincident with acidic fraction I. After removal of terminal sialic acid residues with either neuraminidase or low pH treatment, the resultant terminal sugar on the oligosaccharide side chains is fucose. These results are identical with those observed with mucus glycoproteins secreted by cultured human tracheal explants and purified by these same techniques

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

    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. PMID:26025258

  13. Histology of Somatic Embryogenesis in Cultured Leaf Explants of Pistachio (Pistacia veraL)

    ONAY, Ahmet

    2000-01-01

    The histology of somatic embryo initiation and development in pistachio ( Pistacia were L.) embryogenic masses (EMS) derived from leaf explants was examined using light microscopy. Explants with somatic embryos at different developmental stages were fixed for histological examination, cut into 10 µm thick sections, stained with hematoxylin-eosin and observed microscopically. The histological examination showed that the two types of cell clusters induced originated from explants and were mo...

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

    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.

  15. Chlorophyll Fluorescence as a Tool to Assess the Regeneration Potential of African Violet Leaf Explants

    Norbert KEUTGEN

    2016-06-01

    Full Text Available Micropropagation of many ornamentals has enabled their efficient commercialisation and many problems have been solved by the elaboration of adequate culture protocols. Nevertheless, a non-destructive technique for monitoring the developmental progress of explants would be desirable. The present study focussed on the applicability of chlorophyll fluorescence in leaf explants of African violet (a Saintpaulia ionantha × confusa – hybrid explanted onto Murashige and Skoog basic medium. The explants that survived on the medium without additional phytohormones had the capacity to develop further into two different kinds of explants: light green explants, characterized by a non-regular size growth and stiffer appearance, and dark green explants capable of organogenesis. Compared to the source leaves of African violet plants, explants were characterized by reduced chlorophyll (Chl and carotenoid (Car contents as well as a tendency towards a higher Car/Chl ratio. The Chl a/b ratio decreased significantly in the light green explants. A reduction of maximum quantum efficiency of photosystem II (Fv/Fm accompanied by a high percentage (> 50% of thermal energy dissipation as a consequence of an elevated light intensity (800 µmol m-2 s-1 quanta indicated photoinhibition in the light green explants, whereas in the dark green explants the largest percentage (> 50% of the light energy was dissipated into the fraction of photon energy trapped by ‘closed’ photosystem II reaction centres. These results suggest that the capacity of organogenesis of leaf explants of African violet can be monitored using chlorophyll fluorescence, because it is related to modifications of the photosynthetic system.

  16. Florid pustular dermatitis of breast: A case report on a unusual complication from acellular dermal matrix use

    Justin James; Lee Jackson; Christobel Saunders

    2016-01-01

    Introduction: Idiopathic erythematous reaction of the breast (Red breast syndrome) is a known complication following breast reconstruction with acellular dermal matrix. However pustular dermatitis like presentation is not previously known. Presentation of case: We present a 42-year-old lady who developed bilateral pustular dermatitis like appearance following breast reconstruction with acellular dermal matrix slings. Though surgical washout was done, both expanders and flex HD could be pre...

  17. Prosthetic Breast Reconstruction With Acellular Dermal Matrices: Achieving Predictability and Reproducibility.

    Nahabedian, Maurice Y

    2016-05-01

    The use of acellular dermal matrices in the setting of prosthetic breast reconstruction has captured the attention of many plastic surgeons. The regenerative capacity of these materials has provided additional tissue support to the mastectomy skin flaps with the ultimate result of improving surgical and aesthetic outcomes. Despite the benefits, there remains a significant diversity with regard to outcomes with some surgeons reporting increased morbidity. The reasons for this are varied but ultimately related to differences in patient selection and surgical techniques. The purpose of this article is to provide strategies for using acellular dermal matrix to achieve success in a manner that is usually associated with outcomes that are predictable and reproducible. PMID:27579223

  18. DermACELL: Human Acellular Dermal Matrix Allograft A Case Report.

    Cole, Windy E

    2016-03-01

    Diabetes often causes ulcers on the feet of diabetic patients. A 56-year-old, insulin-dependent, diabetic woman presented to the wound care center with a Wagner grade 3 ulcer of the right heel. She reported a 3-week history of ulceration with moderate drainage and odor and had a history of ulceration and osteomyelitis in the contralateral limb. Rigorous wound care, including hospitalization; surgical incision and drainage; intravenous antibiotic drug therapy; vacuum-assisted therapy; and a new room temperature, sterile, human acellular dermal matrix graft were used to heal the wound, save her limb, and restore her activities of daily living. This case presentation involves alternative treatment of a diabetic foot ulcer with this new acellular dermal matrix, DermACELL. PMID:27031550

  19. Pioneering technique using Acellular Dermal Matrix in the rescue of a radiation ulcer

    NASEEM, S.; PATEL, A.D.; DEVALIA, H.

    2016-01-01

    Background Radiotherapy as an adjuvant to mastectomy is integral to the treatment of breast cancer, but can result in skin ulceration. Skin ulceration following radiotherapy is traditionally managed by removing the implant and allowing the skin to heal by secondary intention. Case report A 42-year-old woman underwent radiotherapy following a breast reconstruction. She developed a 2 x 3cm radiation ulcer. The ulcer was managed by removing the implant and performing capsulectomy. A Beckers 50 expander was placed and reinforced with acellular dermal matrix inferolaterally. At follow-up the patient had a good cosmetic outcome. Conclusion Post-radiation skin ulcers present a challenge to treat with no current standardised management. The use of acellular dermal matrix may present a new technique to promote healing in these testing cases. PMID:27142826

  20. Proliferation of Female Inflorescences explants of Date Palm

    This study was conducted to determine the effect of Abscisic acid (ABA) and Ancymidol on proliferation of female inflorescences explants of date palm. In the first experiment two lengths of spath at (5-7 cm) or at (7-10 cm) were cultured on nutrient media which consists of half macro and full micro salts of MS medium supplemented with gradual decreasing in concentration of Abscisic acid (ABA) and Ancymidol from 4.5, 3.0, 1.5 to 0.5 mg-1. In the second experiment two phases of nutrient medium (solid and liquid) and two source of carbon were investigated. Gradual decreasing of ABA concentrations from 4.5 mg-1 to 1.5 mg-1 in culture medium, stimulated the production of direct somatic embryos and accelerated callus initiation, but at last decrement (0.5 mg-1) of Ancymidol concentration few embryos were produced. Callus initiation from inflorescences explants gave high production and well development of somatic embryos when cultured on liquid medium supplemented with 40 g-1 sucrose. All direct or indirect somatic embryos obtained in these experiments were converted successfully to healthy normal plantlets which could be transferred to acclimatization stage.

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

    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.

  2. Bovine explant model of degeneration of the intervertebral disc

    Sivan Sarit

    2008-02-01

    Full Text Available Abstract Background Many new treatments for degeneration of the intervertebral disc are being developed which can be delivered through a needle. These require testing in model systems before being used in human patients. Unfortunately, because of differences in anatomy, there are no ideal animal models of disc degeneration. Bovine explant model systems have many advantages but it is not possible to inject any significant volume into an intact disc. Therefore we have attempted to mimic disc degeneration in an explant bovine model via enzymatic digestion. Methods Bovine coccygeal discs were incubated with different concentrations of the proteolytic enzymes, trypsin and papain, and maintained in culture for up to 3 weeks. A radio-opaque solution was injected to visualise cavities generated. Degenerative features were monitored histologically and biochemically (water and glycosaminoglycan content, via dimethylmethylene blue. Results and Conclusion The central region of both papain and trypsin treated discs was macro- and microscopically fragmented, with severe loss of metachromasia. The integrity of the surrounding tissue was mostly in tact with cells in the outer annulus appearing viable. Biochemical analysis demonstrated greatly reduced glycosaminoglycan content in these compared to untreated discs. We have shown that bovine coccygeal discs, treated with proteolytic enzymes can provide a useful in vitro model system for developing and testing potential new treatments of disc degeneration, such as injectable implants or biological therapies.

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

    Schleifenbaum, Stefan; Prietzel, Torsten; Aust, Gabriela; Boldt, Andreas; Fritsch, Sebastian; Keil, Isabel; Koch, Holger; Möbius, Robert; Scheidt, Holger A.; Wagner, Martin F. X.; Hammer, Niels

    2016-01-01

    Introduction 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. Materials and Methods 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 s...

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

    Rose, Jessica F.; Sarosh N. Zafar; Ellsworth IV, Warren A.

    2016-01-01

    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 an...

  5. Tissue Engineering of Injectable Soft tissue Filler: Using Adipose Stem Cells and Micronized Acellular Dermal Matrix

    Yoo, Gyeol; Lim, Jin Soo

    2009-01-01

    In this study of a developed soft tissue filler, adipose tissue equivalents were constructed using adipose stem cells (ASCs) and micronized acellular dermal matrix (Alloderm). After labeling cultured human ASCs with fluorescent green protein and attaching them to micronized Alloderm (5×105 cells/1 mg), ASC-Alloderm complexes were cultured in adipogenic differentiation media for 14 days and then injected into the dorsal cranial region of nude male mice. The viabilities of ASCs in micronized Al...

  6. Combination of Acellular Nerve Graft and Schwann Cells-Like Cells for Rat Sciatic Nerve Regeneration

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

    2014-01-01

    Objective. To investigate the effect of tissue engineering nerve on repair of rat sciatic nerve defect. Methods. Forty-five rats with defective sciatic nerve were randomly divided into three groups. Rats in group A were repaired by acellular nerve grafts only. Rats in group B were repaired by tissue engineering nerve. In group C, rats were repaired by autogenous nerve grafts. After six and twelve weeks, sciatic nerve functional index (SFI), neural electrophysiology (NEP), histological and tra...

  7. Xenogenic (porcine) acellular dermal matrix is useful for the wound healing of severely damaged extremities

    Zhang, Zhaoxin; Lv, Lei; MAMAT, MASUT; Chen, Zhao; Liu, Lihua; Wang, Zhizhong

    2014-01-01

    This study was conducted to investigate the possibility of improving the success rate of patient treatment and promoting wound healing by utilizing xenogenic (porcine) acellular dermal matrix (XADM) to cover large areas of severely damaged wounds. Patients with severely damaged large-area wounds (56 cases) were enrolled in the study from May 2002 to May 2012. All patients admitted to hospital received a rapid infusion via intravenous access to maintain an effective circulating blood volume an...

  8. Neoinnervation and neovascularization of acellular pericardial-derived scaffolds in myocardial infarcts

    Gálvez-Montón, Carolina; Fernandez-Figueras, M Teresa; Martí, Mercè; Soler-Botija, Carolina; Roura, Santiago; Perea-Gil, Isaac; Prat-Vidal, Cristina; Llucià-Valldeperas, Aida; Raya, Ángel; Bayes-Genis, Antoni

    2015-01-01

    Engineered bioimplants for cardiac repair require functional vascularization and innervation for proper integration with the surrounding myocardium. The aim of this work was to study nerve sprouting and neovascularization in an acellular pericardial-derived scaffold used as a myocardial bioimplant. To this end, 17 swine were submitted to a myocardial infarction followed by implantation of a decellularized human pericardial-derived scaffold. After 30 days, animals were sacrificed and hearts we...

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

    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...

  10. Design and Synthesis of an Artificial Pulmonary Pleura for High Throughput Studies in Acellular Human Lungs

    Wagner, Darcy E; Fenn, Spencer L.; Bonenfant, Nicholas R.; Marks, Elliot R.; Borg, Zachary; Saunders, Patrick; Oldinski, Rachael A.; Weiss, Daniel J

    2014-01-01

    Whole organ decellularization of complex organs, such as lungs, presents a unique opportunity for use of acellular scaffolds for ex vivo tissue engineering or for studying cell-extracellular matrix interactions ex vivo. A growing body of literature investigating decellularizing and recellularizing rodent lungs has provided important proof of concept models and rodent lungs are readily available for high throughput studies. In contrast, comparable progress in large animal and human lungs has b...

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

    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

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

    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

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

    Ahmed M. S. Ibrahim, MD; Pieter G. L. Koolen, MD; Azra A. Ashraf, MD, MPH; Kuylhee Kim, MD; Marc A. M. Mureau, MD, PhD; Bernard T. Lee, MD, MBA; Samuel J. Lin, MD

    2015-01-01

    Background: Acellular dermal matrices (ADMs) in plastic surgery have become increasingly popular particularly for breast reconstruction. Despite their advantages, questions exist regarding their association with a possible increased incidence of complications. We describe a collective experience of plastic surgeons’ use of ADMs in reconstructive breast surgery using an internet-based survey. Methods: Members of the American Society of Plastic Surgeons were recruited through voluntary, anonymo...

  14. Perceptions of Tetanus-diphteria-acellular pertussis (Tdap) Vaccination among Korean Women of Childbearing Age

    Kim, In Seon; Seo, Yu Bin; Hong, Kyung-Wook; Noh, Ji Yun; Choi, Won Suk; Song, Joon Young; Cho, Geum Joon; Oh, Min Jeong; Kim, Hai Joong; Hong, Soon Choul; Sohn, Jang Wook; Kim, Woo Joo; Cheong, Hee Jin

    2013-01-01

    Background The number of cases of pertussis reported has increased gradually in the last decade. Pertussis vaccination is the most effective strategy for the prevention of infection. Despite the fact that young infants are at the highest risk for pertussis, the rate of tetanus-diphtheria-acellular pertussis (Tdap) vaccination is presumed to be very low among women of childbearing age in Korea. The purpose of this study was to investigate the perceptions of women of childbearing age regarding ...

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

    Latty, Tanya; Beekman, Madeleine

    2010-01-01

    Speed–accuracy trade-offs (SATs) are thought to be a fundamental feature of biological information processing, yet most evidence of SATs comes from animals. Here, we examine SATs in the foraging decisions of an acellular, amoeboid organism: the slime mould Physarum polycephalum. Slime moulds were given a simple discrimination task: selecting the highest-quality food item from a set of three options. We investigated the effect of two stressors, light exposure and hunger, on the speed and accur...

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

    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.

  17. Acute Regeneration and Chronic Acellular Transformation of Rabbit Cryopreserved Aortic Allografts

    An analysis of rabbit cryopreserved aortic allografts excised on postoperative days (POD) 2, 5, 11, 60, 210, 360, and 720, as well as controls that were untransplanted native aortas and cryopreserved aortas, was performed. On POD2, the number of medial smooth muscle cells in the allografts was reduced to approximately 50%. Ki-67 analysis revealed that medial smooth muscle cells in the allografts proliferated from the 2nd day. By the 11th day, their proliferation ceased and the number of medial smooth muscle cells was restored to almost at the same level as in the controls. Polymorphic microsatellite DNA marker analysis disclosed that the restored medial smooth muscle cells were of donor origin. From 7 months through 2 years, the media of cryopreserved aortic allografts were transformed into acellular structures, in which the elastic fibers were preserved. On the other hand, newly accumulated smooth muscle cells were observed in the adventitia just outside of acellular media after 7 months. In some cases, scattered lamellar calcium deposition was observed in the same regions. This study presents a comprehensive documentation of regeneration and acellular transformation in cryopreserved aortic allografts based on short and long-term analysis

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

    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.

  19. Bone tumor

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

  20. Bone Grafts

    A bone graft transplants bone tissue. Surgeons use bone grafts to repair and rebuild diseased bones in your hips, knees, spine, and sometimes other bones and joints. Grafts can also repair bone loss caused by some ...

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

    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.

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

    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.

  3. Methylation of inorganic arsenic by murine fetal tissue explants.

    Broka, Derrick; Ditzel, Eric; Quach, Stephanie; Camenisch, Todd D

    2016-07-01

    Although it is generally believed that the developing fetus is principally exposed to inorganic arsenic and the methylated metabolites from the maternal metabolism of arsenic, little is known about whether the developing embryo can autonomously metabolize arsenic. This study investigates inorganic arsenic methylation by murine embryonic organ cultures of the heart, lung, and liver. mRNA for AS3mt, the gene responsible for methylation of arsenic, was detected in all embryonic tissue types studied. In addition, methylated arsenic metabolites were generated by all three tissue types. The fetal liver explants yielded the most methylated arsenic metabolites (∼7% of total arsenic/48 h incubation) while the heart, and lung preparations produced slightly greater than 2% methylated metabolites. With all tissues the methylation proceeded mostly to the dimethylated arsenic species. This has profound implications for understanding arsenic-induced fetal toxicity, particularly if the methylated metabolites are produced autonomously by embryonic tissues. PMID:26446802

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

    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.

  5. Human Acellular Dermis versus Submuscular Tissue Expander Breast Reconstruction: A Multivariate Analysis of Short-Term Complications

    Armando A. Davila

    2013-01-01

    Full Text Available Background Acellular dermal matrix (ADM allografts and their putative benefits have beenincreasingly described in prosthesis based breast reconstruction. There have been a myriadof analyses outlining ADM complication profiles, but few large-scale, multi-institutionalstudies exploring these outcomes. In this study, complication rates of acellular dermis-assistedtissue expander breast reconstruction were compared with traditional submuscular methodsby evaluation of the American College of Surgeon’s National Surgical Quality ImprovementProgram (NSQIP registry.Methods Patients who underwent immediate tissue expander breast reconstruction from2006-2010 were identified using surgical procedure codes. Two hundred forty tracked variablesfrom over 250 participating sites were extracted for patients undergoing acellular dermisassistedversus submuscular tissue expander reconstruction. Thirty-day postoperative outcomesand captured risk factors for complications were compared between the two groups.Results A total of 9,159 patients underwent tissue expander breast reconstruction; 1,717using acellular dermis and 7,442 with submuscular expander placement. Total complicationsand reconstruction related complications were similar in both cohorts (5.5% vs. 5.3%, P=0.68and 4.7% vs. 4.3%, P=0.39, respectively. Multivariate logistic regression revealed body massindex and smoking as independent risk factors for reconstructive complications in both cohorts(P<0.01.Conclusions The NSQIP database provides large-scale, multi-institutional, independentoutcomes for acellular dermis and submuscular breast reconstruction. Both thirty-daycomplication profiles and risk factors for post operative morbidity are similar between thesetwo reconstructive approaches.

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

    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.

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

    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.

  8. Bone Biopsy

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

  9. Constructing Human Skin Equivalents on Porcine Acellular Peritoneum Extracellular Matrix for In Vitro Irritation Testing.

    Tsai, Pei-Chin; Zhang, Zheng; Florek, Charles; Michniak-Kohn, Bozena B

    2016-01-01

    The irritancy of topical products has to be investigated to ensure the safety and compliance. Although several reconstructed human epidermal models have been adopted by the Organization for Economic Cooperation and Development (OECD) to replace in vivo animal irritation testing, these models are based on a single cell type and lack dermal components, which may be insufficient to reflect all of the components of irritation. In our study, we investigated the use of acellular porcine peritoneum extracellular matrix as a substrate to construct full-thickness human skin equivalents (HSEs) for use as irritation screening tool. The acellular peritoneum matrix (APM) exhibited excellent skin cell attachment (>80%) and proliferation for human dermal fibroblasts (HDF) and immortalized human keratinocytes (HaCaT). APM-HSEs based on coculture of HDF and HaCaT were prepared. Increased HDF seeding density up to 5 × 10(4)/cm(2) resulted in APM-HSEs with a thicker and more organized epidermis. The epidermis of APM-HSEs expressed keratin 15, a keratinocyte proliferation marker, and involucrin, a differentiation marker, respectively. To assess the use of APM-HSEs for irritation testing, six proficiency chemicals, including three nonirritants (phosphate-buffered saline, polyethylene glycol 400, and isopropanol) and three irritants (1-bromohexane, heptanol, and sodium dodecyl sulfate) were applied. The APM-HSEs were able to discriminate nonirritants from irritants based on the viability. Levels of cytokines (interleukin [IL]-1α, IL-1ra, IL-6, IL-8, and granulocyte macrophage colony-stimulating factor [GM-CSF]) in these treatment groups further assisted the irritancy ranking. In conclusion, we have developed partially differentiated full-thickness APM-HSEs based on acellular porcine peritoneum matrix, and these APM-HSEs demonstrated utility as an in vitro irritation screening tool. PMID:26415037

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

    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.

  11. Preparation and characterization of genipin-crosslinked rat acellular spinal cord scaffolds

    Jiang, Tao [Department of Orthopedics, Xinqiao Hospital, The Third Military Medical University, Chongqing (China); Ren, Xian-Jun, E-mail: ren_xianjun@sina.com [Department of Orthopedics, Xinqiao Hospital, The Third Military Medical University, Chongqing (China); Tang, Jin-Liang [Department of Pathology, Xinqiao Hospital, The Third Military Medical University, Chongqing (China); Yin, Hong; Wang, Kai-Jian; Zhou, Chang-Li [Department of Orthopedics, Xinqiao Hospital, The Third Military Medical University, Chongqing (China)

    2013-08-01

    The feasibility of rat acellular spinal cord scaffolds for tissue engineering applications was investigated. Fresh rat spinal cords were decellularized and crosslinked with genipin (GP) to improve their structural stability and mechanical properties. The GP-crosslinked spinal cord scaffolds possessed a porous structure with an average pore diameter of 31.1 μm and a porosity of 81.5%. The resultant scaffolds exhibited a water uptake ratio of 229%, and moderate in vitro degradation rates of less than 5% in phosphate-buffered saline (PBS) and slightly more than 20% in trypsin-containing buffer, within 14 days. The ultimate tensile strength and elastic modulus of GP-crosslinked spinal cord scaffolds were determined to be 0.193 ± 0.064 MPa and 1.541 ± 0.082 MPa, respectively. Compared with glutaraldehyde (GA)-crosslinked acellular spinal cord scaffolds, GP-crosslinked scaffolds demonstrated similar microstructure and mechanical properties but superior biocompatibility as indicated by cytotoxicity evaluation and rat mesenchymal stem cell (MSC) adhesion behavior. Cells were able to penetrate throughout the crosslinked scaffold due to the presence of an interconnected porous structure. The low cytotoxicity of GP facilitated cell proliferation and extracellular matrix (ECM) secretion in vitro on the crosslinked scaffolds over 7 days. Thus, these GP-crosslinked spinal cord scaffolds show great promise for tissue engineering applications. - Highlights: • We prepared a modified acellular spinal cord scaffold by crosslinking with genipin. • Genipin-crosslinked scaffold possessed a good 3-dimension porous structure. • Structural stability and mechanical property of scaffold were enhanced by genipin-crosslinking. • Genipin-crosslinked scaffold demonstrated superior biocompatibility. • Genipin-crosslinked scaffold show great promise for tissue engineering applications.

  12. Life Cycle of Heterodera zeae Koshy, Swarup, and Sethi on Zea mays L. Axenic Root Explants

    Lauritis, J. A.; Rebois, R. V.; Graney, L. S.

    1983-01-01

    Monoxenic cultures of Heterodera zeae, the corn cyst nematode (CCN), were established on root explants of corn Zea mays L., cv. Kenworthy. The life cycle of H. zeae was determined from light anti scanning electron microscopic observations of the root explants grown in the dark at 29.5 ± .5 C under gnotobiotic conditions. The life cycle, from the time the explants were inoculated with second-stage larvae (L2) to the first appearance of newly hatched second-generation L2, required 22 days. The ...

  13. Polyphenol Oxidase (PPO) in Early Stage of Browning of Phalaenopsis Leaf Explants

    Zhiwei Ru; Yanyan Lai; Chuangjun Xu; Ling Li

    2013-01-01

    Explant browning is one of the most common problems during the early stage of tissue culture. The aim of this study was to explore the roles of polyphenol oxidase (PPO, EC 1.10.3.2) in the browning process of leaf explants of Phalaenopsis (Doritaenopsis Queen Bee “Red Sky”) under a routine culture condition. Along with the observation of the time course of browning and PPO distribution, PPO gene expression in leaf explants were investigated. In addition, the expression and activity of PPO wer...

  14. Current opinions on indications and algorithms for acellular dermal matrix use in primary prosthetic breast reconstruction.

    Vu, Michael M; Kim, John Y S

    2015-06-01

    Acellular dermal matrix (ADM) is widely used in primary prosthetic breast reconstruction. Many indications and contraindications to use ADM have been reported in the literature, and their use varies by institution and surgeon. Developing rational, tested algorithms to determine when ADM is appropriate can significantly improve surgical outcomes and reduce costs associated with ADM use. We review the important indications and contraindications, and discuss the algorithms that have been put forth so far. Further research into algorithmic decision-making for ADM use will allow optimized balancing of cost with risk and benefit. PMID:26161304

  15. Interposition Porcine Acellular Dermal Matrix Xenograft Successful Alternative in Treatment for Massive Rotator Cuff

    Neumann, Julie; Zgonis, Miltiadis H.; Reay, Kathleen Dolores; Mayer, Stephanie W.; Boggess, Blake; Toth, Alison P.

    2016-01-01

    Objectives: Despite advances in the surgical techniques of rotator cuff repair (RCR), the management of massive rotator cuff tears in shoulders without glenohumeral arthritis poses a difficult problem for surgeons. Failure of massive rotator cuff repairs range from 20-90% at one to two years postoperatively using arthrography, ultrasound, or magnetic resonance imaging. Additionally, there are inconsistent outcomes reported with debridement alone of massive rotator cuff tears as well as limitations seen with other current methods of operative intervention including arthroplasty and tendon transfers. The purpose of this prospective, comparative study was to determine if the repair of massive rotator cuff tears using an interposition porcine acellular dermal matrix xenograft improves subjective function, pain, range of motion, and strength at greater than two years follow-up. To our knowledge, this is the largest prospective series reporting outcomes of using porcine acellular dermal matrix xenograft as an interposition graft. Methods: Thirty-seven patients (37 shoulders) with an average age of 66 years (range 51-80 years) were prospectively followed for 33 months (range 23-48) following massive RCR using porcine acellular dermal matrix interposition xenograft. Subjective outcomes were measured using the Visual Analog Scale (VAS) pain score (0-10, 0 = no pain), Modified American Shoulder and Elbow Score (M-ASES), and Short-Form12 (SF-12) scores. Preoperative and postoperative objective outcome measures included active range of motion and supraspinatus and infraspinatus manual muscle strength. Postoperative outcome measures included quantitative muscle strength using a dynamometer and static and dynamic ultrasonography to assess the integrity of the repair. Results: Average VAS pain score decreased from 4.5 to 1.1 (Pacellular dermal matrix xenografts, patients had significant improvement in pain, range of motion, strength and reported good subjective function based on

  16. Reproducible simulation of respiratory motion in porcine lung explants

    Purpose: To develop a model for exactly reproducible respiration motion simulations of animal lung explants inside an MR-compatible chest phantom. Materials and Methods: The materials included a piston pump and a flexible silicone reconstruction of a porcine diaphragm and were used in combination with an established MR-compatible chest phantom for porcine heart-lung preparations. The rhythmic inflation and deflation of the diaphragm at the bottom of the artificial thorax with water (1-1.5 L) induced lung tissue displacement resembling diaphragmatic breathing. This system was tested on five porcine heart-lung preparations using 1.5T MRI with transverse and coronal 3D-GRE (TR/TE=3.63/1.58, 256 x 256 matrix, 350 mm FOV, 4 mm slices) and half Fourier T2-FSE (TR/TE=545/29, 256 x 192, 350 mm, 6 mm) as well as multiple row detector CT (16 x 1 mm collimation, pitch 1.5, FOV 400 mm, 120 mAs) acquired at five fixed inspiration levels. Dynamic CT scans and coronal MRI with dynamic 2D-GRE and 2D-SS-GRE sequences (image frequencies of 10/sec and 3/sec, respectively) were acquired during continuous 'breathing' (7/minute). The position of the piston pump was visually correlated with the respiratory motion visible through the transparent wall of the phantom and with dynamic displays of CT and MR images. An elastic body splines analysis of the respiratory motion was performed using CT data. Results: Visual evaluation of MRI and CT showed three-dimensional movement of the lung tissue throughout the respiration cycle. Local tissue displacement inside the lung explants was documented with motion maps calculated from CT. The maximum displacement at the top of the diaphragm (mean 26.26 [SD 1.9] mm on CT and 27.16 [SD 1.5] mm on MRI, respectively [p=0.25; Wilcoxon test]) was in the range of tidal breathing in human patients. Conclusion: The chest phantom with a diaphragmatic pump is a promising platform for multi-modality imaging studies of the effects of respiratory lung motion. (orig.)

  17. Reproducible simulation of respiratory motion in porcine lung explants

    Biederer, J. [Dept. of Diagnostic Radiology, Univ. Hospital Schleswig-Holstein, Campus Kiel (Germany); Dept. of Radiology, German Cancer Research Center, Heidelberg (Germany); Plathow, C. [Dept. of Diagnostic Radiology, Eberhard-Karls-Univ. Tuebingen (Germany); Dept. of Radiology, German Cancer Research Center, Heidelberg (Germany); Schoebinger, M.; Meinzer, H.P. [Dept. of Medical and Biological Informatics, German Cancer Research Center, Heidelberg (Germany); Tetzlaff, R.; Puderbach, M.; Zaporozhan, J.; Kauczor, H.U. [Dept. of Radiology, German Cancer Research Center, Heidelberg (Germany); Bolte, H.; Heller, M. [Dept. of Diagnostic Radiology, Univ. Hospital Schleswig-Holstein, Campus Kiel (Germany)

    2006-11-15

    Purpose: To develop a model for exactly reproducible respiration motion simulations of animal lung explants inside an MR-compatible chest phantom. Materials and Methods: The materials included a piston pump and a flexible silicone reconstruction of a porcine diaphragm and were used in combination with an established MR-compatible chest phantom for porcine heart-lung preparations. The rhythmic inflation and deflation of the diaphragm at the bottom of the artificial thorax with water (1-1.5 L) induced lung tissue displacement resembling diaphragmatic breathing. This system was tested on five porcine heart-lung preparations using 1.5T MRI with transverse and coronal 3D-GRE (TR/TE=3.63/1.58, 256 x 256 matrix, 350 mm FOV, 4 mm slices) and half Fourier T2-FSE (TR/TE=545/29, 256 x 192, 350 mm, 6 mm) as well as multiple row detector CT (16 x 1 mm collimation, pitch 1.5, FOV 400 mm, 120 mAs) acquired at five fixed inspiration levels. Dynamic CT scans and coronal MRI with dynamic 2D-GRE and 2D-SS-GRE sequences (image frequencies of 10/sec and 3/sec, respectively) were acquired during continuous 'breathing' (7/minute). The position of the piston pump was visually correlated with the respiratory motion visible through the transparent wall of the phantom and with dynamic displays of CT and MR images. An elastic body splines analysis of the respiratory motion was performed using CT data. Results: Visual evaluation of MRI and CT showed three-dimensional movement of the lung tissue throughout the respiration cycle. Local tissue displacement inside the lung explants was documented with motion maps calculated from CT. The maximum displacement at the top of the diaphragm (mean 26.26 [SD 1.9] mm on CT and 27.16 [SD 1.5] mm on MRI, respectively [p=0.25; Wilcoxon test]) was in the range of tidal breathing in human patients. Conclusion: The chest phantom with a diaphragmatic pump is a promising platform for multi-modality imaging studies of the effects of respiratory lung

  18. In vitro Flowering of Shoots Regenerated from Cultured Nodal Explants

    Kantamaht KANCHANAPOOM

    2011-05-01

    Full Text Available A protocol for the regeneration of Gypsophila paniculata L. using nodal explants from 2-month-old field grown plants was established. The induction of multiple shoots was best obtained on Murashige and Skoog (MS medium supplemented with 13.3 μM BA. Callus growth was observed on MS medium containing 44.3 μM BA. Calluses were transferred to MS medium supplemented with 2, 4-D (4.5, 13.5, 22.6 μM, NAA (5.3, 16.1, 26.8 μM or BA (4.4, 13.3, 22.1 μM for 2 months to induce shoot formation. After 6 weeks of initial culture, multiple shoots were regenerated from calluses cultured on MS medium supplemented with 13.3 μM BA. All regenerated shoots produced roots on 16.1 μM NAA containing MS medium within 4 weeks. Rooted plantlets were hardened and established in pots at 100% survival. For induction of in vitro flowering, regenerated shoots could be induced to flower efficiently when cultured on MS medium containing 13.3 μM BA and 50 g/l sucrose.

  19. Proteomic Profile of Brucella abortus-Infected Bovine Chorioallantoic Membrane Explants

    Mol, Juliana P. S.; Pires, Simone F.; Chapeaurouge, Alexander D.; Perales, Jonas; Santos, Renato L.; Andrade, Hélida M.; Lage, Andrey P.

    2016-01-01

    Brucella abortus is the etiological agent of bovine brucellosis, a zoonotic disease that causes significant economic losses worldwide. The differential proteomic profile of bovine chorioallantoic membrane (CAM) explants at early stages of infection with B. abortus (0.5, 2, 4, and 8 h) was determined. Analysis of CAM explants at 0.5 and 4 h showed the highest differences between uninfected and infected CAM explants, and therefore were used for the Differential Gel Electrophoresis (DIGE). A total of 103 spots were present in only one experimental group and were selected for identification by mass spectrometry (MALDI/ToF-ToF). Proteins only identified in extracts of CAM explants infected with B. abortus were related to recognition of PAMPs by TLR, production of reactive oxygen species, intracellular trafficking, and inflammation. PMID:27104343

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

    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

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

    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......,12-dimethylbenz[alpha]anthracene, aflatoxin B1, dimethylnitrosamine, 1,2-dimethylhydrazine, and methylazoxymethanol acetate into chemical species that bind to cellular DNA and protein....

  2. Molecular Profiling of Multiplexed Gene Markers to Assess Viability of Ex Vivo Human Colon Explant Cultures

    Drew, Janice E.; Andrew J Farquharson; Vase, Hollie; Carey, Frank A.; Steele, Robert J C; Ross, Ruth A; Bunton, David C.

    2015-01-01

    Abstract Human colon tissue explant culture provides a physiologically relevant model system to study human gut biology. However, the small (20–30 mg) and complex tissue samples used present challenges for monitoring tissue stability, viability, and provision of sufficient tissue for analyses. Combining molecular profiling with explant culture has potential to overcome such limitations, permitting interrogation of complex gene regulation required to maintain gut mucosa in culture, monitor res...

  3. Sterile acellular dermal collagen as a treatment for rippling deformity of breast.

    Busse, Brittany; Orbay, Hakan; Sahar, David E

    2014-01-01

    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. PMID:25610697

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

    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.

  5. Combination of Acellular Nerve Graft and Schwann Cells-Like Cells for Rat Sciatic Nerve Regeneration

    Songtao Gao

    2014-01-01

    Full Text Available Objective. To investigate the effect of tissue engineering nerve on repair of rat sciatic nerve defect. Methods. Forty-five rats with defective sciatic nerve were randomly divided into three groups. Rats in group A were repaired by acellular nerve grafts only. Rats in group B were repaired by tissue engineering nerve. In group C, rats were repaired by autogenous nerve grafts. After six and twelve weeks, sciatic nerve functional index (SFI, neural electrophysiology (NEP, histological and transmission electron microscope observation, recovery ratio of wet weight of gastrocnemius muscle, regenerated myelinated nerve fibers number, nerve fiber diameter, and thickness of the myelin sheath were measured to assess the effect. Results. After six and twelve weeks, the recovery ratio of SFI and wet weight of gastrocnemius muscle, NEP, and the result of regenerated myelinated nerve fibers in groups B and C were superior to that of group A (P0.05. Conclusion. The tissue engineering nerve composed of acellular allogenic nerve scaffold and Schwann cells-like cells can effectively repair the nerve defect in rats and its effect was similar to that of the autogenous nerve grafts.

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

    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

  7. Endometrial explant culture to study the response of equine endometrium to insemination.

    Nash, D M; Sheldon, I M; Herath, S; Lane, E A

    2010-08-01

    Mating-induced endometritis (MIE) is ubiquitous in the horse after natural mating and artificial insemination with frozen/thawed semen causing the most aggressive response. The majority of mares eliminate MIE 24-48 h after insemination. An endometrial explant culture was tested as a potential in vitro exemplar for sperm-induced MIE. Endometrial prostaglandin F(2alpha) (PGF(2alpha)) secretion and expression of interleukin-8 (IL-8) were used as markers of inflammation. Endometrial explants were cultured from uteri collected from follicular phase mares. Explants were challenged with 1 or 10 x 10(6) sperm/ml frozen/thawed semen, chilled semen, washed sperm or seminal plasma. Medium was collected 24 and 72 h after challenge and assayed for PGF(2alpha) by radioimmunoassay. Treatment of endometrial explants with frozen/thawed, chilled semen or washed sperm did not change the secretion of PGF(2alpha) compared with untreated controls. However, 24 h after challenge cultured explants expressed IL-8. The in vitro endometrial explant system did not represent the in vivo response to semen when PGF(2alpha) was used as a marker of inflammation, yet the use of gene expression as an inflammatory marker warrants further investigation. PMID:19144039

  8. Effects of laser irradiation on immature olfactory neuroepithelial explants from the rat

    Mester, A.F.; Snow, J.B. Jr.

    1988-07-01

    The photobiological effect of low-output laser irradiation on the maturation and regeneration of immature olfactory bipolar receptor cells of the rat was studied. The maturation and regeneration of the receptor cells of rat fetuses were quantified in neuroepithelial explants with morphometric analysis. The number of explants with outgrowth and the number and length of neuritic outgrowths were determined on a regular basis for 12 days. Explants in the experimental group were irradiated with a helium-neon laser using different incident energy densities (IED). Explants in the fluorescent light control group were exposed to fluorescent light for the same periods of time as those in the experimental group were exposed to laser irradiation. Explants in another control group were not exposed to laser or fluorescent light irradiation. The IED of 0.5 J/cm2 laser irradiation has been found to increase significantly the number of explants with outgrowth and the number and length of the outgrowths. Other laser IEDs or fluorescent light irradiation did not influence maturation or regeneration.

  9. Effects of hydroxyapatite nanostructure on channel surface of porcine acellular dermal matrix scaffold on cell viability and osteogenic differentiation of human periodontal ligament stem cells

    Ge S

    2013-05-01

    Full Text Available Shaohua Ge,1 Ning Zhao,1 Lu Wang,1 Hong Liu,2 Pishan Yang11Shandong Provincial Key Laboratory of Oral Biomedicine, Department of Periodontology, Shandong University; 2State Key Laboratory of Crystal Materials, Center of Bio and Micro/Nano Functional Materials, Shandong University, Jinan, People's Republic of ChinaAbstract: A new nanostructured hydroxyapatite-coated porcine acellular dermal matrix (HAp-PADM was fabricated by a biomimetic mineralization method. Human periodontal ligament stem cells were seeded on HAp-PADM and the effects of this scaffold on cell shape, cytoskeleton organization, cell viability, and osteogenic differentiation were examined. Periodontal ligament stem cells cultured on HAp-PADM exhibited different cell shape when compared with those on pure PADM. Moreover, HAp-PADM promoted cell viability and alkaline phosphatase activity significantly. Based on quantitative real-time polymerase chain reaction, the expression of bone-related markers runt-related transcription factor 2 (Runx2, osteopontin (OPN, and osteocalcin (OCN upregulated in the HAp-PADM scaffold. The enhancement of osteogenic differentiation of periodontal ligament stem cells on the HAp-PADM scaffold was proposed based on the research results. The results of this study highlight the micro-nano, two-level, three-dimensional HAp-PADM composite as a promising scaffold for periodontal tissue engineering.Keywords: hydroxyapatite, scaffold, nanostructure, proliferation, differentiation, tissue engineering

  10. Galectin-1-induced skeletal muscle cell differentiation of mesenchymal stem cells seeded on an acellular dermal matrix improves injured anal sphincter.

    Ding, Zhao; Liu, Xiangui; Ren, Xianghai; Zhang, Qiulei; Zhang, Tingtao; Qian, Qun; Liu, Weicheng; Jiang, Congqing

    2016-05-01

    According to recent studies, mesenchymal stromal cells (MSCs) transplanted via local or tail vein injection can improve healing after anal sphincter injury (ASI) in animal models. However, the transplanted MSCs do not generate skeletal muscle that completely resembles the natural anal sphincter structure. In the present study, we investigated whether bone marrow (BM)-derived MSCs could be induced by Galectin-1 (Gal-1) to differentiate into skeletal muscle and whether the recellularization of an acellular dermal matrix (ADM) with skeletal muscle-differentiated MSCs represents a promising approach to restore ASI in a rat model. BM-MSCs subjected to adenovirus-mediated transfection with Gal-1-GFP (Ad-GFP-Gal-1) displayed increased Gal-1 and desmin expression and differentiated into skeletal muscle cells. MSCs transfected with Ad-GFP-Gal-1 (MSC-Gal-1) were seeded onto an ADM (ADM-MSC-Gal-1) via co-culture, and fusion was observed using a confocal laser scanning microscope. ADM-MSC-Gal-1, ADM-MSC, ADM-MSC-Ad, ADM, or a saline control was applied to a rat ASI model, and injury healing was evaluated via histological examination 6 weeks following treatment. ADM-MSC-Gal-1 treatment promoted significant healing after ASI and improved external anal sphincter contraction curves compared with the other treatments and also led to substantial skeletal muscle regeneration and neovascularization. Our results indicate that repair using ADMs and differentiated MSCs may improve muscle regeneration and restore ASI. PMID:27355329

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

    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.

  12. Bone Grafts

    ... repair and rebuild diseased bones in your hips, knees, spine, and sometimes other bones and joints. Grafts can also repair bone loss caused by some types of fractures or cancers. Once your body accepts the bone ...

  13. Stimulation of bone marrow cells and bone formation by nacre: in vivo and in vitro studies.

    Lamghari, M; Almeida, M J; Berland, S; Huet, H; Laurent, A; Milet, C; Lopez, E

    1999-08-01

    There is frequently a loss of vertebral bone due to disease or aging. Nacre (mother of pearl from the oyster Pinctada maxima) stimulates bone cell differentiation and bone formation in vitro and in vivo. Experimental bone defects were prepared in the vertebrae of sheep and used to test the suitability of nacre as an injectable osteogenic biomaterial for treating vertebral bone loss. Twenty-one cavities were prepared in the first four upper lumbar vertebrae of 11 sheep and filled with nacre powder. The lumbar vertebrae were removed after 1 to 12 weeks, embedded undecalcified in methacrylate, and processed for histological studies. The nacre slowly dissolved and the experimental cavities contained a large active cell population. By 12 weeks, the experimental cavity was occupied by newly matured bone trabeculae in contact with or adjacent to the dissolving nacre. The functional new bone trabeculae were covered with osteoid lined with osteoblasts, indicating continuing bone formation. The in vitro study on rat bone marrow explants cultured with a water-soluble extract of the nacre organic matrix also resulted in the stimulation of osteogenic bone marrow cells with enhanced alkaline phosphatase activity. Thus, both the in vivo and in vitro findings suggest that nacre contains one or more signal molecules capable of activating osteogenic bone marrow cells. PMID:10458284

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

    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.

  15. Effect of Hormones on Direct Shoot Regeneration in Hypocotyl Explants of Tomato

    Rizwan RASHID

    2010-03-01

    Full Text Available This study was conducted for developing a high frequency regeneration system in two genotypes of tomato (Lycopersicon esculentum Mill., �Punjab Upma� and �IPA-3� for direct shoot regeneration from hypocotyl explants. The explants were excised from in vitro tomato seedlings and cultured on MS medium supplemented with different concentrations and combinations of hormones. Direct regeneration was significantly influenced by the genotype hormones combination and concentrations. The MS medium supplemented with (Kinetin 0.5 mg/l and (BAP 0.5 mg/l was found optimum for inducing direct shoot regeneration and number of shoots per explant from hypocotyl explants on this medium. Shoot regeneration per cent in �Punjab Upma� and �IPA-3� per cent was recorded to be highest i.e (86.02 and (82.57 respectively. Besides this, average number shoots per explant was also highest i.e (3.16 in case of �Punjab Upma� and (2.93 in case of �IPA-3�. A significant decline was observed in percent shoot regeneration and average number of shoots per explant with increase in the hormonal concentration. Shoots were obtained and transferred to the elongation medium (MS + BAP 0.3 mg/l. Hundred per cent rooting was induced in separated shoots upon culturing on MS and � MS basal media. Hardening on moist cotton showed maximum plantlet survival rate in case of both genotypes. After hardening, plants were transferred to soil. Thus, a tissue culture base line was established in tomato for obtaining direct regeneration using hypocotyl as explants.

  16. Ventricular Recovery and Pump Explantation in Patients Supported by Left Ventricular Assist Devices: A Systematic Review.

    Phan, Kevin; Huo, Ya Ruth; Zhao, Dong Fang; Yan, Tristan D; Tchantchaleishvili, Vakhtang

    2016-01-01

    Several studies have reported that a portion of patients who exhibit cardiac recovery during left ventricular assist device (LVAD) support can have their device explanted with reasonable long-term survival. The aim of this systematic review is to assess the survival and cardiac function in patients with explanted LVADs from the current literature. Electronic search was performed to identify all studies in English literature assessing LVAD explantation. All identified articles were systematically assessed using the inclusion and exclusion criteria. Selected studies were subjected to quantitative assessment. From 5 electronic databases, 11 studies (213 patients) were included. Pooled mean perioperative mortality rate of those explanted was 9.2% (95% CI, 5.0-14.5%; I = 0). Pooled mean late mortality rate was 15% (95% CI, 9.0-22.1%; I = 31%). The pooled 1, 5, and 10 year survival postexplant was 91, 76, and 65.7%, respectively. Pooled postweaning freedom from heart failure (HF) recurrence reached 81.3%. Subset analysis demonstrated that patients explanted from a continuous-flow LVAD versus pulsatile LVAD had a lower rate of HF recurrence (6.6 vs. 28.3%, p = 0.03) and LVAD reimplantation (7.5 vs. 37%, p = 0.001). Before LVAD explantation, overall mean left ventricular ejection fraction (LVEF) was 49%. Weighted pooled early and late postexplantation mean LVEF was 47.3 and 41.2%, respectively. Late postexplantation LVEF was significantly higher in the continuous-flow versus pulsatile LVAD subgroup (41.5 vs. 24%, p = 0.001). This review shows encouraging safety and 10 year survival outcomes after explantation of LVADs in carefully selected patients, with rates better than expected after a heart transplant. Recovery of the native heart is the most desirable clinical outcome in patients supported with LVADs and should be actively sought. PMID:26735559

  17. Histological changes during the adventitious shoot formation in seedling explants of pepper (Capsicum annuum L. cultured in vitro

    Andrzej Gatz

    2014-02-01

    Full Text Available Adventitious shoots differentiated directly from explant tissue without intermediate callus on all types of examined explants (shoot tip, cotyledonary node, cotyledon and hypocotyl of Capsicum annuum L. cv. Bryza. First cell divisions took place as early as after 3 days of explant culture within epidermal and subepidermal layers of explants, and in the case of cotyledon also within mesophyll cells located near epidermis. Mitotic activity in these layers led to the formation of meristemoids (meristematic centres. In all types of studied explants, meristematic centres appeared approximately at the same time (after about 7 days of culture. In the second week bud primordia began to differentiate from meristematic centres. Subsequently some of shoot primordia developed into leaves and leaf-like structures (mainly on cotyledon explants, and also into adventitious buds with well developed apical meristem and leaf primordia.

  18. TGF beta suppresses casein synthesis in mouse mammary explants and may play a role in controlling milk levels during pregnancy

    1993-01-01

    Mammary explants from 14-15-d-pregnant mice synthesize and secrete milk proteins in culture in response to insulin, hydrocortisone, and prolactin. Here we demonstrate that transforming growth factor beta (TGF beta) treatment suppresses, in a dose dependent and reversible manner, the ability of explants to synthesize and secrete milk caseins. TGF beta does not affect the level of casein mRNA within explants but inhibits casein synthesis posttranscriptionally. We also show increased expression ...

  19. Direct delayed breast reconstruction with TAP flap, implant and acellular dermal matrix (TAPIA)

    Børsen-Koch, Mikkel; Gunnarsson, Gudjon L; Udesen, Ann;

    2015-01-01

    combining the use of a propeller thoracodorsal artery perforator (TAP) flap with an acellular dermal matrix (ADM) and an implant. METHODS: The paper presents 43 delayed breast reconstructions in 38 women using a modified technique for harvesting the TAP flap in combination with an ADM and an implant for......BACKGROUND: The latissimus dorsi (LD) flap is considered one of the working horses within the field of breast reconstruction and it offers several advantages. However, donor-site morbidity may pose a problem. This article describes a new and modified technique for delayed breast reconstruction...... total breast reconstruction. The focus of this paper is the refinements of our technique and short-term outcome in complication rates. The data presented were collected retrospectively. RESULTS: Three patients experienced major complications including hematoma, partial flap necrosis, and venous...

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

    Block, I.

    1992-01-01

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

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

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

    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

  3. Targeting Bone Alleviates Osteoarthritis in Osteopenic Mice and Modulates Cartilage Catabolism

    Funck-Brentano, Thomas; Lin, Hilène; Hay, Eric; Ah Kioon, Marie-Dominique; Schiltz, Corinne; Hannouche, Didier; Nizard, Rémy; Lioté, Frédéric; Orcel, Philippe; de Vernejoul, Marie-Christine; Cohen-Solal, Martine Esther

    2012-01-01

    Objective Subchondral bone modifications occur early in the development of osteoarthritis (OA). The level of bone resorption might impact cartilage remodeling. We therefore assessed the in vivo and in vitro effects of targeting bone resorption in OA and cartilage metabolism. Methods OA was induced by meniscectomy (MNX) in ovariectomized osteopenic mice (OP) treated with estradiol (E2), pamidronate (PAM), or phosphate buffered saline (PBS) for 6 weeks. We assessed the subchondral bone and cartilage structure and the expression of cartilage matrix proteases. To assess the involvement of bone soluble factors in cartilage metabolism, supernatant of human bone explants pre-treated with E2 or PAM were transferred to cartilage explants to assess proteoglycan release and aggrecan cleavage. OPG/RANKL mRNA expression was assessed in bone explants by real-time quantitative PCR. The role of osteoprotegerin (OPG) in the bone-cartilage crosstalk was tested using an OPG neutralizing antibody. Results Bone mineral density of OP mice and osteoclast number were restored by E2 and PAM (p<0.05). In OP mice, E2 and PAM decreased ADAMTS-4 and -5 expression, while only PAM markedly reduced OA compared to PBS (2.0±0.63 vs 5.2±0.95; p<0.05). OPG/RANKL mRNA was increased in human bone explants treated with both drugs (2.2–3.7-fold). Moreover, supernatants from bone explants cultured with E2 or PAM reduced aggrecan cleavage and cartilage proteoglycan release (73±8.0% and 80±22% of control, respectively, p<0.05). This effect was reversed with osteoprotegerin blockade. Conclusion The inhibition of bone resorption by pamidronate in osteopenic mice alleviates the histological OA score with a reduction in the expression of aggrecanases. Bone soluble factors, such as osteoprotegerin, impact the cartilage response to catabolic factors. This study further highlights the importance of subchondral bone in the regulation of joint cartilage damage in OA. PMID:22432033

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

    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.

  5. Analysis of the National Surgical Quality Improvement Program Database in 19,100 Patients Undergoing Implant-Based Breast Reconstruction: Complication Rates With Acellular Dermal Matrix

    Shuster, Marina

    2015-01-01

    Background: The use of acellular dermal matrices has become increasingly popular in immediate and delayed tissue expander/implant–based breast reconstruction. However, it is unclear whether their use is associated with increased postoperative complication rates. Using the American College of Surgeons National Surgical Quality Improvement Program database, the authors assessed baseline differences in demographics and comorbidities with and without acellular dermal matrix and determined whether...

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

    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.

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

    Holme, Inger Bæksted; Petersen, Karen Koefoed

    1996-01-01

    Different explants of Miscanthus x ogiformis Honda 'Giganteus' were tested in order to develop an efficient tissue culture system. Shoot apices, leaf and root sections from in vitro-propagated plants, and leaf and immature inflorescence sections from 6-month-old greenhouse-grown plants were used...... developmental stage of the explants. Shoot apices formed the highest percentage of embryogenic callus. There was a difference in the formation of embryogenic callus between leaf explants from in vitro-propagated shoots and greenhouse-grown plants. The best results were obtained from newly formed leaves of in...... vitro-propagated shoots and older leaves of greenhouse-grown plants. Immature inflorescences smaller than 2.5 cm produced a higher percentage of embryogenic callus than larger more mature inflorescences. Embryogenic callus derived from immature inflorescences had the highest regeneration capacity...

  8. Plant regeneration via direct shoot organogenesis from cotyledon explants of Bambara groundnut, Vigna subterranea (L. Verdc.

    Koné, M.

    2013-01-01

    Full Text Available Bambara groundnut (Vigna subterranea [L.] Verdc. is mainly grown for human consumption. However, several factors limit a wider adoption of the crop including the presence of antinutritional factors in the seeds that lower product quality and protein availability but also the plant susceptibility to pests and diseases. Tissue culture techniques are very scanty in Bambara groundnut and should be developed before carrying out genetic transformation for the crop improvement. Therefore, here, an efficient system for in vitro shoot induction from cotyledons derived from mature seeds has been established. Different types and concentrations of plant growth regulators were used to induce buds in embryo-free cotyledon explants. Cotyledons were cut transversally or longitudinally into three segments: proximal, middle and distal part. The influence of explant orientation on the medium, the type of segment and landrace has then been studied. Benzylaminopurine (3 mg·l-1 alone or combined with α-naphthaleneacetic acid (0.05 mg·l-1 induced multiple shoot formations. The organogenic potential was restricted to the proximal segment of cotyledons. Frequency of bud induction (30% and average number of buds per explant (12 were higher when the adaxial side of the proximal segment was in contact with the medium. Shoot regeneration from cotyledon explants of ten Bambara landraces revealed that the response is genotype-dependent with varieties Ci6, Ci2, Ci4 and Ci15 exhibiting 20 to 30% shoot regeneration and six to ten buds per explant. Regenerated shoot buds excised from explants were elongated and rooted on MS basal medium devoid of plant growth regulators. All rooted plantlets survived to the transfer on a sand soil mixture, and morphologically normal plants were hardened and transferred to greenhouse for further growth to maturity and seed set.

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

    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.

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

    U R Vishwakarma; Gurav, A M; Sharma, P.C

    2009-01-01

    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 mix...

  11. Efficient Regeneration of �Caralis� Alstroemeria Cultivar from Rhizome Explants

    Amir Ghaffar SHAHRIARI; Bagheri, Abdolreza; Sharifi, Ahmad; Nasrin MOSHTAGHI

    2012-01-01

    In this paper, the effects of a number of growth regulators as well as supplements to the Murashige and Skoog (MS) basal medium were evaluated on the regeneration of Alstroemeria rhizome explants. In the first experiment the effects of three cytokinins (BA, TDZ and 2IP each at 0.5, 1 and 2 mg/l) in combination with NAA (0.2 mg/l), followed by another PGR combination of 2IP (at 0.5, 1 and 2 mg/l) with NAA (0 and 0.2 mg/l), on regeneration of rhizome-derived explants, was investigated. Through ...

  12. Direct axillary shoot regeneration from the mature seed explant of the hairy vetch (Vicia villosa Roth)

    Aasim Muhammad; Sahin-Demirbag Nurdan; Khawar Mahmood Khalid; Kendir Hayrettin; Özcan Sebahattin

    2011-01-01

    The hairy vetch (Vicia villosa Roth) is a climbing, prostrate or trailing legume grown as forage.It fixes atmospheric nitrogen, reduces soil erosion and provides an instant mulch. Multiple axillary shoot regeneration from a mature seed explant (zygotic embryo with two cotyledons) was obtained on MS medium containing 0.05 - 1.6 mg/l TDZ with or without 0.10 mg/l IBA. The frequency (%) of shoot regeneration ranged from 45.83-75.00% with a maximum number of 28.6 shoots per explant on MS me...

  13. MICROPROPAGATION OF BLACK TURMERIC (CURCUMA CAESIA ROXB.) THROUGH IN VITRO CULTURE OF RHIZOME BUD EXPLANTS

    Muhammad Shahinozzaman; Molla Ferdous; Muhammad Faruq; Mustafa Azad; Muhammad Amin

    2013-01-01

    In the present study, preliminary in vitro shoots propagation of Curcuma caesia Roxb. was investigated. Rhizome buds were used as explants and were cultured on Murashige and Skoog MS medium containing 6 Benzyl adenine BA alone or in combination with a Naphthalene acetic acid NAA. The results showed that the optimum shoot proliferation was obtained from MS medium containing 3.0 micro M BA + 0.5 micro M NAA. In this growth regulator combination, maximum 99.97 per cent explants produced 10.38 sh...

  14. Bone Cancer

    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 ...

  15. Bone Cancer

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

  16. Bone Diseases

    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 ...

  17. Ventral hernia: retrospective cost analysis of primary repair, repair with synthetic mesh, and repair with acellular xenograft implant

    Willis, Sharon

    2013-01-01

    George DeNoto III,1 Nancy Reaven,2 Susan Funk2 1Division of General Surgery, St Francis Hospital, Roslyn, and Hofstra North Shore-LIJ School of Medicine, Manhasset, NY, USA; 2Strategic Health Resources, La Cañada, CA, USA Background: The purpose of this study was to evaluate resource utilization and costs of repair of potentially contaminated/infected complex ventral hernias using primary repair, synthetic mesh, or acellular xenograft. Methods: We used 2008–2009 insuranc...

  18. Ventral hernia: retrospective cost analysis of primary repair, repair with synthetic mesh, and repair with acellular xenograft implant

    DeNoto G III; Reaven N; Funk S

    2013-01-01

    George DeNoto III,1 Nancy Reaven,2 Susan Funk2 1Division of General Surgery, St Francis Hospital, Roslyn, and Hofstra North Shore-LIJ School of Medicine, Manhasset, NY, USA; 2Strategic Health Resources, La Cañada, CA, USA Background: The purpose of this study was to evaluate resource utilization and costs of repair of potentially contaminated/infected complex ventral hernias using primary repair, synthetic mesh, or acellular xenograft. Methods: We used 2008–2009 insurance claims ...

  19. A CpG-containing oligodeoxynucleotide adjuvant for acellular pertussis vaccine improves the protective response against Bordetella pertussis

    Asokanathan, Catpagavalli; Corbel, Michael; Xing, Dorothy

    2013-01-01

    We investigated the adjuvant effect of CpG ODN alone or in combination with aluminum hydroxide on the immune response to the three main antigens presented in current acellular pertussis vaccines: pertussis toxoid, filamentous haemagglutinin and pertactin. The development of protection in mice was investigated for the intra-peritoneal and intra-nasal immunisation routes. The results showed that CpG ODN alone, or in combination with aluminum hydroxide, gave enhancement in anti-pertussis toxin, ...

  20. Root coverage using a coronally advanced flap with or without acellular dermal matrix: a meta-analysis

    Guan, Wei; Liao, Haiqing; Guo, Li; Wang, Changning; Cao, Zhengguo

    2016-01-01

    Purpose Gingival recession is a major esthetic concern and may lead to root sensitivity during periodontal treatment. Coronally advanced flaps (CAFs) with and without acellular dermal matrix (ADM) are widely used in root coverage procedures. The aim of this study was to analyze the efficacy of CAF in combination with ADM in the treatment of gingival recession. Methods PubMed, The Cochrane Library, and Embase were used to identify relevant articles. The articles were screened, data were extrac...

  1. Somatic embryogenesis and massive shoot regeneration from immature embryo explants of tef.

    Gugsa, Likyelesh; Kumlehn, Jochen

    2011-01-01

    Tef (Eragrostis tef) provides a major source of human nutrition in the Horn of Africa, but biotechnology has had little impact on its improvement to date. Here, we report the elaboration of an in vitro regeneration protocol, based on the use of immature zygotic embryos as explant. Explant size was an important determinant of in vitro regeneration efficiency, as was the formulation of the culture medium. Optimal results were obtained by culturing 0.2-0.35 mm embryo explants on a medium containing KBP minerals, 9.2-13.8 μM 2,4-dichlorophenoxyacetic acid, 6 mM glutamine, and 0.5% Phytagel. Although this protocol was effective for both the improved cultivar "DZ-01-196" and the landrace "Fesho", the former produced consistently more embryogenic tissue and a higher number of regenerants. An average of more than 2,800 shoots could be obtained from each "DZ-01-196" explant after 12 weeks of in vitro culture. These shoots readily formed roots, and plantlets transferred to soil were able to develop into morphologically normal, fertile plants. This regeneration and multiplication system should allow for the application of a range of biotechnological methods to tef. PMID:22028975

  2. Effect of Pre-culture Irradiation and Explant Types on Efficiency of Brassica napus Genetic Transformation

    The irradiated seeds of canola cv. Drakkar ( Brassica napus l. ) were germinated under aspect conditions, cotyledonary petioles and hypocotyl of 6 days old seedlings were used for Agrobacterium-mediated transformation. Agrobacterium tumefaciens has construct with the selectable marker gene (NPT II) and the desirable gene (HPPD). Direct and indirect shoot organogenesis were obtained from the both explants. Cotyledonary petioles was higher responded than hypocotyl with respective 26% and 14% of the explants producing NPT II-positive shoots after the selection on 50mg/l kanamycin. Calli might develop on and not in the agar medium were un transformation. This explains the higher number of escapes detected in hypocotyl explants than in experiments with cotyledons. The frequency of transformation plants as a function of indirect organogenesis was more than direct shoot regeneration from explants. The pre- irradiation with 75 Gy of gamma rays enhanced the genetic transformation frequencies by about 10 % as compared to that of the un-irradiated material. The obtained shoots were rooted and regenerated mature plants

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

    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.

  4. Somatic Embryogenesis in Peach-Palm (Bactris gasipaes) Using Different Explant Sources.

    Steinmacher, Douglas A; Heringer, Angelo Schuabb; Jiménez, Víctor M; Quoirin, Marguerite G G; Guerra, Miguel P

    2016-01-01

    Peach palm (Bactris gasipaes Kunth) is a member of the family Arecaceae and is a multipurpose but underutilized species. Nowadays, fruit production for subsistence and local markets, and heart-of-palm production for local, national, and international markets are the most important uses of this plant. Conventional breeding programs in peach palm are long-term efforts due to the prolonged generation time, large plant size, difficulties with controlled pollination and other factors. Although it is a caespitose palm, its propagation is currently based on seeds, as off-shoots are difficult to root. Hence, tissue culture techniques are considered to be the most likely strategy for efficient clonal plantlet regeneration of this species. Among various techniques, somatic embryogenesis offers the advantages of potential automated large-scale production and putative genetic stability of the regenerated plantlets. The induction of somatic embryogenesis in peach palm can be achieved by using different explant sources including zygotic embryos, immature inflorescences and thin cell layers from the young leaves and shoot meristems. The choice of a particular explant depends on whether clonal propagation is desired or not, as well as on the plant conditions and availability of explants. Protocols to induce and express somatic embryogenesis from different peach palm explants, up to acclimatization of plantlets, are described in this chapter. PMID:26619867

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

    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.

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

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

    2016-01-01

    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. PMID:27023602

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

    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.

  8. [Establishment of high frequency regeneration via leaf explants of 'Red Sun' kiwifruit (Actinidia chinensis)].

    Zhao, Xupeng; Luo, Keming; Zhou, Yue; Wu, Xiuhua; Yang, Li; Tang, Shaohu

    2013-11-01

    A high efficient in vitro regeneration protocol was developed from leaf explants of the female 'Red Sun' kiwifruit (Actinidia chinensis) and the multiplication coefficient and rooting rate of adventitious buds were also optimized. This method does not require formation of callus tissues which leads to somaclonal variations. The results show that the adventitious buds developing directly from explants tissue were noticed after 30 d of culture. The maximum regeneration frequency of adventitious buds is 100% and 18.67 shoots was observed in each leaf explants when MS medium was supplemented with 3.0 mg/L BA+1.0 mg/L NAA. The optimal culture medium for bud multiplication is MS+2.0 mg/L BA+1.0 mg/L NAA+0.1 mg/L GA3 and the multiplication coefficient reached 8.63. On the rooting medium with 1/2 MS+0.8 mg/L IBA for 15 d, the adventitious plantlets were transferred into matrix perlite supplied with 1/2 MS liquid medium for 15 d and the rooting rate reached 100%. 95 out of 98 plantlets (96.94%) survived acclimatization, producing healthy plants in the greenhouse. Taken together, a highly efficient regeneration method via leaf explants of 'Red Sun' kiwifruit was successfully established. This protocol may be useful for micropropagation and genetic transformation studies of 'Red Sun' kiwifruit. PMID:24701825

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

    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.

  10. Characterisation of coral explants: a model organism for cnidarian-dinoflagellate studies

    Gardner, S. G.; Nielsen, D. A.; Petrou, K.; Larkum, A. W. D.; Ralph, P. J.

    2015-03-01

    Coral cell cultures made from reef-building scleractinian corals have the potential to aid in the pursuit of understanding of the cnidarian-dinoflagellate symbiosis. Various methods have previously been described for the production of cell cultures in vitro with a range of success and longevity. In this study, viable tissue spheroids containing host tissue and symbionts (coral explants) were grown from the tissues of Fungia granulosa. The cultured explants remained viable for over 2 months and showed morphological similarities in tissue structure and internal microenvironment to reef-building scleractinian corals. The photophysiology of the explants (1 week old) closely matched that of the parent coral F. granulosa. This study provides the first empirical basis for supporting the use of coral explants as laboratory models for studying coral symbioses. In particular, it highlights how these small, self-sustaining, skeleton-free models can be useful for a number of molecular, genetic and physiological analyses necessary for investigating host-symbiont interactions at the microscale.

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

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

    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. Improved explant method to isolate umbilical cord-derived mesenchymal stem cells and their immunosuppressive properties.

    Mori, Yuka; Ohshimo, Jun; Shimazu, Takahisa; He, Haiping; Takahashi, Atsuko; Yamamoto, Yuki; Tsunoda, Hajime; Tojo, Arinobu; Nagamura-Inoue, Tokiko

    2015-04-01

    The umbilical cord (UC) has become one of the major sources of mesenchymal stem cells (MSCs). The common explant method of isolating UC-derived MSCs (UC-MSCs) involves mincing the UCs into small fragments, which are then attached to a culture dish bottom from which the MSCs migrate. However, the fragments frequently float up from the bottom of the dish, thereby reducing the cell recovery rate. To overcome this problem, we demonstrate an improved explant method for UC-MSC isolation, which involves the use of a stainless steel mesh (Cellamigo(®); Tsubakimoto Chain Co.), to protect the tissue from floating after the minced fragments are aligned at regular intervals in culture dishes. The culture medium was refreshed every 3 days and the adherent cells and tissue fragments were harvested using trypsin. The number of UC-MSCs isolated from 1 g of UC using the explant method with Cellamigo was 2.9 ± 1.4 × 10(6)/g, which was significantly higher than that obtained without Cellamigo (0.66 ± 0.53 × 10(6)/g) (n = 6, p < 0.01) when cells reached 80-90% confluence. In addition, the processing and incubation time required to reach 80-90% confluence was reduced in the improved explant method compared with the conventional method. The UC-MSCs isolated using the improved method were positive for CD105, CD73, CD90, and HLA class I expression and negative for CD45 and HLA class II expression. The isolated UC-MSCs efficiently inhibited the responder T cells induced by allogeneic dendritic cells in a mixed lymphocyte reaction. Conclusively, we demonstrated that the use of Cellamigo improves the explant method for isolating UC-MSCs. PMID:25220032

  14. Efficient Regeneration of �Caralis� Alstroemeria Cultivar from Rhizome Explants

    Amir Ghaffar SHAHRIARI

    2012-05-01

    Full Text Available In this paper, the effects of a number of growth regulators as well as supplements to the Murashige and Skoog (MS basal medium were evaluated on the regeneration of Alstroemeria rhizome explants. In the first experiment the effects of three cytokinins (BA, TDZ and 2IP each at 0.5, 1 and 2 mg/l in combination with NAA (0.2 mg/l, followed by another PGR combination of 2IP (at 0.5, 1 and 2 mg/l with NAA (0 and 0.2 mg/l, on regeneration of rhizome-derived explants, was investigated. Through the second experiment, the effects of a number of supplements, including glucose (30 g/l as the alternative for sucrose, casein hydrolysate (1 g/l, asparagine and glutamine, (each at 30 mg/l added to MS medium, containing 1 mg/l BA and 0.2 mg/l NAA, was examined on rhizome explants� regeneration. Among the tested cytokinins, BA induced better regeneration of rhizome explants, resulting in a higher number of shoots compared to the other cytokinins. A medium supplemented with 1 mg/l BA and 0.2 mg/l NAA proved to be the most effective, with an average of 4.16 regenerated shoots per explant. In the second PGR combination, addition of NAA at 0.2 mg/l improved regeneration, compared to NAA-free treatments. In the second experiment, glucose substitution for sucrose improved regeneration with an average of 5.10 regenerated shoots per explant, compared to 4.16 shoots in sucrose-containing medium; whereas glutamine and asparagine (with 2.66 shoots and casein hydrolysate (with 3.80 shoots showed a negative influence on rhizome explants� regeneration.

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

    Rose, Jessica F; Zafar, Sarosh N; Ellsworth Iv, Warren A

    2016-01-01

    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 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. PMID:27190645

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

    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 (pbreast 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. PMID:27579264

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

    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.

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

    2016-01-01

    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.

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

    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.

  20. Preparation and characterization of an advanced collagen aggregate from porcine acellular dermal matrix.

    Liu, Xinhua; Dan, Nianhua; Dan, Weihua

    2016-07-01

    The objective of this study was to extract and characterize an advanced collagen aggregate (Ag-col) from porcine acellular dermal matrix (pADM). Based on histological examination, scanning electron microscopy (SEM) and atomic force microscope (AFM), Ag-col was composed of the D-periodic cross-striated collagen fibrils and thick collagen fiber bundles with uneven diameters and non-orientated arrangement. Fourier transform infrared (FTIR) spectra of pADM, Ag-col and Col were similar and revealed the presence of the triple helix. Circular dichroism (CD) analysis exhibited a slightly higher content of α-helix but inappreciably less amount of random coil structure in Ag-col compared to Col. Moreover, imino acid contents of pADM, Ag-col and Col were 222.43, 218.30 and 190.01 residues/1000 residues, respectively. From zeta potential analysis, a net charge of zero was found at pH 6.45 and 6.11 for Ag-col and Col, respectively. Differential scanning calorimetry (DSC) study suggested that the Td of Ag-col was 20°C higher than that of Col as expected, and dynamic mechanical analysis (DMA) indicated that Ag-col possessed a higher storage modulus but similar loss factor compared to Col. Therefore, the collagen aggregate from pADM could serve as a better alternative source of collagens for further applications in food and biological industries. PMID:27039117

  1. Cellular Response to a Novel Fetal Acellular Collagen Matrix: Implications for Tissue Regeneration

    Robert C. Rennert

    2013-01-01

    Full Text Available Introduction. PriMatrix (TEI Biosciences Inc., Boston, MA, USA is a novel acellular collagen matrix derived from fetal bovine dermis that is designed for use in partial- and full-thickness wounds. This study analyzes the cellular response to PriMatrix in vivo, as well as the ability of this matrix to facilitate normal tissue regeneration. Methods. Five by five mm squares of rehydrated PriMatrix were implanted in a subcutaneous fashion on the dorsum of wild-type mice. Implant site tissue was harvested for histology, immunohistochemistry (IHC, and flow cytometric analyses at multiple time points until day 28. Results. PriMatrix implants were found to go through a biological progression initiated by a transient infiltrate of inflammatory cells, followed by mesenchymal cell recruitment and vascular development. IHC analysis revealed that the majority of the implanted fetal dermal collagen fibers persisted through day 28 but underwent remodeling and cellular repopulation to form tissue with a density and morphology consistent with healthy dermis. Conclusions. PriMatrix implants undergo progressive in vivo remodeling, facilitating the regeneration of histologically normal tissue through a mild inflammatory and progenitor cell response. Regeneration of normal tissue is especially important in a wound environment, and these findings warrant further investigation of PriMatrix in this setting.

  2. Glycerolized Reticular Dermis as a New Human Acellular Dermal Matrix: An Exploratory Study.

    Ferrando, Pietro Maria; Balmativola, Davide; Cambieri, Irene; Scalzo, Maria Stella; Bergallo, Massimiliano; Annaratone, Laura; Casarin, Stefania; Fumagalli, Mara; Stella, Maurizio; Sapino, Anna; Castagnoli, Carlotta

    2016-01-01

    Human Acellular Dermal Matrices (HADM) are employed in various reconstructive surgery procedures as scaffolds for autologous tissue regeneration. The aim of this project was to develop a new type of HADM for clinical use, composed of glycerolized reticular dermis decellularized through incubation and tilting in Dulbecco's Modified Eagle's Medium (DMEM). This manufacturing method was compared with a decellularization procedure already described in the literature, based on the use of sodium hydroxide (NaOH), on samples from 28 donors. Cell viability was assessed using an MTT assay and microbiological monitoring was performed on all samples processed after each step. Two surgeons evaluated the biomechanical characteristics of grafts of increasing thickness. The effects of the different decellularization protocols were assessed by means of histological examination and immunohistochemistry, and residual DNA after decellularization was quantified using a real-time TaqMan MGB probe. Finally, we compared the results of DMEM based decellularization protocol on reticular dermis derived samples with the results of the same protocol applied on papillary dermis derived grafts. Our experimental results indicated that the use of glycerolized reticular dermis after 5 weeks of treatment with DMEM results in an HADM with good handling and biocompatibility properties. PMID:26918526

  3. Glycerolized Reticular Dermis as a New Human Acellular Dermal Matrix: An Exploratory Study.

    Pietro Maria Ferrando

    Full Text Available Human Acellular Dermal Matrices (HADM are employed in various reconstructive surgery procedures as scaffolds for autologous tissue regeneration. The aim of this project was to develop a new type of HADM for clinical use, composed of glycerolized reticular dermis decellularized through incubation and tilting in Dulbecco's Modified Eagle's Medium (DMEM. This manufacturing method was compared with a decellularization procedure already described in the literature, based on the use of sodium hydroxide (NaOH, on samples from 28 donors. Cell viability was assessed using an MTT assay and microbiological monitoring was performed on all samples processed after each step. Two surgeons evaluated the biomechanical characteristics of grafts of increasing thickness. The effects of the different decellularization protocols were assessed by means of histological examination and immunohistochemistry, and residual DNA after decellularization was quantified using a real-time TaqMan MGB probe. Finally, we compared the results of DMEM based decellularization protocol on reticular dermis derived samples with the results of the same protocol applied on papillary dermis derived grafts. Our experimental results indicated that the use of glycerolized reticular dermis after 5 weeks of treatment with DMEM results in an HADM with good handling and biocompatibility properties.

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

    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. PMID:27372388

  5. The biocompatibility of silk fibroin and acellular collagen scaffolds for tissue engineering in the ear

    Recent experimental studies have shown the suitability of silk fibroin scaffold (SFS) and porcine-derived acellular collagen I/III scaffold (ACS) as onlay graft materials for tympanic membrane perforation repair. The aims of this study were to further characterize and evaluate the in vivo biocompatibility of SFS and ACS compared with commonly used materials such as Gelfoam and paper in a rat model. The scaffolds were implanted in subcutaneous (SC) tissue and middle ear (ME) cavity followed by histological and otoscopic evaluation for up to 26 weeks. Our results revealed that SFS and ACS were well tolerated and compatible in rat SC and ME tissues throughout the study. The tissue response adjacent to the implants evaluated by histology and otoscopy showed SFS and ACS to have a milder tissue response with minimal inflammation compared to that of paper. Gelfoam gave similar results to SFS and ACS after SC implantation, but it was found to be associated with pronounced fibrosis and osteoneogenesis after ME implantation. It is concluded that SFS and ACS both were biocompatible and could serve as potential alternative scaffolds for tissue engineering in the ear. (paper)

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

    Beekman, Madeleine; Latty, Tanya

    2015-11-20

    Because of its peculiar biology and the ease with which it can be cultured, the acellular slime mould Physarum polycephalum has long been a model organism in a range of disciplines. Due to its macroscopic, syncytial nature, it is no surprise that it has been a favourite amongst cell biologists. Its inclusion in the experimental tool kit of behavioural ecologists is much more recent. These recent studies have certainly paid off. They have shown that, for an organism that lacks a brain or central nervous system, P. polycephalum shows rather complex behaviour. For example, it is capable of finding the shortest path through a maze, it can construct networks as efficient as those designed by humans, it can solve computationally difficult puzzles, it makes multi-objective foraging decisions, it balances its nutrient intake and it even behaves irrationally. Are the slime mould's achievements simply "cute", worthy of mentioning in passing but nothing to take too seriously? Or do they hint at the fundamental processes underlying all decision making? We will address this question after reviewing the decision-making abilities of the slime mould. PMID:26189159

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

    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

  8. Bone Densitometry (Bone Density Scan)

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

    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.

  10. Low Bone Density

    ... Density Exam/Testing › Low Bone Density Low Bone Density Low bone density is when your bone density ... people with normal bone density. Detecting Low Bone Density A bone density test will determine whether you ...

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

    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 ad......), particularly macrophages, in inguinal adipose tissue resulting in stronger responses in terms of hepatotoxicity and insulin-resistance....... 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......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...

  12. Light-Activated Sealing of Acellular Nerve Allografts following Nerve Gap Injury.

    Fairbairn, Neil G; Ng-Glazier, Joanna; Meppelink, Amanda M; Randolph, Mark A; Valerio, Ian L; Fleming, Mark E; Kochevar, Irene E; Winograd, Jonathan M; Redmond, Robert W

    2016-07-01

    Introduction Photochemical tissue bonding (PTB) uses visible light to create sutureless, watertight bonds between two apposed tissue surfaces stained with photoactive dye. In phase 1 of this two-phase study, nerve gaps repaired with bonded isografts were superior to sutured isografts. When autograft demand exceeds supply, acellular nerve allograft (ANA) is an alternative although outcomes are typically inferior. This study assesses the efficacy of PTB when used with ANA. Methods Overall 20 male Lewis rats had 15-mm left sciatic nerve gaps repaired using ANA. ANAs were secured using epineurial suture (group 1) or PTB (group 2). Outcomes were assessed using sciatic function index (SFI), gastrocnemius muscle mass retention, and nerve histomorphometry. Historical controls from phase 1 were used to compare the performance of ANA with isograft. Statistical analysis was performed using analysis of variance and Bonferroni all-pairs comparison. Results All ANAs had signs of successful regeneration. Mean values for SFI, muscle mass retention, nerve fiber diameter, axon diameter, and myelin thickness were not significantly different between ANA + suture and ANA + PTB. On comparative analysis, ANA + suture performed significantly worse than isograft + suture from phase 1. However, ANA + PTB was statistically comparable to isograft + suture, the current standard of care. Conclusion Previously reported advantages of PTB versus suture appear to be reduced when applied to ANA. The lack of Schwann cells and neurotrophic factors may be responsible. PTB may improve ANA performance to an extent, where they are equivalent to autograft. This may have important clinical implications when injuries preclude the use of autograft. PMID:26878685

  13. Neoinnervation and neovascularization of acellular pericardial-derived scaffolds in myocardial infarcts.

    Gálvez-Montón, Carolina; Fernandez-Figueras, M Teresa; Martí, Mercè; Soler-Botija, Carolina; Roura, Santiago; Perea-Gil, Isaac; Prat-Vidal, Cristina; Llucià-Valldeperas, Aida; Raya, Ángel; Bayes-Genis, Antoni

    2015-01-01

    Engineered bioimplants for cardiac repair require functional vascularization and innervation for proper integration with the surrounding myocardium. The aim of this work was to study nerve sprouting and neovascularization in an acellular pericardial-derived scaffold used as a myocardial bioimplant. To this end, 17 swine were submitted to a myocardial infarction followed by implantation of a decellularized human pericardial-derived scaffold. After 30 days, animals were sacrificed and hearts were analyzed with hematoxylin/eosin and Masson's and Gallego's modified trichrome staining. Immunohistochemistry was carried out to detect nerve fibers within the cardiac bioimplant by using βIII tubulin and S100 labeling. Isolectin B4, smooth muscle actin, CD31, von Willebrand factor, cardiac troponin I, and elastin antibodies were used to study scaffold vascularization. Transmission electron microscopy was performed to confirm the presence of vascular and nervous ultrastructures. Left ventricular ejection fraction (LVEF), cardiac output (CO), stroke volume, end-diastolic volume, end-systolic volume, end-diastolic wall mass, and infarct size were assessed by using magnetic resonance imaging (MRI). Newly formed nerve fibers composed of several amyelinated axons as the afferent nerve endings of the heart were identified by immunohistochemistry. Additionally, neovessel formation occurred spontaneously as small and large isolectin B4-positive blood vessels within the scaffold. In summary, this study demonstrates for the first time the neoformation of vessels and nerves in cell-free cardiac scaffolds applied over infarcted tissue. Moreover, MRI analysis showed a significant improvement in LVEF (P = 0.03) and CO (P = 0.01) and a 43 % decrease in infarct size (P = 0.007). PMID:26205795

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

    Z. Moheb Mohamadi; M. Talebi; B.E. Sayed-Tabatabaei; G. Khaksar

    2014-01-01

    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 intr...

  15. Prolonged viability of human organotypic skin explant in culture method (hOSEC)*

    Frade, Marco Andrey Cipriani; de Andrade, Thiago Antônio Moretti; Aguiar, Andréia Fernanda Carvalho Leone; Guedes, Flávia Araújo; Leite, Marcel Nani; Passos, Williane Rodrigues; Coelho, Eduardo Barbosa; Das, Pranab Kummar

    2015-01-01

    BACKGROUND: Currently, the cosmetic industry is overwhelmed in keeping up with the safety assessment of the increasing number of new products entering the market. To meet such demand, research centers have explored alternative methods to animal testing and also the large number of volunteers necessary for preclinical and clinical tests. OBJECTIVES: This work describes the human skin ex-vivo model (hOSEC: Human Organotypic Skin Explant Culture) as an alternative to test the effectiveness of co...

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

    BABAOĞLU, Mehmet

    2000-01-01

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

  17. Shoot regeneration from leaf explants of Cydonia oblonga cultivars in vitro

    Zalunskaitė, Irena; Kavaliauskaitė, Danguolė; Vinskienė, Jurgita; Revin, Viktor V.; Stanys, Vidmantas

    2007-01-01

    Regeneration experiment was started with aim to develop an efficient plant regeneration system for Cydonia oblonga Mill. cultivars. Variation of organogenic capacity of leaves depending on their position on the microshoot, photoperiod influence on differences in regeneration and interaction of these two factors was investigated in this experiment. Shoots were regenerated from leaf obtained from microshoots culture in vitro. Leaf explants of three genotypes were collected from microshoots and ...

  18. Comparative effects of plant growth regulators on leaf and stem explants of Labisia pumila var. alata

    Ling, Anna Pick Kiong; Tan, Kinn Poay; Hussein, Sobri

    2013-01-01

    Objective: Labisia pumila var. alata, 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. Metho...

  19. The Slice Culture Method for Following Development of Tooth Germs In Explant Culture

    Alfaqeeh, Sarah A; Tucker, Abigail S.

    2013-01-01

    Explant culture allows manipulation of developing organs at specific time points and is therefore an important method for the developmental biologist. For many organs it is difficult to access developing tissue to allow monitoring during ex vivo culture. The slice culture method allows access to tissue so that morphogenetic movements can be followed and specific cell populations can be targeted for manipulation or lineage tracing.

  20. Cells that emerge from embryonic explants produce fibers of type IV collagen

    1985-01-01

    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....

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

    Vanderlocht Joris; Lumeij Stijn BJ; Tonnaer Siebe; van der Stegen Sjoukje JC; Huijskens Mirelle JAJ; Van Elssen Catharina HMJ; Meek Bob; Kirkland Mark A; Hesselink Reinout; Germeraad Wilfred TV; Bos Gerard MJ

    2011-01-01

    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 th...

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

    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.

  3. Electrical and synaptic properties of embryonic luteinizing hormone-releasing hormone neurons in explant cultures.

    Kusano, K.; Fueshko, S; Gainer, H; Wray, S

    1995-01-01

    Voltage- and ligand-activated channels in embryonic neurons containing luteinizing hormone-releasing hormone (LHRH) were studied by patch-pipette, whole-cell current and voltage clamp techniques. LHRH neurons were maintained in explant cultures derived from olfactory pit regions of embryonic mice. Cells were marked intracellularly with Lucifer yellow following recording. Sixty-two cells were unequivocally identified as LHRH neurons by Lucifer yellow and LHRH immunocytochemistry. The cultured ...

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

    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. PMID:26876144

  5. Repair of giant omphalocele in a premature neonate with non-cross-linked porcine acellular dermal matrix (Strattice Tissue Matrix

    Helene Engstrand Lilja

    2016-09-01

    Full Text Available The management of giant omphalocele (GO is a major challenge in pediatric surgery and there are many different surgical strategies described. Here we report a complicated case in which the abdominal wall in a premature neonate (gestational age 33 + 2 weeks and 1700 g with GO was reconstructed with a non-cross-linked acellular porcine dermal matrix (Strattice™ combined with vacuum therapy. This strategy can be an alternative method in the repair of GO in premature neonates with high risk of infection, underdeveloped abdominal cavity and insufficient native tissue.

  6. Root Coverage in Smokers with Acellular Dermal Matrix Graft and Enamel Matrix Derivative: A 12-Month Randomized Clinical Trial.

    Costa, Priscila Paganini; Alves, Luciana Bastos; Souza, Sérgio Luís; Grisi, Márcio Fernando; Palioto, Daniela Bazan; Taba, Mario; Novaes, Arthur Belém

    2016-01-01

    This study investigated whether enamel matrix derivative (EMD) contributes to root coverage of gingival recessions performed with acellular dermal matrix graft (ADMG) in smokers during a 12-month follow-up. A sample of 19 smokers presenting bilateral Miller Class I or II gingival recessions were included. Selected sites randomly received both ADMG and EMD (test) or ADMG alone (control). Probing depth, clinical attachment level, gingival recession height, keratinized tissue, and root coverage were evaluated. Mean gain in recession height (P < .05), sites with complete root coverage (P < .05), and percentage of root coverage (59.7% and 52.8%, respectively) favored the test group compared with the control group. PMID:27333010

  7. Synthesis and physicochemical characterization of a series of hemoglobin-based oxygen carriers: objective comparison between cellular and acellular types.

    Sakai, H; Yuasa, M; Onuma, H; Takeoka, S; Tsuchida, E

    2000-01-01

    A series of hemoglobin (Hb)-based O(2) carriers, acellular and cellular types, were synthesized and their physicochemical characteristics were compared. The acellular type includes intramolecularly cross-linked Hb (XLHb), polyoxyethylene (POE)-conjugated pyridoxalated Hb (POE-PLP-Hb), hydroxyethylstarch-conjugated Hb (HES-XLHb), and glutaraldehyde-polymerized XLHb (Poly-XLHb). The cellular type is Hb-vesicles (HbV) of which the surface is modified with POE (POE-HbV). Their particle diameters are 7 +/- 2, 22 +/- 2, 47 +/- 17, 68 +/- 24, and 224 +/- 76 nm, respectively, thus all the materials penetrate across membrane filters with 0.4 microm pore size, though only the POE-HbV cannot penetrate across the filter with 0.2 microm pore size. These characteristics of permeability are important to consider an optimal particle size in microcirculation in vivo. POE-PLP-Hb ([Hb] = 5 g/dL) showed viscosity of 6.1 cP at 332 s(-1) and colloid osmotic pressure (COP) of 70.2 Torr, which are beyond the physiological conditions (human blood, viscosity = 3-4 cP, COP = ca. 25 Torr). XLHb and Poly-XLHb showed viscosities of 1.0 and 1.5 cp, respectively, which are significantly lower than that of blood. COP of POE-HbV is regulated to 20 Torr in 5% human serum albumin (HSA). HES-XLHb and POE-HbV/HSA showed comparable viscosity with human blood. Microscopic observation of human red blood cells (RBC) after mixing blood with POE-PLP-Hb or HES-XLHb disclosed aggregates of RBC, a kind of sludge, indicating a strong interaction with RBC, which is anticipated to modify peripheral blood flow in vivo. On the other hand, XLHb and POE-HbV showed no rouleaux or aggregates of RBC. The acellular Hbs (P(50) = 14-32 Torr) have their specific O(2) affinities determined by their structures, while that of the cellular POE-HbV is regulated by coencapsulating an appropriate amount of an allosteric effector (e.g., P(50) = 18, 32 Torr). These differences in physicochemical characteristics between the acellular

  8. Bone tumor

    ... physical exam. Tests that may be done include: Alkaline phosphatase blood level Bone biopsy Bone scan Chest x- ... also affect the results of the following tests: Alkaline phosphatase isoenzyme Blood calcium level Parathyroid hormone Blood phosphorus ...

  9. Encapsulation of in vitro-derived explants: an innovative tool for nurseries.

    Standardi, Alvaro; Micheli, Maurizio

    2013-01-01

    The encapsulation technology consists of the inclusion of some millimeter-long plant portions in a nutritive and protective matrix. This technology represents a further and promising tool for exchange of plant material between private and public plant tissue culture laboratories, for short- and medium-term storage of valuable plant material and for use of in vitro-derived or micropropagated propagules directly in farm or in nurseries. After encapsulation, transport, storage and sowing in aseptic conditions, the enclosed explants (capsules) may evolve in shoots (regrowth) and be employed for subsequent micropropagation or culture in vitro. When the encapsulated explant evolves in plantlet (conversion) in in vitro or in vivo conditions, the product of the encapsulation is defined as synthetic seed or artificial seed or synseed. The different evolution of the encapsulated plant material depends on tissue or plant material, genotype, nutritive and culture conditions, and treatments before or after encapsulation. In order to make economical the application of the encapsulation technology in the commercial nursery, research is looking for efficient automation or mechanization of the procedure and for preparation of the encapsulable explants. PMID:23179716

  10. In vitro maintenance of spermatogenesis in Xenopus laevis testis explants cultured in serum-free media

    Spermatogenesis has been maintained for extended periods in Xenopus laevis testis explants cultured in serum-free media supplemented with bovine serum albumin, insulin, transferrin, follicle-stimulating hormone, dihydrotestosterone, testosterone, retinol, ascorbate, and tocopherol. The organization of the testis fragments was maintained for 28 days, and all stages of development were present throughout the culture period. 3H-Thymidine-labeled secondary (Type B) spermatogonia developed in 28 days into spermatids at the acrosomal vesicle stage whereas labeled zygotene spermatocytes became mature spermatids in 28 days. Spermatogonial proliferation also continued in vitro for 28 days. Germ cell differentiation was not dependent upon exogenous testosterone, ascorbate, or tocopherol since 3H-labeled spermatogonia became mature spermatids in testes cultured 35 days in media lacking these supplements. Autoradiography demonstrated that 55% of the luminal sperm present in explants cultured 10 days had differentiated in vitro. Sperm from testes cultured 10-35 days were similar to sperm from freshly dissected testes with regard to motility and fecundity, and eggs fertilized with sperm from explant cultures developed normally into swimming tadpoles. The results demonstrate the feasibility of maintaining vertebrate spermatogenesis in culture and suggest that in vitro analysis of Xenopus spermatogenesis using defined media may provide important insights into the evolution of regulatory mechanisms in spermatogenesis

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

    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.

  12. Spinal cord explants use carbon nanotube interfaces to enhance neurite outgrowth and to fortify synaptic inputs.

    Fabbro, Alessandra; Villari, Ambra; Laishram, Jummi; Scaini, Denis; Toma, Francesca M; Turco, Antonio; Prato, Maurizio; Ballerini, Laura

    2012-03-27

    New developments in nanotechnology are increasingly designed to modulate relevant interactions between nanomaterials and neurons, with the aim of exploiting the physical properties of synthetic materials to tune desired and specific biological processes. Carbon nanotubes have been applied in several areas of nerve tissue engineering to study cell behavior or to instruct the growth and organization of neural networks. Recent reports show that nanotubes can sustain and promote electrical activity in networks of cultured neurons. However, such results are usually limited to carbon nanotube/neuron hybrids formed on a monolayer of dissociated brain cells. In the present work, we used organotypic spinal slices to model multilayer tissue complexity, and we interfaced such spinal segments to carbon nanotube scaffolds for weeks. By immunofluorescence, scanning and transmission electronic microscopy, and atomic force microscopy, we investigated nerve fiber growth when neuronal processes exit the spinal explant and develop in direct contact to the substrate. By single-cell electrophysiology, we investigated the synaptic activity of visually identified ventral interneurons, within the ventral area of the explant, thus synaptically connected, but located remotely, to the substrate/network interface. Here we show that spinal cord explants interfaced for weeks to purified carbon nanotube scaffolds expand more neuronal fibers, characterized by different mechanical properties and displaying higher growth cones activity. On the other hand, exploring spontaneous and evoked synaptic activity unmasks an increase in synaptic efficacy in neurons located at as far as 5 cell layers from the cell-substrate interactions. PMID:22339712

  13. Heparin exerts anti-apoptotic effects on uterine explants by targeting the endocannabinoid system.

    Salazar, Ana Inés; Vercelli, Claudia; Schiariti, Victoria; Davio, Carlos; Correa, Fernando; Franchi, Ana María

    2016-09-01

    Miscarriage caused by Gram-negative bacteria infecting the female genital tract is one of the most common complications of human pregnancy. Intraperitoneal administration of LPS to 7-days pregnant mice induces embryo resorption after 24 h. Here, we show that LPS induced apoptosis on uterine explants from 7-days pregnant mice and that CB1 receptor was involved in this effect. On the other hand, heparin has been widely used for the prevention of pregnancy loss in women with frequent miscarriage with or without thrombophilia. Besides its anticoagulant properties, heparin exerts anti-inflammatory, immunomodulatory and anti-apoptotic effects. Here, we sought to investigate whether the administration of heparin prevented LPS-induced apoptosis in uterine explants from 7-days pregnant mice. We found that heparin enhanced cell survival in LPS-treated uterine explants and that this effect was mediated by increasing uterine FAAH activity. Taken together, our results point towards a novel mechanism involved in the protective effects of heparin. PMID:27364950

  14. Effect of Macrophage Migration Inhibitory Factor (MIF) in Human Placental Explants Infected with Toxoplasma gondii Depends on Gestational Age

    de Oliveira Gomes, Angelica; de Oliveira Silva, Deise Aparecida; Silva, Neide Maria; de Freitas Barbosa, Bellisa; Franco, Priscila Silva; Angeloni, Mariana Bodini; Fermino, Marise Lopes; Roque-Barreira, Maria Cristina; Bechi, Nicoletta; Paulesu, Luana Ricci; dos Santos, Maria Célia; Mineo, José Roberto; Ferro, Eloisa Amália Vieira

    2011-01-01

    Because macrophage migration inhibitory factor (MIF) is a key cytokine in pregnancy and has a role in inflammatory response and pathogen defense, the objective of the present study was to investigate the effects of MIF in first- and third-trimester human placental explants infected with Toxoplasma gondii. Explants were treated with recombinant MIF, IL-12, interferon-γ, transforming growth factor-β1, or IL-10, followed by infection with T. gondii RH strain tachyzoites. Supernatants of cultured...

  15. In vitro morphogenic response of different explants of Gentiana kurroo Royle from Western Himalayas—an endangered medicinal plant

    Sharma, Anshu; Kaur, Rajinder; Sharma, Neha

    2014-01-01

    Micropropagation offers a great potential to produce millions of clonal individuals through tissue culture via induction of morphogenesis. The aim of this work was to obtain an efficient protocol for callus regeneration for Gentiana kurroo Royle. The morphogenic response of different explants (leaves, petioles, roots) varied and responded differently for regeneration according to combinations of growth regulators. The petiole explants were best responding for callus induction and subsequently...

  16. Selection of valine-resistance in callus culture of Arabidopsis thaliana (L.) Heynh. derived from leaf explants

    Małgorzata D. Gaj; Grzegorz Czaja; Małgorzata Nawrot

    2014-01-01

    The selection of valine-resistant mutants was carried out in leaf explant cultures of three Arabidopsis thaliana (L.) Heynh. ecotypes: C-24, RLD and Columbia. The valine concentration used for in vitro selection, lethal for seed-growing plants, has not affected callus formation and growth. However, strong inhibition of shoot regeneration ability of calli growing under selection pressure was noticed. In total, 1043 explants were cultured on valine medium and 18 shoots were regenerated with an ...

  17. PATTERN OF PLANT REGENERATION FROM SHOOT TIP EXPLANTS OF PIGEONPEA (CAJANUS CAJAN L MILLLSP VAR LRG-41

    T. Raghavendra

    2014-02-01

    Full Text Available An efficient direct shoot bud differentiation and multiple shoot induction from shoot tip explants of pigeon pea (Cajanus cajan L. has been achieved. The frequency of shoot bud regeneration was influenced by the type of explants, genotype and concentrations of cytokinin. Explants viz. shoot tip isolated from 10 day old seedlings showed better explants response Explants were cultured on Murashige and skoog (MS medium augmented with different concentrations of BAP and NAA. Among the various concentrations tested, 2.0mg/l BAP (Benzyl amino purine and 0.1 mg/l Napthalene acetic acid (NAA were found to be the best for maximum shoot bud differentiation. Percentage, as well as the number of shoots per explant showing differentiation of shoot buds was higher on MS media supplement with BAP and optimal BAP concentration for shoot regeneration was 2mg/l. The elongated shoots were successfully rooted on MS medium containing different concentrations of auxins. Among them indole buteric acid (IBA at 1.0mg/l induced maximum frequency of rooting. Regenerated plants were successfully established in soil where 91% of them have been developed into morphologically normal and fertile plants. This method can thus be advantageously applied in the production of transgenic pigeon pea plants.

  18. PATTERN OF PLANT REGENERATION FROM SHOOT TIP EXPLANTS OF PIGEONPEA (CAJANUS CAJAN L MILLLSP VAR LRG-41

    T. Raghavendra

    2014-03-01

    Full Text Available An efficient direct shoot bud differentiation and multiple shoot induction from shoot tip explants of pigeon pea (Cajanus cajan L. has been achieved. The frequency of shoot bud regeneration was influenced by the type of explants, genotype and concentrations of cytokinin. Explants viz. shoot tip isolated from 10 day old seedlings showed better explants response Explants were cultured on Murashige and skoog (MS medium augmented with different concentrations of BAP and NAA. Among the various concentrations tested, 2.0mg/l BAP (Benzyl amino purine and 0.1 mg/l Napthalene acetic acid (NAA were found to be the best for maximum shoot bud differentiation. Percentage, as well as the number of shoots per explant showing differentiation of shoot buds was higher on MS media supplement with BAP and optimal BAP concentration for shoot regeneration was 2mg/l. The elongated shoots were successfully rooted on MS medium containing different concentrations of auxins. Among them indole buteric acid (IBA at 1.0mg/l induced maximum frequency of rooting. Regenerated plants were successfully established in soil where 91% of them have been developed into morphologically normal and fertile plants. This method can thus be advantageously applied in the production of transgenic pigeon pea plants.

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

    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

  20. In Vitro Shoot Regeneration of NAA-Pulse Treated Plumular Leaf Explants of Cowpea

    Muhammad AASIM

    2010-06-01

    Full Text Available Cowpea (Vigna unguiculata L. is an economically important grain legume crop and is an important source of dietary protein in many of the developing countries. The present study reports the effect of pulse treatment duration, concentration of NAA and presence of NAA in the culture medium on shoot regeneration from plumular leaf explant of Turkish cowpea cv. �Akkiz� and �Karagoz�. Pulse treatment of mature embryos with 20 mg l-1 NAA for 1 and 3 weeks followed by culturing of plumular leaf explant on MS medium containing 0.25, 0.50 and 1.0 BAP with 1.0, 2.0 and 4.0 mg l-1 NAA promoted somatic embryogenesis in both cultivars. Longer duration of pulse treatment was deleterious resulting in browning and consequently death of the embryos on explants. Pulse treatment with 20 mg l-1 NAA for one week was less deleterious and developed two plantlets after the explants were transferred to MS0 medium after 6 weeks through somatic embryogenesis in cv. �Akkiz�. Pulse treatment with 10 mg l-1 NAA for 1 week showed 33.33-50.00 % and 25.00-50.00% shoot regeneration frequency in cv. �Akkiz� and �Karagoz� respectively on MS medium containing 0.25-1.00 mg l-1 BAP. Maximum number of 2.50 shoots each per explant were recorded in cv. �Akkiz� and �Karagoz� on MS medium containing 1.00 and 0.50 mg l-1 BAP respectively. Contrarily, maximum shoot length of 8.98 cm of cv. �Akkiz� and 9.42 cm of cv. �Karagoz� was recorded on MS medium containing 0.50 mg l-1 BAP and 1.00 mg l-1 BAP respectively. Regenerated shoots were rooted on MS medium containing 0.5 mg l-1 IBA and and acclimatized in growth room at room temprature where they produced viable seeds.

  1. In Vivo Bone Formation Within Engineered Hydroxyapatite Scaffolds in a Sheep Model.

    Lovati, A B; Lopa, S; Recordati, C; Talò, G; Turrisi, C; Bottagisio, M; Losa, M; Scanziani, E; Moretti, M

    2016-08-01

    Large bone defects still represent a major burden in orthopedics, requiring bone-graft implantation to promote the bone repair. Along with autografts that currently represent the gold standard for complicated fracture repair, the bone tissue engineering offers a promising alternative strategy combining bone-graft substitutes with osteoprogenitor cells able to support the bone tissue ingrowth within the implant. Hence, the optimization of cell loading and distribution within osteoconductive scaffolds is mandatory to support a successful bone formation within the scaffold pores. With this purpose, we engineered constructs by seeding and culturing autologous, osteodifferentiated bone marrow mesenchymal stem cells within hydroxyapatite (HA)-based grafts by means of a perfusion bioreactor to enhance the in vivo implant-bone osseointegration in an ovine model. Specifically, we compared the engineered constructs in two different anatomical bone sites, tibia, and femur, compared with cell-free or static cell-loaded scaffolds. After 2 and 4 months, the bone formation and the scaffold osseointegration were assessed by micro-CT and histological analyses. The results demonstrated the capability of the acellular HA-based grafts to determine an implant-bone osseointegration similar to that of statically or dynamically cultured grafts. Our study demonstrated that the tibia is characterized by a lower bone repair capability compared to femur, in which the contribution of transplanted cells is not crucial to enhance the bone-implant osseointegration. Indeed, only in tibia, the dynamic cell-loaded implants performed slightly better than the cell-free or static cell-loaded grafts, indicating that this is a valid approach to sustain the bone deposition and osseointegration in disadvantaged anatomical sites. PMID:27075029

  2. The initial attachment of cemental fibrils to the root dentin surface in acellular and cellular cementogenesis in rat molars.

    Yamamoto, T; Domon, T; Takahashi, S; Islam, M N; Suzuki, R

    2001-03-01

    To elucidate the initial attachment mechanism of cemental fibrils to the root dentin surface in acellular and cellular cementogenesis, developing rat molars were observed by light microscopy and scanning electron microscopy combined with NaOH maceration. The NaOH maceration was used to observe details of the positional association of cemental and dentinal fibrils during cementogenesis. An initial hematoxylin stained, cementum layer began to form on the root dentin surface with the first dentin mineralization in both acellular and cellular cementogenesis. The initial attachment of cemental fibrils to the dentin surface also began at this point. At the initial attachment the intermingling of cemental and dentinal fibrils occurred only in places. With advanced cementogenesis the initial cementum layer became the fibril-poor cemento-dentinal junction. This suggests that cemental fibrils attach on the initial cementum layer, and not directly on dentinal fibrils, so that the layer results in the fibril-poor cemento-dentinal junction. The present study suggests that an intervening adhesive is necessary for the cemento-dentinal attachment at any stage of cementogenesis in rat molars. PMID:11325058

  3. A dynamic distention protocol for whole-organ bladder decellularization: histological and biomechanical characterization of the acellular matrix.

    Consolo, F; Brizzola, S; Tremolada, G; Grieco, V; Riva, F; Acocella, F; Fiore, G B; Soncini, M

    2016-02-01

    A combined physical-chemical protocol for whole full-thickness bladder decellularization is proposed, based on organ cyclic distention through repeated infusion/withdrawal of the decellularization agents through the urethra. The dynamic decellularization was intended to enhance cell removal efficiency, facilitating the delivery of detergents within the inner layers of the tissue and the removal of cell debris. The use of mild chemical detergents (hypotonic solution and non-ionic detergent) was employed to limit adverse effects upon matrix 3D ultrastructure. Inspection of the presence of residual DNA and RNA was carried out on decellularized matrices to verify effective cell removal. Histological investigation was focused on assessing the retention of adequate structural and functional components that regulate the biomechanical behaviour of the acellular tissue. Biomechanical properties were evaluated through uniaxial tensile loading tests of tissue strips and through ex vivo filling cystometry to evaluate the whole-organ mechanical response to a physiological-like loading state. According to our results, a dynamic decellularization protocol of 17 h duration with a 5 ml/min detergent infusion flow rate revealed higher DNA removal efficiency than standard static decellularization, resulting in residual DNA content < 50 ng/mg dry tissue weight. Furthermore, the collagen network and elastic fibres distribution were preserved in the acellular ECM, which exhibited suitable biomechanical properties in the perspective of its future use as an implant for bladder augmentation. Copyright © 2013 John Wiley & Sons, Ltd. PMID:23737121

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

    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.

  5. Surgical Outcomes of Deep Superior Sulcus Augmentation Using Acellular Human Dermal Matrix in Anophthalmic or Phthisis Socket.

    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. PMID:27258711

  6. In Vivo Confocal Microscopic Observation of Lamellar Corneal Transplantation in the Rabbit Using Xenogenic Acellular Corneal Scaffolds as a Substitute

    Yun Feng; Wei Wang

    2015-01-01

    Background:The limiting factor to corneal transplantation is the availability of donors.Research has suggested that xenogenic acellular corneal scaffolds (XACS) may be a possible alternative to transplantation.This study aimed to investigate the viability of performing lamellar corneal transplantation (LCT) in rabbits using canine XACS.Methods:Fresh dog corneas were decellularized by serial digestion,and LCT was performed on rabbit eyes using xenogeneic decellularized corneal matrix.Cellular and morphological changes were observed by slit-lamp,light,and scanning electron microscopy at 7,30 and 90 days postoperatively.Immunocytochemical staining for specific markers such as keratin 3,vimentin and MUC5AC,was used to identify cells in the graft.Results:Decellularized xenogenic corneal matrix remained transparent for about 1-month after LCT.The recipient cells were able to survive and proliferate into the grafts.Three months after transplantation,grafts had merged with host tissue,and graft epithelialization and vascularization had occurred.Corneal nerve fibers were able to grow into the graft in rabbits transplanted with XACS.Conclusions:Xenogenic acellular corneal scaffolds can maintain the transparency of corneal grafts about 1-month and permit growth of cells and nerve fibers,and is,therefore,a potential substitute or carrier for a replacement cornea.

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

    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

  8. Mandibular Jaw Bone Regeneration Using Human Dental Cell-Seeded Tyrosine-Derived Polycarbonate Scaffolds.

    Zhang, Weibo; Zhang, Zheng; Chen, Shuang; Macri, Lauren; Kohn, Joachim; Yelick, Pamela C

    2016-07-01

    Here we present a new model for alveolar jaw bone regeneration, which uses human dental pulp cells (hDPCs) combined with tyrosine-derived polycarbonate polymer scaffolds [E1001(1k)] containing beta-tricalcium phosphate (β-TCP) [E1001(1k)/β-TCP]. E1001(1k)/β-TCP scaffolds (5 mm diameter × 1 mm thickness) were fabricated to fit a 5 mm rat mandibular ramus critical bone defect. Five experimental groups were examined in this study: (1) E1001(1k)/β-TCP scaffolds seeded with a high density of hDPCs, 5.0 × 10(5) hDPCs/scaffold (CH); (2) E1001(1k)/β-TCP scaffolds seeded with a lower density of hDPCs, 2.5 × 10(5) hDPCs/scaffold (CL); (3) acellular E1001(1k)/β-TCP scaffolds (SA); (4) acellular E1001(1k)/β-TCP scaffolds supplemented with 4 μg recombinant human bone morphogenetic protein-2 (BMP); and (5) empty defects (EDs). Replicate hDPC-seeded and acellular E1001(1k)/β-TCP scaffolds were cultured in vitro in osteogenic media for 1 week before implantation for 3 and 6 weeks. Live microcomputed tomography (μCT) imaging at 3 and 6 weeks postimplantation revealed robust bone regeneration in the BMP implant group. CH and CL groups exhibited similar uniformly distributed mineralized tissue coverage throughout the defects, but less than the BMP implants. In contrast, SA-treated defects exhibited sparse areas of mineralized tissue regeneration. The ED group exhibited slightly reduced defect size. Histological analyses revealed no indication of an immune response. In addition, robust expression of dentin and bone differentiation marker expression was observed in hDPC-seeded scaffolds, whereas, in contrast, BMP and SA implants exhibited only bone and not dentin differentiation marker expression. hDPCs were detected in 3-week but not in 6-week hDPC-seeded scaffold groups, indicating their survival for at least 3 weeks. Together, these results show that hDPC-seeded E1001(1k)/β-TCP scaffolds support the rapid regeneration of osteo

  9. Primary vaccination of adults with reduced antigen-content diphtheria-tetanus-acellular pertussis or dTpa-inactivated poliovirus vaccines compared to diphtheria-tetanus-toxoid vaccines.

    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

  10. Dermal fat graft from simultaneous abdominoplasty as an adjunct to revision aesthetic and reconstructive breast surgery: A poor man's acellular dermal matrix?

    F. Xie

    2014-01-01

    CONCLUSION: We herein report the use of free dermal fat graft in revision aesthetic and reconstructive surgery in a manner akin to recent acellular dermal matrix use. The comparable enhanced aesthetic outcomes, minimal complication rate and substantial cost savings merit dissemination to a global audience and encourage surgeons to consider this economic alternative.

  11. Clinical Outcomes for Breast Cancer Patients Undergoing Mastectomy and Reconstruction with Use of DermACELL, a Sterile, Room Temperature Acellular Dermal Matrix

    Christopher Vashi

    2014-01-01

    Full Text Available Background. Decellularized human skin has been used in a variety of medical applications, primarily involving soft tissue reconstruction, wound healing, and tendon augmentation. Theoretically, decellularization removes potentially immunogenic material and provides a clean scaffold for cellular and vascular in growth. The use of acellular dermal matrix in two-stage postmastectomy breast reconstruction is described. Methods. Ten consecutive breast cancer patients were treated with mastectomies and immediate reconstruction from August to November 2011. There were 8 bilateral and 1 unilateral mastectomies for a total of 17 breasts, with one exclusion for chronic tobacco use. Reconstruction included the use of a new 6 × 16 cm sterile, room temperature acellular dermal matrix patch (DermACELL soaked in a cefazolin bath. Results. Of the 17 breasts, 15 reconstructions were completed; 14 of them with expander to implant sequence and acellular dermal matrix. Histological analysis of biopsies obtained during trimming of the matrix at the second stage appeared nonremarkable with evidence of normal healing, cellularity, and vascular infiltration. Conclusion. Postoperative observations showed that this cellular dermal matrix appears to be an appropriate adjunct to reconstruction with expanders. This acellular dermal matrix appeared to work well with all patients, even those receiving postoperative chemotherapy, postoperative radiation, prednisone, or warfarin sodium.

  12. Design and in vivo evaluation of a molecularly defined acellular skin construct: reduction of early contraction and increase in early blood vessel formation

    Nillesen, S.T.M.; Lammers, G.; Wismans, R.; Ulrich, M.M.; Middelkoop, E.; Spauwen, P.H.M.; Faraj, K.A.; Schalkwijk, J.; Daamen, W.F.; Kuppevelt, A.H.M.S.M. van

    2011-01-01

    Skin substitutes are of great benefit in the treatment of patients with full thickness wounds, but there is a need for improvement with respect to wound closure with minimal contraction, early vascularisation, and elastin formation. In this study we designed and developed an acellular double-layered

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

    Małgorzata Podwyszyńska

    2011-04-01

    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.

  14. Influência do cloreto de cálcio no crescimento de explantes de Gypsophila paniculata L. (Caryophyllaceae, cultivados in vitro Influence of calcium chloride on the growth of Gypsophila paniculata L. (Caryophyllaceae explants, cultivated in vitro

    R. Jun Takane

    1994-08-01

    Full Text Available Este trabalho teve por finalidade verificar a influência do Cloreto de Cálcio (CaCl12 no crescimento de explantes de Gypsophila paniculata L., cultivados em meio de cultura a fim de fornecer subsídios para a micropropagação desta cultura. Foram utilizados como explantes somente as gemas apicais das plantas em fase de crescimento vegetativo. Os explantes foram inoculados em meio de cultura MS (Murashigue & Skoog modificado com diferentes concentrações de CaCl2: a 440; b 880; c 1.760; d 2.640 e e 3.520 mg/1. Os explantes foram deixados em câmara de crescimento sob uma temperatura constante de 25°C ± 2°C sob fotoperíodo de 16 horas. Os explantes do meio de cultura MS com 1.760 mg/1 de CaCl2 foram os de melhor resultado, com crescimento vigoroso e presença de coloração verde intensa nas folhas. Os explantes do meio de cultura MS com 440 mg/1 de CaCl2 não apresentaram um crescimento satisfatório, com estiolamento e coloração verde pouco intensa nas folhas; os explantes do meio de cultura MS com 3.520 mg/1 de CaCl2, também não apresentaram um crescimento satisfatório, pois com 10 dias de inoculação já se percebia uma menor indução no crescimento, podendo tal efeito ser conseqüência de dois fatores: excesso de cálcio no meio de cultura ou a toxidez ocasionada pelo cloro.Shoot tips from plants still in the vegetative growth phase were used as explants. They were sterilised with tetraciclin 1% for four minutes, sodium hypochloridre (commercial sanitary water with 5% of active cloride 20% (v/v for 15 minutes and alcohol 70 GL for 2 minutes. The explants were inoculated in a growth medium MS (Murashigue & Skoog, with different concentrations of CaCl2: a 440; b 880; c 1,760; d 2,640 and e 3,520 mg/1. Explants were kept in a growth chamber at constant temperature (25°C ± 2°C with a photo period of 16 hours. The explants in the medium modified with 1,760 mg/1 of CaCl2, presented the best results, with a vigorous growth and

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

    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.

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

    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.

  17. Rapid in vitro multiplication of the ethnomedicinal shrub, Acacia caesia (L.) Willd. (Mimosaceae) from leaf explants

    Thambiraj J; Paulsamy S

    2012-01-01

    Objective: To develop an efficient protocol for in vitro multiplication of the ethnomedicinal shrub Acacia caesia (A. caesia) L. Willd., Methods: Leaf explants were inoculated on MS medium supplemented with TDZ and NAA for callus induction. Subculturing experiments were conducted by using leaf derived calli for shoot proliferation on MS medium fortified with various growth regulators like IBA, TDZ, BAP and GA3. The regenerated shoots were rooted in half strength MS medium supplemented with various concentrations of IBA, IAA and Kn. After roots were developed, the plantlets were transplanted to pots filled with garden soil, sand and vermicompost and kept in growth chamber with 70%-80%humidity under16h photoperiod. After acclimatization, the plantlets were transferred to the garden and survival percentage was calculated. Data were statistically analyzed and means were compared using Duncan’s Multiple Range Test (P<0.05). Results: An in vitro multiplication protocol was developed for the locally demanded medicinal plant species, A. caesia by using leaf explant. The study revealed that the callus formation was effective in MS medium containing TDZ and NAA at 1.5 and 0.3 mg/L respectively. Shoot induction was most successful in MS medium supplemented with combination of the auxin, IBA and cytokinin, TDZ at 2.0 and 0.5 mg/L respectively. A single leaf explant was capable of producing 12 shoots/callus after 30 days of culture. The other supplementation in MS medium with IBA and Kn at 2.0 and 0.4 mg/L respectively produced higher rooting frequency, roots/shoot and root length. The survivability rate of leaf callus derived plantlets was significantly higher (84%) in the hardening medium composed by garden soil, sand and vermicompost (1:1:1) by volume. Conclusions: A significant progress has been made in the in vitro regeneration system of this medicinally important plant species, A.caesia.

  18. [Bone diseases].

    Uebelhart, Brigitte; Rizzoli, René

    2016-01-13

    Calcium intake shows a small impact on bone mineral density and fracture risk. Denosumab is a more potent inhibitor of bone resorption than zoledronate. Abaloparatide, PTHrP analog, increases bone mineral density and decreases fracture incidence. Teriparatide could be delivered via a transdermic device. Romosozumab and odanacatib improve calculated bone strength. Sequential or combined treatments with denosumab and teriparatide could be of interest, but not denosumab followed by teriparatide. Fibrous dysplasia, Paget disease and hypophosphatasia are updated, as well as atypical femoral fracture and osteonecrosis of the jaw. PMID:26946704

  19. Micropropagation of Origanum acutidens (HAND.-MAZZ.) IETSWAART Using Stem Node Explants

    Mehmet Ugur Yildirim

    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 prolifer...

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

    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.

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

    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.

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

    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

  3. IgG4-positive plasma cell infiltration in explanted livers for primary sclerosing cholangitis

    Zen, Yoh; Quaglia, Alberto; Portmann, Bernard

    2011-01-01

    Abstract Aims: To explore whether IgG4-related sclerosing cholangitis (IgG4-SC) contributes to end-stage primary sclerosing cholangitis (PSC) in UK. Methods and Results: This study consisted of 41 patients who underwent liver transplantation for advanced PSC. Explanted livers were histologically examined with an emphasis on IgG4-positive plasma cell infiltration. Thirty-nine cases (95%) had minimal or mild infiltration of IgG4-positive plasma cells (? 30 cells/high power field)...

  4. Adventitious shoot regeneration in Sesame (Sesamum indicum L.) (Pedaliaceae) via deembryonated cotyledonary explants

    Shilpa V. Malaghan; Ramappa Lokesha; Revadi Savitha; Ajjanavara R. G. Ranganatha

    2013-01-01

    Adventitious shoot regeneration in sesame, via de-embryonated cotyledonary explants, was attempted using six diverse genotypes; two were land races (Kankapura Local – KNL and Tamil Nadu Local- TNL), three were released varieties (DS-1, DSS-9 and WII) and a breeding line (RT-273). Three media compositions (½ MS basal media supplemented with 20 μM TDZ + 2.5 μM IAA and with or without 25 μM BAP and full MS basal media supplemented with only 25 μM BAP) were used of which highest shoot regeneratio...

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

    Alireza Bonyanpour; Morteza Khosh-Khui

    2013-01-01

    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 o...

  6. Radiation-induced outgrowth inhibition in explant cultures from surgical specimens of five human organs

    Mothersill, Carmel; Cusack, Anne; Seymour, C.B.

    1988-03-01

    An explant outgrowth technique to determine the radiation response of five different human organs (bladder, oesophagus, colon, breast and thyroid) is described. In each case except thyroid, where malignancies are rare, data are presented for normal and malignant tissue. Results show that variations in response, consistent with those observed in vivo, can be measured. Tumours were in all cases highly resistant to radiation relative to their corresponding normal tissue. Possible reasons for this are discussed. The method may prove useful in the prediction of the radiobiological response for tumour and surrounding normal tissue where post-operative therapy is planned.

  7. Progesterone regulates the activity of collagenase and related gelatinases A and B in human endometrial explants.

    Marbaix, Etienne; Donnez, Jacques; Courtoy, Pierre J; Eeckhout, Yves

    1992-01-01

    Explants of human endometrium were cultured to study the release of matrix metalloproteinases (MMPs). Analysis of conditioned media by zymography revealed latent and active forms of collagenase (MMP-1, EC 3.4.24.7), 72-kDa gelatinase A (MMP-2, EC 3.4.24.24), and 92-kDa gelatinase B (MMP-9, EC 3.4.24.35). These proteinases were identified by their M(r), their inhibition by tissue inhibitor of metalloproteinases, and the activation of their zymogens by trypsin or aminophenylmercuric acetate. In...

  8. Boron neutron capture therapy for an explanted organ: The logistical challenges

    Single liver metastases of colorectal cancer can be cured by surgery; disseminated liver metastases are incurable. A research group in Pavia, Italy, used BNCT as an experimental method to irradiate in curative intention the explanted liver of patients suffering from disseminated hepatic metastases. The situation in Pavia, where a reactor with a specially adapted thermal column and the hospital are close by, is unique. For the purpose of the present study, it was necessary to investigate how the Pavia experience can be repeated with transplantation centers located at distance from a reactor. Some basic investigations of the logistics of such a procedure are reported.

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

    Owk ANIEL KUMAR; Sape SUBBA TATA

    2010-01-01

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

  10. Boron neutron capture therapy for an explanted organ: The logistical challenges

    Wittig, A. [Department of Radiation Oncology, University Hospital Essen, University Duisburg-Essen, Hufelandstrasse 55, Essen (Germany)], E-mail: andrea.wittig@uni-due.de; Moss, R. [HFR Unit, Institute for Energy, Joint Research Centre, European Commission, Westerduinweg 3, Petten (Netherlands); Kaiser, G.M. [Department of General-, Visceral- and Transplantation Surgery, University Hospital Essen, University Duisburg-Essen, Hufelandstrasse 55, Essen (Germany); Malago, M. [Department of Surgery-UCL Division of Surgical and Interventional Sciences, University College London-University of London, 74 Huntley Street, London (United Kingdom); Nievaart, V. [HFR Unit, Institute for Energy, Joint Research Centre, European Commission, Westerduinweg 3, Petten (Netherlands); Sauerwein, W.A. [Department of Radiation Oncology, University Hospital Essen, University Duisburg-Essen, Hufelandstrasse 55, Essen (Germany)

    2009-07-15

    Single liver metastases of colorectal cancer can be cured by surgery; disseminated liver metastases are incurable. A research group in Pavia, Italy, used BNCT as an experimental method to irradiate in curative intention the explanted liver of patients suffering from disseminated hepatic metastases. The situation in Pavia, where a reactor with a specially adapted thermal column and the hospital are close by, is unique. For the purpose of the present study, it was necessary to investigate how the Pavia experience can be repeated with transplantation centers located at distance from a reactor. Some basic investigations of the logistics of such a procedure are reported.