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Objective: To study the effect of chemical extraction of allogeneic tendon and allogeneic chondrocytes for reconstruction of anterior labrum of shoulder joint in rabbits. Methods: The body weight of 45 adult New Zealand white rabbits ranged from 2.5 to 3.0 kg. The Achilles tendons of 15 rabbits were taken and the allogeneic tendons were prepared by chemical extraction with antigen inactivation. The extracted tendons were compared with untreated tendons by HE and Masson stainings. Chondrocytes were isolated and cultured by trypsin method and identified by immunohistochemical staining of collagen type Ⅱ. The remaining 30 rabbits were used to prepare the model of anterior labrum defect of shoulder joint. After the allogeneic tendon was transplanted to the damaged labrum, the rabbits was randomly divided into two groups (15 in each group). In group A, the allogeneic chondrocytes were injected into the joint immediately after transplantation, while in group B, no treatment was made. At 4, 6, and 8 weeks after operation, 5 transplanted tendons of each group were taken. After general observation, HE staining was used to observe the number of nuclei, Masson staining was used to observe the expression of collagen fibers in muscle fiber tissues, and AB staining was used to detect the glycosaminoglycan level after transplantation, to evaluate the cell growth in the tissues of the two groups of allogeneic tendon. Results: By HE and Masson stainings, the allogeneic tendon antigen prepared by chemical extraction method was inactivated and the fibrous tissue structure was intact; collagen type Ⅱ immunohisto-chemistry staining showed that the cultured cells were chondrocytes. After tendon transplantation, the content of glycosaminoglycan in group A was significantly higher than that in group B ( P<0.05). At 6 weeks after operation, HE staining showed that the nuclear in tendon tissue of group A was significantly more than that of group B ( t=20.043, P=0.000). Masson staining showed that the number of nuclei in tendon tissue of group A was significantly increased, the muscle fibers and collagen fibers were interlaced, the tissue structure was more compact, and the tendon tissue was mainly blue stained; while the number of nuclei in group B was less, mainly collagen fibers of the original graft. Conclusion: The allogeneic tendon inactivated by chemical extraction can be used to reconstruct the defect of anterior labrum of shoulder joint in rabbits, and the combination of allogeneic chondrocytes can promote the healing of tendon transplantation.
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The chemical extraction method was used to prepare the rat uterine decellularized scaffolds, and to investigate the feasibility of preparing the extracellular matrix (ECM) hydrogel. The rat uterus were collected and extracted by 1%sodium dodecyl sulfate (SDS), 3% TritonX-100 and 4% sodium deoxycholate (SDC) in sequence. Scanning electron microscopy, histochemical staining and immunohistochemistry was used to assess the degree of decellularization of rat uterine scaffold. The prepared decellularized scaffold was digested with pepsin to obtain a uterine ECM hydrogel, and the protein content of ECM was determined by specific ELISA kit. Meanwhile, the mechanical characteristic of ECM hydrogel was measured. The results showed that the chemical extraction method can effectively remove the cells effectively in the rat uterine decellularized scaffold, with the ECM composition preserved completely. ECM hydrogel contains a large amount of ECM protein and shows a good stability, which provides a suitable supporting material for the reconstruction of endometrium .
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Objective To explore a method of preparing the acellular tendons of human with chemical approaches.Methods From April 2011 to June 2012,several treatments were performed on human tendons using 1.00％ tri(n-butyl) Phosphate (TnBP) combinded with Triton X-100 at the concentration of 0,0.25％,0.50％,1.00％ respectively.Specimens were examined using histological observation,scanning electron microscopy,biomechanical testing and hydroxyproline quantitation.The results were then compared with fresh tendons of control group,so as to value the characteristics of decellularized human tendon scaffold.Results Acellular human tendons had glossy surface,intact aponeurotic membrane and satisfactory flexibility.A small number of disrupted cells remained in the 1.00％ TnBP + 0％ Triton X-100 treated tissue,while other three experimental groups successfully eliminate all cells.Intact and regular collagen architecture was retained in 1.00％ TnBP +0.25％ Triton X-100 treated tissue.1.00％ TnBP + 0.50％ Triton X-100 and 1.00％ TnBP + 1.00％ Triton X-100 treated tissue were nearly identical to 1.00％ TnBP +0.25％ Triton X-100 treated tissue,but the interval of collagen was slightly wider than the control group,the maximum load (385.22 ± 80.32N,398.22 ± 127.20N),ultimate tensile strength(46.69 ± 16.30Mpa,46.20 ±5.52Mpa) and hydroxyproline content(0.282663 ± 0.0110109 μg/mg,0.279355 ± 0.0102129 μg/mg) were statistically lower (P ＜ 0.05) compared with those of the control group,maximum load(533.28 ± 135.77N),ultimate tensile strength (65.56 ± 14.40Mpa) and hydroxyproline content (0.292882 ± 0.0100988 μg/mg) respectively.Conclusion The decellularization treatment with 1.00％ TnBP + 0.25％ Triton X-100 could be optimized for preparing acellular human tendons.
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0.05).There were better effect of removal of myelin(P2
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Objective To look for an ideal substance to repair large gap of nerve defect after injuries by culture,population of Schwann cells(Scs)and preparation of acellular allogenous nerve grafts with chemical extraction. Method The double adhesion culture and Arab c to prohibit the fibroblast growth were used to achieve high purified Scs;detergents:Triton X 100 and sodium deoxycholate were used to achieve acellural nerve grafts (ANG);and finally the Scs were micro injected into the acellural nerve grafts and the consequence studied Result By the way above,high purified Scs and the ANG was acquired,which can integrated each other well Scs can survived and transformed to aline in vitro Conclusion ANG populated isogenous Scs may be an ideal substance to repair the large gap of nerve defect after injuries