【Abstract】Objective To investigate the effects of human interlukin-13 (hIL-13) on the expression of E-selectin and intercellular adhesion molecule-1(ICAM-1) on bovine aortic endothelial cells(BAECs) stimulated by tumor necrosis factor alpha(TNF-α), and to provide experimental basis for hIL-13 inducing immunity endure and relieving the repulsion reaction of xenograft. Methods BAECs were co-cultured with different concentrations of hIL-13 for 2 h and followed by co-cultured with 4 ng/ml TNF-α for 6 h or 18 h. The expressions of E-selectin and ICAM-1 on BAECs were detected by Cell-ELISA. The effect of hIL-13 on activity of BAECs was detected by MTT colorimetry.Results BAECs pretreateded with hIL-13 could inhibit the expression of E-selectin and ICAM-1 induced by TNF-α, and showed a doesdependent manner from 5 ng/ml to 20 ng/ml of hIL-13 (P＜0.01). The experimental result of BAECs activity measured by MTT proved no significant difference in the activities of BAECs in every experimental groups compared with control group’s. Conclusion hIL-13 could inhibit the expression of E-selectin and ICAM-1 on BAECs induced by TNF-α, which may contribute to the xenotransplant immune tolerance.
Objective To compare the effects of the denudedfreeze-dried-amniotic-membrane and the denuded freeze-dried bovine corneal stroma when they were explanted to repair the corneal defect of rabbits. Methods The amnia from healthy human placentae were prepared with the method reported by LUO Jingcong, which were freeze-dried and sterilized. The bovine cornea was also denuded by typsin, rinsed, freeze-dried, and sterilized. Twenty Japan rabbits weredivided into group A(the amniontic group) and group B(the bovine-corneal-stroma group) at random. The defect was made, which was 7.5 mm in diameter and 1/3 ofthe thickness of the cornea, and the two kinds of materials were explanted to repair the defect. The vascularization and the changes of the operated eye were observed. The samples were taken at 2, 4 and 8 weeks for histologicalexamination. Results The explanted materials were not melted or excluded. There were visible neovessels in both groups, yet there was no significant difference between them. According to the histological observation, there was severe inflammation in both groups 2 weeks after operation, the fibroblasts were proliferated, and the collagen fibers were disorganized; however,the reactions became milder from 4 weeks after operations,andthe neovessles could be seen in groups A and B; at 8 weeks, the collagen fibers were more organized in groups A and B; however,there was still a small area of disorganized fibers left. Conclusion The two materials can lead to rejection to some extent, and so they need to be improved.
Thirteen patients with intractable nonunions of fractures of long bones were sucessfully treated by a combination of internal fixation and implantation of bBMP. There was an average of 1.5 operative procedures per patient in an attempting to establish reunion prior to bBMP implantation. Union was obtained in 12 of the 13 patients exapt in one who gained success from establish the second attempt. The average time requited to union was 4.7 months. No complication was seen.
Purpose To investigate whether experimental autoimmune uveitis can be induced equally in different rats by urea soluble fraction of bovine melanin-associated antigen(USF-BMAA),and,if so,difference among them. Methods Lewis rats,F344 rats,Wistar rats were immunized with USF-BMAA emulsified with complete Freud is adijuvant and Bordelella pertussis to induce experimental autoimmune uveitis.The animal models were investigated clinically and histopathologically and compared with each other. Rusults Experimental autoimmune uveitis could be induced in Lewis rats,F344 rats and Wistar rats with US-BMAA.Clinical and histopathalogical examination showed that bilateral ocular inflammation developed after immunization 9-13 days.Although inflammation was mainly located in anterior uvea,a mild focal choroiditis was noted in those with severe anterior inflammation.No inflammation was observed in the retina and pineal gland.Experimental autoimmune uveieis induced with USF-BMAA was similar to experimental autoimmune anterior uveitis incited with BMAA presented by other authors.Inflammation induced with USF-BMAA in F344 rats and in Lewis rats was quite similar in the severity and course of the model.But the inflammation was less in Wistar rats compared with that in Lewis rats and F344 rats. Conclusion Experimental autoimmune anterior uveitis was successfully induced with USF-BMAA in Lewis rats,F344 rats and Wistar rats.The difference with regard to the severity among these aminals were propably attributed to their genetic bankground. (Chin J Ocul Fundus Dis,1998,14:149-152)
Objective To verify the technics of inactivating/removing virus in collagen sponge derived from bovine Achilles tendon. Methods Possible pathogen species were determined according to the raw material of bovine Achilles tendon used in production, then vesicular stomatitis virus (VSV), theiler’s mouse encephalomyelitis virus (TEMV), pseudorabies virus (PRV), and simian vacuolating virus 40 (SV40) were selected as indicator virus. Virus suspension was prepared in accordance with Technical Standard for Disinfection. 60Co radiation 25 kGy of collagen sponge was determined as inactivating/removing virus process according to the analysis of the manufacture process, the virus inactivation/removal effect was verified by the measurement of median tissue culture infective dose (TCID50) and showed by virus reduction factor (sample average values of numerical difference before and after processing). Results Reduction factors of VSV, TEMV, PRV, and SV40 after 60Co radiation 25 kGy were 5.646, 4.792, 5.042, and 5.292 logTCID50/0.1 mL (logs), respectively. Reduction factor of each indicator virus was greater than 4 logs, showing that 60Co irradiation 25 kGy can effectively inactivate and remove viruses. Conclusion 60Co radiation 25 kGy of collagen sponge derived from bovine Achilles tendon can be used as the technics of inactivating/removing virus during the preparation process of collagen sponge to guarantee the safety of the product.
ObjectiveTo assess clinical results of aortic cusps replacement with bovine pericardium for bicuspid aortic valve (BAV) and severe aortic regurgitation (AR). MethodsClinical data of 79 patients with BAV and severe AR who underwent aortic cusps replacement with bovine pericardium in Wuhan Asia Heart Hospital from June 2008 to December 2013 were retrospectively analyzed. There were 60 male and 19 female patients with their age of 38±14 years (ranged 12-78 years). All the patients were in NYHA class Ⅱ. There were 26 patients with ascending aorta and sinotubular expanding. ResultsNo early death or major complication was recorded. Intraoperative transesophageal echocardiography showed successful repair with normal coaptation of the aortic leaflets in all the patients. AR grade was less than mild in all the patients with peak aortic valve pressure gradients of 14.2±2.8 mm Hg. All the patients were discharged from the hospital within 15 postoperative days without any adverse symptom, and were followed up for 50±16 months (ranged 9-64 months). During follow-up, all the patients were in NYHA classⅠ. There were 57 patients without AR, 16 patients with mild AR, 5 patients with moderate AR and 1 patients with severe AR. The peak of aortic valve pressure gradient was 12.4±3.2 mm Hg. The average diameter of ascending aorta was 2.7 cm in the patients with ascending aorta and sinotubular expanding. The shape of sinotubular kept normal. The height of coaptation of aortic valve was 0.58 cm by echocadiography. None of the patients died or required reoperation. The structural valve degeneration was not observed during the follow-up. ConclusionThree aortic cusps replacement with bovine pericardium can produce good hemodynamics and midterm results for patients with BAV and severe AR. The ascending aorta and sinotubular should be reduced and fixed in the patients with ascending aorta and sinotubular expanding.
Objective To investigate the effect of repeated freezing and thawing combining nuclease treatment on the decellularization of bovine tendons, and the morphology, structure, biochemical compositions, and mechanical properties of the decellularized tendons. Methods A total of 48 fresh 1-day-old bovine Achilles tendons were randomly divided into 3 groups (n=16): fresh normal tendons (group A), repeated freezing and thawing for 5 times (liquid nitrogen refrigeration/37℃ thawing, group B), and repeated freezing and thawing combining nuclease processing for 24 hours (group C). In each group, 2 tendons were used for scanning electron microscope (SEM), 3 tendons for histological and immunohistochemical observations, 3 tendons for DNA content detection, and 8 tendons for biomechanical testing. Results SEM observation indicated the intact, aligned, and densely packed collagen fibers with no disruption in groups A and B, and the slightly loose collagen fibers with little disruption in group C. The alcian blue staining, sirius red staining, and immunohistochemical staining showed that the most of glycosaminoglycan, collagen type I, collagen type III, and fibronectin in group C were retained after decellularization treatment. HE and DAPI staining showed that the cell nuclei between the collagen fibers were clearly visible in groups A and B; however, the cell nuclei between collagen fibers almost were invisible with a few residual nuclei on the endotendineum in group C. DNA quantitative detection confirmed that DNA content in group C [(0.05 ± 0.02) μg/mg] was significantly lower than those in group A [(0.24 ± 0.12) μg/mg] and group B [(0.16 ± 0.07) μg/mg] (P lt; 0.05). Biomechanical testing showed that the values of tensile strength, failure strain, stiffness, and elastic modulus were different among 3 groups, but no significant difference was found (P gt; 0.05). Conclusion Repeated freezing and thawing combining nuclease processing can effectively remove the component of cells, and simultaneously retain the original collagen fibrous structure, morphology, most of the extracellular matrix compositions, and mechanical properties of the bovine tendons.
Abstract: Objective To evaluate a new type of treatment that reduces calcification of glutaraldehydetreated bovine jugular venous conduit (BJVC). Methods Fresh bovine jugular veins were treated with glutaraldehyde, followed by Triton X-100 and epoxy chloropropane (EC+Tr group). We compared the group’s appearance, histology, shrinkage temperature, tensile strength, and elongation at break with those of a fresh group, and with a group treated with glutaraldehyde only (GA group). We then implanted the EC+Tr and GA group BJVCs subcutaneously into the backs of SD rats and left them for eight weeks (n=8). The morphologic properties and inflammatory response of the test specimens were evaluated by HE staining. The tissue calcium content was determined by atomic absorption spectrophotometer. Results The shrinkage temperature, tensile strength, and elongation at break of the EC+Tr group were significantly higher than those of the fresh group (86.15±0.92 ℃ vs. 69.94±0.92 ℃,t=35.239, P=0.000; 5.31±0.14 mPa vs.3.15±0.95 mPa,t=6.362, P=0.000; 265.11%±27.80% vs. 16521%±25.06%,t=7.550, P=0.000) and of the GA group (86.15±0.92 ℃ vs. 82.73±1.28 ℃, t=6.137, P=0.000; 5.31±0.14 mPa vs. 4.52±0.56 mPa,t=3.871, P=0.002; 265.11%±27.80% vs.237.85%±17.41%,t=2.351,P=0.034). The tissue structure of the subcutaneously implanted EC+Tr veins remained intact;degradation was slight and they contained few inflammatory cells. The calcium content of the EC+Tr group was lower than that of the GA group (51.22±2.69 μg/mg vs. 73.24±3.82 μg/mg, t=11.545，P=0.000). Conclusion Treatment with Triton X-100 and epoxy chloropropane modification with glutaraldehydetreated bovine jugular venous conduit was an effective way to prepare BJVC that avoided calcification.
Objective To compare the biological and biomechanical characteristics of decellularized bovine jugular venous tissue-engineered valved conduit scaffolds with that of fresh bovine jugular veins. Methods Fortyeight fresh bovine jugular veins were divided into control group and experimental group with random number table method, 24 veins in each group. There were fresh bovine jugular veins in control group, decellularized bovine jugular veins in experimental group. The veins of experimental group were treated with sodium deoxyeholate plus Triton-X-100 to decellularize the cells in valves and vessel walls. The thickness, water absorption rate, water maintenance rate, destroying strength, stretch rate of valves and vessel walls in two groups were detected. Results The endothelial cell and fibroblast of valves and vessel walls in experimental group were completely decellularized, no cell fragments were retained within the matrix scaffold; collagen fiber and elastin fiber had been preserved with intact structure and wavily arrayed; deoxyribonucleic acid content of valves and vessel walls in experimental group were decreased by 97.58%, 97.25% compared with that of control group. The thickness, water absorption rate and water maintenance rate of valves and vessel walls in experimental group were lightly increased than those of control group, but there were no significant differences between them （P 〉 0. 05）. There were no significant differences in destroying strength and stretch rate of valves and vessel walls between two groups （P〉0. 05）. Conclusion Decellularized bovine jugular vein scaffold has stable biological and biomechanical characteristics and it may be ideal natural fibrous matrix for developing the tissue-engineered valved conduit by host recellularization.
Objective To observe clinical effects of burn wounds treatment with bovine amnion and to screen the best method of preparing and storing of bovine amnion. Methods From January 2004 to January 2005,We selected randomly 58 patients with superficial Ⅱ° wound, deepⅡ° wound, autografting area for removal of eschars and tangential excision, fetching skin area or residual burn wound . Using auto-control, every burn wound was divided into 3 parts and was treated with 3 dressings: bovine amnion dealt with by 0.1% chlorhexidine（group A）, bovine amnion dealt with by 0.4% glutaraldehyde（group B） and vaseline gauze dressing（group C as control）. The clinical effects were compared between different groupsand the method of preparing and storing bovine amnion was evaluated. Results The dressing texture of group A was softer than that of group B, and its flexibility was fine. The pretreatment was not necessary for dressing in group A. When the dressing was used on burn wounds in groups A and B, painwas slight, but pain was obvious in group C; healing time in groups A and B was much less than that in group C, showing statistically significant difference(P＜0.01). There was no statistically significant difference in healing time between groups A and B (P＞0.05). The infection ratio of burn wound in deepⅡ° wound and residual burn wound of groups A and B is much lower than that of group C, showing statistically significant difference (P＜0.05); in theother burn wounds there was no significant difference (P＞0.05). There was no statistically significant difference between groups A and B (P＞0.05). Conclusion Bovine amnion could make benefit on burn wounds healing, reduce infection ratio of burn wounds, could be used on different kinds of burn wounds. The clinical effect between bovine amnion dealt with by glutaraldehyde and by chlorhexidine is similar. Whereas the latter is more easy to be popularized.