摘要
背景:目前,组织工程瓣膜去细胞支架的制备方法和效果各不相同,但总的来说,酶结合去垢剂法显示出一定的优势作用。目的:探讨脱氧胆酸钠、十二烷基硫酸钠、曲拉通3种去垢剂结合酶消化法对于猪主动脉瓣膜脱细胞的效果以及去细胞后对支架的影响。方法:将新鲜猪主动脉瓣膜用抗生素溶液浸泡12h后,置于胰蛋白酶和EDTA溶液中12h,再分别用脱氧胆酸钠、十二烷基硫酸钠、曲拉通进行处理,最后转入核酸酶溶液中浸泡12h,以达到去除内皮细胞和间质细胞的目的,常规苏木精-伊红染色观察内皮细胞是否除完全,Masson染色观察胶原纤维和弹力纤维损害程度,电子扫描显微镜观察纤维结构以评价效果。结果与结论:3种去垢剂结合酶消化法均能彻底去细胞,但脱氧胆酸钠对支架胶原纤维和弹力纤维的损害较轻微,十二烷基硫酸钠次之,曲拉通对支架的损害作用最大。提示脱氧胆酸钠结合酶消化法是一种较合理的组织工程瓣膜支架构建方法。
BACKGROUND：The method and effect of construction of tissue-engineering heart valve scaffold with cell detergents are different,and the method of detergent combining with the trypsase and nuclease is more suitable than others.OBJECTIVE：To investigate the effect of different detergents（sodium deoxycholate,sodium dodecylsulfate,and triton）combined with trypsase on decellularized porcine heart valve and the influence on the accellular scaffold.METHODS：Porcine aortic valve leaflets were sterilized by antibiotics for 12 hours,maintained in the solution of the trysin and the EDTA for 12 hours,and treated with sodium deoxycholate,sodium dodecylsulfate,and triton.Finally,the sample was dip in nuclease solution for 12 hours to remove endothelial cells and interstitial cells.HE staining was used to detect whether the endothelial cells were removed completely,Masson staining was used to evaluate damage level of collagen fiber and elastic fiber,and electronic scanning was used to observe the microstructure.RESULTS AND CONCLUSION：All the three detergents completely removed the endothelial cells;however,the effect of sodium deoxycholate on collagen fiber and elastic fiber was light,and then sodium dodecylsulfate and triton.This suggested that the method of DOA combining with the enzyme digestion was a suitable technique to construct tissue-engineering heart valve scaffold.
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2010年第8期1349-1352,共4页
Journal of Clinical Rehabilitative Tissue Engineering Research
