应用小牛皮制备软骨修复载体的脱毛-脱细胞-结构改建工艺探讨

发布时间：2019-08-08 12:16

【摘要】：[目的]软骨修复载体的制备是软骨组织工程的重要课题。本实验以小牛皮为原料,通过脱毛-脱细胞-结构改建的早期工艺探讨,掌握最佳制备流程,为后续工艺做准备。[方法]将小牛皮分别浸入0.5%dispase溶液和0.25%胰蛋白酶溶液,4℃脱毛72 h,比较脱毛效果。乙酸溶胀,刮除/切除表皮和皮下组织,获得真皮组织。真皮组织分别浸入0.5%SDS、1%SDS与0.5%TritonX-100进行脱细胞,充分冲洗之后冻干,HE染色比较脱细胞效果,MTT法进行细胞毒性检测。应用0.25%胰蛋白酶浸泡、0.25%胰蛋白酶浸泡联合220 V-250 W-40%功率超声振荡、0.5%胰蛋白酶浸泡、0.5%胰蛋白酶浸泡联合220 V-250 W-40%功率超声振荡四种方法进行结构改建,扫描电镜观察、MASSON染色、孔隙率检测比较结构改建效果。[结果]0.5%dispase溶液和0.25%胰蛋白酶溶液均能够良好的去除小牛皮的毛发。0.5%SDS、1%SDS脱细胞效果基本一致,TritonX-100似有将胶原纤维破坏的征兆,由于低浓度SDS更易于冲洗,后续实验采用0.5%SDS脱细胞。细胞毒性检测为0级。0.5%胰蛋白酶浸泡联合220 V-250 W-40%功率超声是最佳的结构改建方法,改建后的载体胶原纤维光滑、立体感明显增强,孔隙率显著高于未改建的载体(P0.05)。[结论]0.25%胰蛋白酶溶液脱毛-0.5%SDS脱细胞-0.5%胰蛋白酶浸泡联合220 V-250 W-40%功率超声进行结构改建,是应用小牛皮制备软骨修复载体的最佳工艺。
[Abstract]:[objective] the preparation of cartilage repair carrier is an important subject in cartilage tissue engineering. In this experiment, calf skin was used as raw material, and the early process of hair removal, acellular and structural reconstruction was discussed, and the best preparation process was grasped in order to prepare for the follow-up process. [methods] the calf skin was immersed in 0.5%dispase solution and 0.25% trypsin solution respectively, and the hair removal effect was compared at 4 鈩，

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