脱细胞肠系膜制备生物膜支架的实验研究

目的应用肠系膜脱细胞基质制备生物膜支架,探讨其理化特性和生物学特性。方法应用反复冻融肠系膜组织后,将肠系膜置于胰蛋白酶消化脱肠系膜脱细胞,并分为肠系膜基质组(A组)和去细胞肠系膜基质组(B组),通过HE染色、电镜、DNA检测、细胞毒性实验、拉伸力学测试,检测两组肠系膜基质的理化特性;制备覆膜支架,植入兔髂血管,术后1周、1月、2月超声检测血管血流情况,血管取材病理检测。结果 HE染色、电镜检测结果表明,B组基质组织疏松,纤维排列较整齐,未见细胞存留; DNA检测结果表明,B组DNA表达水平低,脱细胞较彻底; CCK-8细胞毒性实验检测结果表明,两组均无细胞毒性; FDA-PI荧光染色结果表明,两组细胞存活良好,未见死亡细胞;拉伸力学测试结果表明,两组最大拉伸力、最大力伸长率、屈服强度、屈服点伸长率差异无统计学意义;脱细胞肠系膜支架植入兔血管后超声检测两组早期通畅性良好,支架植入2月内皮增生较明显。结论冻融和酶消化法脱肠系膜细胞,肠系膜基质去除细胞彻底,无细胞毒性,力学特征良好;肠系膜支架植入血管后早期通畅性良好,2月后内皮增生明显。

Experimental Study on Preparation of Biofilm Scaffolds from Acellular Mesentery

Objective To prepare biofilm scaffolds by using mesenteric acellular matrix, so as to investigate their physicochemical and biological characteristics. Methods The mesenteric tissues were subjected to trypsin digestion, and the mesenteric cells were removed after repeated freezing and thawing of mesenteric tissue. Mesenteries were divided into mesenteric matrix group (Group A) and acellular mesenteric matrix group (Group B). The physical and chemical properties of mesenteric matrix in two groups were tested by HE staining, electron microscopy, DNA detection, cytotoxicity test and tensile mechanics test. The blood flow of the vessels was detected by ultrasonography at 1st week, 1st month and 2nd month, and the vessels were observed by pathological examination. Results HE staining and electron microscopy showed that the mesentery of Group B was loose in acellular mesentery matrix, and the arrangement of fibers was neat, with no cells remaining. Compared with Group A, the expression level of DNA in Group B was lower, with more completely decellularized cells. CCK-8 cytotoxicity test showed that there was no cytotoxicity in Group A and Group B. FDA-PI fluorescence staining showed no cytotoxicity of cells in both groups. Cells in Croup A and Group B survived well, and no dead cells were found. Tensile mechanics test showed that there were no significant differences in maximum tensile force, maximum elongation, yield strength, yield point elongation between Group A and Group B. The early patency of acellular mesenteric stent implantation was good, and endothelial hyperplasia was obvious at 2nd month after stent implantation. Conclusion sMesenteric cells were removed by freeze-thaw and enzymatic digestion. Mesenteric stroma was completely removed without cytotoxicity, which showed good mechanical characteristics. Mesenteric stent implantation had good early patency and endothelial proliferation after 2 months.