脱细胞牛心包膜因其具有优良特性而成为组织工程生物瓣膜中理想的支架材料.采用冻融+表面活性剂法对牛心包组织进行脱细胞处理后,用戊二醛或京尼平对其进行表面修饰固定.通过HE染色及扫描电镜观察脱细胞效果,并对交联组织进行厚度检测、含水量和接触角测试、力学检测、交联指数和差示扫描量热(DSC)测定、体外降解、溶血试验以及细胞毒性试验,判断两种交联方法对脱细胞牛心包基质的影响,从而选择出交联脱细胞组织的最好方法.结果显示,冻融+表面活性剂法脱除细胞彻底,戊二醛或京尼平交联后组织厚度明显增加分别约20%,25%,亲水性良好,力学特性稳定,交联指数都高达90%,且28 d降解率分别为5%和3%,交联后组织稳定性高.但京尼平组溶血率(0.37%)远小于戊二醛组溶血率(13.77%),且细胞毒性很低.作为瓣膜组织工程支架材料,京尼平交联脱细胞牛心包膜比戊二醛交联效果具有更好生物相容性,交联效果好,是较好的交联方法.%The acellular bovine pericardium has become an ideal scaffold material for heart valve tissue engineering because of its fine features. In this study,the celluar components of the bovine pericardiums were removed by the method of freeze-thaw-detergent 24 h, and then the acellular bovine pericardium were crosslinked with glutaraldehyde or genipin. The effects of decellularization were tested by HE staining and scanning electron microscope. Properties of the crosslinked acellular matrix were evaluated by tissue thickness measurement, moisture content test, surface water contact angle, mechanical test, fixation index determination,differential scanning calorimetry (DSC) test,in vitro enzymatic degradation,hemolysis test and cytotoxicity test.The results showed that freeze-thaw-detergent 24 h method could remove the cell components of bovine pericardium effectively. And after crosslinking with glutaraldehyde, thickness of the tissue were increased by 20％ ,while that of genipin were about 25％. Both crosslinked tissues were of strong hydrophilicity and good mechanical properties with crosslinking index more than 90％ ,and the degradation ratio after degrading 28 days were only 5％ and 3％ respectively, with stable tructure. However, the hemolysis rate of genipin crosslinked tissue were much lower than that of glutaraldehyde (0. 37％ vs 13.77％ ) ,and with lower cytotoxicity as well.Acellular bovine pericardium crosslinked with genipin was a better one for its better biocompatibility and good crosslinking effects.
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