去细胞猪主动脉瓣叶的获取和内皮细胞的种植

PREPARATION OF ACELLULARIZED PORCINE HEART VALVE AND SEEDING OF BOVINE AORTIC ENDOTHELIAL CELLS

目的 探讨猪主动脉瓣叶去细胞后作为组织工程心脏瓣膜支架的可行性。 方法 经胰酶 - EDTA、表面活性剂和核酸酶处理 ,去除猪主动脉瓣叶的细胞成分 ,测定瓣叶去细胞前、后的生物力学特性 ,并在其表面种植新生牛主动脉内皮细胞 (BAECs) ;分别行大鼠皮下包埋实验。 结果 猪主动脉瓣叶中的细胞成分能完全去除 ,获得完整无细胞的纤维网状支架 ,断裂强度和断裂伸长率无明显变化 ;种植的 BAECs在去细胞瓣叶表面可形成一层连续的细胞层 ,其分泌前列环素 (PGI2 )的能力同直接种植在 2 4孔板中的比较 ,差异无统计学意义 (P>0 .0 5 )。 结论 猪主动脉瓣去细胞后获得的纤维支架可以用来构建组织工程瓣膜 ,适宜于血管内皮细胞的生长。

Objective To explore the possibility of detergent acellularized porcine heart valve serving as a scaffold for tissue engineering valve. Methods The porcine aortic valves were acellularized by use of trypsin-EDTA. Triton X-100, RNase and DNase treatment. Biomechanical characteristics of fresh valves and acellularized valve were tested; also fresh valves, acellularized valve and valves treated with method of bioprothetic treatment were implanted subcutaneously in rats; frequently seeded with bovine aortic endothelial cells(BAECs), and then cultured for 7 days. Results The acellularization procedure resulted in complete removal of the cellular components while the construction of matrix was maintained. The matrix could be successfully seeded with in vitro expanded BAECs, which formed a continuous monolayer on the surface.There is no significant difference of PGI 2 secretion of BAECs between cells seeded onto the acellular leaflets and that anto the wells of 24-wells plate (P>0.05). Conclusion Acellularied porcine aortic valve can be applied as a scaffold to develop tissue engineering heart valve.