摘要
目的体外构建组织工程心脏瓣膜（TEHV），初步探讨内皮细胞黏附生长的分子机制。方法猪主动脉瓣膜经胰酶-EDTA、表面活性剂、核酸酶处理，去除猪主动脉瓣叶的细胞成分，测定瓣叶脱细胞后的生物力学特性；将扩增的人脐静脉血管内皮细胞（HUVECs）种植在瓣叶上，体外静态构建TEHV，观察内皮细胞的生长状态。消化瓣膜内皮细胞，半定量RT-PCR检测内皮细胞整合素＆mRNA的表达，Western-Blot检测内皮细胞膜上整合素＆蛋白的表达。结果猪主动脉瓣膜中的细胞成分能完全去除，脱细胞瓣叶的生物力学特性同新鲜瓣叶相比无明显变化；种植的HUVECs在瓣叶表面生长状态良好，长成一层连续的细胞层。瓣膜内皮细胞可检测到整合素岛mRNA和蛋白的表达。结论脱细胞猪主动脉瓣膜作为支架，HUVECs做种子细胞可以成功构建TEHV，瓣膜内皮细胞可以表达整合素岛。
Objective To create the tissue engineering heart valve（TEHV） by seeding human umbilical vein endothelial cells （HUVECs） on decellar porcine aortic valve and investigate its mechanism of endothelial cells in adhesion and proliferation. Methods The porcine aortic valve was acellularized by using trypsin,EDTA,Triton X-100,RNase and DNase treatment. Biomechanical characteristics of fresh valves and aceUularized valves were tested, HUVECs were cultured and seeded on decelluar valve to create TEHV in vitro. Histology and growth condition of cells were observed. The integrin β1 expression of HUVECs was tested by RT-PCR and Western blot. Results The acellularization resulted in complete removal of the cellular components. There was no significant difference of biomeehanical characteristics between acellular valves and fresh valves. In vitro TEHV by seeding HUVECs on decellular porcine aortic valve was created successfully. HUVECs formed a continuous monolayer on the surface of valve . The integrin β1 mRNA and protein expression was identified. Conclusion TEHV can be created using HUVECs as seeding cells on decellular porcine aortic vaive.
出处
《中华胸心血管外科杂志》
CSCD
北大核心
2007年第1期36-39,共4页
Chinese Journal of Thoracic and Cardiovascular Surgery
基金
本课题受军队“十五”医学科研规划重点课题（01Z060）资助
