体外构建的同种生物组织工程瓣脉动流培养的实验研究  

Pulsatile-flow-cultivation recellularization of homograft bioprosthetic valve

作　　者：冯滨[1] 刘迎龙[1] 晏明全 朱晓东[1] 谢宁[1] 阮英卯[3] 

机构地区：[1]中国医学科学院中国协和医科大学心血管病研究所阜外心血管病医院外科,北京现在100037 [2]四川省南充市中心医院心胸外科 [3]中国医学科学院中国协和医科大学心血管病研究所阜外心血管病医院病理科,北京100037

出　　处：《中华胸心血管外科杂志》2006年第1期35-38,共4页Chinese Journal of Thoracic and Cardiovascular Surgery

基　　金：国家高技术研究发展计划(863计划)(2001AA216061);国家自然科学基金(30271289);中国博士后科学基金(2003033224)资助

摘　　要：目的对静态培养条件下构建的同种生物组织工程瓣(TEHV)进行脉动流培养,增强种植的内皮细胞(ECs)抗切应力,改良其生物学性能,为今后动物实验和临床的实际应用提供技术路线。方法将静态培养的、再内皮化TEHV置入自行设计构建的脉动流培养装置中进行培养,在培养介质流量和压力逐渐增加的条件下培养14d,不同时点取瓣膜片样本进行硝酸银染色立体显微镜以及扫描电镜观察、摄片。瓣膜表面内皮细胞(ECs)覆盖面积用百分率(%)表示,确定其ECs黏附率。结果在脉动流培养前,组织工程瓣内皮化程度为92%,而脉动流培养的第7、10、14d,其ECs黏附率分别为36%、23%和11%,说明种植的ECs脱落严重。结论TEHV脉动流培养的生物性能改良效果不佳,可能与多种因素有关。在进行动物实验以及今后的临床应用之前还有许多问题亟待解决。Objective Using a bioreactor to culture reendothelializing homograft bioprosthetic valve （I-IBV） in order to ultimately form viable tissue in vitro, so as to provide basic material for animal experiment and clinical application. Methods The reendothelializign HBV was placed in a pulse duplicator system （＂bioreactor＂）, designed and fabricated by us. These grown under gradually increasing nutrient media flow and pressure for additional 14 days： 125 ml/min al 30 mm Hg （days 1-4）, 250 ml/min al 40mm Hg （daya 5-7 ）, 500 ml/min at 50mm Hg （days 8-10 ）, 750 ml/min at 75 mm Hg （days 11 to 14）. Throughout the studies, the flow device was placed in an incubator with 95% humidity and 5% carbon dioxide. The morphologic structure was observed and photographed by stereomicroscope with 0.5 % silver nitrate （AgNO3） staining and SEM on the 7th, 10th and 14th day. The retention rate of the ECs overlaid onto the HBV was evaluated with the percentage （ % ）. Results The reendothelization level on the surface of the leaflets was 92% before pulsatile-flow-cultivation. But the rate of ECs retention on the scaffold exposured to pulsatile flow was only 36%, 23% and 11% on the 7th, 10th and 14th day, respectively. Conclusion The retention rate of ECs on the scaffold was low for pulsatile-flow-cultivation in vitro. Problems, including deficiency of mature stress fibers in the endothelial cells from MSC, s deficiency of the adherence and growth factors still need improvement.

关 键 词：心脏瓣膜 人工生物假体 移植 干细胞 组织培养 组织工程 

分 类 号：R318.0[医药卫生—生物医学工程]