转化生长因子-β_1基因转染构建组织工程瓣膜  

Experiments on constructing tissue engineering heart valve by transforming growth factor-β_1 gene transfected cells

作　　者：史嘉玮[1] 仇玉明[1] 董念国[1] 孙宗全[1] 

机构地区：[1]华中科技大学协和医院心外科华中科技大学同济医学院心血管病研究所,武汉430022

出　　处：《临床心血管病杂志》2009年第1期60-63,共4页Journal of Clinical Cardiology

基　　金：国家自然科学基金(No:30571839)

摘　　要：目的:转化生长因子(TGF)-β1基因转染细胞,研究对天然支架组织工程瓣膜(TEHV)构建作用。方法:脂质体介导pcDNA3.0/TGF-β1基因载体进入大鼠肌成纤维细胞(MFB),48h和4周后G418筛选法培养,SABC法检测TGF-β1表达。A组将转基因细胞种植去细胞猪主动脉瓣叶,B组种植未转基因细胞,C组单纯支架,体外培养2周构建TEHV。苏木精-伊红染色、扫描电镜观察,测定羟脯氨酸、DNA含量和最大负荷力。结果:基因转染48h和4周,MFB瞬时和稳定表达TGF-β1蛋白。A组较B组细胞生长旺盛,胶原分泌较多,力学性能增强。结论:TGF-β1基因转染MFB作为种子细胞,能较好发挥TGF-β1活性,促进TEHV构建。Objective：To investigate the effects of transforming growth factor （TGF） -β1 gene transfection on tissue engineering heart valves （TEHV） construction. Method： Rat myofibroblasts were transfected with peDNA3.0/TGF-β1 gene plasmid mediated by lipofectamine 2000 after 48 hours and 4 weeks, and cultivated by G418 selection for 3 weeks. Immunohistochemistry method of SABC was performed to observe expression of TGF-β1 protein in cells. They were seeded onto decellularized valve scaffold as group A. Non-transfected myofibroblasts were seeded onto scaffold with normal culture medium as group B. Decellularized valve scaffold was performed in group C. Hematoxylin eosin staining and scanning electronic microscopy were performed to observe cell proliferation of valves in three groups. Cell DNA assay, hydroxyproline content and Max strain of leaflets were measured. Result： pcDNA3/TGF-β1 gene transfected myofibro blasts could express TGF-β1instantly and stably. Compared with group B and C, histological investigation showed that cells and extracellular matrix were more bloomy in valve of group A. And DNA assay for cell sum, hydroxyproline con tent for collagen production, Max-strain of leaflets for mechanical characteristics also increased in group A. Conclusion： The transfected myofibroblasts could be regarded as one kind of new seed cells and were beneficial for constructing TEHV in vitro with the help of TGF-β1.

关 键 词：转化生长因子-Β1 组织工程心脏瓣膜 基因转染 

分 类 号：R512.62[医药卫生—内科学]