转化生长因子-β_1对体外构建组织工程瓣膜的实验研究  

Effects of Transforming Growth Factor β_1 on Constructing Tissue Engineering Heart Valves in Vitro

作　　者：仇玉明[1] 董念国[1] 史嘉玮[1] 叶晓峰[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院心外科,湖北省武汉市430022

出　　处：《中国全科医学》2006年第6期469-471,共3页Chinese General Practice

基　　金：国家自然科学基金资助项目(C30371414)

摘　　要：目的研究转化生长因子-β1(TGF-β1)在组织工程心脏瓣膜(TEHV)体外构建中的作用。方法经胰酶/EDTA、去污剂Triton X-100和核酸酶处理,去除猪主动脉瓣叶的细胞成分,然后在其表面种植大鼠肌成纤维细胞,构建TEHV。实验组培养基中添加TGF-β1(10ng/ml),对照组采用普通培养基。体外培养2周后取瓣叶,行HE染色和扫描电镜观察,并检测羟脯氨酸、DNA含量及瓣叶最大负荷力,应用逆转录-聚合酶链反应(RT-PCR)方法检测瓣膜细胞MMP-13和TIMP-1mRNA的表达。结果HE染色和扫描电镜显示实验组较对照组的细胞联合紧密且细胞外基质更丰富,羟脯氨酸含量和DNA含量增高,最大负荷力提高,MMP-13mRNA和TIMP-1mRNA的表达增强。结论TGF-β1在TEHV体外构建过程中具有积极作用,能为种子细胞生长提供有利因素,促进细胞增殖和细胞外基质分泌,提高瓣膜的最大负荷力。Objective To investigate effects of transforming growth factor - β1 （TGF - β1 ） on tissue engineering heart valves （TEHV）. Methods Porcine aortic valves were decellulafized with trypsin and TritonX - 100, and then were seeded by myofibroblasts harvested from rats. They were cultivated in culture medium including TGF - β1 at concentration of 10 ng/ ml. Two weeks later, light and electron microscopy observation was performed. And cell DNA assay, hydroxyproline content together with max - strain of leaflets were measured, RT - PCR technique were used to analyse expressions of MMP - 13 and TIMP - 1 mRNA. Results Histological investigation and showed that cells and extracellualr matrix were more bloomy in the valves treated with TGF - β1 than normal culture medium. And DNA assay, hydroxyproline content, max - strain of leaflets and expression of MMP - 13 and TIMP - 1 mRNA were also increased with the help of TGF - β1. Condusion TGF - β1 was an important and effective factor for cell proliferation and extracellualr matrix growth. It plays an important role in constructing tissue engineering heart valve in vitro.

关 键 词：组织工程心脏瓣膜 转化生长因子-Β1 细胞外基质 

分 类 号：R654.2[医药卫生—外科学]