自聚肽纳米纤维支架承载骨髓源性心肌干细胞移植治疗心肌梗死的实验研究  

Self-assembled peptide nanofiber scaffold and marrow-derived cardiac stem cells transplantation in treatment of myocardial infarction

作　　者：崔晓军[1] 谭玉珍[2] 谢华[1] 国海东[2] 王存[2] 张剑凯[1] 郭金华[1] 

机构地区：[1]广东医学院基础医学院人体解剖学教研室,广东东莞524808 [2]复旦大学上海医学院人体解剖学与组织学胚胎学系,上海200032

出　　处：《生物医学工程与临床》2011年第4期303-307,F0003,共6页Biomedical Engineering and Clinical Medicine

基　　金：湛江市科技攻关计划项目(2010C3102004)

摘　　要：目的探讨自聚肽纳米纤维支架承载骨髓源性心肌干细胞(MCSC)移植对大鼠心肌梗死后心功能恢复的影响,寻找一种更适合承载心肌干细胞治疗心肌梗死的可注射性生物材料。方法通过单细胞克隆培养技术,从骨髓间充质干细胞中筛选具有心肌特异分化潜能的MCSC,固相合成自聚多肽。将雌性大鼠心肌梗死模型随机分为3组:对照组、MCSC移植组、支架承载MCSC移植组。细胞移植后4周,用M型超声心动图检测心功能恢复情况;对心脏组织切片进行Masson染色,利用图像分析检测胶原纤维比率;用原位荧光杂交法标记心肌组织中Y染色体细胞,并检测其心肌肌钙蛋白T(cTnT)的表达。结果支架承载MCSC移植组与MCSC移植组相比,大鼠的心功能指数明显提高,胶原纤维比率明显减小,Y染色体阳性细胞数明显增多,差异均具有统计学意义。结论支架承载MCSC移植治疗心肌梗死,有利于移植细胞在受损心肌内的存活和向功能性心肌分化,并促进心功能恢复,其疗效优于单纯干细胞移植。Objective To explore the effect of the self-assembled peptide nanofiber scaffold（NFS） and marrow-derived cardiac stem cells（MCSC） transplantation on cardiac function after myocardial infarction（MI）,and provide theoretical and experimental basis for new nano-material in clinical stem cell transplantation.Methods The MCSC were isolated from marrow mesenchymal stem cells with single-cell cloning culture,and identified by c-kit immunoflourence staining and Nkx 2.5 expression analysis.The self-assembled peptides were designed and synthesized.Thirty minutes after the establishment of acute MI model of female rats,the rats were divided into 3 groups and injected at the edge of infracted area with phosphate buffered solution（PBS）,MCSC,and NFS ＋ MCSC,respectively.Four weeks after MI,the cardiac function was analyzed by M-mode echocardiography,and the organizational structure was analyzed by Masson staining.The donor cells in myocardial tissue were marked with Y-chromosome through fluorescence in situ hybridization and the expression of cTnT of Y-chromosome-positive cells was detected by immunohistochemistry.Results The functional recovery of rat hearts,myocardial scar size reduction and the Y-chromosome positive cells in the NFS ＋ MCSC transplantation group were superior to that in the MCSC transplantation group,and the difference was statistically significant.The distributions of Y-chromosome positive cells were concentrated in the NFS ＋ MCSC transplantation group,and the Y-chromosome-positive cells expressed cTnT.Conclusion It is demonstrated the self-assembled peptide build a three-dimensional environment which is conductive to the survival and myocardial differentiation of MCSC in the ischemic myocardium,which in turn leads to enhanced angiogenesis,reduced scar area and improved cardiac function.

关 键 词：自聚肽纳米纤维支架 骨髓源性心肌干细胞 心肌梗死 移植 

分 类 号：R542.22[医药卫生—心血管疾病]