应用骨髓干细胞和自制胶原-壳聚糖构建组织工程心瓣膜  

Construction of tissue engineering heart valve with rabbit bone mesenchymal stem cells and self-made collage-chitosan complex scaffold

作　　者：付建华[1,2] 赵曼[3] 杜桂英 杨达宽[2] 黄云超[5] 

机构地区：[1]聊城市人民医院胸心外科,山东聊城252000 [2]昆明医学院第二附属医院胸心外科,昆明650031 [3]聊城市人民医院信息科,山东聊城252000 [4]聊城市疾控中心艾防所,山东聊城252000 [5]昆明医学院第三附属医院胸心外科,昆明650031

出　　处：《第三军医大学学报》2011年第2期152-155,共4页Journal of Third Military Medical University

摘　　要：目的探讨应用骨髓基质干细胞(bone mesenchymal stem cells,BMSCs)诱导分化的内皮细胞为种子细胞,以胶原-壳聚糖复合物为支架材料,运用组织工程学的原理和方法体外构建组织工程心脏瓣膜(tissue engineering heart valve,TEHV)的可行性。方法兔主动脉壁体外分离、培养、传代平滑肌细胞、成纤维细胞,将培养的成纤维细胞、平滑肌细胞与兔骨髓基质干细胞诱导分化的内皮细胞按顺序种植在预湿处理的胶原-壳聚糖复合支架上,通过组织学及倒置显微镜、扫描电镜观察内皮细胞、成纤维细胞、平滑肌细胞在支架上生长、增殖和基质分泌情况。结果 HE染色示内皮细胞能够在胶原-壳聚糖复合支架黏附、伸展、增殖,在TEHV表面长成连续的细胞层;扫描电子显微镜示TEHV表面光滑,细胞生长良好,细胞层完整,且瓣膜表面的内皮细胞具有合成和分泌前列环素(PGI2)的功能。结论以兔主动脉壁体外分离、培养、传代的平滑肌细胞、成纤维细胞和BMSCs诱导分化的内皮细胞为种子细胞,依次种植在胶原-壳聚糖复合支架上可以构建TEHV,且内皮细胞能够分泌PGI2。Objective To explore the feasibility of constructing a tissue engineering heart valve（TEHV） using endothelial cells induced and differentiated from（bone mesenchymal stem cells,BMSCs） as seeded cells and collagen-chitosan complex as scaffold material.Methods Smooth muscle cell and fibroblasts were isolated and cultured from rabbit aortic wall and Fibroblast,smooth muscle cells and endothelial cell induced and differentiated from BMSCs were seeded in pre-moisture collagen-chitosan complex scaffold in turn.The cell adhesion,growth,proliferation and stroma secretion were observed by using histochemistry,inverted and SEM scanning microscopy.Results The differented endothelial cells could adhere to,grow and proliferate on collagen-chitosan complex scaffold by HE staining observation,and cells on TEHV surface grew to form a serial cell layer.TEHV surface was smoothd and cells grew better.SEM scanning observation showed that cell layer was intact.Endothelial cells on TEHV surface synthesized and secreted PGI2.Conclusion Smooth muscle cells,fibroblasts cultured from aortic wall and endothelial cells induced and differentiated from BMSCs can seed on collagen-chitosan complex scaffold in turn,to construct TEHV where endothelial cells can secrete PGI2.

关 键 词：组织工程心脏瓣膜 瓣膜构建 骨髓基质干细胞 诱导分化 内皮细胞 

分 类 号：R318.08[医药卫生—生物医学工程] R318.11[医药卫生—基础医学]