骨髓间充质干细胞联合可降解支架体外构建组织工程带瓣静脉（英文）  

Preliminary fabrication of tissue engineered veins containing valves using bone marrow mesenchymal stem cells and biodegradable scaffolds in vitro

作　　者：刘池拽[1,2] 殷恒讳[1] 吕伟明[1] 曾晨光[3] 刘畅[4] 王文见[1] 全大萍[3] 项鹏[4] 王深明[1] 

机构地区：[1]中山大学附属第一医院血管外科,广东省广州市510080 [2]中山大学附属中山医院(中山市人民医院)血管外科,广东省中山市528400 [3]中山大学化学与化学工程学院高分子研究所,广东省广州市510275 [4]中山大学干细胞与组织工程研究中心,中山大学教育部干细胞与组织工程重点实验室,广东省广州市510080

出　　处：《中国组织工程研究》2014年第3期348-356,共9页Chinese Journal of Tissue Engineering Research

基　　金：the National Natural Science Foundation of China,No.30700804~~

摘　　要：背景:临床上治疗慢性静脉功能不全的主要方法是静脉瓣膜修复及带瓣静脉段移植,但这些方法创伤较大,且带瓣静脉来源有限。组织工程学和再生医学在修复病变血管方面取得的进步,而以自体来源的内皮细胞为种子细胞的组织工程带瓣静脉也见于了报道,但存在排出反应。目的:构建一个有可自我更新、修复、类似天然瓣膜结构并具有功能的带瓣静脉。方法:麻醉取Beagle犬的骨髓获取骨髓间充质干细胞,采用密度梯度离心和贴壁法获取骨髓间充质干细胞,并进行细胞的传代、冻存复苏、流式细胞仪检测和定向诱导分化。采用热致相分离技术,以聚(乳酸-乙醇酸)共聚物为基材,利用自制带瓣静脉模具制备三维组织工程带瓣静脉支架,制备组织工程带瓣静脉支架,并研究其形态结构。将骨髓间充质干细胞种植在支架上构建可降解的带瓣静脉,在体外培养2周。结果与结论:扫描电镜观察显示支架孔隙率高。培养的细胞符合骨髓间充质干细胞的形态特征,培养的细胞大部分表达CD29和CD44,不表达CD34和CD45。细胞毒性实验显示支架无毒性,有利于细胞增殖和迁移。将细胞种植在支架表面上培养后可形成单层细胞层。体外实验验证细胞支架复合物的瓣膜有一定的开闭功能。利用三维聚(乳酸-乙醇酸)共聚物支架和骨髓间充质干细胞可成功构建组织工程带瓣静脉,组织工程带瓣静脉将有可能作为静脉瓣膜的的替代物治疗静脉瓣膜疾病。BACKGROUND： Chronic venous insufficiency is a major health problem worldwide. Clinical treatments include venous valve repair and venous segment containing valve transplantation. However, these are invasive procedures, and the supply of vein containing valves is limited. Significant progress in the fields of tissue engineering and regenerative medicine has been made towards the creation of tissue engineered vascular grafts for the repair of damaged or malformed vessels. It has been reported that using tissue engineering, a tissue engineered vein containing valves constructed with self-derived endothelial cells and allogeneic acellular matrices can provide the complex physiological valve structure and mechanical stability, but this elicited an immunogenic response. OBJECTIVE： To create a viable and functional vein containing valves, which has the ability to grow, repair, and imitate natural tissues. METHODS： Bone marrow mesenchymal stem cells were obtained from Beagle dogs by density gradient centrifugation and adherence methods. Bone marrow mesenchymal stem cells were cultured in vitro. Following isolation and culture the cells were examined using flow cytometry and identified by direct induction towards the osteogenic and adipogenic lineages. We fabricated biodegradable venous scaffold containing valves using the method of injection molding combined with thermally induced phase separation. Based on the self-made cast, a three-dimensional biodegradable vein scaffold containing valves was constructed from poly（lactic-co-glycolic acid）. Morphological structure was tested. Bone marrow mesenchymal stem cells were used as seed cells to be seeded onto the lumen of the tissue engineered vein scaffold containing valves in vitro and then incubated for 2 weeks. RESULTS AND CONCLUSION： Scanning electron microscopy images showed that the scaffold demonstrated sufficient porosity. Cultured cells expressed mesenchymal cell markers, CD44 and CD29, but did not express hematopoietic cell markers, CD34 and CD45 at

关 键 词：生物材料 材料相容性 可降解吸收材料 静脉瓣膜 骨髓间充质干细胞 聚(乳酸-乙醇酸)共聚物 慢性静脉功能不全 组织构建 干细胞 国家自然科学基金 

分 类 号：R318[医药卫生—生物医学工程]