骨髓间充质干细胞与复合Ⅰ型胶原和骨形态发生蛋白2的聚乳酸乙醇酸构建组织工程骨  

Construction of tissue engineering bone by bone marrow mesenchymal stem cells on poly-lactide-co- glycolic acid modified with type I collagen and recombinant human bone morphogenetic protein 2

作　　者：查振刚[1] 李劼若[1] 周长忍[2] 姚平[3] 吴昊[1] 刘宁[1] 黄春华[1] 杨淑野[1] 王双利[1] 

机构地区：[1]暨南大学附属第一医院骨科,广州510630 [2]暨南大学理工学院 [3]暨南大学医学院生理教研室

出　　处：《中华生物医学工程杂志》2008年第3期171-174,共4页Chinese Journal of Biomedical Engineering

基　　金：广东省科技计划（2006860501009）;广州市科技攻关（2006Z3-E5211）

摘　　要：目的探讨骨髓间充质干细胞（BMSC）体外分离培养后种植到复合Ⅰ型胶原和重组人类骨形态发生蛋白2（rhBMP-2）的聚乳酸乙醇酸（PLGA）生物支架上，构建组织工程骨的可行性。方法密度梯度离心法提取分离BMSC，倒置显微镜观察细胞形态，流式细胞分析法对细胞表面抗原进行鉴定。相分离法制备多孔三维PLGA生物支架，支架材料上复合Ⅰ型胶原和rhBMP-2，扫描电镜观察其超微结构。将第3代的BMSC接种于复合支架上，扫描电镜观察材料的细胞黏附性，将培养6h后的细胞-支架复合体植入sD大鼠肌袋内，于2个月后取材进行HE染色，观察其构建组织工程骨的情况。结果BMSC可在体外分离扩增，表达CD29、CD44，不表达CD34和CIM5。制备的PLGA支架孔隙率为90％，平均孔径为100μm，与BMSC有较好的黏附性。2个月后动物体内细胞-支架复合体的大体观察和HE染色显示，BMSC种植到复合Ⅰ型胶原和rhBMP-2的PLGA生物支架上可构建骨组织。结论BMSC可在体外长期、稳定培养，是理想的组织工程种子细胞。PLGA与干细胞有较好的黏附性，可用来做组织工程生物材料。BMSC种植到复合Ⅰ型胶原和rhBMP-2的PLGA生物支架上后，在动物体内可构建组织工程骨。Objective To construct the tissue engineering bone by bone marrow mesenchymal stem cells （BMSCs） cultured on poly-lactide-co-glycolie acid （PLGA） modified with typeⅠ collagen and recombinant human bone morphogenetic protein 2 （rhBMP-2）. Methods BMSCs were isolated and cultured in vitro. The cellular configuration was observed continually under inverted phase-contrast microscope, and the cell exterior antigen was analyzed by flow cytometry method. PLGA was made by phase separation and modified with type Ⅰcollagen and rhBMP-2. The biomaterial structure was examined by scan electron microscope. The third generation BMSCs were cultured on PLGA. The adhesion of cell to biomaterial was examined by scan electron microscope as well. The complex of BMSCs-PLGA was then cultured for 6 h in SD rat muscle and was observed two months later by HE staining. Results BMSCs could be isolated and cultured in vitro, and the cell morphology was spindle. They expressed CD29, CD44, but did not express CD34, CD45. The average pore diameter of PLGA modified with type Ⅰ collagen and rhBMP-2 was 100μm, the interval porosity was 90%, and PLGA had good adhesion with BMSCs. Positive staining and strong reaction for HE staining of the bone tissue in the complex was observed. Conclusions BMSCs can be cultured stably in vitro for long time, and are good seminal cell of tissue engineering. The PLGA modified with type Ⅰ collagen and rhBMP-2 has good adhesion with BMSCs. BMSCs cultured on PLGA modified with type Ⅰ collagen and rhBMP-2 can construct the tissue engineering bone in animal body.

关 键 词：骨髓 间质干细胞 聚乳酸乙醇酸 组织工程骨 

分 类 号：R318.08[医药卫生—生物医学工程]