聚乙醇酸-乳酸共聚物复合Ⅱ型胶原和生长因子体内构建组织工程软骨(英文)

Construction of tissue engineered cartilage in vivo with poly(lactide-co-glycolic acid)composited with collagen Ⅱ and growth factors

背景:软骨组织工程的发展为处理关节软骨损伤提供了新的思路和方法,使体内构建组织工程软骨得以实现。目的:观察骨髓基质干细胞种植到复合胶原和生长因子的聚乙醇酸-乳酸共聚物(polylactide-co-glycolicacid,PLGA)生物材料,再种植到大鼠体内构建组织工程软骨组织的可行性。方法:相分离法制作PLGA,复合Ⅱ型胶原和碱性成纤维细胞生长因子,转化生长因子β1。将第3代的骨髓基质干细胞种植到复合材料上。36只SD大鼠随机分为实验组、对照组、空白组,分别于肌袋内植入骨髓基质干细胞/复合生长因子和胶原的PLGA、复合生长因子和胶原的PLGA、复合胶原的PLGA,于术后第4,8,12周取材观察细胞的定向分化及生长情况,包括大体观察、苏木精-伊红染色、甲苯胺蓝染色、Ⅱ型胶原染色、扫描电镜观察。结果与结论:大体观察可见实验组材料有类软骨样组织生长,而对照组和空白组则仅见纤维组织生长。各种染色及电镜观察显示:实验组复合物内可见多的成软骨细胞及少量的破骨细胞。实验组甲苯胺蓝染色和Ⅱ型胶原染色为阳性,对照组和空白组均为阴性。从而证明胶原修饰的PLGA生物材料具有较好的细胞相容性;骨髓基质干细胞种植到复合胶原和生长因子的PLGA生物材料上在大鼠体内可构建组织工程软骨复合组织。

BACKGROUND：The development of cartilage tissue engineering provides novel ideas for treatment of articular cartilage defects and implements construction of tissue-engineered cartilage in vivo.OBJECTIVE：To investigate the feasibility of constructing tissue-engineered osteochondral composite through bone marrow stem cells（BMSCs）cultured on the poly（lactide-co-glycolic acid）（PLGA）,which was modified with collagen and cellular growth factors.METHODS：PLGA was made by phase separation technique,composited with collagen Ⅱ,basic fibroblast growth factor,and transforming growth factor-β1.The BMSCs of passage 3 were cultured on the above scaffolds.Thirty-six SD rats were randomly divided into experimental,control,and blank groups.These three groups received implantation of BMSCs composited with growth factors and collagen-PLGA,implantation of BMSCs composited with collagen-PLGA,and implantation of collagen-PLGA into the muscle,respectively.At 4,8,and 12 weeks after surgery,cell directional differentiation and growth were examined by gross observation,hematoxylin-eosin staining,toluidine blue staining,collagen Ⅱ staining,and scanning electron microscope.RESULTS AND CONCLUSION：Gross observation showed that there were many chondroid tissues in the experimental group and fibrous tissues in the control and black groups.Stainings and electron microscope revealed that many chondroblasts and a few osteoclasts appeared in the composite of the experimental group.Toluidine blue and collagen Ⅱ stainings were positive in the experimental group and negative in the control and blank groups.These findings demonstrate that PLGA modified with collagen had a good cellular compatibility.BMSCs cultured on PLGA,which was modified with collagen and cellular growth factors,can construct the tissue-engineered osteochondral composite in rats.