体外诱导骨髓间充质干细胞构建纤维蛋白胶软骨模块

目的:探讨体外构建骨髓间充质干细胞、改良纤维蛋白胶及生长因子的组织工程软骨模块的可行性。方法:分离兔骨髓间充质干细胞,种植在改良纤维蛋白胶支架内。实验组培养液中加入TGF-β1和BMP-6,对照组未加入诱导因子。体外进行形态组织学观察。1、2、3和4周取材作Masson染色、Ⅱ型胶原染色和扫描电镜观察。结果:细胞在支架上贴附,增殖。实验组中Masson染色可见胞浆及细胞周围有胶原形成;Ⅱ型胶原免疫组化染色阳性。结论:TGF-β1和BMP-6可诱导骨髓间充质干细胞成软骨分化,在改良纤维蛋白胶支架内体外成功构建组织工程软骨模块。

Experimental study on reconstruction of cartilage compound with rabbit bone mesenchymal stem cells in improved fibrin glue bracket

Objective To study the feasibility of the reconstruction in vitro of cartilage tissue engineered compound using rabbit bone mesenchymal stem cells(BMSCs)with growth factors in improved fibrin glue bracket.Methods BMSCs were extracted from New Zealand rabbit bone marrow,purified and proliferated in vitro.Then the cells were seeded in improved fibrin glue bracket to construct the compound in vitro.In experimental group,TGF-β1 and BMP-6 were applied to induce chondrogenic transformation while in control group only DMEM was used.The compounds were observed and were harvested in 1,2,3 and 4 weeks respectively and examined by histology,type II collagen immunohistochemical test(S-P test) and scanning electronic microscopy.Results When seeded in improved fibrin glue bracket,the number of BMSCs in each group was significantly increased along with the cultured time prolonged.BMSCs were deduced to excrete extracellular matrix,which is positive with toluidine and masson staining.From the type II collagen immunohistochemical test,the brown granules were seen surround the cell in the blank bubble after cultured for two weeks and four weeks.Under the scanning electron microscopy,BMSCs could be attached to and extended on improved fibrin glue bracket,and normally grown,proliferated,had active function.Conclusion BMSCs can be induced to express chondrocytic phenotype by the synergistic action of TGF-β1 and BMP-6 in vitro.Improved fibrin glue bracket is fitted for the demand of the cartilage tissue engineering.