摘要
目的探索用骨形态发生蛋白(BMP)-2和高分子透明质酸在体外诱导骨髓基质细胞(MSC)向软骨细胞分化的可能性。方法自兔双侧股骨粗隆处抽取4～6ml骨髓后在体外行原代和传代培养,实验分为两组,实验组培养液中含BMP-2(100ng/ml),培养瓶底预涂高分子透明质酸;对照组常规培养及传代。传代细胞玻片行苏木精-伊红染色、甲苯胺蓝染色、碱性磷酸酶染色(ALP)、免疫组织化学染色;培养细胞与聚乳酸/聚乙醇酸(PLGA)复合后植入自体兔股部肌内,大体及组织学观察形成的结节。结果实验组传代细胞,尤其是第3代细胞甲苯胺蓝染色阳性,免疫组织化学染色S-100蛋白阳性,兔股部植入PLGA-细胞后3周形成软骨结节;而对照组免疫组织化学染色个别细胞Ⅰ型和Ⅲ型胶原染色阳性,植入PLGA-细胞后主要形成纤维组织。结论应用BMP-2和高分子透明质酸有效诱导MSC向软骨细胞分化,MSC源性软骨细胞可作为软骨组织工程修复关节软骨损伤的较理想的种子细胞。
Aim To explore the possibility of the differentiation from marrow stromal cells(MSC) to chondrocytes induced by BMP 2 and high polymer hyaluronic acid in vitro.Methods About 4- 6 ml bone marrow aspirated from 14 New Zealand rabbits with 16 gauge puncture needles from bilateral femoral tuberosities,were primarily cultured and sub cultured in vitro.There were 2 groups of experiments: experimental group and control group.In the experimental group,BMP 2( 100 ng/ml) was contained in culture medium and high polymer hyaluronic acid was spread on the bottom of culture flasks.In the control group,the cells were cultivated and subcultured as usual.HE staining,toluidine blue staining,alkaline phosphatase staining and immunohistochemical staining were done for subcultured cells.After PLGA and cells had been compounded,they were implanted into thigh muscle of rabbits.The nodules were observed histologically and microscopically. Results In the experimental group,toluidine blue staining and immunohistochemical staining(S 100 protein) were positive for the passage cells,especially the third passage cells.After 3 weeks,chondral nodules were formed in the thigh muscle.But in the control group,immunohistochemical staining for collagenⅠ and collagen Ⅲ were positive in individual cells.Fibrous tissues were mainly formed in the thigh muscle after implanting PLGA cell compounds.Conclusion BMP 2 and high polymer hyaluronic acid are used to induce marrow stromal cells into chondrocytes successfully.MSC derived chondrocytes are used as seed cells to repair articular cartilage injury in cartilage tissue engineering.<P>
出处
《中国临床康复》
CSCD
2003年第17期2397-2399,T002,共4页
Chinese Journal of Clinical Rehabilitation
基金
全军重大课题基金资助(01Z079)~~
