制备附载rhTGF-β1的PLGA缓释支架及其对MSCs细胞形态的影响

Preparation of PLGA scaffold containing rhTGF-β1 delivery and its effect on morphology of MSCs in vitro

为探讨自制的附载转化生长因子-β1(rhTGF-β1)的D,L-乳酸-CO-乙醇酸(PLGA)体外缓释生物支架对人骨髓间充质干细胞(MSCs)细胞形态的影响,制备了附载rhTGF-β1的PLGA生物支架,并检测了在PLGA的降解过程中rhTGF-β1的释药规律;同时分离培养人骨髓间充质干细胞,体外培养后分别接种于附载和未附载rhTGF-β1的PLGA支架上.通过扫描电镜观察不同时间段MSC在支架上的生长情况,实验结果表明,rhTGF-β1能被包裹进PLGA支架中,而且可在PLGA支架降解过程中持续释放出来.通过扫描电镜还观察到实验组支架上细胞呈多角或梭形,且有较多基质分泌,而对照组细胞较少.因此附载转化生长因子-β1的PLGA复合载体是一种较为理想的新型生物支架.

In order to prepare the biodegradable poly （lactic-co-glycolicacid） （PLGA） scaffolds containing recombinant human transforming growth factor-β1 （rhTGF-β1） delivery and to investigate its bioactivity on the morphology of mesenchymal stem cells （MSCs）in vitro, the growth-impregnated PLGA scaffolds were prepared by combination of porogen-leaching and freeze-drying. In vitro drug release kinetics, as measured by enzyme-linked immunosorbent assay （ELISA）, was monitored. MSCs were isolated from the bone marrow of human and cultured. Then, MSCs were seeded onto PLGA scaffolds. PLGA pieces without rhTGF-β1 were used as controls. Scanning electron microscopy（SEM） was used to observe the morphology of MSCs. Results show that rhTGF-β1 can be encapsulated into PLGA scaffolds and can be released from the scaffolds continuously. SEM shows the distribution of spindle and polygonal cells in the investigated group of PLGA scaffolds and less of these cells can be found in the control group. PLGA scaffolds containing rhTGF-β1 is a new tissue engineering scaffold.