BMSCs在PLGA-[ASP-PEG]基质材料表面粘附及增殖的研究

目的：探讨大鼠骨髓间充质干细胞BMSCs在聚丙交酯/乙交酯/天冬氨酸-聚乙二醇三嵌段多元共聚物 PLGA-ASP-PEG]表面粘附、增殖的情况，为组织工程学体外诱导种子细胞生长提供新的生物材料。方法：在PLGA支架材料中引入聚乙二醇（PEG）和含有多个功能位点的天冬氨酸（ASP），制成PLGA-ASP-PEG]高分子支架材料。 将PLGA-ASP-PEG]支架材料与BMSCs复合培养，以未改性的PLGA支架材料作对照，通过沉淀法、MTT法和考马斯亮蓝法分别检测BMSCs的粘附和增殖变化；扫描电镜观察黏附细胞的形态。结果 BMSCs在PLGA-ASP-PEG]材料表面帖壁生长，细胞数目明显多于单纯PLGA组。细胞粘附率检测显示：改性后的PLGA-ASP-PEG]表面BMSCs的粘附性能和增殖能力明显高于对照组，P＜0.05。MTT比色试验，BMSCs在三嵌段材料上培养20d后，吸光值A=1.336，约为对照组0.780的两倍。细胞内蛋白总量间接反映细胞黏附及增殖情况。培养12d时，在PLGA-ASP-PEG]材料组细胞的蛋白含量为66.44μg/孔，单纯PLGA组为41.23μg/孔，间接说明了三嵌段材料生物相容性好，细胞黏附力强的特点。结论PLGA-ASP-PEG]能促进组织工程种子细胞在骨基质材料表面的黏附、增殖并能较好地保持细胞的形态。

Adhesion and proliferation of bone marrow stromal cells with PLGA-ASP-PEG polymer scaffolds

Objectives: To investigate the effects of adhesion and proliferation of bone mesenchymal stem cells (BMSCs) in the surface of lactic acid/glycolic acid/asparagic acid-co-polyethylene glycol PLGA-ASP-PEG] tri-block polymer scaffolds, try to find a new biomaterial to induce seed cells in vitro for bone tissue engineering. Methods: Modified PLGA with polyethylene glycol (PEG) and asparagic acid (ASP) that has many ligands, and synthesis PLGA-ASP-PEG] polymer material. BMSCs were cultured in PLGA-ASP-PEG] polymer material and PLGA used as control group. Through precipitation method, MTT assay and total cellular protein detection to test the adhersion and proliferation of BMSCs. Scanning electron microscope is used to observe cells appearance. Results: BMSCs on the surface of PLGA-ASP-PEG] polymer scaffolds are adherention to the culture flask, the number of cell's is much higher than PLGA's. The precipitation method suggest that adhesion and proliferation of BMSCs on the surface of PLGA-ASP-PEG] is much higher then the control group. P＜0.05. MTT assay show: after culture on PLGA-ASP-PEG] polymer scaffolds for 20 days, the absorbance A is 1.336, and PLGA as control group is 0.780. Total cellular protein can image the adhersion and proliferation of BMSCs indirectly. After 12 days culture, the total cellular protein of PLGA-ASP-PEG] is 66.44μg/pore, and PLGA is 41.23μg/pore, that is to say PLGA-ASP-PEG] polymer scaffolds have well biocompatibility and cells adhersion. Conclusion: PLGA-ASP-PEG] can improve the seed cell's adhesion and proliferation, and keep cells in good condition.