通过室温模压/粒子浸出方法制备得到聚乙交酯丙交酯(PLGA)多孔支架,每个质量50mg、孔径200~300μ,zm、孔隙率略大于90%的PLGA85/15多孔支架在10mL磷酸盐缓冲液(PBS)中37℃体外降解24周.降解液每周换一次,不同时间点的降解液被收集、并加入骨髓基质干细胞(MSC)的培养液或者成骨诱导液中,利用胞外乳酸脱氢酶含量检测、细胞死活染色、四唑盐检测、碱性磷酸酶染色和定量检测的方法考察降解液对MSC的活力和成骨分化能力的影响.实验结果表明,PLGA多孔支架材料在PBS中逐渐降解,其质量、尺寸、孔径、孔与孔的连通性、分子量有不同程度的降低;其降解液在本研究的实验条件下未发现对MSC有明显的细胞毒性,对MSC的活力、增殖以及成骨分化均无显著的负面影响.
Poly(lactide-co-glycolide) (PLGA) porous scaffolds were prepared via room-temperature molding/ particle leaching method, and in vitro degradations of those porous scaffolds were examined in phosphate buffer saline (PBS) solution at 37 ℃ for 24 weeks. Each PLGA85/15 scaffold of porosity a bit above 90% and weight about 50 mg was immersed in 10 mL PBS. The degradation media were replaced by fresh ones weekly. Part of degradation media were collected at series of degradation time points, and then added into the culture medium of mesenchymal stem cells (MSCs). Cell viability and osteogenic differentiation were detected via lactate dehydrogenase assay, Live/Dead staining, 3-( 4, 5-dimethylthiazol-2-yl )-2, 5-diphenyltetrazolium bromide assay,alkaline phosphatase staining and quantification. The results indicated that molecular weight of remaining PLGA, and weight, dimension, pore size, and interpore connectivity of scaffolds changed with different degrees. The degradation media at all of degradation times up to 24 weeks exhibited no severe cytotoxicity, and no significant negative influences on viability, proliferation and osteogenic differentiation of MSC were observed under the present conditions and examined period.