摘要:
目的 评估聚D,L-乳酸(PDLLA)三维多孔支架的体外细胞相容性,探讨其作为骨组织工程支架类材料的可行性,为进一步进行体内试验提供依据.方法 将分离、获取的第3代骨髓间充质干细胞(BMSCs)按随机数字表法分成5组(0,3,6,9,12周组),分别用含体积分数20%降解液(0,3,6,9,12周时的PDLLA降解液)的细胞培养液和成骨诱导分化培养液进行细胞培养(以不含有降解液的细胞培养液或者成骨诱导分化培养液培养组为对照组,即0周组),从BMSCs的毒性、活力、成骨分化能力等方面定量评估PDLLA支架材料的细胞相容性;将分离、获取的第3代BMSCs种植于PDLLA支架材料上,定期进行扫描电子显微镜观察,从不同时相点细胞在PDLLA材料表面的生长情况定性评估PDLLA支架材料的细胞相容性. 结果 各组与对照组比较均未发现对细胞具有明显的毒性作用;各组均未发现对细胞的活力具有明显的负面影响(t3=-0.441,P=0.671;t6=1.596,P=0.154;t9=-0.492,P=0.636;t12=-1.135,P=0.283);各组细胞ALP染色和钙结节染色结果均表现为阳性,ALP活性定量检测和Ⅰ型胶原蛋白表达的定量检测与对照组之间差异均无统计学意义.扫描电镜结果显示,BMSCs在PDLLA三维多孔支架材料上呈现较好的贴壁生长.结论 PDLLA支架材料对BMSCs具有较好的体外相容性,可以作为一种骨组织工程支架类材料进行后续的体内试验.%Objective To investigate the in vitro cytocompatibility of three-dimensional porous scaffolds of poly-D,L-lactic acid (PDLLA) and discuss the feasibility of PDLLA as a scaffold for bone tissue engineering.Methods BMSCs of the third passage were seeded on osteogenetic differentiation medium or culture medium containing 20% volume fraction degraded liquid (PDLLA degradation liquid of 0,3,6,9,and 12 weeks) according to the random number table.Osteogenetic differentiation medium or culture medium without PDLLA was used as controls.Cell viability,cytotoxicity,and osteogenic differentiation were detected for study on cytocompatibility of PDLLA.Scanning electron microscopy was used to observe the growth of BMSCs on the surface of PDLLA scaffolds.Results PDLLA scaffolds presented no significant cytotoxic on the growth of BMSCs.PDLLA scaffolds had no negative effect on cell viability compared with the controls (t3 =-0.441,P =0.671; t6 =1.596,P =0.154; t9 =-0.492,P =0.636; t12 =-1.135,P=0.283).ALP staining and calcium nodule staining were positive and there were no significant differences in ALP and collagen Ⅰ protein quantitative detection compared with the controls.BMSCs grew well on the inner surface of the PDLLA three-dimensional porous scaffolds.Conclusion Three-dimensional porous scaffolds of PDLLA present good cytocompatibility in vitro and can be used as bone tissue engineering scaffolds for subsequent in vivo research.