目的 构建装载基质细胞衍生因子-1α(SDF-1α)的Pluronic F-127新型复合材料,并研究其对骨髓间充质干细胞(BMSCs)的体外生物效应.方法 构建装载不同浓度SDF-1α(200、50 ng/ml)的Pluronic F-127复合水凝胶,采用Transwell小室进行复合水凝胶对BMSCs的体外趋化实验.将BMSCs与复合材料均匀混合,加成骨细胞诱导液进行培养,并对其行碱性磷酸酶(AKP)活性测定及茜素红染色,由结果可观察到BMSCs的成骨诱导分化情况,并用MTT法检测复合培养后细胞的活性状况.结果 趋化实验结果显示,含SDF-1α浓度为200 ng/ml的复合水凝胶对BMSCs的募集效果显著(P<0.05).MTT法证实了Pluronic F-127对细胞增殖不产生影响(P>0.05).BMSCs-复合材料、单纯BMSCs在成骨诱导培养条件下,两组细胞的AKP表达量均明显增高,并且都高于非成骨诱导组(P<0.05),同时诱导组镜下可见明显的钙结节形成.结论 成功构建装载SDF-1α的Pluronic F-127新型复合材料,其在体外对BMSCs有明显的趋化作用,在细胞材料复合培养的三维支架中,BMSCs能够存活并被成功诱导分化为成骨细胞.
Objective To detect the effect of Pluronic F-127 hydrogel combined with stromal cell derived factor-1α(SDF-1 α) on bone mesenchymal stem cells (BMSCs) ' biological effect in vitro.Methods Transwell chamber was used to detect the effect of Pluronic F-127 hydrogel combined with various concentration of SDF-1α (200,50 ng/ml) on BMSCs' migration.The BMSCs were osteogenesis cultured in 3-dimensional scaffold SDF-1α-Pluronic F127 hydrogel,then alkaline phosphatase (AKP) activity and alizarin red was evaluated,and MTT was used to explore the cytoactive of BMSCs after cultured in SDF-1 α-gel.Results The result of chemotaxis displayed that the concentration of SDF-1α 200 ng/ml in hydrogel had obvious effect on migration of BMSCs(P < 0.05).MTT confirmed Pluronic F-127 had no effect on cell proliferation (P > 0.05).Compared with control group,BMSCs-gel group and pure BMSCs group' s AKP had significantly increased in the circumstances of osteogenesis induction(P < 0.05),and calcium nodule could be found in the induction group.Conclusion The composite material F-127 loaded with SDF-1 α significantly promotes chemotactic migration of BMSCs in vitro,BMSCs can survive and be induced differentiation into osteoblast in the 3-dimensional scaffold SDF-1 α-gel.