目的:探索人牙髓细胞条件培养液(HDPSCs-CM)对人牙囊细胞(HDFSCs)向成骨分化的作用.方法:利用胶原酶消化法获得HDFSCs,经纯化鉴定后培养于HDPSCs-CM中;CCK-8检测HDFSCs增殖活性,观察细胞形态改变;碱性磷酸酶(ALP)染色、茜素红S染色定性分析细胞成骨能力的改变.qRT-PCR检测HDFSCs中骨膜蛋白(POSTN)、Ⅰ型胶原(Col-Ⅰ)、碱性磷酸酶(ALP)、骨涎蛋白(BSP)以及骨桥蛋白(OPN)的mRNA表达情况.结果:经HDPSCs-CM诱导后,HDFSCs形态发生成牙骨质或成骨样改变;诱导组HDFSCs增殖活性受到明显抑制(P<0.05或P<0.01);诱导组ALP染色强于对照组,茜素红S染色矿化结节多于对照组;诱导组POSTN、Col-Ⅰ、ALP、BSP、OPN的表达量明显增高(P<0.05或P<0.01).结论:HDPSCs-CM能促进HDFSCs成骨分化.%Objective:To evaluate the effects of human dental pulp stem cells-conditioned medium(HDPSCs-CM)on the osteogenic differentiation of human dental follicle stem cells(HDFSCs) in vitro.Methods:HDFSCs were in vitro cultured,purified and identified.CCK-8 assay was applied to evaluate the HDFSCs viability after 7 days cultured by HDPSCs-CM;the morphological changes of HDFSCs were observed;the osteogenic differentiation was studied by alkaline phosphatase(ALP) staining and alizarin red staining.The mRNA expression of POSTN,Col-Ⅰ,ALP,BSP and OPN was detected by Real-Time PCR.Results:Induced by HDPSCs-CM,HDFSCs exhibited several characteristics of cementoblast or osteoblast lineages.In the induction group the viability of HDFSCs was inhibited(P<0.05 or P<0.01),ALP staining was stronger and there were more mineralized nodules,the expression levels of POSTN,Col-Ⅰ,ALP,BSP and OPN were upregulated(P<0.05 or P<0.01).Conclusion:HDPSCs-CM can promote the osteogenic differentiation of HDFSCs.
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