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目的观察脱矿牙本质基质(demineralized dentin matrix,DDM)与人牙髓干细胞(human dental pulp stem cells,hDP-SCs)的生物相容性及其作为支架材料的牙向诱导作用。方法体外分离培养DPSCs,分别用四唑盐比色法(MTS)、茜素红染色法、碱性磷酸酶(alkaline phosphatase,ALP)和逆转录聚合酶链式反应(reverse transcription-polymerase chain reaction,RT-PCR)方法观察DDM生物材料对DPSCs增殖活性和牙向分化能力的影响。结果 DDM显著地刺激体外培养的DPSCs增殖,诱导了细胞的矿化和提高细胞ALP活性。RT-PCR结果显示DDM诱导后细胞mRNA水平表达牙本质涎磷蛋白(dentin sialophosphoprotein,DSPP)和牙本质基质蛋白(dentin matrix protein1,DMP-1)。结论体外培养条件下,DDM与DPSCs有良好的生物相容性,作为支架材料对于DPSCs有较好牙向诱导作用。
OBJECTIVE: To observe the biocompatibility of demineralized dentin matrix (hDM-hs) and human dental pulp stem cells (hDP-SCs) and to investigate their dental effects as scaffold materials. METHODS: DPSCs were isolated and cultured in vitro. The cells were stained with MTS, alizarin red staining, alkaline phosphatase (ALP) and reverse transcription-polymerase chain reaction RT-PCR) method was used to observe the effects of DDM biomaterials on proliferation activity and dental differentiation ability of DPSCs. Results DDM significantly stimulated the proliferation of DPSCs cultured in vitro, induced the mineralization of cells and increased the cellular ALP activity. RT-PCR results showed that mRNA expression of dentin sialophosphoprotein (DSPP) and dentin matrix protein1 (DMP-1) in DDM-induced cells. Conclusion DDM and DPSCs have good biocompatibility under in vitro culture conditions. As a scaffold material, DDMCs have better dentin induction effect.