脱矿牙本质基质对人牙髓干细胞体外增殖、牙向分化能力的影响

摘要

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,RTPCR)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 adhesion and growth of human dental pulp stem cells seeded on demineralized dentin matrix ( DDM) in vitro and the adontogenic induction of DOM. Methods OPSCs were successfully isolated and cultured. The compatibility of the biomaterial was evaluated by MTS assay. The odontogenic differentiation of DPSCs induced by DDM was measured using Alizarin red staining, the alkaline phosphatase (ALP) activity and reverse transcription-polymerase chain reaction (RT-PCR) assay. Results The proliferation of DPSCs in DDM groups was obviously higher than that in the control groups. The higher levels of ALP activity and larger mineralizing nods were also detected in the experimental groups. RT-PCR demonstrated mRNA expression of dentin sialophosphoprotein ( DSPP ) and dentin matrix protein 1 (DMP-1) in DPSCs induced by DDM. Conclusions DDM possesses a good cellular compatibility with DPSCs, As an engineering scaffold.it can induce the odontogenic differentiation of DPSCs.