脱矿牙本质基质诱导骨髓间充质干细胞向成牙本质样细胞分化的体外实验研究

摘要

AIM; To observe the adhesion and growth of rat bone marrow mesenehymal stem ceils (rBMSCs)seeded on demineralized dentin matrix (DDM) in vitro and the odontogenic induction of DDM. METHODS; BMSCs were successfully isolated and cultured. The compatibility of the biomaterial was evaluated by MTT assay. The odontogenic differentiation of BMSCs 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 BMSCs in DDM groups was obviously higher than that in the control groups. The higher levels of ALP activity and larger mineralizing nodes were also detected in the experiment groups. RT-PCR demonstrated mRNA expression of dentin sialophospho-protein ( DSPP ) and dentin matrix proteinl ( DMP-1) in BMSCs induced by DDM. CONCLUSION; DDM possesses a good cellular compatibility with BMSCs. It can induce the odontogenic differentiation of BMSCs.%目的:观察脱矿牙本质基质(demineralized dentin matrix,DDM)与大鼠骨髓间充质干细胞(rat bone marrow mesenehymal stem ceils,rBMSCs)的生物相容性及其作为支架材料对BMSCs的牙向诱导作用.方法:分离培养BMSCs,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BMSCs增殖活性和牙向分化能力的影响.结果:DDM能显著促进体外培养的BMSCs的增殖、诱导细胞的矿化和提高细胞ALP活性；同时还能诱导BMSCs在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 proteinl,DMP-1).结论:体外培养条件下,DDM与BMSCs有良好的生物相容性,能够诱导BMSCs牙向分化.