【摘要】 目的:研究Vitapex糊剂对根尖牙乳头干细胞(SCAPs)成牙/骨向分化的影响，初步讨论其潜在机制。方法:酶消化法分离并培养SCAPs;制备不同浓度Vitapex条件培养基，通过碱性磷酸酶(ALP)活性测定选取最适浓度；ALP染色、实时荧光定量PCR和Western blot检测成牙/骨向相关基因和蛋白的变化情况；Western blot检测自噬相关蛋白表达水平。结果:0.2 g/L Vitapex条件培养基组ALP活性最高，选取为最适浓度进行后续实验。与对照组比较，该浓度Vitapex条件培养基能显著促进SCAPs细胞ALP、核心结合蛋白因子2(RUNX2)、成骨相关转录因子(OSX)、牙本质涎磷蛋白(DSPP)基因和蛋白表达(P<0.05)。同时，0.2 g/L Vitapex组中微管相关蛋白1轻链3(LC3)Ⅱ/LC3Ⅰ及Beclin-1蛋白表达水平升高，P62蛋白表达水平降低(P<0.01)。结论:Vitapex可促进SCAPs成牙/骨向分化，并激活自噬。更多还原

【Abstract】 Objective: To investigate the effect of Vitapex on the odonto/osteogenic differentiation of stem cells from apical papilla(SCAPs) and its potential mechanism. Methods: SCAPs were harvested by enzymatic digestion and cultured in medium with different concentrations of Vitapex. The optimal concentration was determined by the alkaline phosphatase(ALP) activity assay. ALP staining, qRT-PCR and Western blot were used to investigate the expression of odonto/osteogenic related markers of SCAPs. The expression level of autophagy related markers was analyzed by Western blot. Results: 0.2 g/L Vitapex conditioned medium group expressed the highest level of ALP activity, and it was selected as the optimal concentration. When treated with 0.2 g/L Vitapex conditioned medium, the expression of odonto/osteogenic markers(ALP, RUNX2, OSX, DSPP) in SCAPs was significantly upregulated(P<0.05). LC3 Ⅱ/Ⅰ ratio and Beclin-1 protein level increased, while P62 decreased(P<0.01). Conclusion: Vitapex can potentially promote the odonto/osteogenic capacity of SCAPs and activate autophagy.更多还原