Vitapex对根尖牙乳头干细胞成牙/骨向分化及自噬的影响

口腔生物医学 ›› 2022, Vol. 13 ›› Issue (4): 214-218.

Vitapex对根尖牙乳头干细胞成牙/骨向分化及自噬的影响

张筱岚¹,马艳霞²,陈路遥²,方玉心²,李泽汉³,闫明¹,于金华⁴

1. 南京医科大学口腔医学研究所
2. 无
3. 南京医科大学附属口腔医院牙体牙髓科
4. 南京医科大学

收稿日期:2022-08-09 修回日期:2022-11-18 出版日期:2022-12-25 发布日期:2022-12-24
通讯作者: 于金华 E-mail:yujinhua@njmu.edu.cn
基金资助:
国家自然科学基金面上项目

Effect of Vitapex conditioned medium on the odonto/osteogenic differentiation and autophagy of stem cells from apical papilla

Received:2022-08-09 Revised:2022-11-18 Online:2022-12-25 Published:2022-12-24

摘要/Abstract

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/骨向相关指标（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））基因和蛋白的变化情况；Western blot检测自噬相关指标（LC3 II/LC3 I、Beclin-1、P62）的蛋白表达水平。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 ALP、RUNX、OSX、DSPP基因和蛋白表达（P＜0.05）。同时，0.2 g/L Vitapex组中LC3 II/LC3 I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。

关键词: Vitapex, 根尖牙乳头干细胞, 成牙/骨向分化, 自噬

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, RUNX, 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.

Key words: Vitapex, stem cells from apical papilla, odonto/osteogenic differentiation, autophagy

张筱岚马艳霞陈路遥方玉心李泽汉闫明于金华. Vitapex对根尖牙乳头干细胞成牙/骨向分化及自噬的影响[J]. 口腔生物医学, 2022, 13(4): 214-218.