过表达MMP1对根尖牙乳头干细胞骨/牙向分化能力的影响

›› 2018, Vol. 9 ›› Issue (2): 63-68.

过表达MMP1对根尖牙乳头干细胞骨/牙向分化能力的影响

许艳¹,闫明²,汪延秋³,于金华¹,庞希瑶⁴

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

收稿日期:2018-03-06 修回日期:2018-05-29 出版日期:2018-06-25 发布日期:2018-06-28
通讯作者: 于金华 E-mail:yujinhua@njmu.edu.cn
基金资助:
国家自然科学基金资助项目;江苏高校优势学科建设工程资助项目

Effect of MMP1 overexpression on the osteo/odontogenic differentiation of stem cells from apical papilla

Received:2018-03-06 Revised:2018-05-29 Online:2018-06-25 Published:2018-06-28

摘要/Abstract

摘要： 目的：研究过表达基质金属蛋白酶1（MMP1）对根尖牙乳头干细胞（SCAPs)骨/牙向分化能力的影响。方法：从牙根未发育完全的第三磨牙上摘取根尖牙乳头组织，采用酶消法和组织块法结合的方法分离纯化得到SCAPs。设计构建MMP1过表达质粒，转染SCAPs。通过碱性磷酸酶（ALP）活性检测试剂盒、茜素红染色、Western blot和实时定量RT-PCR检测MMP1过表达组和空载质粒组的骨/牙向分化相关指标的变化。结果：成功构建SCAPs MMP1过表达模型；ALP活性检测和茜素红染色结果显示MMP1过表达组的ALP活性降低和矿化能力减弱，Western bolt结果显示MMP1过表达组的Runt相关转录因子2（RUNX2）、成骨细胞特异性转录因子Osterix（OSX）、牙本质涎蛋白（DSP）、骨钙素（OCN）的表达降低，实时定量RT-PCR结果显示骨钙素基因（OCN）、Ⅰ型胶原蛋白基因（COL1）、OSX、RUNX2、DSPP表达降低。结论：过表达MMP1抑制SCAPs的骨/牙向分化潜能。

Abstract: Objective：To research the effect of matrix metalloproteinase 1 (MMP1) overexpression on the osteo/odontogenic differentiation of human stem cells from the apical papilla (SCAPs). Methods： SCAPs were isolated and purified from human third molars with developing roots. Retrovirus expression plasmid PMSCV/MMP1 was constructed. The osteo/odontogenic differentiation capacity was tested by alkaline phosphatase (ALP) activity, alizarin red staining, Western blot assay, and real-time RT-PCR. Results： The ALP activity assay and alizarin red staining showed that ALP activity and mineralization capacity of MMP1 overexpression were decreased. The results of RT-PCR and Western bolt indicated that the mRNA levels of COL1, OCN, RUNX2, OSX and DSPP were downregulated and the protein levels of DSP, RUNX2, OSX and OCN were decreased. Conclusions：MMP1 overexpression downregulates the osteo/odontogenic differentiation potential of stem cells from apical papilla.

许艳闫明汪延秋于金华庞希瑶. 过表达MMP1对根尖牙乳头干细胞骨/牙向分化能力的影响[J]. , 2018, 9(2): 63-68.

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