摘要
目的:基于骨形态发生蛋白2(BMP2)/母亲信号蛋白同源物通路(Smads)通路,分析富血小板纤维蛋白(PRF)对人牙髓干细胞成骨分化的作用。方法:分离纯化牙髓干细胞,流式细胞仪检测并鉴定牙髓干细胞表面抗原标记物,随机分为对照组、PRF组、BMP激活剂组和BMP抑制剂组(简称激活剂组和抑制剂组),检测培养7d、14d后的碱性磷酸酶(ALP)活性;RT-qPCR检测牙髓干细胞胶原蛋白(I COL-I)、Runt相关转录因子2(RUNX2)、骨钙素(OCN)mRNA相对表达水平;茜素红染色观察矿化结节形成;Western-Blot检测牙髓干细胞BMP2、Smad1、Smad5、p-Smad1/5、RUNX2蛋白相对表达水平。结果:STRO-1、CD29、CD90表达阳性,符合牙髓干细胞的表型。各组ALP活性随时间延长而升高(P<0.05),从对照组到抑制剂组、PRF组、激活剂组,牙髓干细胞ALP活性及COL-Ⅰ、RUNX2、OCNmRNA相对表达水平依次升高,红色矿化结节数目、A值依次增加(P<0.05)。从抑制剂组到对照组、PRF组、激活剂组,牙髓干细胞BMP2、p-Smad1/5、RUNX2蛋白相对表达水平依次升高(P<0.05)。结论:PRF可能通过激活BMP2/Smads信号通路促进人牙髓干细胞成骨分化。
Objective:To observe the effect of platelet-rich fibrin(PRF)on the osteogenic differentiation ofhuman dental pulp stem cells,and to explore the role of bone morphogenetic protein 2(BMP2)/maternal signalprotein homolog(Smads)pathway.Methods:The dental pulp stem cells were isolated and purified,and the surfaceantigen markers of the dental pulp stem cells were deltected by flow cytometry.The identified dental pulp stem cellswere randomly divided into thecontrol group,PRF group,BMP activator group and BMP inhibitor group(hereinafter referred to as activator group and inhibitor group).Each group of dental pulp stem cells in thelogarithmic growth phase were detected for the alkaline phosphatase(ALP)activity after 7 days and 14 days ofculture;RT-qPCR was conducted for detection of the osteogenic differentiation marker collagen I(COL-1).Runt-related transcription factor 2(RUNX2),osteocalcin(OCN)mRNA relative expression levels;Alizarin redstaining was performed to observe the formation of mineralized nodules;Western-Blot detection of BMP2,maternalsignal protein homolog 1(Smad1),Smad5,P-Smad1/5,relative expression level of RUNX2 protein.Results:Flowcytometry detection of cell surface antigens showed that STRO-1,CD29,and CD90 were positively expressed.which was consistent with the phenotype of dental pulp stem cells.Compared with the control group,the ALPactivity of dental pulp stem cells in the PRF group,activator group,and inhibitor group increased for 7 days and 14 days,and the activator group>PRF group>inhibitor group(P<0.05).ALP activity in each group were elevated withthe increase of culture time,the ALP activity of the control group,PRF group,activator group and inhibitor groupincreased(P<0.05).Compared with the control group,the relative expression levels of COL-I,RUNX2 and OCN inthe PRF group,activator group,and inhibitor group increased,and the number of red mineralized nodules alsoincreased,and the activator group>PRF group>inhibitor group(P<0.05).The relative expression levels of BMP2,p-smad1/5 and RUNX2 protein
作者
孙琛
朱青青
王佳琼
张雪
SUN Chen;ZHU Qing-qing;WANG Jia-qiong;ZHANG Xue(Zhengzhou University First Affiliated Hospital Oral Clinic 3,Crossing,Zhengzhou 450000,China;Dept.Of Oral Implant,Zhengzhou University First Affiliated Hospital University Road,Crossing,Zhengzhou 450000,China)
出处
《口腔颌面修复学杂志》
2021年第5期333-337,共5页
Chinese Journal of Prosthodontics
基金
河南省医学科技攻关计划联合共建项目(项目编号:LHGJ20190191)。
关键词
富血小板纤维蛋白
牙髓干细胞
成骨分化
骨形态发生蛋白2
platelet-rich fibrin
dental pulp stem cells
osteogenic differentiation
bone morphogenetic protcin 2lmaternal signal protein homolog pathway
