低频脉冲电磁场增强骨形态发生蛋白9诱导牙周膜干细胞成骨分化作用  

Low frequency pulsed electromagnetic fields enhance bone morphogenetic protein 9-induced osteogenic differentiation of periodontal ligament stem cells

作　　者：王彦蒽 汪沛 王晟磊 吴贤慧 孙学兰 刘军[1] 叶晖[1] 冯琴 俞佳颖 曹志中[1] 唐卫忠[1] WANG Yan-en;WANG Pei;WANG Sheng-lei;WU Xian-hui;SUN Xue-lan;LIU Jun;YE Hui;FENG Qin;YU Jia-ying;CAO Zhi-zhong;TANG Wei-zhong(Department of Stomatology,Changhai Hospital,Naval Medical University(Second Military Medical University),Shanghai 200433,China;Department of Oral Implant and Maxillofacial Surgery,Hongkou Branch of Changhai Hospital,Naval Medical University(Second Military Medical University),Shanghai 200081,China)

机构地区：[1]海军军医大学(第二军医大学)长海医院口腔科,上海200433 [2]海军军医大学(第二军医大学)长海医院虹口院区口腔种植与颌面外科,上海200081

出　　处：《第二军医大学学报》2020年第8期855-863,共9页Academic Journal of Second Military Medical University

基　　金：上海市科学技术委员会自然科学基金面上项目(09ZR1400200);海军军医大学(第二军医大学)长海医院青年启动基金(2019QN16)。

摘　　要：目的探讨低频脉冲电磁场能否增强骨形态发生蛋白9(BMP9)诱导人牙周膜干细胞(PDLSC)成骨分化的作用。方法培养健康人前磨牙牙周膜组织细胞,通过免疫磁珠分选后得到CD146+/STRO-1+细胞,即PDLSC。用携带过表达BMP9基因的重组腺病毒(Ad-GFP-BMP9)感染PDLSC,并暴露于频率为15 Hz、磁场强度分别为0.6、1.2、1.8、2.4、3.0 mT的脉冲电磁场进行干预(每12 h辐照1 h)。通过qRT-PCR和蛋白质印迹法检测Runt相关转录因子2(Runx2)、碱性磷酸酶(ALP)、骨桥蛋白(OPN)、骨钙蛋白(OCN)等成骨基因及蛋白表达量。结果过表达BMP9基因可以促进PDLSC中成骨标志物Runx2、ALP、OCN、OPN的表达(P<0.05)。给予磁场强度分别为1.2、1.8、2.4、3.0 mT的脉冲电磁场干预后,PDLSC内的成骨标志物表达水平均较单独BMP9时增高(P<0.05),并且在磁场强度为2.4 mT时达到最高峰(P<0.05),表明低频脉冲电磁场可以增强BMP9诱导PDLSC成骨分化并且存在“窗口效应”。结论磁场强度为1.8~3.0 mT的15 Hz脉冲电磁场可以体外增强BMP9诱导人PDLSC的成骨分化。Objective To investigate whether the low-frequency pulsed electromagnetic fields(LF-PEMFs)can enhance the osteogenic differentiation of human periodontal ligament stem cells(hPDLSCs)induced by bone morphogenetic protein 9(BMP9).Methods We isolated CD146+/STRO-1+cells(namely PDLSC)from periodontal ligament cells of healthy human premolars,transfected the PDLSC with BMP9-overexpressing recombinant adenoviruses(Ad-GFP-BMP9),exposed the cells to the different intensities of PEMF stimulation(15 Hz,0.6,1.2,1.8,2.4,3.0 mT,1 h/12 h),and then detected the expression of runt-related transcription factor 2(Runx2),alkaline phosphatase(ALP),osteopontin(OPN)and osteocalcin(OCN)by qRT-PCR and Western blotting.Results Overexpression of BMP9 significantly promoted the expression of osteogenic markers(Runx2,ALP,OCN and OPN)in PDLSC(P<0.05).After the intervention with 1.2,1.8,2.4 and 3.0 mT PEMF,the expression levels of osteogenic markers in PDLSC were significantly higher than those exposed to BMP9 alone(P<0.05),and reached the peak at 2.4 mT(P<0.05),indicating that LF-PEMFs enhanced BMP9-induced osteogenic differentiation of PDLSC,and there was a“window effect”.Conclusion LF-PEMFs stimulation(15 Hz,1.8 to 3.0 mT)can enhance BMP9-induced osteogenesis of hPDLSCs in vitro.

关 键 词：低频脉冲电磁场 牙周膜干细胞 骨形态发生蛋白9 成骨分化 窗口效应 

分 类 号：R781.4[医药卫生—口腔医学] R329.29[医药卫生—临床医学]