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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 |
王彦蒽1△,汪沛2△,王晟磊2,吴贤慧1,孙学兰1,刘军1,叶晖1,冯琴1,俞佳颖1,曹志中1,唐卫忠1* |
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(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 △共同第一作者 *通信作者) |
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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15Hz、磁场强度分别为0.6、1.2、1.8、2.4、3.0mT的脉冲电磁场进行干预(每12h辐照1h)。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、3.0mT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2.4mT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1.8~3.0mT的15Hz脉冲电磁场可以体外增强BMP9诱导人PDLSC的成骨分化。 |
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 |
DOI:10.16781/j.0258-879x.2020.08.0855 |
投稿时间:2020-02-12修订日期:2020-04-29 |
基金项目: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). |
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Low frequency pulsed electromagnetic fields enhance bone morphogenetic protein 9-induced osteogenic differentiation of periodontal ligament stem cells |
WANG Yan-en1△,WANG Pei2△,WANG Sheng-lei2,WU Xian-hui1,SUN Xue-lan1,LIU Jun1,YE Hui1,FENG Qin1,YU Jia-ying1,CAO Zhi-zhong1,TANG Wei-zhong1* |
(1. Department of Stomatology, Changhai Hospital, Naval Medical University(Second Military Medical University), Shanghai 200433, China; 2. Department of Oral Implant and Maxillofacial Surgery, Hongkou Branch of Changhai Hospital, Naval Medical University(Second Military Medical University), Shanghai 200081, China △Co-first authors. * Corresponding author) |
Abstract: |
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. |
Key words: low frequency pulsed electromagnetic fields periodontal ligament stem cells bone morphogenetic protein 9 osteogenic differentiation window effect |