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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 |
