口腔医学研究

口腔医学研究 ›› 2018, Vol. 34 ›› Issue (8): 857-861.DOI: 10.13701/j.cnki.kqyxyj.2018.08.014

• 牙周病学研究 • 上一篇    下一篇

重组人骨形态发生蛋白-9对人牙周膜成纤维细胞成骨分化的影响

陆蓓   

  1. 成都市第二人民医院口腔科 四川 成都 610017
  • 收稿日期:2018-01-25 出版日期:2018-08-28 发布日期:2018-08-23
  • 作者简介:陆蓓(1973~ ),女,浙江余姚人,学士,副主任医师,研究方向:牙髓治疗,E-mail:tap2463@163.com
  • 基金资助:
    四川省科技厅基金(编号:2015SZ0158)

Effect of Recombinant Human BMP-9 on Osteogenesis of Human Periodontal Ligament Fibroblasts

LU Bei   

  1. Department of Stomatology, Second People's Hospital of Chengdu, Chengdu 610017, China
  • Received:2018-01-25 Online:2018-08-28 Published:2018-08-23

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摘要： 目的:研究重组人骨形态发生蛋白-9(recombinant human bone morphogenetic protein-9, rhBMP-9)对体外人牙周膜成纤维细胞成骨细胞分化的影响。方法:体外培养人牙周膜成纤维细胞,成骨分化培养基(osteogenic differentiation medium,ODM)诱导成骨分化,加入不同浓度rhBMP-9或p38和胞外信号调节激酶1/2(extracellular signalregulated kinase1/2,ERK1/2)抑制剂SB203580和U0126处理。采用LabAssayTM碱性磷酸酶(alkaline phosphatase,ALP)检测试剂盒检测ALP活性。提取总RNA,实时荧光定量聚合酶链反应(Real time fluorescence quantitative PCR,qPCR)检测RUNT相关转录因子2(Runt-related transcription factor 2,RUNX2)、分化抑制因子1(inhibitor of differentiation factor 1,ID1)、骨唾液酸蛋白(bone sialoprotein,BSP)和骨桥蛋白(osteopontin,OPN)表达情况。结果:rhBMP-9可以显著增加人牙周膜成纤维细胞ALP活性,且呈浓度和时间依赖性。成骨分化相关转录因子RUNX2和ID1以及晚期成骨标志物BSP和OPN的表达在rhBMP-9的刺激下也显著升高。与rhBMP-9单独刺激组比较,加入SB203580和U0126共刺激后可完全抑制rhBMP-9促人牙周膜成纤维细胞成骨分化的作用。结论:rhBMP-9能够显著诱导人牙周膜成纤维细胞向成骨细胞样细胞分化,且其可能机制为调节p38和ERK1/2通路。

关键词: 重组人骨形态发生蛋白-9, 成纤维细胞, 成骨细胞分化, 胞外信号调节激酶1/2

Abstract: Objective: To investigate the effect of recombinant human BMP-9 (rhBMP-9) on the differentiation of human periodontal ligament fibroblasts into osteoblasts. Methods: Human periodontal ligament fibroblasts were cultured in vitro and osteogenic differentiation was induced by osteogenic differentiation medium (ODM). Then the cells were treated with different concentrations of rhBMP-9 in the presence or absence of p38 and extracellular signal-regulated kinase 1/2 (ERK1/2). Alkaline phosphatase (ALP) activity was measured using LabAssayTM ALP assay kit. The total RNA was extracted and real time fluorescence quantitative (qPCR) was used to detect the expressions of Runt-related transcription factor 2 (RUNX2),inhibitor of differentiation factor 1 (ID1),bone sialoprotein (BSP) and osteopontin (OPN). Results: RhBMP-9 stimulation significantly increased the ALP activity of human periodontal ligament fibroblasts in a concentration and time-dependent manner. Osteogenic differentiation-related transcription factors RUNX2 and ID1, as well as the expression of late osteogenic markers BSP and OPN, were also significantly increased by stimulation with rhBMP-9. Compared with rhBMP-9 alone, the effect of rhBMP-9 on osteogenic differentiation of human periodontal ligament fibroblasts was completely abolished by adding SB203580 and U0126 costimulation. Conclusion: RhBMP-9 could induce the osteogenic differentiation of human periodontal ligament fibroblasts, which might relate to the adjustment of p38 and ERK1/2 pathways.

Key words: Recombinant human bone morphogenetic protein-9, Fibroblasts, Osteoblasts, Extracellular signal-regulated kinase 1/2