摘要
目的:研究重组人骨形态发生蛋白(rhBMP-2)和重组人碱性成纤维细胞生长因子(FGF-2)对成骨细胞(MC3T3-E1 Subclone 14)矿化及焦磷酸合成酶(ENPP1)、跨膜蛋白(ANK)和组织非特异性碱性磷酸酶(TNAP)表达的影响,探讨生长因子影响细胞矿化的机制。方法:将MC3T3-E1 Subclone 14细胞分成3组:成骨细胞诱导液(OS)成骨诱导培养组(对照组),OS与rhBMP-2培养组(rhBMP-2组),OS与FGF-2培养组(FGF-2组)。培养12 d后进行ALP活性检测及茜素红染色,实时荧光定量PCR检测矿化相关基因ENPP1、ANK和TNAP表达的差异。结果:rhBMP-2组ALP活性以及钙化结节明显高于对照组,ENPP1、ANK和TNAP均高表达;FGF-2组ALP活性以及钙化结节明显低于对照组,ENPP1和ANK呈高表达,TNAP低表达。结论:rhBMP-2和FGF-2通过调节ENPP1、ANK和TNAP的表达变化来影响骨的矿化。
Objective： The aim of this study was to test the effects of rhBMP-2 and FGF-2 on osteoblast mineralization, and ENPP1, ANK and TNAP gene expression on osteoblast-like cell line MC3T3-E1 Subclone 14 in vitro. Methods： The MC3T3-E1 Subclone 14 cells were divided into 3 groups. Group 1, osteoblast-like cells were cultured only with osteogenic medium as control group. Group 2, cells were cultured with rhBMP-2 ＋ osteogenic medium. Group3, cells were cultured with FGF-2＋ osteogeuic medium. Alkaline phosphatase activities were determined and specific mineralization detection was performed with Alizarin red staining. Osteoblast mineralization-related gene expression of ENPP1, ANK and TNAP were evaluated by real -time PCR. Analysis of variance was performed by SPSS 13.0 software. Data were expressed as the ^-＋s of triplicate measurements. P〈0.05 was accepted as significant. Results： In rhBMP-2 intervention group, ALP activities, calcium deposition and the mineralization-related genes expression were markedly higher than those of control group. In FGF-2 intervention group, ALP activities and calcium deposition were markedly lower than those of control group, the mRNA of ENPP1 and ANK was highly expressed, the mRNA of TNAP was marked lowly expressed than those of control group. Conclusion： rhBMP-2 and FGF-2 may regulate ENPP1, ANK and TNAP gene expression on osteoblast mineralization.
出处
《口腔颌面外科杂志》
CAS
2012年第5期333-336,共4页
Journal of Oral and Maxillofacial Surgery
