Pan K, Yan S, Ge S, Li S, Zhao Y, Yang P
Department of Periodontology, School of Dentistry, Shandong University, Jinan, China.
Cell Prolif. 2009 Jun;42(3):364-72. doi: 10.1111/j.1365-2184.2009.00599.x. Epub 2009 Apr 24.
Bone morphogenic protein-2 (BMP-2) has long been used to promote bone and periodontal regeneration, while core binding factor alpha1 (CBFA1) plays important roles in both osteogenic differentiation and tooth morphogenesis. The aim of this study was to evaluate the effects of CBFA1 or BMP-2 overexpression on osteoblast/cementoblast-related gene expressions in NIH3T3 cells and dental follicle cells (DFCs).
CBFA1 or BMP-2 overexpression in NIH3T3 and DFCs was achieved by infection with retroviral vectors containing CBFA1 or BMP-2 cDNA. Cells stably integrated with CBFA1 or BMP-2 cDNA were selected with G418 for 14 days. Western blotting, real-time reverse transcriptase-polymerase chain reaction, and in vitro mineralization assay were performed to evaluate effects of CBFA1 or BMP-2 overexpression in cells undergoing osteoblast/cementoblast differentiation.
Our results demonstrated that osteoblast/cementoblast-related gene expression levels in CBFA1-overexpressing NIH3T3 cells were higher than those in BMP-2-overexpressing cells. More mineral nodules were observed in CBFA1-overexpressing NIH3T3 cells than in BMP-2-overexpressing cells. CBFA1 overexpression in DFCs also increased osteoblast/cementoblast-related gene expression and promoted mineral nodule formation. However, no significant changes in gene expression levels nor mineral nodule formation were found in BMP-2-overexpressing DFCs when compared with empty vector transduced DFCs.
CBFA1 overexpression up-regulated expression levels of osteoblast/cementoblast-related genes and enhanced in vitro osteogenic differentiation more efficiently than BMP-2 in both NIH3T3 cells and DFCs.
骨形态发生蛋白-2(BMP-2)长期以来一直用于促进骨和牙周组织再生,而核心结合因子α1(CBFA1)在成骨细胞分化和牙齿形态发生中均发挥重要作用。本研究旨在评估CBFA1或BMP-2过表达对NIH3T3细胞和成牙骨质细胞相关基因表达以及对牙囊细胞(DFCs)的影响。
通过用含有CBFA1或BMP-2 cDNA的逆转录病毒载体感染来实现NIH3T3细胞和DFCs中CBFA1或BMP-2的过表达。用G418筛选稳定整合有CBFA1或BMP-2 cDNA的细胞14天。进行蛋白质免疫印迹法、实时逆转录-聚合酶链反应和体外矿化试验,以评估CBFA1或BMP-2过表达对成骨细胞/成牙骨质细胞分化过程中细胞的影响。
我们的结果表明,过表达CBFA1的NIH3T3细胞中,成骨细胞/成牙骨质细胞相关基因表达水平高于过表达BMP-2的细胞。过表达CBFA1的NIH3T3细胞中观察到的矿化结节比过表达BMP-2的细胞更多。DFCs中CBFA1过表达也增加了成骨细胞/成牙骨质细胞相关基因表达并促进了矿化结节形成。然而,与转导空载体的DFCs相比,过表达BMP-2的DFCs中基因表达水平和矿化结节形成均未发现显著变化。
在NIH3T3细胞和DFCs中,CBFA1过表达上调了成骨细胞/成牙骨质细胞相关基因的表达水平,并且比BMP-2更有效地增强了体外成骨分化。