Alveolar bone osteoblast differentiation and Runx2/Cbfa1 expression
Alveolar bone osteoblast differentiation and Runx2/Cbfa1 expression
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DOI：
10.1016/j.archoralbio.2005.09.004
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发表时间：
2006-05-01
影响因子：
3
通讯作者：
Dziak, Rosemary
中科院分区：
文献类型：
--
作者：
Perinpanayagam, Hiran;Martin, Thomas;Dziak, Rosemary
牙槽骨细胞具有独特的起源和功能，但可能类似于骨骼成骨细胞。成骨细胞分化和基因表达受Runx2/Cbfa1转录因子调控。然而，大多数关于Runx2/Cbfa1表达的研究都是在啮齿动物细胞上进行的，而对人类成骨细胞的少数研究则有不同的结果。本研究的目的是表征Runx2/Cbfa1在人牙槽骨原代细胞培养物中的表达。将牙槽骨芯片在补充有胎牛血清（10%FCS）的α-最低必需培养基（α-MEM）中孵育。在长出3 - 4周后收获外植体培养物，并在含有FCS的a-MEM中生长。该培养基补充有抗坏血酸盐、P-甘油磷酸盐和地塞米松以促进成骨细胞分化超过14天。RT-PCR分析和蛋白质印迹显示Runx2/Cbfa1 mRNA（2.1倍）和蛋白质（2.3倍）水平在3天内迅速增加，随后略有下降。骨唾液酸蛋白表达在3天内也迅速增加（2.9倍），随后在14天进一步增加（3.6倍）。碱性磷酸酶表达（1.6倍）和活性（3.1倍）在7天内缓慢增加，随后逐渐下降。相比之下，胶原mRNA水平在14天内几乎没有变化。这些发现证实了来源于人牙槽骨的原代细胞培养物的成骨潜力。人牙槽骨中的成骨细胞分化涉及Runx2/Cbfa1表达的增加，这可能是分化过程的重要组成部分。(c)2005爱思唯尔有限公司保留所有权利。
Alveolar bone cells have a unique origin and functionality, but may resemble skeletal osteoblasts. Osteoblast differentiation and gene expression are regulated by the Runx2/Cbfa1 transcription factor. However, most studies on Runx2/Cbfa1 expression have been on rodent cells and the few studies on human osteoblasts have had differing results. The purpose of this study was to characterize Runx2/Cbfa1 expression in primary cell cultures derived from human alveolar bone. An alveolar bone chip was incubated in a-minimum essential medium (alpha-MEM) supplemented with fetal calf serum (10% FCS). Explant cultures were harvested after 3-4 weeks of outgrowth and grown in a-MEM with FCS. This media was supplemented with ascorbate, P-glycerophosphate and dexamethasone to promote osteoblast differentiation over 14 days. RT-PCR analysis and Western blots showed a rapid increase in Runx2/Cbfa1 mRNA (2.1-fold) and protein (2.3-fold) levels in 3 days, followed by a slight decline. There was also a rapid increase in bone sialoprotein expression (2.9-fold) in 3 days, followed by a further increase (3.6-fold) at 14 days. There was a slower increase in alkaline phosphatase expression (1.6-fold) and activity (3.1-fold) over 7 days, followed by a gradual decline. In contrast, collagen mRNA levels showed little change over 14 days. These findings attest to the osteogenic potential of primary cell cultures derived from human alveolar bone. Osteoblastic differentiation in human alveolar bone involves an increase in Runx2/Cbfa1 expression that may be an important component of the differentiation process. (c) 2005 Elsevier Ltd. All rights reserved.