The sequential expression profiles of growth factors from osteroprogenitors to osteoblasts In vitro
The sequential expression profiles of growth factors from osteroprogenitors to osteoblasts In vitro
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DOI：
10.1089/ten.2006.0423
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发表时间：
2007-09-01
影响因子：
--
通讯作者：
Goodman, Stuart B.
中科院分区：
文献类型：
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作者：
Huang, Zhinong;Nelson, Ehren Robert;Goodman, Stuart B.
在这项研究中，我们描绘了体外骨形成相关的生长因子的顺序表达。测定矿化、骨钙素和碱性磷酸酶（ALP-2）以监测从C57 BL小鼠收集的骨祖细胞的分化和成熟。选择骨相关生长因子，包括转化生长因子β（TGF-β）、成纤维细胞生长因子2（FGF-2）、血小板衍生生长因子（PDGF）、胰岛素样生长因子（IGF）-1、血管内皮生长因子（VEGF）、骨形态发生蛋白（BMP）-2和BMP-7。采用酶联免疫吸附试验（ELISA）和逆转录聚合酶链反应（RT-PCR）分别在蛋白和信使核糖核酸（mRNA）水平上检测生长因子。结果发现，ALP-2的表达随着时间的推移逐渐增加，而矿化和骨钙素并没有变得明显，直到培养第14天。VEGF和IGF-1在增殖早期上调。PDGF和TGF-β mRNA表达呈双峰型。FGF-2和BMP-2 mRNA仅在分化后期表达。FGF-2 mRNA信号水平在第14天最高，并在培养的第28天保持显著。BMP-2显示出与FGF-2相似的特征。使用RT-PCR或ELISA检测不到BMP-7。几种早期反应生长因子（VEGF、TGF-β和IGF-1）之间的表达模式存在强相关性，几种晚期反应生长因子（BMP-2、PDGF和FGF-2）也存在明显相关性。随着骨祖细胞体外成熟的进行，在骨特异性标志物的时间获得过程中发生生长因子分组集的差异表达。
In this study, we delineate the sequential expression of selected growth factors associated with bone formation in vitro. Mineralization, osteocalcin, and alkaline phosphatase (ALP-2) were measured to monitor the differentiation and maturation of osteoprogenitor cells collected from C57BL mice. Bone-related growth factors, including transforming growth factor beta (TGF-beta), fibroblast growth factor 2 (FGF-2), plateletderived growth factor (PDGF), insulinlike growth factor (IGF)-1, vascular endothelial growth factor (VEGF), bone morphogenetic protein (BMP)-2, and BMP-7, were selected. Enzyme-linked immunosorbent assay (ELISA) and reverse transcriptase polymerase chain reaction (RT-PCR) were used to measure growth factors at the protein and messenger ribonucleic acid (mRNA) level, respectively. The results found that ALP-2 expression increased progressively over time, whereas mineralization and osteocalcin did not become evident until culture day 14. VEGF and IGF-1 were upregulated early during proliferation. PDGF and TGF-beta mRNA expression was bimodal. FGF-2 and BMP-2 mRNAs were expressed only later in differentiation. FGF-2 mRNA signal levels were highest at day 14 and remained prominent through day 28 of culture. BMP-2 showed a similar profile as FGF-2. BMP-7 was not detectable using RT-PCR or ELISA. Strong correlations existed for the expression patterns between several early-response growth factors (VEGF, TGF-beta, and IGF-1) and were also evident for several late-response growth factors (BMP-2, PDGF, and FGF-2). Differential expression for grouped sets of growth factors occurs during the temporal acquisition of bone-specific markers as osteoprogenitor cell maturation proceeds in vitro.