The sequential expression profiles of growth factors from osteoprogenitors [correction of osteroprogenitors] to osteoblasts in vitro.

Zhinong Huang, Ehren Robert Nelson, R Lane Smith, Stuart B Goodman
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引用次数: 187

Abstract

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), platelet-derived 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.

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生长因子在体外从骨祖细胞到成骨细胞的序列表达谱[纠正骨祖细胞]。
在这项研究中,我们描述了与体外骨形成相关的选定生长因子的顺序表达。通过矿化、骨钙素和碱性磷酸酶(ALP-2)的测定来监测C57BL小鼠骨祖细胞的分化和成熟。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。采用酶联免疫吸附法(ELISA)和逆转录聚合酶链反应法(RT-PCR)分别检测生长因子蛋白和信使核糖核酸(mRNA)水平。结果发现,随着时间的推移,ALP-2的表达逐渐增加,而矿化和骨钙素直到培养第14天才变得明显。在细胞增殖早期,VEGF和IGF-1表达上调。PDGF和tgf - β mRNA呈双峰表达。FGF-2和BMP-2 mrna仅在分化后期表达。FGF-2 mRNA信号水平在培养第14天最高,并在培养第28天保持显著。BMP-2表现出与FGF-2相似的特征。RT-PCR和ELISA均未检测到BMP-7。几种早期反应生长因子(VEGF、tgf - β和IGF-1)的表达模式之间存在很强的相关性,而几种晚期反应生长因子(BMP-2、PDGF和FGF-2)的表达模式也很明显。成组生长因子的差异表达发生在骨特异性标记物的时间获取过程中,骨祖细胞在体外成熟。
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Tissue engineering
Tissue engineering CELL & TISSUE ENGINEERING-BIOTECHNOLOGY & APPLIED MICROBIOLOGY
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