Inhibitory effect of alcohol on osteogenic differentiation in human bone marrow-derived mesenchymal stem cells
Inhibitory effect of alcohol on osteogenic differentiation in human bone marrow-derived mesenchymal stem cells
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DOI：
10.1097/01.alc.0000118315.58404.c1
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发表时间：
2004-03-01
影响因子：
3.2
通讯作者：
Wezeman, FH
中科院分区：
文献类型：
--
作者：
Gong, ZD;Wezeman, FH
背景：酒精引起的骨质疏松症的特征是骨生成受到相当大的抑制。本研究的目的是确定酒精对基因表达，蛋白质，合成和矿化的影响，在人骨髓间充质干细胞诱导向成骨分化在vitro.Methods：人骨髓间充质干细胞诱导向成骨培养在存在或不存在的50 mM酒精。通过使用针对细胞表面抗原CD 105/内皮糖蛋白的SH 2抗体来表征干细胞，并且定量它们在醇存在下的增殖。通过北方分析定量成骨谱系的早期、中期和晚期标志物的基因表达，并测定骨基质蛋白合成。酒精对细胞介导的基质矿化在终末分化culture.Results的影响进行了测定von Kossa染色：与Percoll梯度分离的人骨髓间充质干细胞的活化细胞分选分析证明99%的同质性通过使用SH 2抗体的表面抗原CD 105。这些干细胞的增殖的剂量依赖性抑制发生在浓度大于50 mM的酒精。成骨细胞特异性，因子2/核心结合因子a1（Osf 2/Cbfa 1），I型胶原蛋白，碱性磷酸酶和骨钙素（早期，中期和晚期标记成骨，分别）的基因表达进行了分析，有和没有成骨诱导和治疗50 mM酒精。诱导后，Osf 2/Cbfa 1水平对酒精无反应。为了确定酒精对人间充质干细胞进展的影响，沿着成骨途径，在成骨诱导后检测I型胶原、碱性磷酸酶和骨钙素的信使RNA（mRNA）水平。成骨诱导后，酒精下调I型胶原基因表达，并显着减少其合成。酒精并没有改变碱性磷酸酶的mRNA表达，骨生成的中期标志物，但显着降低其活性相比，单独的成骨诱导。诱导后，骨钙素在mRNA和蛋白水平上均未被酒精改变。组织化学显示碱性磷酸酶染色减少，酒精处理的人骨髓间充质干细胞培养物中碱性磷酸酶阳性细胞较少。von Kossa染色显示，在酒精treatment.Conclusions后，干细胞培养物中矿化结节的数量减少：总的来说，数据表明，酒精alters-osteogenic分化在人骨髓间充质干细胞培养过程中的谱系进展，并提供进一步了解酒精诱导减少骨形成。
Background: Alcohol-induced osteoporosis is characterized by a considerable suppression of osteogenesis. The objective of this investigation was to determine the effect of alcohol on gene expression, protein, synthesis, and mineralization in human bone marrow-derived mesenchymal stem cells induced toward osteogenic differentiation in vitro.Methods: Human bone marrow-derived mesenchymal stem cells induced toward osteogenesis were cultured in the presence or absence of 50 mM alcohol. Stem cells were characterized by using SH2 antibody to the cell-surface antigen CD105/endoglin, and their proliferation in the presence of alcohol was quantified . The expression of genes for early, middle, and late markers of the osteogenic lineage was quantified by Northern analysis, and bone matrix protein synthesis was assayed. The effect of alcohol on cell-mediated matrix mineralization in terminally differentiated cultures was determined by von Kossa staining.Results: Fluorescence-activated cell sorting analysis of human mesenchymal stem cells separated with a Percoll gradient proved 99% homogeneity by using SH2 antibody to the surface antigen CD105. Dose-dependent inhibition of proliferation of these stem cells occurred at concentrations greater than 50 mM alcohol. Gene expression of osteoblast-specific, factor 2/core binding factor a1 (Osf2/Cbfa1), type I collagen, alkaline phosphatase, and osteocalcin (early, middle; and late markers for osteogenesis, respectively) was analyzed with and without osteogenic induction and treatment with 50 mM alcohol. After induction, Osf2/Cbfa1 levels were unresponsive to alcohol. To determine the effect of alcohol on human mesenchymal stem cell progression,along the osteogenic pathway, messenger RNA (mRNA) levels for type I Collagen, alkaline phosphatase, and osteocalcin were examined after osteogenic induction. After osteogenic induction, alcohol down-regulated the gene expression of type I collagen and significantly reduced its synthesis. Alcohol did not alter mRNA expression of alkaline phosphatase, a midstage marker for osteogenesis, but significantly decreased its activity compared with osteogenic induction alone. After induction, osteocalcin remained unchanged by alcohol at both the mRNA and protein levels. Histochemistry revealed decreased alkaline phosphatase staining and fewer alkaline phosphatase-positive cells in alcohol-treated human mesenchymal stem cell cultures. von Kossa staining revealed a reduction in the number of mineralizing nodules in stem cell cultures after alcohol treatment.Conclusions: Collectively, the data suggest that alcohol alters-osteogenic differentiation in human bone marrow-derived mesenchymal stem cell cultures during-lineage progression and provide further insight into alcohol-induced reduced bone formation.