Effects of tricalcium silicate cements on osteogenic differentiation of human bone marrow-derived mesenchymal stem cells in vitro.
Effects of tricalcium silicate cements on osteogenic differentiation of human bone marrow-derived mesenchymal stem cells in vitro.
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DOI：
10.1016/j.actbio.2014.04.006
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发表时间：
2014-07
影响因子：
9.7
通讯作者：
Tay, Franklin R.
中科院分区：
文献类型：
--
作者：
Eid, Ashraf A.;Hussein, Khaled A.;Niu, Li-na;Li, Guo-hua;Watanabe, Ikuya;Al-Shabrawey, Mohamed;Pashley, David H.;Tay, Franklin R.
关键词：
硅酸三钙骨水泥作为生物活性骨和牙本质替代物已成功应用于生物医学领域，具有广泛的骨活性。本研究分析了不同硅酸三钙骨水泥配方对人骨髓间充质干细胞（hMW-MSCs）时间骨活性的影响。将这些细胞暴露于成骨分化培养基中的4种市售硅酸三钙骨水泥制剂。分别于术后1、3、7、10 d，采用真实的时间-聚合酶链反应（RT-PCR）和Western blotting检测成骨标志物ALP、RUNX 2、OSX、OPN、MSX 2、OCN的表达。在3、7、14和21天后，进行碱性磷酸酶测定以检测细胞内酶水平的变化。28天后进行茜素红S测定以检测细胞外基质矿化。在硅酸三钙骨水泥的存在下，目标成骨标志物在mRNA和蛋白质水平在所有时间点下调。细胞内碱性磷酸酶水平和细胞外矿化实验组与未经处理的对照组没有显着差异。定量聚合酶链反应结果显示，随着硅酸三钙骨水泥暴露时间的增加，RUNX 2、OSX、MSX 2和OCN的下调增加，而ALP在第7天出现峰值下调。对于蛋白质印迹，OSX、OPN、MSX 2和OCN显示随着暴露于测试的骨水泥的时间增加而下调增加。碱性磷酸酶水平一般在第7天后下降。基于这些结果，可以得出结论，硅酸三钙水泥不诱导hBM-MSCs在体外成骨分化。
Tricalcium silicate cements have been successfully employed in the biomedical field as bioactive bone and dentin substitutes, with widely acclaimed osteoactive properties. This research analyzed the effects of different tricalcium silicate cement formulations on the temporal osteoactivity profile of human bone marrow-derived mesenchymal stem cells (hMW-MSCs). These cells were exposed to 4 commercially-available tricalcium silicate cement formulations in osteogenic differentiation medium. After 1, 3, 7 and 10 days, quantitative real time-polymerase chain reaction and Western blotting were performed to detect the expression of target osteogenic markers ALP, RUNX2, OSX, OPN, MSX2, and OCN. After 3, 7, 14 and 21 days, alkaline phosphatase assay was performed to detect changes in intracellular enzyme level. Alizarin Red S assay was performed after 28 days to detect extracellular matrix mineralization. In the presence of tricalcium silicate cements, target osteogenic markers were downregulated at the mRNA and protein levels at all time-points. Intracellular alkaline phosphatase enzyme levels and extracellular mineralization of the experimental groups were not significantly different from the untreated control. Quantitative polymerase chain reaction results showed increases in downregulation of RUNX2, OSX, MSX2 and OCN with increase in time of exposure to the tricalcium silicate cements, while ALP showed peak downregulation at day 7. For Western blotting, OSX, OPN, MSX2 and OCN showed increased downregulation with increased exposure time to the tested cements. Alkaline phosphatase enzyme levels generally declined after day 7. Based on these results, it is concluded that tricalcium silicate cements do not induce osteogenic differentiation of hBM-MSCs in vitro.
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DOI：
10.4049/jimmunol.0900803
发表时间：
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期刊：
Journal of immunology (Baltimore, Md. : 1950)
影响因子：
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影响因子：
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影响因子：
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