The effect of purmorphamine and sirolimus on osteogenic differentiation of human bone marrow-derived mesenchymal stem cells
The effect of purmorphamine and sirolimus on osteogenic differentiation of human bone marrow-derived mesenchymal stem cells
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DOI：
10.1016/j.biopha.2012.10.004
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发表时间：
2013-02-01
影响因子：
7.5
通讯作者：
Salekdeh, G. H.
中科院分区：
文献类型：
--
作者：
Faghihi, F.;Eslaminejad, M. Baghaban;Salekdeh, G. H.
小分子已经作为较便宜的生物活性化合物引入，可以调节不同的发育现象。Purmorphamine和西罗莫司是两种小分子，根据一些研究，具有一定的骨调节作用。本研究旨在强调这些小分子在人骨髓间充质干细胞从早期到中期和晚期分化阶段中促进成骨的适当剂量和响应时间。碱性磷酸酶活性，基质矿化和成骨细胞基因的表达进行了定量评估在体外。对于体内研究，我们将基于干细胞的构建体皮下移植到大鼠体内，并每天用最有希望的小分子剂量对其进行治疗。通过实时PCR和组织学染色分析构建体。我们的研究结果表明，西罗莫司通过在诱导后14天和21天降低100 nM剂量的碱性磷酸酶活性和基质矿化来降低间充质干细胞的成骨分化。在第14天，Purmorphamine诱导碱性磷酸酶活性和RUNX-2表达的上调。在诱导后第14天，在3和5 μ M剂量的purmorphamine下检测到骨钙素的上调。在存在或不存在嘌呤吗啡的情况下，基质矿化保持不变。该剂量的小分子还加速了体内碱性磷酸酶转录物的表达。总之，西罗莫司对人骨髓间充质干细胞的成骨分化具有抑制作用;而purmorphamine，特别是在3 mM的剂量下，在体外和体内显示出促进作用。(C)2012年Elsevier Masson SAS。All rights reserved.
Small molecules have been introduced as less expensive biologically active compounds that can regulate different developmental phenomena. Purmorphamine and sirolimus are two small molecules that, according to some studies, possess certain osteomodulatory effects. This study was set out to highlight the appropriate dose and response time of these small molecules on enhancement of osteogenesis in human bone marrow-derived mesenchymal stem cells from early to mid and late stages of differentiation. Alkaline phosphatase activity, matrix mineralization and expression of osteoblast genes were quantitatively assessed in vitro. For the in vivo study, we transplanted stem cell-based constructs subcutaneously into rats, and treated them daily with the most promising doses of the small molecule. The constructs were analyzed by real-time PCR and histological staining. Our results showed that Sirolimus reduced osteogenic differentiation of mesenchymal stem cells by decreasing alkaline phosphatase activity at dose of 100 nM after 14 days and mineralization of the matrix at 14 and 21 days post-induction. Purmorphamine induced up-regulation of alkaline phosphatase activity and expression of RUNX-2 at day 14. Up-regulation of osteocalcin was detected at the 3 and 5 mu M doses of purmorphamine on day 14 post-induction. Matrix mineralization remained unchanged in the presence or absence of purmorphamine. This dose of small molecule also accelerated expression of Alkaline phosphatase transcripts in vivo. In conclusion, sirolimus had an inhibitory effect on osteogenic differentiation of human bone marrow-derived mesenchymal stem cells; while purmorphamine, particularly at a dose of 3 mM, showed a promotive effect in vitro and in vivo. (C) 2012 Elsevier Masson SAS. All rights reserved.