Hyaluronic acid promotes osteogenic differentiation of human amniotic mesenchymal stem cells through the TGF-β/Smad signaling pathway
Hyaluronic acid promotes osteogenic differentiation of human amniotic mesenchymal stem cells through the TGF-β/Smad signaling pathway
复制标题
透明质酸通过TGF-β/Smad信号通路促进人羊膜间充质干细胞的成骨分化
DOI：
10.1016/j.lfs.2019.116669
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发表时间：
2019
期刊：
影响因子：
6.1
通讯作者：
Jian-Hui Xiao
中科院分区：
文献类型：
--
作者：
Ling-Tao Zhang;Ru-Ming Liu;Yi Luo;Yu-Jie Zhao;Dai-Xiong Chen;Chang-Yin Yu;Jian-Hui Xiao
目的研究常用成骨介质透明质酸(HA)及其联合应用对人羊膜间充质干细胞(HAMSCs)成骨分化的影响，并探讨其作用机制。碱性磷酸酶(ALP)和钙沉积实验检测hAMSC的成骨分化。分别用定量逆转录聚合酶链式反应(qRT-PCR)和Western blotting检测成骨相关基因和蛋白的表达。结果表明，与DAG组相比，CG组能显著刺激hAMSC碱性磷酸酶活性和钙沉积，而HA组作用不明显。成骨相关分子和干性相关分子在mRNA和蛋白水平均有表达上调，其中CG组上调最为显著。此外，与DAG处理相比，CG处理显著增加了参与调节转化生长因子-β/Smad信号通路的基因表达。此外，当转化生长因子-β/Smad通路的抑制剂SB431542预先处理细胞时，CG处理组的成骨分化效应和基因表达上调被显著抑制。这些结果表明，HA与DAG联合应用可能通过转化生长因子-β/Smad信号通路促进HAMSCs的成骨分化。
AimsThis study investigated the effects of hyaluronic acid (HA), a commonly used osteogenic medium referred to as DAG, and the combined administration of HA and DAG (CG) on the osteogenic differentiation of human amniotic mesenchymal stem cells (hAMSCs), and the underlying mechanism.Main methodsThe phenotype of hAMSCs was detected by flow cytometry and immunocytochemical staining. Alkaline phosphatase (ALP) and calcium deposition assays were employed for evaluating the osteogenic differentiation of hAMSCs. The expression of osteogenesis-related genes and proteins was determined by quantitative reverse transcription PCR (qRT-PCR) and Western blotting, respectively. Meanwhile, the molecular mechanism of osteogenic differentiation of hAMSCs was detected by PCR array and qRT-PCR.Key findingsThe results showed that treatment with CG could significantly stimulate hAMSC ALP activity and calcium deposition compared to treatment with DAG, while HA had little effect. The expression of osteogenesis-related molecules and stemness-related molecules was up-regulated at the mRNA and protein levels in all three groups, and this up-regulation was most significant in the CG group. In addition, treatment with CG significantly increased the gene expressions involved in regulation of the TGF-β/Smad signalling pathway compared to treatment with DAG. Furthermore, the pro-osteogenic differentiation effects as well as the up-regulated expression of genes observed in the CG treatment group were significantly inhibited when the cells were pre-treated with SB431542, an inhibitor of the TGF-β/Smad pathway.SignificanceThese results suggest that HA in combination with DAG could significantly enhance the osteogenic differentiation of hAMSCs, potentiallyviathe TGF-β/Smad signalling pathway.