Osteogenic effect of bone marrow mesenchymal stem cell-derived exosomes on steroid-induced osteonecrosis of the femoral head
Osteogenic effect of bone marrow mesenchymal stem cell-derived exosomes on steroid-induced osteonecrosis of the femoral head
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DOI：
10.2147/dddt.s178698
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发表时间：
2019-01-01
影响因子：
4.8
通讯作者：
Chen, Peng
中科院分区：
文献类型：
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作者：
Fang, Shanhong;Li, Yongfeng;Chen, Peng
坏境：动物实验已经证实了间充质干细胞（mesenchymal stem cells，MSCs）对成骨的治疗作用，但对骨髓间充质干细胞（bone MSCs，BMSCs）释放的外泌体（exosomes，Exos）的功能知之甚少。在这里，我们研究了BMSC Exos对激素诱导的股骨头坏死（SFHN）的影响，并探讨了参与这一过程的重要基因。材料和方法：从健康和SFHN大鼠中分离BMSC。使用外泌体沉淀试剂盒分离BMSC外泌体，并通过透射电子显微镜和蛋白质印迹进行表征。将SFHN BMSC与来自健康BMSC的Exos孵育。油红O染色和茜素红染色评价成骨能力。使用成骨RT 2 Profiler PCR阵列筛选由Exos诱导的差异表达基因（DEG）。结果：体外培养的BMSC Exos大小为100-150 nm，表达CD 63。此外，与模型细胞相比，BMSC Exo处理的SFHN细胞表现出抑制的脂肪形成。PCR阵列显示，与模型组相比，BMSC Exo处理的SFHN细胞中分别有11个和9个基因上调和下调。在Dais中，成骨相关基因，包括Bmp 2、Bmp 6、Bmpr 1b、Mmp 9和Sox 9，可能在SHIN中起重要作用。此外，DEG主要参与免疫应答、成骨细胞分化和转化生长因子-β/骨形态发生蛋白信号通路。结论：我们的数据表明，BMSC来源的Exos主要影响SFHN的成骨作用，这一发现可以进一步研究，以开发一种新的治疗SFHN的药物。
Badcground: Animal studies have demonstrated the therapeutic effect of mesenchymal stem cells (MSCs) on osteogenesis, but little is known about the functions of exosomes (Exos) released by bone MSCs (BMSCs). Here, we investigated the effect of BMSC Exos on steroid-induced femoral head necrosis (SFHN) and explored the vital genes involved in this process.Materials and methods: BMSCs were isolated from healthy and SFHN rats. BMSC Exos were isolated using the Exosome Precipitation Kit and characterized by transmission electron microscopy and Western blotting. SFHN BMSCs were incubated with Exos from healthy BMSCs. Osteogenic ability was assessed by oil red O staining and alizarine red staining. Differentially expressed genes (DEGs) induced by Exos were screened using the Osteogenesis RT2 Profiler PCR Array. The effect of upregulated Sox9 was examined using lentivirus-mediated siRNA.Results: The results revealed that BMSC Exos were 100-150 nm in size and expressed CD63. Moreover, BMSC Exo-treated SFHN cells exhibited suppressed adipogenesis compared to model cells. PCR array showed that eleven and nine genes were upregulated and downregulated, respectively, in the BMSC Exo-treated SFHN cells compared to the model group. Among the Dais, osteogenesis-related genes, including Bmp2, Bmp6, Bmpr1b, Mmp9, and Sox9, may play important roles in SHIN. Furthermore, the DEGs were mainly involved in immune response, osteoblast differentiation, and in the transforming growth factor-beta/bone morphogenetic protein signaling pathway. The level of the SOX9 protein was upregulated by Exos, and Sox9 silencing significantly decreased the osteogenic effect of BMSC Exos.Conclusion: Our data suggest that Exos derived from BMSCs mainly affect SFHN osteogenesis, and this finding can be further investigated to develop a novel therapeutic agent for SFHN.