Paracrine effect of inflammatory cytokine-activated bone marrow mesenchymal stem cells and its role in osteoblast function
Paracrine effect of inflammatory cytokine-activated bone marrow mesenchymal stem cells and its role in osteoblast function
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炎症细胞因子激活的骨髓间充质干细胞的旁分泌作用及其对成骨细胞功能的作用
DOI：
10.1016/j.jbiosc.2015.05.017
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发表时间：
2016-02-01
影响因子：
2.8
通讯作者：
Zhou, Haibin
中科院分区：
文献类型：
--
作者：
Li, Cheng;Li, Guoqiang;Zhou, Haibin
间充质干细胞（MSCs）在骨再生中具有重要的功能。炎症是成骨微环境的一个有充分证据的组成部分。在本研究中，我们研究了炎症细胞因子刺激MSCs是否通过旁分泌介质促进骨生成。用炎症因子ifn - γ和tnf - α预先刺激MSCs。预刺激后，骨髓间充质干细胞IL-6、HGF、VEGF、tgf - β分泌水平显著升高（p < 0.01）；而IL-2、IL-4、IL-10的产生无明显变化（p < 0.05）。MG63是一种成骨细胞样细胞系，在不同的msc条件培养基中培养。经细胞因子预处理的MSCs条件培养基处理后，MG63细胞的增殖和迁移明显改善，成骨细胞分化标志物ALP、COLT、OCN和OPN的表达水平显著升高（p < 0.05）。此外，免疫荧光染色显示更多的MG63细胞呈opn阳性，而茜素红染色显示ifn - γ和tnf - α联合预处理组钙结节的形成增加。结果表明，炎症细胞因子激活MSCs的条件培养基可以显著促进成骨细胞的增殖、迁移、分化和矿化，并最终通过旁分泌机制促进成骨。这些发现为MSCs在骨缺损修复中的临床应用提供了新的方向。(C) 2015，生物技术学会，日本。版权所有。
Mesenchymal stem cells (MSCs) have a crucial function in bone regeneration. Inflammation is a well-documented component of the osteogenic microenvironment. In the present study, we investigated whether stimulation of MSCs with inflammatory cytokines promotes osteogenesis through a paracrine mediator. MSCs were pre-stimulated with the inflammatory factors IFN-gamma and TNF-alpha. After pre-stimulation, the MSC secretion levels of IL-6, HGF, VEGF, and TGF-beta were significantly elevated (p < 0.01); however, the production of IL-2, IL-4, and IL-10 was not changed (p > 0.05). MG63, an osteoblast-like cell line, was cultured in different MSC-conditioned media. After treatment with conditioned media collected from MSCs pre-treated with cytokines, the proliferation and migration of MG63 cells were significantly improved, and the expression levels of the osteoblast differentiation markers ALP, COLT, OCN and OPN were significantly increased as revealed by a quantitative PCR analysis (p < 0.05). Furthermore, an immunofluorescence staining assay showed that more MG63 cells were OPN-positive, while an Alizarin red staining indicated the increased formation of calcium nodules in the IFN-gamma and TNF-alpha combined pretreatment group. The results indicated that conditioned medium from inflammatory cytokine-activated MSCs can significantly promote osteoblast proliferation, migration, differentiation, and mineralization and ultimately enhance osteogenesis through paracrine mechanisms. These findings present a new direction for the clinical application of MSCs in the repair of bone defects. (C) 2015, The Society for Biotechnology, Japan. All rights reserved.