Secretomes from mesenchymal stem cells participate in the regulation of osteoclastogenesis in vitro
Secretomes from mesenchymal stem cells participate in the regulation of osteoclastogenesis in vitro
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DOI：
10.1007/s00784-016-1986-x
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发表时间：
2017-07-01
影响因子：
3.4
通讯作者：
Hibi, Hideharu
中科院分区：
文献类型：
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作者：
Ogata, Kenichi;Katagiri, Wataru;Hibi, Hideharu
目的核因子kappa B受体激活剂配体(RANKL)抑制剂是一种新型的临床有效药物，通过与RANKL结合抑制破骨细胞的分化、功能和存活。药物相关性颌骨坏死(MRONJ)是一种新出现的骨坏死类型，其确切发病机制尚不清楚。最近的一些研究表明，静脉注射间充质干细胞(MSCs)改善了颌骨的骨坏死，推测MSCs分泌体的旁分泌作用是其主要成分。本研究旨在探讨人骨髓间充质干细胞无血清条件培养液(MSC-CM)和RANKL抑制剂对破骨细胞分化的影响。材料与方法应用细胞因子阵列分析鉴定MSC-CM中的细胞因子。在含有RANKL抑制剂的大鼠破骨细胞前体细胞培养液中加入MSC-CM。采用破骨细胞分化实验、免疫组织化学、实时定量逆转录聚合酶链式反应(RT-PCR)分析和凹坑形成实验。结果MSC-CM中含有多种细胞因子，如细胞募集、成骨、血管生成、细胞增殖等。MSC-CM促进破骨细胞分化和破骨细胞生成的主要调控转录因子的表达。结论MSC-CM中的分泌体参与了破骨细胞分化的调节，这可能降低了RANKL抑制剂的作用。
Objectives The receptor activator of nuclear factor kappa-B ligand (RANKL) inhibitors are novel clinically effective agents that inhibit osteoclast differentiation, function, and survival by binding to RANKL. Medication-related osteonecrosis of the jaw (MRONJ), caused as a result of treatment using denosumab, is a newly emerging type of bone necrosis, the exact pathogenesis of which is unknown. Several studies recently showed that the intravenous administration of mesenchymal stem cells (MSCs) improved the osteonecrosis of the jaw, and it was hypothesized that paracrine effects by secretomes from MSCs are the main constituent. Our aim was to investigate the effects of serum-free conditioned media from human MSCs (MSC-CM) and RANKL inhibitors on osteoclast differentiation.Materials and methods Cytokines included in MSC-CM were identified using the cytokine array analysis. MSC-CM was added to the culture medium of rat osteoclast precursors containing RANKL inhibitor. Osteoclast differentiation assays, immunohistochemistry, real-time reverse-transcriptase polymerase chain reaction (RT-PCR) analysis, and pit formation assays were performed.Results MSC-CM included various cytokines such as the recruitment of cell osteogenesis angiogenesis and cell proliferation. MSC-CM promoted osteoclast differentiation and expression of master regulatory transcriptional factors for osteoclastogenesis. In addition, MSC-CM showed function maintenance in osteoclasts despite the presence of RANKL inhibitors.Conclusions Our findings suggest that secretomes in MSC-CM were related to the regulation of osteoclast differentiation, which may reduce the effect of RANKL inhibitors.