Proinflammatory cytokines inhibit osteogenic differentiation from stem cells: implications for bone repair during inflammation
Proinflammatory cytokines inhibit osteogenic differentiation from stem cells: implications for bone repair during inflammation
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DOI：
10.1016/j.joca.2008.11.011
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发表时间：
2009-06-01
影响因子：
7
通讯作者：
Hamilton, J. A.
中科院分区：
文献类型：
--
作者：
Lacey, D. C.;Simmons, P. J.;Hamilton, J. A.
目的：由于间充质干细胞（MSC）的分离困难，炎症对其骨发育的影响尚不清楚。本研究的目的是建立一种MSC分离方法，并确定白细胞介素-1 β (IL-1 β)和肿瘤坏死因子α (TNF α)在体外对其成骨分化的影响。方法：采用骨胶原酶消化法从C57/BI6小鼠四肢分离小鼠间充质干细胞，富集为干细胞抗原（Sca-1）（+） CD31(-) CD45（-）群体，然后采用荧光活化细胞分选（FACS）。在IL-1 β和TNF α存在或不存在的情况下，它们沿着成骨细胞谱系分化。通过定量聚合酶链反应（PCR）测量矿化和一些成骨基因的表达。结果：我们发现MSC群体的成骨分化受到IL-1 β和TNF α的抑制。除了抑制骨矿化外，这两种细胞因子还抑制分化相关的碱性磷酸酶（ALP）活性的增加以及ALP、α 1(I)前胶原、矮子相关转录因子2 （Runx2）和骨甾体的基因表达。然而，只有TNF α抑制骨连接蛋白和骨桥蛋白mRNA的表达，只有IL-1 β抑制细胞增殖。结论：便捷的分离技术可方便地生成足够的间充质干细胞，以便对其分化进行分子分析。因此，我们能够证明，促炎细胞因子，IL-1 β和TNF α，可以损害这些原发性MSC群体的骨发育，尽管存在一些显着差异。如果要利用间充质干细胞实现最佳的骨修复和其他组织修复，则需要考虑特定炎症介质的潜在参与。(C) 2008国际骨关节炎研究学会。Elsevier Ltd.出版。版权所有。
Objective: The effects of inflammation on bone development from mesenchymal stem cells (MSC) are unclear due to the difficulty in isolating MSC. The aim of this study was to develop a MSC isolation method and to determine the in vitro effects of interleukin-1 beta (IL-1 beta) and tumor necrosis factor alpha (TNF alpha) on their osteogenic differentiation.Methods: Murine MSC were isolated from the limbs of C57/BI6 mice through collagenase digestion of bone and enriched as the Stem cell antigen (Sca-1)(+) CD31(-) CD45(-) population, using lineage immunodepletion, followed by fluorescence-activated cell sorting (FACS). They were differentiated along the osteoblast linage in the presence or absence of IL-1 beta and TNF alpha. Mineralization was measured as was the expression of a number of osteogenic genes by quantitative polymerase chain reaction (PCR).Results: We show that osteogenic differentiation from the MSC population is suppressed by IL-1 beta and TNF alpha. In addition to suppression of bone mineralization, both cytokines inhibited the differentiation-associated increases in alkaline phosphatase (ALP) activity and the gene expression for ALP, alpha 1(I) procollagen, runt-related transcription factor 2 (Runx2) and osterix. However, only TNF alpha inhibited osteonectin and osteopontin mRNA expression and only IL-1 beta reduced cell proliferation.Conclusions: The convenient isolation technique enables the easy generation of sufficient MSC to permit the molecular analysis of their differentiation. We were thus able to show that the proinflammatory cytokines, IL-1 beta and TNF alpha, can compromise bone development from this primary MSC population, although with some significant differences. The potential involvement of specific inflammatory mediators needs to be taken into account if optimal bone repair and presumably that of other tissues are to be achieved with MSC. (C) 2008 Osteoarthritis Research Society International. Published by Elsevier Ltd. All rights reserved.