Indomethacin induces differential effects on in vitro endochondral ossification depending on the chondrocyte's differentiation stage
Indomethacin induces differential effects on in vitro endochondral ossification depending on the chondrocyte's differentiation stage
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DOI：
10.1002/jor.23324
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发表时间：
2017-04-01
影响因子：
2.8
通讯作者：
Welting, Tim J. M.
中科院分区：
文献类型：
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作者：
Caron, Marjolein M. J.;Emans, Pieter J.;Welting, Tim J. M.
异位骨化（HO）是软组织骨的异常形成，是髋关节置换术中常见的并发症。异位骨化是通过软骨内成骨形成的，标准治疗方法是使用吲哚美辛。目前尚不清楚吲哚美辛如何在分子水平上影响异位骨化；因此，我们的目的是确定吲哚美辛是否可能通过损害软骨内成骨的软骨形成阶段来影响异位骨化。在软骨谱系中分化的祖细胞模型（ATDC5，原代人骨髓干细胞和离体骨膜琼脂糖培养物）用增加浓度的吲哚美辛处理，观察到软骨和增生性标志物的基因和蛋白质表达减少（通过RT-qPCR和免疫印迹检测）以及糖胺聚糖含量减少（通过阿利新蓝组织化学）。即使引起增生性分化，吲哚美辛的加入也导致增生性标志物表达降低。有趣的是，当用吲哚美辛处理成熟软骨细胞时，观察到2型胶原蛋白表达明显增加。同样，当ATDC5细胞和骨髓干细胞预分化获得软骨细胞表型时，从这个时间点开始添加吲哚美辛，低浓度的吲哚美辛也会导致软骨细胞分化增加。吲哚美辛对体外软骨内成骨的影响不同，取决于软骨细胞的分化阶段，其中最明显的作用是完全抑制软骨细胞分化。这一观察结果可能为吲哚美辛治疗异位骨化的难以捉摸的作用模式提供了一个合理的解释。(C) 2016骨科研究学会。
Heterotopic ossification (HO) is the abnormal formation of bone in soft tissues and is a frequent complication of hip replacement surgery. Heterotopic ossifications are described to develop via endochondral ossification and standard treatment is administration of indomethacin. It is currently unknown how indomethacin influences heterotopic ossification on a molecular level; therefore, we aimed to determine whether indomethacin might influence heterotopic ossification via impairing the chondrogenic phase of endochondral ossification. Progenitor cell models differentiating in the chondrogenic lineage (ATDC5, primary human bone marrow stem cells and ex vivo periosteal agarose cultures) were treated with increasing concentrations of indomethacin and a decrease in gene- and protein expression of chondrogenic and hypertrophic markers (measured by RT-qPCR and immunoblotting) as well as decreased glycosamino-glycan content (by alcian blue histochemistry) was observed. Even when hypertrophic differentiation was provoked, the addition of indomethacin resulted in decreased hypertrophic marker expression. Interestingly, when mature chondrocytes were treated with indomethacin, a clear increase in collagen type 2 expression was observed. Similarly, when ATDC5 cells and bone marrow stem cells were pre-differentiated to obtain a chondrocyte phenotype and indomethacin was added from this time point onward, low concentrations of indomethacin also resulted in increased chondrogenic differentiation. Indomethacin induces differential effects on in vitro endochondral ossification, depending on the chondrocyte's differentiation stage, with complete inhibition of chondrogenic differentiation as the most pronounced action. This observation may provide a rational behind the elusive mode of action of indomethacin in the treatment of heterotopic ossifications. (C) 2016 Orthopaedic Research Society.