Matrikine stimulation of equine synovial fibroblasts and chondrocytes results in an in vitro osteoarthritis phenotype.

IF 2.1 3区 医学 Q2 ORTHOPEDICS
Rachel Gagliardi, Drew W Koch, Richard Loeser, Lauren V Schnabel
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Abstract

Osteoarthritis (OA) is a debilitating disease that impacts millions of individuals and has limited therapeutic options. A significant hindrance to therapeutic discovery is the lack of in vitro OA models that translate reliably to in vivo preclinical animal models. An alternative to traditional inflammatory cytokine models is the matrikine stimulation model, in which fragments of matrix proteins naturally found in OA tissues and synovial fluid, are used to stimulate cells of the joint. The objective of this study was to determine if matrikine stimulation of equine synovial fibroblasts and chondrocytes with fibronectin fragments (FN7-10) would result in an OA phenotype. We hypothesized that FN7-10 stimulation of equine articular cells would result in an OA phenotype with gene and protein expression changes similar to those previously described for human chondrocytes stimulated with FN7-10. Synovial fibroblasts and chondrocytes isolated from four horses were stimulated in monolayer culture for 6 or 18 h with 1 µM purified recombinant 42 kD FN7-10 in serum-free media. At the conclusion of stimulation, RNA was collected for targeted gene expression analysis and media for targeted protein production analysis. Consistent with our hypothesis, FN7-10 stimulation resulted in significant alterations to many important genes that are involved in OA pathogenesis including increased expression of IL-1β, IL-4, IL-6, CCL2/MCP-1, CCL5/RANTES, CXCL6/GCP-2, MMP-1, MMP-3, and MMP13. The results of this study suggest that the equine matrikine stimulation model of OA may prove useful for in vitro experiments leading up to preclinical trials.

Matrikine 对马滑膜成纤维细胞和软骨细胞的刺激会导致体外骨关节炎表型。
骨关节炎(OA)是一种使人衰弱的疾病,影响着数百万人,而且治疗方案有限。治疗发现的一大障碍是缺乏能可靠地转化为体内临床前动物模型的体外 OA 模型。替代传统炎症细胞因子模型的一种方法是matrikine刺激模型,在这种模型中,OA组织和滑液中天然存在的基质蛋白片段被用来刺激关节细胞。本研究的目的是确定用纤维连接蛋白片段(FN7-10)刺激马滑膜成纤维细胞和软骨细胞是否会导致 OA 表型。我们假设,FN7-10 对马关节细胞的刺激会导致 OA 表型,其基因和蛋白质表达的变化类似于之前用 FN7-10 刺激人类软骨细胞时所描述的变化。在无血清培养基中,用 1 µM 纯化重组 42 kD FN7-10 刺激单层培养的滑膜成纤维细胞和软骨细胞 6 或 18 小时。刺激结束后,收集 RNA 进行定向基因表达分析,收集培养基进行定向蛋白质生产分析。与我们的假设一致,FN7-10 刺激导致许多参与 OA 发病机制的重要基因发生显著变化,包括 IL-1β、IL-4、IL-6、CCL2/MCP-1、CCL5/RANTES、CXCL6/GCP-2、MMP-1、MMP-3 和 MMP13 的表达增加。本研究结果表明,马蹄筋刺激 OA 模型可用于临床前试验前的体外实验。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Journal of Orthopaedic Research®
Journal of Orthopaedic Research® 医学-整形外科
CiteScore
6.10
自引率
3.60%
发文量
261
审稿时长
3-6 weeks
期刊介绍: The Journal of Orthopaedic Research is the forum for the rapid publication of high quality reports of new information on the full spectrum of orthopaedic research, including life sciences, engineering, translational, and clinical studies.
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