Hamasaki Masanari, Terkawi Mohamad Alaa, Onodera Tomohiro, Homan Kentaro, Iwasaki Norimasa
Department of Orthopedic Surgery, Faculty of Medicine and Graduate School of Medicine, Hokkaido University, Sapporo, Japan.
Global Station for Soft Matter, Global Institution for Collaborative Research and Education (GSS, GI-CoRE), Hokkaido University, Sapporo, Japan.
Cartilage. 2021 Jul;12(3):354-361. doi: 10.1177/1947603519828426. Epub 2019 Feb 1.
Osteoarthritis is a progressive joint disease characterized by cartilage degradation and synovial inflammation. Presence of cartilage fragments in the joint due to degradation of cartilage is thought to be associated with local inflammatory response and progressive osteoarthritic process. Understanding the mechanism by which cartilage fragments elicit this destructive process should aid in designing novel therapeutic approaches. Therefore, objective of current study is to establish an in vitro model to examine the cross-talk between chondrocytes and cartilage fragments-stimulated macrophages.
Cartilage fragments were prepared from femoral head cartilages of mice and analyzed using a scanning electron microscope and particle size analyzer. Bone marrow-derived macrophages were co-cultured with cartilage fragments and chondrocytes using transwell co-culture system. Macrophage inflammatory mediators in supernatant of cultures were determined by ELISA and gene expression of macrophages and chondrocyte were quantified by qRT-PCR.
Shapes of cartilage fragments were irregular with sizes ranged between 0.54 and 55 μm. Macrophages cultured with cartilage fragments released significantly higher concentrations of TNF-α, IL-6, and NO than those of mock and control. Consistently, gene expressions of , and were significantly increased in stimulated macrophages. The elevation in production of pro-inflammatory molecules in stimulated macrophages cultures were coincident with an increase in gene expression of chondrocyte , and .
We developed an co-culture model to study the impact of stimulation of macrophage by cartilage fragments on the expression of chondrocyte carbolic factors. Our results revealed that cartilage fragments triggered macrophages inflammatory response that enhanced the production of chondrocyte catabolic factors.
骨关节炎是一种以软骨降解和滑膜炎症为特征的进行性关节疾病。由于软骨降解导致关节中存在软骨碎片，被认为与局部炎症反应和进行性骨关节炎进程有关。了解软骨碎片引发这一破坏过程的机制应有助于设计新的治疗方法。因此，本研究的目的是建立一个体外模型，以研究软骨细胞与软骨碎片刺激的巨噬细胞之间的相互作用。
从小鼠股骨头软骨制备软骨碎片，并用扫描电子显微镜和粒度分析仪进行分析。使用Transwell共培养系统将骨髓来源的巨噬细胞与软骨碎片和软骨细胞共培养。通过ELISA测定培养上清液中的巨噬细胞炎症介质，并通过qRT-PCR定量巨噬细胞和软骨细胞的基因表达。
软骨碎片形状不规则，大小在0.54至55μm之间。与软骨碎片共培养的巨噬细胞释放的TNF-α、IL-6和NO浓度明显高于模拟组和对照组。一致地，刺激的巨噬细胞中 、 和 的基因表达显著增加。刺激的巨噬细胞培养物中促炎分子产生的升高与软骨细胞 、 和 的基因表达增加一致。
我们建立了一个 共培养模型，以研究软骨碎片刺激巨噬细胞对软骨细胞分解代谢因子表达的影响。我们的结果表明，软骨碎片引发巨噬细胞炎症反应，增强了软骨细胞分解代谢因子的产生。
