Osteoarthritis Research Program, Division of Orthopedic Surgery, Schroeder Arthritis Institute, University Health Network, Canada; Krembil Research Institute, University Health Network, Canada; Institute of Biomedical Engineering, University of Toronto, Canada.
Osteoarthritis Research Program, Division of Orthopedic Surgery, Schroeder Arthritis Institute, University Health Network, Canada; Krembil Research Institute, University Health Network, Canada.
Osteoarthritis Cartilage. 2022 Jan;30(1):147-159. doi: 10.1016/j.joca.2021.09.007. Epub 2021 Sep 20.
There is a need to incorporate multiple tissues into in vitro OA models to evaluate novel therapeutics. This approach is limited by inherent donor variability. We present an optimized research tool: a human OA cartilage-synovium explant co-culture model (OA-EXM) that employs donor-matched lower and upper limit response controls combined with statistical approaches to address variability. Multiple rapid read-outs allow for evaluation of therapeutics while cataloguing cartilage-synovium interactions.
48-h human explant cultures were sourced from OA knee arthroplasties. An OA-like cartilage-synovium co-culture baseline was established relative to donor-matched upper limit supraphysiological pro-inflammatory cytokine and lower limit OA cartilage or synovium alone controls. 100 nM dexamethasone treatment validated possible "rescue effects" within the OA-EXM dual tissue environment. Gene expression, proteoglycan loss, MMP activity, and soluble protein concentrations were analyzed using blocking and clustering methods.
The OA-EXM demonstrates the value of the co-culture approach as the addition of OA synovium increases OA cartilage proteoglycan loss and expression of MMP1, MMP3, MMP13, CXCL8, CCL2, IL6, and PTGS2, but not to the extent of supraphysiological stimulation. Conversely, OA cartilage does not affect gene expression or MMP activity of OA synovium. Dexamethasone shows dual treatment effects on synovium (pro-resolving macrophage upregulation, protease downregulation) and cartilage (pro-inflammatory, catabolic, and anabolic downregulation), and decreases soluble CCL2 levels in co-culture, thereby validating OA-EXM utility.
The OA-EXM is representative of late-stage OA pathology, captures dual interactions between cartilage and synovium, and combined with statistical strategies provides a rapid, sensitive research tool for evaluating OA therapeutics.
为了评估新疗法，需要将多种组织纳入体外 OA 模型中。但这种方法受到供体固有变异性的限制。我们提出了一种优化的研究工具：一种人 OA 软骨-滑膜共培养模型（OA-EXM），它采用与供体匹配的上下限反应对照，并结合统计方法来解决变异性问题。多个快速读出允许评估治疗方法，同时记录软骨-滑膜相互作用。
从 OA 膝关节置换术获得 48 小时的人组织培养物。相对于与供体匹配的上限位超生理促炎细胞因子和下限位 OA 软骨或滑膜单独对照，建立了类似 OA 的软骨-滑膜共培养基线。用 100nM 地塞米松处理验证了 OA-EXM 双组织环境中可能的“挽救作用”。使用阻断和聚类方法分析基因表达、蛋白聚糖损失、MMP 活性和可溶性蛋白浓度。
OA-EXM 证明了共培养方法的价值，因为添加 OA 滑膜会增加 OA 软骨蛋白聚糖的损失和 MMP1、MMP3、MMP13、CXCL8、CCL2、IL6 和 PTGS2 的表达，但不如超生理刺激的程度。相反，OA 软骨不会影响 OA 滑膜的基因表达或 MMP 活性。地塞米松对滑膜（促修复巨噬细胞上调、蛋白酶下调）和软骨（促炎、分解代谢和合成代谢下调）具有双重治疗作用，并降低共培养物中的可溶性 CCL2 水平，从而验证了 OA-EXM 的实用性。
OA-EXM 代表晚期 OA 病理，捕捉软骨和滑膜之间的双重相互作用，并结合统计策略，为评估 OA 治疗方法提供了一种快速、敏感的研究工具。
