Cadelano Francesca, Della Morte Elena, Niada Stefania, Anzano Francesco, Zagra Luigi, Giannasi Chiara, Brini Anna Teresa Maria
Department of Biomedical, Surgical and Dental Sciences, University of Milan, Milan, Italy.
Laboratory of Biotechnological Applications, IRCCS Istituto Ortopedico Galeazzi, Milan, Italy.
Regen Ther. 2024 Jun 28;26:346-353. doi: 10.1016/j.reth.2024.06.010. eCollection 2024 Jun.
Osteoarthritis (OA), a chronic inflammatory joint disorder, still lacks effective therapeutic interventions. Consequently, the development of convenient experimental models is crucial. Recently, research has focused on the plasticity of Mesenchymal Stem/stromal Cells, particularly adipose-derived ones (ASCs), in halting OA progression. This study investigates the therapeutic potential of a cell-free approach, ASC-derived conditioned medium (CM), in reversing cytokine-induced OA markers in an model of human cartilage explants.
4 mm cartilage punches, derived from the femoral heads of patients undergoing total hip replacement, were treated with 10 ng/ml TNFα, 1 ng/ml IL-1β, or a combination of both, over a 3-day period. Analysis of OA-related markers, such as MMP activity, the release of NO and GAGs, and the expression of , allowed for the selection of the most effective inflammatory stimulus. Subsequently, explants challenged with TNFα+IL-1β were exposed to CM, consisting of a pool of concentrated supernatants from 72-h cultured ASCs, in order to evaluate its effect on cartilage catabolism and inflammation.
The 3-day treatment with both 10ng/ml TNFα and 1ng/ml IL-1β significantly increased MMP activity and NO release, without affecting GAG release. The addition of CM significantly downregulated the abnormal MMP activity induced by the inflammatory stimuli, while also mildly reducing , and gene expression. Finally, and were downregulated by the cytokines, and further decreased by CM.
The proposed cartilage explant model offers encouraging evidence of the therapeutic potential of ASC-derived CM against OA, and it could serve as a convenient platform for drug screening.
骨关节炎（OA）是一种慢性炎症性关节疾病，目前仍缺乏有效的治疗干预措施。因此，开发便捷的实验模型至关重要。最近，研究聚焦于间充质干/基质细胞，尤其是脂肪来源的间充质干细胞（ASCs）在阻止OA进展方面的可塑性。本研究调查了一种无细胞方法——ASC来源的条件培养基（CM）在逆转人软骨外植体模型中细胞因子诱导的OA标志物方面的治疗潜力。
从接受全髋关节置换术患者的股骨头获取4毫米的软骨块，在3天时间内用10纳克/毫升肿瘤坏死因子α（TNFα）、1纳克/毫升白细胞介素-1β（IL-1β）或两者组合进行处理。对OA相关标志物进行分析，如基质金属蛋白酶（MMP）活性、一氧化氮（NO）和糖胺聚糖（GAGs）的释放以及[此处原文缺失相关基因名称]的表达，以选择最有效的炎症刺激物。随后，用TNFα + IL-1β刺激的外植体暴露于CM（由72小时培养的ASCs的浓缩上清液池组成），以评估其对软骨分解代谢和炎症的影响。
用10纳克/毫升TNFα和1纳克/毫升IL-1β进行3天治疗显著增加了MMP活性和NO释放，而不影响GAG释放。添加CM显著下调了炎症刺激诱导的异常MMP活性，同时也轻度降低了[此处原文缺失相关基因名称]、[此处原文缺失相关基因名称]和[此处原文缺失相关基因名称]基因的表达。最后，[此处原文缺失相关基因名称]和[此处原文缺失相关基因名称]被细胞因子下调，并被CM进一步降低。
所提出的软骨外植体模型为ASC来源的CM对OA的治疗潜力提供了令人鼓舞的证据，并且它可以作为一个便捷的药物筛选平台。
