Faure Elodie, Wegrzyn Julien, Bernabei Ilaria, Falgayrac Guillaume, Bertheaume Nicolas, Pascart Tristan, Hugle Thomas, Busso Nathalie, Nasi Sonia
Service of Rheumatology, Department of Musculoskeletal Medicine, Centre Hospitalier Universitaire Vaudois and University of Lausanne, Lausanne, Switzerland.
Service of Orthopedics, Department of Musculoskeletal Medicine, Centre Hospitalier Universitaire Vaudois and University of Lausanne, Lausanne, Switzerland.
Rheumatology (Oxford). 2025 Feb 1;64(2):880-885. doi: 10.1093/rheumatology/keae064.
Cartilage pathologic calcification is a hallmark of osteoarthritis (OA). Here, we aimed to describe a new ex vivo human model to study the progression of cartilage calcification.
Cartilage explants (n = 11), as well as primary chondrocytes (n = 3), were obtained from OA patients undergoing knee replacement. Explants and chondrocytes were cultured in control (NT) or calcification (CM) medium (supplemented with ascorbic acid and β-glycerophosphate). Calcification was evaluated by micro-CT scan at day 0 and 21 in explants, and by Alizarin red staining in chondrocyte monolayers. Raman spectrometry allowed characterization of the crystal type. Interleukin-6 (IL-6) secretion in explant and cell supernatants was measured by ELISA. Finally, matrix degradation was evaluated by Safranin-O staining of explant sections and by glycosaminoglycans (GAG) released in supernatants.
Micro-CT scan showed calcifications in all explants at baseline (day 0), which in the CM group increased significantly in number and size after 21 days compared with the NT group. Raman spectrometry revealed that crystals were exclusively basic calcium phosphate crystals (carbonated hydroxyapatite) both in NT and CM. IL-6 secretion was significantly increased in calcifying conditions. Finally, CM significantly increased cartilage catabolism as assessed by decreased Safranin-O staining of tissue explants and increased GAG release in supernatants. CM effects (enhanced calcification, IL-6 secretion and proteoglycans turn-over) were recapitulated in vitro in OA chondrocytes.
We have described a new ex vivo human model of cartilage calcification that can summarize the triad of events seen during osteoarthritis progression, i.e. calcification, inflammation and cartilage degradation. This model will allow the identification of new anti-calcification compounds.
软骨病理性钙化是骨关节炎（OA）的一个标志。在此，我们旨在描述一种新的体外人体模型，以研究软骨钙化的进展。
从接受膝关节置换的OA患者获取软骨外植体（n = 11）以及原代软骨细胞（n = 3）。将外植体和软骨细胞在对照（NT）或钙化（CM）培养基（补充有抗坏血酸和β -甘油磷酸）中培养。通过对外植体在第0天和第21天进行微型计算机断层扫描（micro - CT扫描）评估钙化情况，通过对软骨细胞单层进行茜素红染色评估钙化情况。拉曼光谱法可对晶体类型进行表征。通过酶联免疫吸附测定（ELISA）测量外植体和细胞上清液中白细胞介素 - 6（IL - 6）的分泌。最后，通过对外植体切片进行番红O染色以及测量上清液中释放的糖胺聚糖（GAG）评估基质降解情况。
微型计算机断层扫描显示所有外植体在基线（第0天）时就存在钙化，与NT组相比，CM组在21天后钙化的数量和大小显著增加。拉曼光谱显示，NT组和CM组中的晶体均仅为碱性磷酸钙晶体（碳酸羟基磷灰石）。在钙化条件下，IL - 6分泌显著增加。最后，通过组织外植体番红O染色减少以及上清液中GAG释放增加评估，CM显著增加了软骨分解代谢。OA软骨细胞在体外重现了CM的作用（增强钙化、IL - 6分泌和蛋白聚糖周转）。
我们描述了一种新的软骨钙化体外人体模型，该模型可概括骨关节炎进展过程中出现的一系列事件，即钙化、炎症和软骨降解。该模型将有助于鉴定新的抗钙化化合物。
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