Department of Prosthodontics, School of Dentistry, Faculty of Health Sciences (FHS), Aristotle University of Thessaloniki (AUTh), 54124, Thessaloniki, Greece.
Department of Anatomy, Histology & Embryology, School of Veterinary Medicine, Aristotle University of Thessaloniki (AUTh), 54124, Thessaloniki, Greece.
Stem Cell Rev Rep. 2023 Nov;19(8):2957-2979. doi: 10.1007/s12015-023-10628-9. Epub 2023 Sep 26.
The potential therapeutic role of the Dental Pulp Stem Cells Secretome (SECR) in a rat model of experimentally induced Temporomandibular Joint (TMJ) Osteoarthritis (OA) was evaluated. Proteomic profiling of the human SECR under specific oxygen tension (5% O2) and stimulation with Tumor Necrosis Factor-alpha (TNF-α) was performed. SECR and respective cell lysates (CL) samples were collected and subjected to SDS-PAGE, followed by LC-MS/MS analysis. The identified proteins were analyzed with Bioinformatic tools. The anti-inflammatory properties of SECR were assessed via an in vitro murine macrophages model, and were further validated in vivo, in a rat model of chemically-induced TMJ-OA by weekly recording of the head withdrawal threshold, the food intake, and the weight change, and radiographically and histologically at 4- and 8-weeks post-treatment. SECR analysis revealed the presence of 50 proteins that were enriched and/or statistically significantly upregulated compared to CL, while many of those proteins were involved in pathways related to "extracellular matrix organization" and "immune system". SECR application in vitro led to a significant downregulation on the expression of pro-inflammatory genes (MMP-13, MMP-9, MMP-3 and MCP-1), while maintaining an increased expression of IL-10 and IL-6. SECR application in vivo had a significant positive effect on all the clinical parameters, resulting in improved food intake, weight, and pain suppression. Radiographically, SECR application had a significant positive effect on trabecular bone thickness and bone density compared to the saline-treated group. Histological analysis indicated that SECR administration reduced inflammation, enhanced ECM and subchondral bone repair and regeneration, thus alleviating TMJ degeneration.
本研究评估了牙髓干细胞 secretome（SECR）在实验诱导的颞下颌关节（TMJ）骨关节炎（OA）大鼠模型中的潜在治疗作用。在特定氧张力（5% O2）下对人 SECR 进行蛋白质组学分析，并刺激肿瘤坏死因子-α（TNF-α）。收集 SECR 和相应的细胞裂解物（CL）样本，进行 SDS-PAGE，然后进行 LC-MS/MS 分析。使用生物信息学工具分析鉴定的蛋白质。通过体外小鼠巨噬细胞模型评估 SECR 的抗炎特性，并通过每周记录头部退缩阈值、食物摄入量和体重变化以及在化学诱导的 TMJ-OA 大鼠模型中在 4 和 8 周治疗后进行影像学和组织学评估，进一步在体内验证 SECR 的抗炎特性。SECR 分析显示，与 CL 相比，SECR 中存在 50 种丰富和/或统计学上显著上调的蛋白质，而这些蛋白质中的许多参与与“细胞外基质组织”和“免疫系统”相关的途径。SECR 在体外的应用导致促炎基因（MMP-13、MMP-9、MMP-3 和 MCP-1）的表达显著下调，同时维持 IL-10 和 IL-6 的表达增加。SECR 在体内的应用对所有临床参数均有显著的积极影响，导致食物摄入量、体重和疼痛抑制的改善。影像学分析表明，与盐水处理组相比，SECR 应用对小梁骨厚度和骨密度有显著的积极影响。组织学分析表明，SECR 给药可减轻炎症，增强细胞外基质和软骨下骨修复和再生，从而缓解 TMJ 退变。
