UMR 7365 CNRS-UL, IMoPA (Ingénierie Moléculaire et Physiopathologie Articulaire), Biopôle de l'Université de Lorraine, Campus Brabois-Santé, 9 Avenue de la Forêt de Haye, BP 20199, 54505, Vandœuvre-Lès-Nancy, France.
Service de Chirurgie Orthopédique, Traumatologique et Arthroscopique, CHRU Nancy, 29 Avenue du Maréchal de Lattre de Tassigny CO 60034, F54035, Nancy, France.
Stem Cell Res Ther. 2018 Nov 28;9(1):329. doi: 10.1186/s13287-018-1071-2.
Mesenchymal stem cells (MSCs) are found in synovial fluid (SF) and can easily be harvested during arthrocentesis or arthroscopy. However, SF-MSC characterization and chondrogenicity in collagen sponges have been poorly documented as well as their hypothetical in vivo chondroprotective properties with intra-articular injections during experimental osteoarthritis (OA).
SF-MSCs were isolated from human SF aspirates in patients suffering from advanced OA undergoing total knee joint replacements. SF-MSCs at passage 2 (P2) were characterized by flow cytometry for epitope profiling. SF-MSCs at P2 were subsequently cultured in vitro to assess their multilineage potentials. To assess their chondrogenicity, SF-MSCs at P4 were seeded in collagen sponges for 4 weeks under various oxygen tensions and growth factors combinations to estimate their gene profile and matrix production. Also, SF-MSCs were injected into the joints in a nude rat anterior cruciate ligament transection (ACLT) to macroscopically and histologically assess their possible chondroprotective properties,.
We characterized the stemness (CD73+, CD90+, CD105+, CD34-, CD45-) and demonstrated the multilineage potency of SF-MSCs in vitro. Furthermore, the chondrogenic induction (TGF-ß1 ± BMP-2) of these SF-MSCs in collagen sponges demonstrated a good capacity of chondrogenic gene induction and extracellular matrix synthesis. Surprisingly, hypoxia did not enhance matrix synthesis, although it boosted chondrogenic gene expression (ACAN, SOX9, COL2A1). Besides, intra-articular injections of xenogenic SF-MSCs did exert neither chondroprotection nor inflammation in ACLT-induced OA in the rat knee.
Advanced OA SF-MSCs seem better candidates for cell-based constructs conceived for cartilage defects rather than intra-articular injections for diffuse OA.
间充质干细胞(MSCs)存在于滑液(SF)中,在关节穿刺或关节镜检查期间很容易收获。然而,SF-MSC 的特征及其在胶原海绵中的软骨生成能力以及它们在实验性骨关节炎(OA)期间关节内注射的潜在软骨保护特性记录不佳。
从接受全膝关节置换术的晚期 OA 患者的 SF 抽吸物中分离 SF-MSCs。通过流式细胞术对第 2 代(P2)SF-MSCs 进行表型分析。随后,将 P2 SF-MSCs 在体外培养以评估其多能性。为了评估其软骨生成能力,将 P4 SF-MSCs 接种于胶原海绵中,在不同氧张力和生长因子组合下培养 4 周,以评估其基因谱和基质产生。此外,将 SF-MSCs 注射到裸鼠前交叉韧带切断(ACLT)关节中,以宏观和组织学评估其可能的软骨保护特性。
我们对干细胞特性(CD73+、CD90+、CD105+、CD34-、CD45-)进行了特征描述,并在体外证明了 SF-MSCs 的多能性。此外,这些 SF-MSCs 在胶原海绵中的软骨诱导(TGF-β1±BMP-2)显示出良好的软骨基因诱导和细胞外基质合成能力。令人惊讶的是,缺氧并没有增强基质合成,尽管它增强了软骨基因表达(ACAN、SOX9、COL2A1)。此外,异种 SF-MSCs 的关节内注射在大鼠膝关节 ACLT 诱导的 OA 中既没有发挥软骨保护作用,也没有引发炎症。
晚期 OA SF-MSCs 似乎更适合用于软骨缺损的基于细胞的构建体,而不是用于弥漫性 OA 的关节内注射。