Sports Medicine Center, Department of Orthopedic Surgery and Orthopedic Research Institute, West China Hospital, Sichuan University, Chengdu, Sichuan, China.
Department of Orthopedic Surgery and Orthopedic Research Institute, Laboratory of Stem Cell and Tissue Engineering, State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, Chengdu, Sichuan, China.
Am J Sports Med. 2023 Sep;51(11):3008-3024. doi: 10.1177/03635465231187042. Epub 2023 Aug 2.
Osteoarthritis (OA) is a common disease that causes joint pain and disability. Stem cell therapy is emerging as a promising treatment for OA.
To evaluate the ability of peripheral blood-derived mesenchymal stem cells (PBMSCs) combined with donor-matched platelet-rich plasma (PRP) to treat OA in a rabbit model.
Controlled laboratory study.
PBMSCs and donor-matched PRP were isolated and prepared from the same rabbit. PBMSCs were treated with serum-free medium, fetal bovine serum, and PRP; a series of PBMSC behaviors, including proliferation, migration, and adhesion, were compared among groups. The ability of PBMSCs or PRP alone and PBMSCs+PRP to protect chondrocytes against proinflammatory cytokine (interleukin 1β [IL-1β]) treatment was compared by analyzing reactive oxygen species (ROS)-scavenging ability and apoptosis. Real-time quantitative polymerase chain reaction and immunofluorescence were used to investigate the expression of extracellular matrix (ECM) metabolism genes and proteins, and Western blotting was used to explore the potential mechanism of the corresponding signaling pathway. In vivo, the effect of PBMSCs+PRP on cartilage and inflammation of the synovium was observed in a surgery-induced OA rabbit model via gross observation, histological and immunohistochemical staining, and enzyme-linked immunosorbent assay.
Proliferation, migration, and adhesion ability were enhanced in PBMSCs treated with PRP. Moreover, compared with either PBMSCs or PRP alone, PBMSCs+PRP enhanced ROS-scavenging ability and inhibited apoptosis in IL-1β-treated chondrocytes. PBMSCs+PRP also reversed the IL-1β-induced degradation of collagen type 2 and aggrecan and increased expression of matrix metalloproteinase 13, and this effect was related to increased expression of ECM synthesis and decreased expression of degradation and inflammatory genes and proteins. Mechanistically, PBMSCs+PRP reduced the phosphorylation of inhibitor of nuclear factor-κBα (IκBα), which further inhibited the phosphorylation of downstream nuclear factor-κB (NF-κB) in the NF-κB signaling pathway. In vivo, compared with PBMSCs or PRP alone, intra-articular (IA) injection of PBMSCs+PRP enhanced cartilage regeneration and attenuated synovial inflammation in OA-induced rabbits.
These results demonstrate that PRP could enhance biological activities, including viability, migration, and adhesion, in PBMSCs. PBMSCs+PRP could rescue ECM degeneration by inhibiting inflammatory signaling in IL-1β-treated OA chondrocytes. In addition, IA injection of PBMSCs+PRP effectively attenuated OA progression in a surgery-induced OA rabbit model.
PBMSCs+PRP may provide a promising treatment for knee OA, and this study can advance the related basic research.
骨关节炎（OA）是一种常见的疾病，会导致关节疼痛和功能障碍。干细胞疗法作为 OA 的一种有前途的治疗方法正在出现。
评估外周血源性间充质干细胞（PBMSCs）与供体匹配的富血小板血浆（PRP）联合治疗兔 OA 模型的能力。
对照实验室研究。
从同一只兔子中分离和制备 PBMSCs 和供体匹配的 PRP。将 PBMSCs 用无血清培养基、胎牛血清和 PRP 处理；比较各组 PBMSC 的增殖、迁移和黏附等行为。通过分析活性氧（ROS）清除能力和细胞凋亡，比较单独使用 PBMSCs 或 PRP 以及 PBMSCs+PRP 保护软骨细胞免受促炎细胞因子（白细胞介素 1β[IL-1β]）治疗的能力。实时定量聚合酶链反应和免疫荧光用于研究细胞外基质（ECM）代谢基因和蛋白质的表达，Western blot 用于探讨相应信号通路的潜在机制。在体内，通过大体观察、组织学和免疫组织化学染色以及酶联免疫吸附试验，观察 PBMSCs+PRP 在手术诱导的 OA 兔模型中对软骨和滑膜炎症的影响。
PRP 处理后的 PBMSCs 增殖、迁移和黏附能力增强。此外，与单独使用 PBMSCs 或 PRP 相比，PBMSCs+PRP 增强了 IL-1β 处理的软骨细胞中的 ROS 清除能力并抑制了细胞凋亡。PBMSCs+PRP 还逆转了 IL-1β 诱导的 II 型胶原和聚集蛋白聚糖的降解，并增加了基质金属蛋白酶 13 的表达，这种作用与 ECM 合成的增加和降解及炎症基因和蛋白质的表达减少有关。在机制上，PBMSCs+PRP 降低了核因子-κB 抑制因子-α（IκBα）的磷酸化，从而进一步抑制了 NF-κB 信号通路中下游核因子-κB（NF-κB）的磷酸化。在体内，与单独使用 PBMSCs 或 PRP 相比，关节内（IA）注射 PBMSCs+PRP 增强了 OA 诱导兔的软骨再生并减轻了滑膜炎症。
这些结果表明，PRP 可以增强 PBMSCs 的生物学活性，包括活力、迁移和黏附。PBMSCs+PRP 可通过抑制 IL-1β 处理的 OA 软骨细胞中的炎症信号来挽救 ECM 退变。此外，IA 注射 PBMSCs+PRP 可有效减轻手术诱导的 OA 兔模型中的 OA 进展。
PBMSCs+PRP 可能为膝骨关节炎提供一种有前途的治疗方法，本研究可为相关基础研究提供参考。
