Grupo de Terapia Celular y Medicina Regenerativa, Dpto. de Fisioterapia, Medicina y Ciencias Biomédicas, Facultad de Ciencias de La Salud, Universidade da Coruña, INIBIC-CHUAC, CICA, 15006, A Coruña, Spain.
Cell Mol Life Sci. 2022 Oct 20;79(11):557. doi: 10.1007/s00018-022-04580-z.
Osteoarthritis (OA) is closely linked to the increase in the number of senescent cells in joint tissues, and the senescence-associated secretory phenotype (SASP) is implicated in cartilage degradation. In the last decade, extracellular vesicles (EV) in combination with the use of miRNAs to modify post-transcriptional expressions of multiple genes have shown their utility in new therapies to treat inflammatory diseases. This work delves into the anti-inflammatory effect of extracellular vesicles derived from mesenchymal stem cells (MSC) previously modified to inhibit the expression of miR-21. We compare the efficacy of two treatments, MSC with their miR-21 inhibited through lentiviral transfection and their EV, against inflammation in a new OA animal model. The modified MSC and their EV were intraperitoneally injected in an OA animal model twice. One month after treatment, we checked which therapy was the most effective to reduce inflammation compared with animals untreated. Treated OA model sera were analyzed for cytokines and chemokines. Subsequently, different organs were analyzed to validate the results obtained. EV were the most effective treatment to reduce chemokines and cytokines in serum of OA animals as well as SASP, in their organs checked by proteomic and genomic techniques, compared with MSC alone in a statistically significant way. In conclusion, MSC-miR-21-derived EV showed a higher therapeutic potential in comparison with MSCs-miR-21-. They ameliorate the systemic inflammation through inactivation of ERK1/2 pathway in OA in vivo model. Workflow of the realization of the animal model of OA by injecting cells into the joint cavity of the left knee of the animals, which produces an increase in serum cytokines and chemokines in the animals in addition to the increase in SASP and markers of inflammation. Inhibition of miR-21 in MSCs, from the stroma of the human umbilical cord, by lentivirus and extraction of their EVs by ultracentrifugation. Finally, application of MSC therapy with its miR-21 inhibited or its EVs produces a decrease in serum cytokines and chemokines in the treated animals, in addition to an increase in SASP and markers of inflammation. The cell-free therapy being the one that produces a greater decrease in the parameters studied.
骨关节炎 (OA) 与关节组织中衰老细胞数量的增加密切相关，衰老相关分泌表型 (SASP) 与软骨降解有关。在过去的十年中，细胞外囊泡 (EV) 结合使用 miRNA 来修饰多个基因的转录后表达，已显示出它们在治疗炎症性疾病的新疗法中的应用价值。这项工作深入研究了先前通过慢病毒转染抑制 miR-21 表达的间充质干细胞 (MSC) 衍生的细胞外囊泡的抗炎作用。我们比较了两种治疗方法的疗效，即经慢病毒转染抑制 miR-21 的 MSC 及其 EV，针对新的 OA 动物模型中的炎症。改良的 MSC 和它们的 EV 通过腹腔内注射到 OA 动物模型中两次。治疗一个月后，我们检查了哪种治疗方法最有效，可以与未治疗的动物相比降低炎症。分析治疗性 OA 模型血清中的细胞因子和趋化因子。随后，分析不同器官以验证获得的结果。与单独的 MSC 相比，EV 在统计学上更有效地降低 OA 动物血清中的趋化因子和细胞因子以及 SASP，并用蛋白质组学和基因组学技术检查其器官。总之，与 MSC-miR-21-相比，MSC-miR-21 衍生的 EV 显示出更高的治疗潜力。它们通过在体内 OA 模型中失活 ERK1/2 途径来改善全身性炎症。通过将细胞注射到动物左膝关节腔中来实现 OA 动物模型的工作流程，这会导致动物血清细胞因子和趋化因子增加，以及 SASP 和炎症标志物增加。通过慢病毒抑制人脐带基质细胞中的 miR-21，并通过超速离心提取其 EV。最后，应用 MSC 治疗及其 miR-21 抑制或其 EVs 会导致治疗动物血清细胞因子和趋化因子减少，以及 SASP 和炎症标志物增加。无细胞治疗是产生研究参数下降幅度更大的一种治疗方法。
