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Abstract：
Multiple sclerosis is a chronic inflammatory disease of the central nervous system characterized by autoimmune destruction of the myelin sheath, leading to irreversible and progressive functional deficits in patients. Pre-clinical studies involving the use of neural stem cells (NSCs) have already demonstrated their potential in neuronal regeneration and remyelination. However, the exclusive application of cell therapy has not proved sufficient to achieve satisfactory therapeutic levels. Recognizing these limitations, there is a need to combine cell therapy with other adjuvant protocols. In this context, extracellular vesicles (EVs) can contribute to intercellular communication, stimulating the production of proteins and lipids associated with remyelination and providing trophic support to axons. This study aimed to evaluate the therapeutic efficacy of the combination of NSCs and EVs derived from oligodendrocyte precursor cells (OPCs) in an animal model of multiple sclerosis. OPCs were differentiated from NSCs and had their identity confirmed by gene expression analysis and immunocytochemistry. Exosomes were isolated by differential ultracentrifugation and characterized by Western, transmission electron microscopy and nanoparticle tracking analysis. Experimental therapy of C57BL/6 mice induced with experimental autoimmune encephalomyelitis (EAE) were grouped in control, treated with NSCs, treated with OPC-derived EVs and treated with a combination of both. The treatments were evaluated clinically using scores and body weight, microscopically using immunohistochemistry and immunological profile by flow cytometry. The animals showed significant clinical improvement and weight gain with the treatments. However, only the treatments involving EVs led to immune modulation, changing the profile from Th1 to Th2 lymphocytes. Fifteen days after treatment revealed a reduction in reactive microgliosis and astrogliosis in the groups treated with EVs. However, there was no reduction in demyelination. The results indicate the potential therapeutic use of OPC-derived EVs to attenuate inflammation and promote recovery in EAE, especially when combined with cell therapy.
摘要：
多发性硬化症是一种中枢神经系统的慢性炎症性疾病，其特征是髓鞘的自身免疫破坏，导致患者不可逆和进行性功能缺陷。涉及使用神经干细胞（NSC）的临床前研究已经证明了它们在神经元再生和髓鞘再生中的潜力。然而,细胞疗法的排他性应用还不足以达到令人满意的治疗水平。认识到这些限制，需要将细胞疗法与其他辅助方案相结合.在这种情况下，细胞外囊泡（EV）可以促进细胞间通讯，刺激与髓鞘再生相关的蛋白质和脂质的产生，并为轴突提供营养支持。这项研究旨在评估NSC和源自少突胶质细胞前体细胞（OPCs）的EV组合在多发性硬化症动物模型中的治疗效果。OPCs从NSC分化,并且通过基因表达分析和免疫细胞化学证实了它们的身份。通过差异超速离心分离外泌体，并通过Western，透射电子显微镜和纳米粒子跟踪分析。将实验性自身免疫性脑脊髓炎（EAE）诱导的C57BL/6小鼠的实验治疗分为对照组，用NSC治疗，用OPC衍生的电动汽车治疗，并用两者的组合治疗。使用评分和体重对治疗进行临床评估，通过流式细胞术使用免疫组织化学和免疫学谱进行显微镜检查。随着治疗,动物显示出显著的临床改善和体重增加。然而,只有涉及电动汽车的治疗导致免疫调节，改变从Th1到Th2淋巴细胞的轮廓。治疗后15天，电动汽车治疗组的反应性小胶质细胞增生和星形胶质细胞增生减少。然而,脱髓鞘没有减少。结果表明OPC衍生的EV在减轻炎症和促进EAE恢复方面的潜在治疗用途，特别是与细胞疗法结合时。
