ABSTRACT

Objective

Acquired neurological diseases such as severe traumatic brain or spinal cord injury (SCI) cause irreversible disability. Olfactory ensheathing cell (OEC) transplantation has been trialed as a promising SCI treatment. Extracellular vesicles (EVs), which regulate cell–cell interactions, have recently garnered extensive research interests and emerged as a non-cell-based therapy in neurological disorders, including in SCI animal models. However, there have been no reports of human OEC-EVs and their beneficial effects on neuron regeneration. Here, we investigated the effects of EVs isolated from human OEC on the viability of neuronal cells.

Methods

EVs were isolated from primary human OECs (hOECs) by serial ultracentrifugation. The hOEC-EVs were characterized by transmission electron microscopy, western blotting, and nanoparticle tracking analyses. We conducted CCK8 and lactate dehydrogenase assays to assess the cell proliferation and cytotoxicity of neural progenitor cells (NPCs) exposed to hOEC-EVs. Tert-butyl hydroperoxide (t-BHP) was utilized to mimic oxidative stress-induced cytotoxicity in NPCs.

Results

The modal diameter of hOEC-derived EVs was 113.2 nm. Expressions of EV markers such as CD9, CD63, and CD81 were detected by western blotting. hOEC-derived EVs enhanced the proliferation of NPCs and ameliorated cell cytotoxicity mediated by t-BHP.

Discussion

Our findings reveal a role for hOEC-derived EVs in NPC proliferation and oxidative stress-induced neuronal toxicity model. These results may be useful for developing non-cell therapy OEC-EV-based treatment in acquired nervous system disease.

中文翻译：

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从人类嗅鞘细胞中分离的细胞外囊泡增强了神经祖细胞的活力。

摘要

目标

获得性神经系统疾病，如严重的脑外伤或脊髓损伤 (SCI)，会导致不可逆转的残疾。嗅鞘细胞 (OEC) 移植已作为一种有希望的 SCI 治疗进行试验。调节细胞间相互作用的细胞外囊泡 (EVs) 最近引起了广泛的研究兴趣，并成为神经系统疾病（包括 SCI 动物模型）的非细胞疗法。然而，还没有关于人类 OEC-EV 及其对神经元再生的有益影响的报道。在这里，我们研究了从人类 OEC 中分离的 EV 对神经元细胞活力的影响。

方法

通过连续超速离心从原代人类 OECs (hOECs) 中分离出 EVs。hOEC-EV 的特征在于透射电子显微镜、蛋白质印迹和纳米粒子跟踪分析。我们进行了 CCK8 和乳酸脱氢酶测定，以评估暴露于 hOEC-EV 的神经祖细胞 (NPC) 的细胞增殖和细胞毒性。叔丁基过氧化氢 (t-BHP) 用于模拟 NPC 中氧化应激诱导的细胞毒性。

结果

hOEC 衍生的 EV 的模态直径为 113.2 nm。通过蛋白质印迹检测 EV 标记物（如 CD9、CD63 和 CD81）的表达。hOEC 衍生的 EV 增强了 NPC 的增殖并改善了 t-BHP 介导的细胞毒性。

讨论

我们的研究结果揭示了 hOEC 衍生的 EV 在 NPC 增殖和氧化应激诱导的神经元毒性模型中的作用。这些结果可能有助于在获得性神经系统疾病中开发基于 OEC-EV 的非细胞疗法。