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.
Keywords: Extracellular vesicle; cytotoxicity; exosome; neural progenitor cell; neurogenesis; olfactory ensheathing cell; proliferation.