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Extracellular vesicles (EVs) exhibit potential as functional biomolecules for tissue regeneration and immunomodulation as they play important roles in the physiological communication between cells. EV internal cargo contains miRNAs, proteins, lipids, and so on. Osteoarthritis (OA) is a common joint disease causing disability owing to impaired joint function and pain. EVs originating from animal cells and tissue matrices are also being considered for OA, in addition to research involving non-steroidal therapeutic agents. However, there are no studies on EVs from marine organisms. Hence, we focused on sea cucumber-derived EVs and conducted experiments to set up an extraction protocol and to demonstrate their efficacy to modulate the inflammatory environment.