Enhancing Exosome Production from Adipose-Derived Stem Cells via Arachidonic Acid Priming for Diabetic Wound Healing

Adipose-derived stem cell (ADSC)-derived exosomes are promising for regenerative therapy. However, their clinical translation is hindered by limited secretion yield and reduced functional potency. Here, we tested whether arachidonic acid (AA) priming increases exosome production while preserving exosomal identity, and whether AA-primed exosomes (AA-Exos) enhance diabetic wound repair. ADSCs were treated with AA, and exosome yield was quantified using particle counts and cumulative acetylcholinesterase (AChE) activity. Exosome morphology and canonical markers were assessed to confirm identity. Mechanistically, transcriptomic and functional analyses revealed that AA promoted multivesicular body and intraluminal vesicle formation through CHMP1B upregulation and facilitated exosome release through Rab27b, and knockdown of either gene impaired the corresponding process. Proteomic profiling further showed that AA-Exos were enriched in wound-repair-related cargo, particularly interleukin-8 (IL-8/CXCL8). In high-glucose-challenged endothelial cells, AA-Exos enhanced proliferation and migration and reduced apoptosis by activating FAK/AKT/ERK signaling, whereas IL-8 neutralization attenuated these effects. In a diabetic mouse model, AA-Exos accelerated wound closure and improved tissue perfusion, angiogenesis, and collagen deposition. Taken together, AA priming enhances exosome production while improving their therapeutic function, suggesting a practical strategy to strengthen the translational potential of ADSC-derived exosomes for diabetic wound regeneration.