Electric fields trigger ceramide-dependent vesicle budding and boost the generation of small extracellular vesicles

Small extracellular vesicles (sEVs) hold therapeutic promise, but clinical translation is limited by low yields. Physiological electric fields (EFs) naturally present during wound healing, embryonic development, and cancer progression—processes known to involve elevated production of sEVs. Here, we demonstrate that physiological-strength EFs (direct current, 50-200 mV/mm) dramatically enhance sEV secretion, achieving nearly 100-fold increases strength-dependently. Mechanistically, EFs augmented intraluminal vesicle formation within multivesicular bodies. Using giant plasma membrane vesicles, we observed EF-induced nanoscale vesicle formation, supporting an EF-driven membrane electroassembly and vesiculogenesis hypothesis. Notably, alternating current EFs (50 – 400 Hz) were 80-90% less effective than direct current EFs, highlighting membrane electropolarization-dependent modulation. Lipid raft/ceramide inhibition almost abolished EF-induced sEV secretion. Inhibition of the PI3K abolished approximately half of EF-triggered sEV release. Our findings not only unveil EFs as a potent physiological regulator of sEV secretion but also establish a biomimetic, high-yield production strategy leveraging bioelectric cues. Physiological electric fields dramatically boost small extracellular vesicle secretion via regulating membrane assembly and vesiculogenesis, revealing key regulatory pathways and offering a biomimetic strategy for high-yield sEV production.