Music-inspired acoustic-piezoelectric stimulation accelerates extracellular vesicle production and programs...

Macrophage small extracellular vesicles (sEVs) carry phenotype-linked cargo and bioactivity for immunomodulation and regeneration, but therapeutic translation is limited by low secretion and poor control of function. We introduce a music-activated piezoelectric nanofiber substrate (PES) that converted audible sound into programmable electrical stimulation to enhance sEV biogenesis while tuning macrophage polarization. Adjusting acoustic parameters increased sEV yield, while musically inspired "assemblies" biased macrophage phenotypes: dissonant, low-frequency stimuli promoted M1-like inflammation, whereas consonant, higher-frequency stimuli favored M2-like, regenerative states. These shifts produced distinct sEV cargo and bioactivities. We rationally designed customized music stimulus that maximized both vesicle production and M2 bias, yielding sEVs exhibited regeneration potentials. This work establishes a programmable acoustic-piezoelectric strategy to scale macrophage sEV production while tailoring their therapeutic potency. STATEMENT OF SIGNIFICANCE: Macrophage small extracellular vesicles (sEVs) are promising as a multi-faceted therapeutic tool for regeneration and immunomodulation. Current methods are challenged by low production rate and poor functional control of sEVs. Here, we developed a piezoelectric nanofiber substrate (PES), as a highly controlled stimulative culturing platform for enhancing sEV production and programming macrophages based on the tuned acoustic input. This provides an important platform that allows for scaled sEV production, and tunable cellular phenotypes tailored for various therapeutic applications.