High-intensity ultrasound irradiation promotes the release of extracellular vesicles from C2C12 myotubes

•High-intensity ultrasound strongly induced extracellular vesicle (EV) secretion from cultured myotubes.•High-intensity ultrasound did not change the viability of myotubes.•High-intensity ultrasound is a novel modality for inducing myocellular EV release. Skeletal muscle is an important secretory organ in mammals, producing myriad chemical mediators (“myokines”) with distinct biological action in different tissues, including anti-inflammatory activity. Extracellular vesicles (EVs) have recently been identified as a mode of myokine transport from muscle, facilitating such anti-inflammatory activity. In this report, we have demonstrated that high-intensity ultrasound (US) strongly induces EV secretion from cultured myotubes without a reduction in cell viability. High-intensity US of 3.0 W/cm2 with 20% duty cycle increased the number of EVs by 2-fold compared to control at 6 h. This effect was specific to EVs in the 100–150 nm size range. Thus, high-intensity US is a novel modality for inducing myocellular EV release and may hold therapeutic value.