Non‐Invasive Ultrasound Treatment Enhances the Release of Skeletal Muscle‐Derived Extracellular Vesicles in Mice

Skeletal muscle, the largest secretory organ, regulates distant organs through the secretion of various factors. Among these are extracellular vesicles (EVs), which play a significant role in mediating communication between muscle and other tissues and hold therapeutic potential due to their target specificity and anti-inflammatory properties. Enhancing the release of EVs from skeletal muscle into the circulation is crucial for eliciting their diverse effects, including anti-inflammatory actions; however, effective strategies to increase the levels of muscle-derived EVs in the bloodstream have not yet been developed. While exercise is the most studied method to increase blood EV levels, its efficacy in enhancing skeletal muscle-derived EVs remains unclear. This study reveals that a brief, 5-min ultrasound (US) treatment of skeletal muscle robustly elevates circulating levels of muscle-derived EVs, introducing a novel, non-invasive stategy to modulate EV release. US-induced EVs showed altered miRNA profiles enriched with anti-inflammatory miRNAs that target pathways involved in inflammatory responses. Additionally, transient upregulation of reactive oxygen species and activation of the endoplasmic reticulum stress pathway were observed in US-treated muscle, suggesting a mechanism for enhanced EV release. Our findings establish US as the first non-invasive and rapid method to selectively enhance skeletal muscle-derived EV release into the circulation, highlighting its potential for therapeutic applications through its anti-inflammatory effect.