Direct Cytosolic Delivery of Nanovesicles via Gigahertz Acoustic Streaming.

Abstract

The development of advanced intracellular delivery systems is essential for biopharmaceutical progress, particularly in drug delivery systems, genetic engineering, and cellular therapeutics. While nanovesicles show significant therapeutic potential, challenges remain due to cell membrane barriers and the limitations of endocytosis-mediated pathways. In this study, we introduce an efficient acoustofluidic delivery system that utilizes a gigahertz (GHz)-range bulk acoustic wave (BAW) resonator to generate spatiotemporally controlled acoustic streaming vortices. This system enables rapid (within 10 min) and highly efficient direct cytosolic delivery of nanovesicles by bypassing conventional endosomal entrapment pathways. It effectively delivers both synthetic drug carriers (doxorubicin-loaded small unilamellar vesicles, Dox-SUVs) and biologically active exosomes through GHz-driven hydrodynamic shear forces that induce transient membrane permeability while maintaining cellular viability (>91.5% at 300 mW). Key performances demonstrate 86.5% drug delivery efficiency for Dox-SUVs with near-complete nuclear accumulation, while exosome-mediated delivery exhibits 2.4-fold accelerated migration and 3-fold proliferation enhancement within 24 h. The system's capacity to modulate fluidic shear stresses via BAW power tuning (100-500 mW) allows precise control over membrane permeabilization kinetics and cargo flux. By overcoming endolysosomal sequestration through a noninvasive, physics-driven mechanism, this acoustofluidic approach expands opportunities for next-generation therapeutics, including macromolecular biologic delivery, genome editing, and exosome-mediated intercellular communication.