Abstract
Sonoporation employs ultrasound accompanied by microbubble (MB) cavitation to induce the reversible disruption of cell membranes and has been exploited as a promising intracellular macromolecular delivery strategy. Due to the damage to cells resulting from strong cavitation, it is difficult to balance efficient delivery and high survival rates. In this paper, a traveling surface acoustic wave (TSAW) device, consisting of a TSAW chip and a polydimethylsiloxane (PDMS) channel, was designed to explore single-cell sonoporation using targeted microbubbles (TMBs) in a non-cavitation regime. A TSAW was applied to precisely manipulate the movement of the TMBs attached to MDA-MB-231 cells, leading to sonoporation at a single-cell level. The impact of input voltage and the number of TMBs on cell sonoporation was investigated. In addition, the physical mechanisms of bubble cavitation or the acoustic radiation force (ARF) for cell sonoporation were analyzed. The TMBs excited by an ARF directly propelled cell membrane deformation, leading to reversible perforation in the cell membrane. When two TMBs adhered to the cell surface and the input voltage was 350 mVpp, the cell sonoporation efficiency went up to 83%.
Highlights
The delivery of membrane-impermeant compounds into living cells for molecular biology and gene therapy is a critical step in clinical and research applications, which can be achieved by using carrier-based and membrane disruption-based techniques [1].Carrier-based methods mainly depend on endocytosis and fusogenic activity [2,3], endowing the carrier with the ability to merge directly with the cell membrane
The adhesion of targeted microbubbles (TMBs) to the cell surface is indispensable for the reversible perfora‐
Figurestimulation, 2a, two TMBs adhered to cell 1,showed while nothat fluorescent images cell adhered to cell stimulation, fluorescent images showed that cell 1 and cell 2 emit green fluorescence and no red fluorescence, indicating the good cell via‐
Summary
The delivery of membrane-impermeant compounds into living cells for molecular biology and gene therapy is a critical step in clinical and research applications, which can be achieved by using carrier-based and membrane disruption-based techniques [1]. Due to the relatively low efficiency of carrier-based methods at delivering membrane-impermeant compounds, membrane disruption methods have attracted increasing attention. An acoustofluidic device may help researchers investigate the mechanism of non-cavitation, TMB-mediated single-cell sonoporation. When the input voltage was 350 mVpp and two TMBs were attached to the cells, the sonoporation efficiency went up to 83% This TSAW device may provide a new strategy for intracellular delivery and gene therapy in a non-viral and label-free manner
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