Influence of cell shape on sonoporation efficiency in microbubble-facilitated delivery using micropatterned cell arrays

Abstract

Microbubble-facilitated sonoporation is a rapid, versatile, and non-viral intracellular delivery technique with potential for clinical and ex vivo cell engineering applications. We developed a micropatterning-based approach to investigate the impact of cell shape on sonoporation efficacy. Cationic microbubbles were employed to enhance sonoporation by binding to the cell membrane electrostatically. NIH/3T3 fibroblasts were micropatterned into circle, square, triangle, and rectangle. A two-plate system ensured high-throughput and efficient sonoporation by controlling cationic microbubble-cell attachment. High-speed video microscopy captured the acoustic dynamics of microbubbles under short ultrasound pulses. Our findings reveal that for NIH/3T3 fibroblasts, rectangular cells achieved the highest sonoporation and survival rate, while square-shaped cells demonstrated the greatest propidium iodide uptake. Triangle-shaped NIH/3T3 fibroblasts displayed an initial rise then a plateau in the sonoporation and survival rate as the ultrasound pulse duration increased from 10 cycles to 100 cycles, and then to 200 cycles. Conversely, rectangle-shaped cells showed a decrease followed by a stabilization. Circle-shaped and rectangle-shaped HeLa cells exhibited similar sonoporation outcomes, which were not as effective as NIH/3T3 fibroblasts. This study underscores the significance of cell shape in optimizing sonoporation efficiency and highlights the potential of combining micropatterning with controlled targeting sonoporation to advance intracellular delivery technologies.