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<p>A single cavitation microbubble can transiently disrupt the cellular membrane, providing a chemical-free, targeted drug delivery mechanism. Here, we investigate the spatiotemporal dynamics of membrane permeabilization in an <i>Escherichia coli</i> (<i>E. coli</i>) monolayer exposed to a single cavitation event. Using high-resolution fluorescence microscopy and propidium iodide (PI) uptake as a marker of membrane disruption, we tracked the response of 5565 individual cells around the center of the cavitation event over timescales from microseconds to minutes and spatial scales from 1 to 165&#xa0;µm. PI uptake rates exhibited a strong spatial dependence, with cells closer to the cavitation center showing rapid and extensive permeabilization. A modified Goldman equation describing PI concentrations inside and outside the cells was used and related to the spatiotemporal measurements of fluorescence intensity. The model accurately captured the first-order PI uptake kinetics, which resulted in saturated fluorescence intensity profiles. Additionally, the model predicted an exponential decay of permeability post-cavitation, implicitly suggesting that pore-resealing dynamics were taking place. Membrane permeability decreased with distance as 1/r, with a characteristic decay time of approximately 3.4&#xa0;min. Our model thus predicts cell damage induced by a single cavitation event in both space and time. In the present case, where the cavitation bubble reached maximum radius of 29.7&#xa0;µm, we found that at a distance of 11&#xa0;µm from the cavitation center, ~ 50% of cell membranes are damaged and permeable to PI, but a pore-resealing mechanism reduces this damage to ~ 1% after 10&#xa0;min. Our results are consistent with existing sonoporation studies and offer novel insights for optimizing cavitation-assisted drug delivery and biofilm disruption strategies.</p>

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Spatiotemporal analysis of Escherichia coli membrane permeabilization and uptake kinetics induced by a single microbubble cavitation event

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2025年11月28日

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  • Mitja Drab,
  • Aleš Iglič,
  • David Stopar,
  • Žiga Pandur

摘要

A single cavitation microbubble can transiently disrupt the cellular membrane, providing a chemical-free, targeted drug delivery mechanism. Here, we investigate the spatiotemporal dynamics of membrane permeabilization in an Escherichia coli (E. coli) monolayer exposed to a single cavitation event. Using high-resolution fluorescence microscopy and propidium iodide (PI) uptake as a marker of membrane disruption, we tracked the response of 5565 individual cells around the center of the cavitation event over timescales from microseconds to minutes and spatial scales from 1 to 165 µm. PI uptake rates exhibited a strong spatial dependence, with cells closer to the cavitation center showing rapid and extensive permeabilization. A modified Goldman equation describing PI concentrations inside and outside the cells was used and related to the spatiotemporal measurements of fluorescence intensity. The model accurately captured the first-order PI uptake kinetics, which resulted in saturated fluorescence intensity profiles. Additionally, the model predicted an exponential decay of permeability post-cavitation, implicitly suggesting that pore-resealing dynamics were taking place. Membrane permeability decreased with distance as 1/r, with a characteristic decay time of approximately 3.4 min. Our model thus predicts cell damage induced by a single cavitation event in both space and time. In the present case, where the cavitation bubble reached maximum radius of 29.7 µm, we found that at a distance of 11 µm from the cavitation center, ~ 50% of cell membranes are damaged and permeable to PI, but a pore-resealing mechanism reduces this damage to ~ 1% after 10 min. Our results are consistent with existing sonoporation studies and offer novel insights for optimizing cavitation-assisted drug delivery and biofilm disruption strategies.