In this study, the bioeffects of acoustic droplet vaporization (ADV) on adjacent cells were investigated by evaluating the real-time cell response at the single-cell level in situ, using a combined ultrasound-exposure and optical imaging system. Two imaging modalities, high-speed and fluorescence imaging, were used to observe ADV bubble dynamics and to evaluate the impact on cell membrane permeabilization (i.e., sonoporation) using propidium iodide (PI) uptake as an indicator. The results indicated that ADV mainly led to irreversible rather than reversible sonoporation. Further, the rate of irreversible sonoporation significantly increased with increasing nanodroplet concentration, ultrasound amplitude, and pulse duration. The results suggested that sonoporation is correlated to the rapid formation, expansion, and contraction of ADV bubbles near cells, and strongly depends on ADV bubble size and bubble-to-cell distance when subjected to short ultrasound pulses (1 μs). Moreover, the displacement of ADV bubbles was larger when using a long ultrasound pulse (20 μs), resulting in considerable cell membrane deformation and a more irreversible sonoporation rate. During sonoporation, cell membrane blebbing as a recovery manoeuvre was also investigated, indicating the essential role of Ca²⁺ influx in the membrane blebbing response. This study has helped us gain further insights into the dynamic behavior of ADV bubbles near cells, ADV bubble-cell interactions, and real-time cell response, which are invaluable in the development of optimal approaches for ADV-associated theranostic applications.

在这项研究中，通过评估原位单细胞水平的实时细胞反应，研究了声滴汽化（ADV）对相邻细胞的生物效应。，使用组合的超声波曝光和光学成像系统。使用两种成像方式（高速成像和荧光成像）来观察ADV气泡动力学，并使用碘化丙啶（PI）吸收作为指标来评估对细胞膜通透性（即声纳穿孔）的影响。结果表明，ADV主要导致不可逆而不是可逆的声穿孔。此外，不可逆的声纳穿孔率随着纳米液滴浓度，超声振幅和脉冲持续时间的增加而显着增加。结果表明，声纳穿孔与ADV气泡在细胞附近的快速形成，膨胀和收缩有关，并且在短超声脉冲（1μs）时强烈依赖于ADV气泡的大小和气泡到细胞的距离。而且，当使用较长的超声脉冲（20μs）时，ADV气泡的位移较大，从而导致细胞膜发生较大的形变，并具有更高的不可逆声波穿透率。在声处理过程中，还研究了细胞膜起泡作为恢复手段，这表明钙的基本作用²⁺流入膜起泡反应。这项研究帮助我们进一步了解了ADV气泡在细胞附近的动态行为，ADV气泡与细胞之间的相互作用以及实时细胞反应，这对于开发与ADV相关的治疗方法的最佳方法具有不可估量的价值。