Sonoporation by Single-Shot Pulsed Ultrasound with Microbubbles Adjacent to Cells
Sonoporation by Single-Shot Pulsed Ultrasound with Microbubbles Adjacent to Cells
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DOI：
10.1016/j.bpj.2009.02.072
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发表时间：
2009-06-17
影响因子：
3.4
通讯作者：
Yamamoto, Katsuyuki
中科院分区：
文献类型：
--
作者：
Kudo, Nobuki;Okada, Kengo;Yamamoto, Katsuyuki
本文介绍了单次脉冲短脉冲超声照射内皮细胞后所引起的膜穿孔，并探讨了膜损伤和修复的机制。在超声操作过程中对细胞-气泡相互作用的实时光学观察，以及在知道气泡位置的情况下对膜损伤的连续扫描电子显微镜观察表明，在气泡位置处产生了微米级的膜穿孔。对微泡的高速观察显示了在气泡非均匀收缩过程中产生的液体微射流，表明这些微射流是细胞膜破坏的原因。荧光显微镜下观察到的声孔细胞再封闭过程表明，钙离子不依赖和钙(2+)触发的再封闭机制参与了快速再封闭过程。在几乎所有细胞都有一个相邻气泡的实验条件下，单次脉冲超声对25.4%的细胞造成损伤，15.9%(与损伤细胞的60%)在5 S内再次密封，这些结果表明，当微泡附着在细胞上时，单次脉冲超声足以实现声孔修复。
In this article, membrane perforation of endothelial cells with attached microbubbles caused by exposure to single-shot short pulsed ultrasound is described, and the mechanisms of membrane damage and repair are discussed. Real-time optical observations of cell-bubble interaction during sonoporation and successive scanning electron microscope observations of the membrane damage with knowledge of bubble locations revealed production of micron-sized membrane perforations at the bubble locations. High-speed observations of the microbubbles visualized production of liquid microjets during nonuniform contraction of bubbles, indicating that the jets are responsible for cell membrane damage. The resealing process of sonoporated cells visualized using fluorescence microscopy suggested that Ca(2+)-independent and Ca(2+)-triggered resealing mechanisms were involved in the rapid resealing process. In an experimental condition in which almost all cells have one adjacent bubble, 25.4% of the cells were damaged by exposure to single-shot pulsed ultrasound, and 15.9% (similar to 60% of the damaged cells) were resealed within 5 s. These results demonstrate that single-shot pulsed ultrasound is sufficient to achieve sonoporation when microbubbles are attached to cells.