Journal
ULTRASOUND IN MEDICINE AND BIOLOGY
Volume 39, Issue 12, Pages 2393-2405Publisher
ELSEVIER SCIENCE INC
DOI: 10.1016/j.ultrasmedbio.2013.08.003
Keywords
Sonoporation; Membrane perforation; Membrane recovery; Spatiotemporal dynamics; Real-time confocal imaging; Surface topography
Funding
- Hong Kong Innovation and Technology Fund [ITS/292/11]
- University of Hong Kong [201111159091]
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Transient sonoporation can essentially be epitomized by two fundamental processes: acoustically induced membrane perforation and its subsequent resealing. To provide insight into these processes, this article presents a new series of direct evidence on the membrane-level dynamics during and after an episode of sonoporation. Our direct observations were obtained from anchored fetal fibroblasts whose membrane topography was imaged in situ using real-time confocal microscopy. To facilitate controlled sonoporation at the single-cell level, microbubbles that can passively adhere to the cell membrane were first introduced at a 1: 1 cell-to-bubble ratio. Single-pulse ultrasound exposure (1-MHz frequency, 10-cycle pulse duration, 0.85-MPa peak negative pressure in situ) was then applied to trigger microbubble pulsation/collapse, which, in turn, instigated membrane perforation. With this protocol, five membrane-level phenomena were observed: (i) localized perforation of the cell membrane was synchronized with the instant of ultrasound pulsing; (ii) perforation sites with temporal peak area <30 mu m(2) were resealed successfully; (iii) during recovery, a thickened pore rim emerged, and its temporal progression corresponded with the pore closure action; (iv) membrane resealing, if successful, would generally be completed within 1 min of the onset of sonoporation, and the resealing time constant was estimated to be below 20 s; (v) membrane resealing would fail for overly large pores (>100 mu m(2)) or in the absence of extracellular calcium ions. These findings serve to underscore the spatiotemporal complexity of membrane-level dynamics in sonoporation. (E-mail: alfred.yu@hku.hk) (C) 2013 World Federation for Ultrasound in Medicine & Biology.
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