Sonoporation by single-shot pulsed ultrasound with microbubbles adjacent to cells.
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 Ca2+-independent and Ca2+-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% (approximately 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.
Kudo
Nobuki
N
Graduate School of Information Science and Technology, Hokkaido University, Sapporo, Japan. kudo@bme.ist.hokudai.ac.jp
Okada
Kengo
K
Yamamoto
Katsuyuki
K
D000818:Animals; D002118:Calcium / Q000494:pharmacology; D002417:Cattle; D002462:Cell Membrane / Q000187:drug effects; D002478:Cells, Cultured; D003584:Cytological Techniques / Q000379:methods*; D042783:Endothelial Cells; D045423:Microbubbles; D008855:Microscopy, Electron, Scanning; D014465:Ultrasonics
D002118:Calcium