Stabilizing in vitro ultrasound-mediated gene transfection by regulating cavitation.
It is well known that acoustic cavitation can facilitate the inward transport of genetic materials across cell membranes (sonoporation). However, partially due to the unstationary behavior of the initiation and leveling of cavitation, the sonoporation effect is usually unstable, especially in low intensity conditions. A system which is able to regulate the cavitation level during sonication by modulating the applied acoustic intensity with a feedback loop is implemented and its effect on in vitro gene transfection is tested. The regulated system provided better time stability and reproducibility of the cavitation levels than the unregulated conditions. Cultured hepatoma cells (BNL) mixed with 10 μg luciferase plasmids are exposed to 1-MHz pulsed ultrasound with or without cavitation regulation, and the gene transfection efficiency and cell viability are subsequently assessed. Experimental results show that for all exposure intensities (low, medium, and high), stable and intensity dependent, although not higher, gene expression could be achieved in the regulated cavitation system than the unregulated conditions. The cavitation regulation system provides a better control of cavitation and its bioeffect which are crucial important for clinical applications of ultrasound-mediated gene transfection.
Department of Physical Medicine and Rehabilitation, National Taiwan University Hospital & National Taiwan University College of Medicine, No. 7, Zhongshan S. Rd., Taipei 100, Taiwan.
Chia-Wen
CW
Lo
Cyril
C
Desjouy
Shing-Ru
SR
Chen
Jyun-Lin
JL
Lee
Claude
C
Inserra
Jean-Christophe
JC
Béra
Wen-Shiang
WS
Chen
D001692:Biological Transport; D002460:Cell Line; D004247:DNA / Q000235:genetics / Q000378:metabolism; D010795:Phthalic Acids / Q000737:chemistry; D014162:Transfection / Q000379:methods*; D014465:Ultrasonics / Q000379:methods*
D010795:Phthalic Acids; C011363:terephthalic acid; D004247:DNA
Cavitation; Gene transfection; Sonoporation; Ultrasound