SONOPORATION BY ULTRASOUND-ACTIVATED MICROBUBBLE CONTRAST AGENTS: EFFECT OF ACOUSTIC EXPOSURE PARAMETERS ON CELL MEMBRANE PERMEABILITY AND CELL VIABILITY

被引:268
作者
Karshafian, Raffi [1 ]
Bevan, Peter D. [1 ]
Williams, Ross [2 ]
Samac, Sanya [2 ]
Burns, Peter N. [1 ,2 ]
机构
[1] Univ Toronto, Dept Med Biophys, Toronto, ON M4N 3M5, Canada
[2] Sunnybrook Hlth Sci Ctr, Toronto, ON M4N 3M5, Canada
基金
加拿大健康研究院;
关键词
Sonoporation; Cell membrane permeabilisation; Drug delivery; Microbubble; Ultrasound exposure; DRUG-DELIVERY; IN-VITRO; FOCUSED ULTRASOUND; LOCAL-DELIVERY; GENE DELIVERY; SOLID TUMORS; CAVITATION; BUBBLES; BRAIN; DNA;
D O I
10.1016/j.ultrasmedbio.2008.10.013
中图分类号
O42 [声学];
学科分类号
070206 ; 082403 ;
摘要
This work investigates the effect of ultrasound exposure parameters on the sonoporation of KHT-C cells in suspension by perflutren microbubbles. Variations in insonating acoustic pressure (0.05 to 3.5 MPa), pulse frequency (0.5 to 5.0 MHz), pulse repetition frequency (10 to 3000 Hz), pulse duration (4 to 32 mu s) and insonation time (0.1 to 900 s) were studied. The number of cells permeabilised to a fluorescent tracer molecule (70 kDa FITC-dextran) and the number of viable cells were measured using flow cytometry. The effect of exposure on the microbubble population was measured using a Coulter counter. Cell viability and membrane permeability were found to depend strongly on the acoustic exposure conditions. Cell permeability increased and viability decreased with increasing peak negative pressure, pulse repetition frequency, pulse duration and insonation time and with decreasing pulse centre frequency. The highest therapeutic ratio (defined as the ratio of permeabilised to nonviable cells) achieved was 8.8 with 32 +/- 4% permeabilisation and 96 +/- 1% viability at 570 kPa peak negative pressure, 8 mu s pulse duration, 3 kHz pulse repetition frequency, 500 kHz centre frequency and 12 s insonation time with microbubbles at 3.3% volume concentration. These settings correspond to an acoustic energy density (E(SPPA)) of 3.12 J/cm(2). Cell permeability and viability did not correlate with bubble disruption. The results indicate that ultrasound exposure parameters can be optimized for therapeutic sonoporation and that bubble disruption is a necessary but insufficient indicator of ultrasound-induced permeabilisation. (E-mail: raffik@sri.utoronto.ca) (C) 2009 World Federation for Ultrasound in Medicine & Biology.
引用
收藏
页码:847 / 860
页数:14
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