SONOPORATION BY ULTRASOUND-ACTIVATED MICROBUBBLE CONTRAST AGENTS: EFFECT OF ACOUSTIC EXPOSURE PARAMETERS ON CELL MEMBRANE PERMEABILITY AND CELL VIABILITY
SONOPORATION BY ULTRASOUND-ACTIVATED MICROBUBBLE CONTRAST AGENTS: EFFECT OF ACOUSTIC EXPOSURE PARAMETERS ON CELL MEMBRANE PERMEABILITY AND CELL VIABILITY
复制标题
DOI：
10.1016/j.ultrasmedbio.2008.10.013
复制
发表时间：
2009-05-01
影响因子：
2.9
通讯作者：
Burns, Peter N.
中科院分区：
文献类型：
--
作者：
Karshafian, Raffi;Bevan, Peter D.;Burns, Peter N.
这项工作研究了超声辐照参数对悬浮液中的KHT-C细胞通过全氟微泡进行声学修饰的影响。研究了辐射声压(0.05~3.5兆帕)、脉冲频率(0.5~5.0 MHz)、脉冲重复频率(10~3000 Hz)、脉冲宽度(4~32亩S)和辐射时间(0.1~900 S)的变化。用流式细胞仪检测荧光示踪剂分子(70 kDa FITC-葡聚糖)对细胞的渗透数和活细胞数。使用库尔特计数器测量暴露对微泡数量的影响。细胞存活率和膜通透性强烈依赖于声暴露条件。随着峰值负压、脉冲重复频率、脉冲持续时间和辐照时间的增加以及脉冲中心频率的降低，细胞通透性增加，细胞存活率下降。在峰值负压570kPa、S脉宽8 mU、脉冲重复频率3 kHz、中心频率500 kHz、S微泡照射时间12次、体积浓度3.3%的条件下，获得的最高治疗比为8.8，通透率为32+/-4%，存活率为96+/-1%。这些设置对应于3.12J/cm(2)的声能密度(E(SPPA))。细胞通透性和存活率与气泡破裂无关。结果表明，超声暴露参数可以为治疗性声刀治疗进行优化，气泡破裂是超声诱导通透性的必要但不充分的指标。(电子邮件：raffik@sri.utoronto.ca)(C)2009年世界医学和生物学超声联合会。
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.