The correlation between acoustic cavitation and sonoporation involved in ultrasound-mediated DNA transfection with polyethylenimine (PEI) in vitro
The correlation between acoustic cavitation and sonoporation involved in ultrasound-mediated DNA transfection with polyethylenimine (PEI) in vitro
复制标题
超声介导的聚乙烯亚胺 (PEI) 体外 DNA 转染中声空化与声孔作用之间的相关性。
DOI：
10.1016/j.jconrel.2010.04.010
复制
发表时间：
2010-07-01
影响因子：
10.8
通讯作者：
Tu, Juan
中科院分区：
文献类型：
--
作者：
Qiu, Yuanyuan;Luo, Yi;Tu, Juan
先前的研究已经证明，基因/药物递送的效率可以提高超声（US）暴露下与US造影剂微泡的存在下，由于声空化诱导的声致孔。然而，实现可控声致穿孔结果仍然存在障碍。指导本研究的一般假设是，在US暴露期间积累的惯性空化（IC）活动可以量化为基于被动空化检测（PCD）的IC剂量（ICD），声孔效应结果的评估应与ICD测量相关。在本工作中，MCF-7细胞与PEI：DNA复合物和UCD微泡混合，暴露于具有20个周期脉冲和不同声峰负压的1-MHz超声脉冲（P-; 0（假手术）、0.3、0.75、1.4、2.2或3.0 MPa），总处理时间（0、5、10、20、40或60 s）和脉冲重复频率（PRF; 0、20、100、250、500或1000 Hz）。然后进行了四个系列的实验：（1）使用PCD系统检测IC活性并以ICD定量;（2）使用流式细胞术评估DNA转染效率;（3）使用PI染色检测细胞活力并使用流式细胞术测量;（4）使用扫描电子显微镜观察声致孔对细胞膜的影响。结果表明：（1）美国暴露期间生成的ICD可能会受到美国参数的影响（例如，P-、总处理时间和PRF）;（2）合并的数据分析表明，DNA转染效率最初随着ICD的增加而线性增加，然后当ICD继续上升时，其趋于饱和而不是试图达到最大值;和（3）测量的ICD、声孔孔径和细胞活力彼此之间表现出高度相关性。这些结果表明IC活性在超声介导的DNA声孔转染中起重要作用，ICD可作为超声介导的基因/药物传递效应的有效监测和控制工具。(C)2010 Elsevier B. V.保留所有权利。
Previous studies have demonstrated that the efficiency of gene/drug delivery can be enhanced under ultrasound (US) exposure with the presence of US contrast agent microbubbles, due to the acoustic cavitation-induced sonoporation. However, obstacles still remain to achieve controllable sonoporation outcome. The general hypotheses guiding present studies were that inertial cavitation (IC) activities accumulated during US exposure could be quantified as IC dose (ICD) based on passive cavitation detection (PCD), and the assessment of sonoporation outcome should be correlated with ICD measurements. In current work, MCF-7 cells mixed with PEI:DNA complex and UCD microbubbles were exposed to 1-MHz US pulses with 20-cycle pulse and varied acoustic peak negative pressure (P-; 0 (sham), 0.3, 0.75, 1.4, 2.2 or 3.0 MPa), total treatment time (0, 5, 10, 20, 40 or 60 s), and pulse-repetition-frequency (PRF; 0, 20, 100, 250, 500, or 1000 Hz). Then, four series experiments were conducted: (1) the IC activities were detected using a PCD system and quantified as ICD; (2) the DNA transfection efficiency was evaluated with flow cytometry; (3) the cell viability was examined by PI dying then measured using flow cytometry; and (4) scan electron microscopy was used to investigate the sonoporation effects on the cell membrane. The results showed that: (1) the ICD generated during US exposure could be affected by US parameters (e.g., P-, total treatment time, and PRF); (2) the pooled data analyses demonstrated that DNA transfection efficiency initially increased linearly with the increasing ICD, then it tended to saturate instead of trying to achieve a maximum value while the ICD kept going up; and (3) the measured ICD, sonoporation pore size, and cell viability exhibited high correlation among each other. All the results indicated that IC activity should play an important role in the US-mediated DNA transfection through sonoporation, and ICD could be used as an effective tool to monitor and control the US-mediated gene/drug delivery effect. (C) 2010 Elsevier B.V. All rights reserved.