Improving ultrasound gene transfection efficiency by controlling ultrasound excitation of microbubbles.
Improving ultrasound gene transfection efficiency by controlling ultrasound excitation of microbubbles.
复制标题
DOI：
10.1016/j.jconrel.2013.05.039
复制
发表时间：
2013-09-28
期刊：
影响因子：
--
通讯作者：
Deng CX
中科院分区：
文献类型：
--
作者：
Fan Z;Chen D;Deng CX
在微泡存在下的超声应用已经显示出通过瞬时破坏细胞膜（声致穿孔）进行非病毒基因转染的巨大潜力。然而，其效率的提高在很大程度上依赖于经验的方法，没有一致的和可翻译的结果。本研究的目的是开发一个合理的策略，使用新的实验技术和分析获得的新结果的基础上，以提高声孔基因转染。我们使用附着在细胞膜上的靶向微泡进行实验，以促进声孔作用。我们量化了暴露于脉冲超声的微泡的动态活动和由此产生的声致穿孔结果，并确定了特征微泡行为的不同制度：稳定空化，合并和平移，以及惯性空化。我们发现，惯性空化产生的膜穿孔率最高。通过建立超声诱导的气泡活动与细胞内摄取和孔径的直接相关性，我们设计了一种斜坡脉冲曝光方案，用于优化微泡激发以改善声孔基因转染。我们实现了一种新的声致孔基因转染系统，使用一个水两相系统（ATPS）的试剂和高通量操作的有效利用。使用编码绿色荧光蛋白（GFP）的质粒，我们在大鼠主动脉平滑肌细胞（RASMCs）中获得了6.9% ± 2.2%（n = 9）的声孔转染效率，与脂质体转染（7.5% ± 0.8%，n = 9）相当。我们的研究结果揭示了负责声致穿孔的特征微泡行为，并证明了一个合理的策略，以提高声致穿孔基因转染。
Ultrasound application in the presence of microbubbles has shown great potential for non-viral gene transfection via transient disruption of cell membrane (sonoporation). However, improvement of its efficiency has largely relied on empirical approaches without consistent and translatable results. The goal of this study is to develop a rational strategy based on new results obtained using novel experimental techniques and analysis to improve sonoporation gene transfection. We conducted experiments using targeted microbubbles that were attached to cell membrane to facilitate sonoporation. We quantified the dynamic activities of microbubbles exposed to pulsed ultrasound and the resulting sonoporation outcome and identified distinct regimes of characteristic microbubble behaviors: stable cavitation, coalescence and translation, and inertial cavitation. We found that inertial cavitation generated the highest rate of membrane poration. By establishing direct correlation of ultrasound-induced bubble activities with intracellular uptake and pore size, we designed a ramped pulse exposure scheme for optimizing microbubble excitation to improve sonoporation gene transfection. We implemented a novel sonoporation gene transfection system using an aqueous two phase system (ATPS) for efficient use of reagents and high throughput operation. Using plasmid coding for the green fluorescence protein (GFP), we achieved a sonoporation transfection efficiency in rate aortic smooth muscle cells (RASMCs) of 6.9% ± 2.2% (n = 9), comparable with lipofection (7.5% ± 0.8%, n = 9). Our results reveal characteristic microbubble behaviors responsible for sonoporation and demonstrated a rational strategy to improve sonoporation gene transfection.
登录
查看更多内容
影响因子：
3.5
作者：
Chomas, JE;Dayton, P;Ferrara, K
通讯作者：
Ferrara, K
影响因子：
2.9
作者：
Karshafian, Raffi;Bevan, Peter D.;Burns, Peter N.
通讯作者：
Burns, Peter N.
DOI：
10.1109/58.656630
发表时间：
1997-11-01
影响因子：
3.6
作者：
Dayton, PA;Morgan, KE;Ferrara, KW
通讯作者：
Ferrara, KW
影响因子：
3.5
作者：
Garbin, Valeria;Overvelde, Marlies;Versluis, Michel
通讯作者：
Versluis, Michel
影响因子：
18.3
作者：
Ferrara, Katherine W.;Borden, Mark A.;Zhang, Hua
通讯作者：
Zhang, Hua