In Situ Transfection by Controlled Release of Lipoplexes Using Acoustic Droplet Vaporization.
In Situ Transfection by Controlled Release of Lipoplexes Using Acoustic Droplet Vaporization.
复制标题
DOI：
10.1002/adhm.201600008
复制
发表时间：
2016-07
影响因子：
10
通讯作者：
Fabiilli ML
中科院分区：
文献类型：
--
作者：
Juliar BA;Bromley MM;Moncion A;Jones DC;O'Neill EG;Wilson CG;Franceschi RT;Fabiilli ML
将核酸局部递送至靶位点（例如患病组织）对于安全有效的基因治疗至关重要。一种称为声学液滴汽化（ADV）的超声技术已被用于时空控制声敏乳剂中所含治疗性小分子和蛋白质的释放。在此，ADV 用于控制 lipoplex（含有带有增强型绿色荧光蛋白 (eGFP) 报告基因的质粒 DNA）从声敏乳液中的释放。聚焦超声（3.5 MHz，机械指数（MI）≥ 1.5）产生荧光素（即替代有效负载）和脂质体从乳液中的强烈释放。使用 ADV（MI=1.5，30 个循环）从乳液中原位释放 lipoplex，释放效率为 55%，从而导致 C3H/10T1/2 细胞的转染效率为 43%，活力为 95%。在不暴露于超声波的情况下，释放和转染效率分别为 5% 和 7%，存活率为 99%。 ADV 释放的 Lipoplex 保留了其生物活性，而 ADV 过程没有产生任何可测量的转染声孔增强。将 Ficoll PM 400 共封装在 lipoplex 负载乳液中，并随后使用 ADV 释放，比单独使用 lipoplex 产生更高的转染效率。结果表明，ADV 可用于基因传递的时空控制。
Localized delivery of nucleic acids to target sites (e.g., diseased tissue) is critical for safe and efficacious gene therapy. An ultrasound-based technique termed acoustic droplet vaporization (ADV) has been used to spatiotemporally control the release of therapeutic small molecules and proteins contained within sonosensitive emulsions. Here, ADV was used to control the release of lipoplex – containing plasmid DNA with an enhanced green fluorescent protein (eGFP) reporter - from a sonosensitive emulsion. Focused ultrasound (3.5 MHz, mechanical index (MI) ≥ 1.5) generated robust release of fluorescein (i.e., surrogate payload) and lipoplex from the emulsion. In situ release of the lipoplex from the emulsion using ADV (MI=1.5, 30 cycles) yielded a 55% release efficiency, resulting in 43% transfection efficiency and 95% viability with C3H/10T1/2 cells. Without exposure to ultrasound, the release and transfection efficiencies were 5% and 7%, respectively, with 99% viability. Lipoplex released by ADV retained its bioactivity while the ADV process did not yield any measureable sonoporative enhancement of transfection. Co-encapsulation of Ficoll PM 400 within the lipoplex loaded emulsion, and its subsequent release using ADV, yielded higher transfection efficiency than the lipoplex alone. The results demonstrate that ADV could have utility in the spatiotemporal control of gene delivery.