Micro/Nanophysics Research Laboratory, School of Engineering, RMIT University, Melbourne, VIC 3000, Australia.
RMIT Microscopy and Microanalysis Facility, RMIT University, Melbourne, VIC 3000, Australia.
Nanoscale. 2018 Jul 13;10(27):13165-13178. doi: 10.1039/c8nr02898b.
Recent breakthroughs in gene editing have necessitated practical ex vivo methods to rapidly and efficiently re-engineer patient-harvested cells. Many physical membrane-disruption or pore-forming techniques for intracellular delivery, however, result in poor cell viability, while most carrier-mediated techniques suffer from suboptimal endosomal escape and hence cytoplasmic or nuclear targeting. In this work, we show that short exposure of cells to high frequency (>10 MHz) acoustic excitation facilitates temporal reorganisation of the lipid structure in the cell membrane that enhances translocation of gold nanoparticles and therapeutic molecules into the cell within just ten minutes. Due to its transient nature, rapid cell self-healing is observed, leading to high cellular viabilities (>97%). Moreover, the internalised cargo appears to be uniformly distributed throughout the cytosol, circumventing the need for strategies to facilitate endosomal escape. In the case of siRNA delivery, the method is seen to enhance gene silencing by over twofold, demonstrating its potential for enhancing therapeutic delivery into cells.
近年来，基因编辑技术取得了突破性进展，这就需要实际的离体方法来快速有效地对患者采集的细胞进行重新设计。然而，许多用于细胞内递药的物理细胞膜破坏或成孔技术会导致细胞活力不佳，而大多数载体介导的技术则存在内体逃逸不理想的问题，因此无法实现细胞质或核靶向。在这项工作中，我们表明，细胞短时间暴露于高频（>10 MHz）声激发下会促进细胞膜中脂质结构的暂时重排，从而在十分钟内促进金纳米颗粒和治疗分子进入细胞。由于其瞬态特性，观察到快速的细胞自我修复，导致细胞活力高（>97%）。此外，内化的货物似乎均匀分布在整个细胞质中，从而无需采用策略来促进内体逃逸。在 siRNA 递药的情况下，该方法被证明可以将基因沉默提高两倍以上，这表明其在增强治疗性药物向细胞内递送方面具有潜力。
