多孔硅纳米管束作为纳米载体输送小干扰 RNA

Nano Select Pub Date : 2024-02-20 DOI:10.1002/nano.202300111
Nguyen T. Le, G. Akkaraju, J. Coffer
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引用次数: 0

摘要

我们在这项工作中描述了对中空介孔硅纳米管束的评估,这种纳米管束有助于使用小干扰 RNA 转染 HeLa 细胞,从而抑制增强型绿色荧光蛋白(eGFP)的表达。这些实验需要使用扫描电子显微镜(SEM)和共聚焦荧光成像技术直接观察与细胞相关的纳米管束。这些纳米管束是用氨基丙基三乙氧基硅烷(APTES)对纳米管阵列进行表面改性,然后在水中进行超声处理而产生的,其胺端结构能够静电连接 siRNA,效率为 23%-50%(取决于初始 siRNA 浓度)。通过荧光成像定量验证了向 HeLa 细胞的递送和转染;eGFP 的平均敲除率达到了 50%。由于纳米级给药载体在临床使用中预计会被吸收,我们还使用扫描和透射电子显微镜(TEM)评估了 SiNT 束在上述体外时间范围内的降解情况。最后,我们简要讨论了该平台未来实验所面临的挑战和机遇。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Porous silicon nanotube bundles as nanocarriers for small interfering RNA delivery
We describe in this work an evaluation of bundles of hollow mesoporous silicon nanotubes to facilitate transfection of HeLa cells using small interfering RNA designed to knock down expression of Enhanced Green Fluorescent Protein (eGFP). These experiments entail direct visualization of the nanotube bundles associated with the cells using both scanning electron microscopy (SEM) and confocal fluorescence imaging. These nanotube bundles are generated by surface modification of nanotube arrays with aminopropyl‐triethoxysilane (APTES), followed by their ultrasonication in water, to create the amine‐terminated structures capable of electrostatic conjugation of siRNA at an efficiency of 23%–50% (depending on initial siRNA concentration). Delivery and transfection to HeLa cells are verified by quantification of fluorescence imaging; an average percent knockdown of ∼50% eGFP is achieved. As nanoscale drug delivery vehicles are expected to be resorbed in clinical use, we also assess SiNT bundle degradation during the above in vitro timescale using scanning and transmission electron (TEM) microscopies. We conclude with a brief discussion of challenges and opportunities in future experiments involving this platform.
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