UCL, School of Pharmacy, 29-39 Brunswick Square, London WC1N 1AX, UK.
Int J Pharm. 2013 Oct 15;455(1-2):241-7. doi: 10.1016/j.ijpharm.2013.07.024. Epub 2013 Jul 20.
To explore the potential of crosslinked chitosan nanoparticles as carriers for delivery of siRNA using a jet nebuliser.
Nanoparticles encapsulating siRNA were prepared using an ionic crosslinking technique at chitosan to siRNA weight/weight ratios of 10:1, 30:1 and 50:1. Particles were characterised for their size, charge, morphology, pH stability and siRNA encapsulation efficiency. Gel electrophoresis was used to assess the association and stability of siRNA with nanoparticles, including after aerosolisation using a Pari LC Sprint jet nebuliser. The aerosolisation properties of FITC labelled chitosan nanoparticles were investigated using a two-stage impinger. Cell viability was performed with H-292 cells using a WST-1 assay.
Positively charged spherical nanoparticles were produced with mean diameters less than 150 nm, at all chitosan to siRNA ratios. Nanoparticles were non-aggregated at the pH of the airways and showed high siRNA encapsulation efficiency (>96%). Complete binding of siRNA to chitosan nanoparticles was observed when the w/w ratio was 50:1. Nebulisation produced fine particle fractions of 54±11% and 57.3±1.9% for chitosan and chitosan:siRNA (10:1 w/w) nanoparticles respectively. The stability of chitosan-encapsulated siRNA was maintained after nebulisation. Cell viability was high (>85%) at the highest chitosan concentration (83 μg/ml).
The results suggest that crosslinked chitosan nanoparticles have potential for siRNA delivery to the lungs using a jet nebuliser.
探索使用喷射雾化器将交联壳聚糖纳米粒子作为 siRNA 传递载体的潜力。
使用离子交联技术，在壳聚糖与 siRNA 的重量/重量比为 10:1、30:1 和 50:1 的情况下制备包封 siRNA 的纳米粒子。对粒子的粒径、电荷、形态、pH 稳定性和 siRNA 包封效率进行了表征。凝胶电泳用于评估 siRNA 与纳米粒子的结合和稳定性，包括使用 Pari LC Sprint 喷射雾化器进行雾化后。使用两步冲击器研究了 FITC 标记壳聚糖纳米粒子的雾化特性。使用 WST-1 测定法对 H-292 细胞进行细胞活力测定。
在所有壳聚糖与 siRNA 比例下，均产生了平均直径小于 150nm 的带正电荷的球形纳米粒子。纳米粒子在气道 pH 值下未聚集，并显示出高的 siRNA 包封效率（>96%）。当 w/w 比为 50:1 时，观察到 siRNA 与壳聚糖纳米粒子的完全结合。雾化产生了 54±11%和 57.3±1.9%的细颗粒分数，分别为壳聚糖和壳聚糖：siRNA（10:1 w/w）纳米粒子。雾化后壳聚糖包封的 siRNA 稳定性得以维持。细胞活力在最高壳聚糖浓度（83μg/ml）时>85%。
结果表明，交联壳聚糖纳米粒子具有通过喷射雾化器将 siRNA 递送到肺部的潜力。
