Gene Silencing in a Mouse Lung Metastasis Model by an Inhalable Dry Small Interfering RNA Powder Prepared Using the Supercritical Carbon Dioxide Technique
| Autor: | Daiki Hira, Michiko Fukushima, Ai Oiwa, Hirokazu Okamoto, Tomoyuki Okuda, Daisuke Kito |
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| Rok vydání: | 2013 |
| Předmět: |
Male
Small interfering RNA Biodistribution Lung Neoplasms Surface Properties Drug Compounding Pharmaceutical Science Nanotechnology Chitosan Mice chemistry.chemical_compound Drug Stability Luciferases Firefly Cell Line Tumor medicine Animals Gene silencing Tissue Distribution Gene Silencing Particle Size RNA Small Interfering Pharmacology Drug Carriers Mice Inbred BALB C Supercritical carbon dioxide technology industry and agriculture RNA Dry Powder Inhalers General Medicine Carbon Dioxide Molecular biology chemistry Microscopy Electron Scanning Mannitol Drug carrier medicine.drug |
| Zdroj: | Biological and Pharmaceutical Bulletin. 36:1183-1191 |
| ISSN: | 1347-5215 0918-6158 |
| DOI: | 10.1248/bpb.b13-00167 |
| Popis: | In this study, a novel dry small interfering RNA (siRNA) powder for inhalation, containing chitosan and mannitol, was prepared using the supercritical carbon dioxide (CO2) technique. Although the siRNA/chitosan powder was difficult to disperse because of a long needle-like structure, it could be reduced to fragments of 10-20 µm by manual grinding, which allowed for administration into mice. Electrophoresis revealed that the supercritical CO2 technique and manual grinding didn't greatly affect the integrity of the siRNA. Furthermore, the siRNA was more stable in the lungs than in blood, suggesting the utility of pulmonary delivery. Biodistribution experiments using Cy5.5-labeled siRNA demonstrated that pulmonary administration of the powder achieved a prolonged exposure of the siRNA/chitosan complex on the lung epithelial surface at a higher concentration. For the evaluation of the in-vivo gene silencing effect of the siRNA/chitosan powder, mice bearing colon26/Luc cells were used. The powder significantly inhibited the increase in luminescence intensity in the lungs, but the siRNA/chitosan solution and a non-specific dry siRNA/chitosan powder didn't, indicating the effective and specific gene silencing against the tumor cells metastasized in the lungs of mice by the siRNA/chitosan powder. These results strongly indicate that inhalable dry siRNA powders have the possibility of effective pulmonary gene silencing and that the supercritical CO2 technique can be applied to the production. |
| Databáze: | OpenAIRE |
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