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1.
Pharm Res ; 27(12): 2520-7, 2010 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-20824309

RESUMEN

PURPOSE: This work describes the production and application of an aerosolised formulation of chitosan nanoparticles for improved pulmonary siRNA delivery and gene silencing in mice. METHODS: Aerosolised chitosan/siRNA nanoparticles were pneumatically formed using a nebulising catheter and sized by laser diffraction. In vitro silencing of aerosolised and non-aerosolised formulations was evaluated in an EGFP endogenous-expressing H1299 cell line by flow cytometry. Non-invasive intratracheal insertion of the catheter was used to study nanoparticle deposition by histological detection of Cy3-labeled siRNA and gene silencing in transgenic EGFP mouse lungs using a flow cytometric method. RESULTS: Flow cytometric analysis demonstrated minimal alteration in gene silencing efficiency before (68%) and after (62%) aerosolisation in EGFP-expressing H1299 cells. Intratracheal catheter administration in mice resulted in nanoparticle deposition throughout the entire lung in both alveoli and bronchiolar regions using low amounts of siRNA. Transgenic EGFP mice dosed with the aerosolised nanoparticle formulation showed significant EGFP gene silencing (68% reduction compared to mismatch group). CONCLUSIONS: This work provides a technology platform for effective pulmonary delivery and gene silencing of RNAi therapeutics with potential use in preclinical studies of respiratory disease treatment.


Asunto(s)
Quitosano , Silenciador del Gen , Proteínas Fluorescentes Verdes/genética , Aerosoles , Animales , Secuencia de Bases , Línea Celular , Citometría de Flujo , Técnicas de Silenciamiento del Gen , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Interferencia de ARN
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