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In vitro reporter gene transfection via plasm ID DNA delivered by metered dose inhaler

Bains, Baljinder Kaur, Birchall, James Caradoc, Toon, Richard and Taylor, Glyn 2010. In vitro reporter gene transfection via plasm ID DNA delivered by metered dose inhaler. Journal of Pharmaceutical Sciences 99 (7) , pp. 3089-3099. 10.1002/jps.22085

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Abstract

Aerosolised DNA administration could potentially advance the treatment of inheritable lung diseases, lung malignancies and provide genetic immunisation against infection. Jet nebulisation, the current standard for introducing DNA formulations into the lung, is inherently inefficient. Pressurised metered dose inhalers (pMDIs) offer a potentially more efficacious and convenient alternative, especially for repeat administration. We aim to modify a novel low-energy nanotechnology process to prepare surfactant-coated pDNA nanoparticles for pulmonary gene delivery via a pMDI. Water-in-oil microemulsions containing green fluorescent protein reporter plasmid were snap-frozen and lyophilised. Lyophilised pDNA, in some cases following a surfactant wash, was incorporated into pMDIs with hydrofluoroalkane 134a (HFA134a) propellant and ethanol as cosolvent. To assess biological functionality, A549 human lung epithelial cells were exposed to aerosolised pDNA particles in the presence of dioleoyl-trimethylammonium propane (DOTAP). Transfection studies demonstrated that pDNA biological functionality was maintained following aerosolisation. In vitro toxicity assays (MTT) showed no significant cell viability loss following aerosolised pDNA treatment. We have demonstrated that pDNA particles can be incorporated into an HFA134a formulation and aerosolised using a standard valve and actuator. Particles prepared by this novel process have potential for stable and efficient delivery of pDNA to the lower respiratory tract via standard pMDI technology.

Item Type: Article
Date Type: Publication
Status: Published
Schools: Pharmacy
Subjects: R Medicine > RM Therapeutics. Pharmacology
Uncontrolled Keywords: pulmonary drug delivery; pressurised metered dose inhaler; aerosol; plasmid DNA; nonviral gene delivery; nanotechnology; formulation; microemulsion
Publisher: Wiley-Blackwell
ISSN: 1520-6017
Last Modified: 04 Jun 2017 04:05
URI: http://orca-mwe.cf.ac.uk/id/eprint/32372

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