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- W2972145391 abstract "Objectives A number of respiratory diseases could potentially benefit from genetransfer to the lung, including cystic fibrosis, asthma and emphysema. Jet nebulisation,the current standard for introducing gene therapy formulations into the lung, ishighly inefficient. While newer nebuliser technologies are under development, pressurisedmetered dose inhalers (pMDIs) offer an alternative with potential advantagesof more efficacious and rapid administration. As both solution and suspensionpMDI formulations have drawbacks relating to stability and dose reproducibility, we aim to explore the potential of modifying a novel low-energy nanotechnologyprocess (Dickinson et al 2001) to prepare surfactant-coated plasmid DNA (pDNA)nanoparticles for pulmonary gene delivery via a dispersion pMDI.Methods Water-in-oil microemulsions containing pEGFP-N1 reporter plasmidwere prepared from sucrose solution (as cryoprotectant and aqueous phase), lecithin:propan-2-ol (as stabilising surfactant) and iso-octane (as organic phase).Resultant microemulsions were snap frozen in liquid nitrogen and lyophilised.Excess surfactant was removed by repeated washes with iso-octane and centrifugation.Scanning electron microscopy (SEM) and gel electrophoresis were used tocharacterise surface morphology and deduce pDNA integrity, respectively.Results A ternary phase diagram was constructed to identify optimised microemulsioncompositions (Figure 1). Microemulsions with a surfactant to water ratioof 1.5 and above formed stable water in oil isotropic systems. Unstable biphasicsystems were formed when the surfactant to water ratio fell below 1.5. Optimisedformulations resulted in effective incorporation of pDNA into the aqueous pool ofreverse micelles. Controlled lyophilisation enabled the formation of novel surfactant-coated pDNA nanoparticles. A qualitative analysis performed using gelelectrophoresis showed that the freeze-dried particles retained pDNA structuralintegrity. SEM images conferred aggregates of DNA-cryoprotectant particles.Conclusions Freeze-drying pDNA microemulsions produced surfactant coatedpDNA particles whilst successfully maintaining the integrity of the pDNA. Thenanotechnology process used offers the potential for the incorporation of pDNAnanoparticles into pMDI systems for pulmonary delivery of gene vectors." @default.
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- W2972145391 date "2007-09-01" @default.
- W2972145391 modified "2023-09-26" @default.
- W2972145391 title "Novel nanoparticles for pulmonary gene delivery [Abstract]" @default.
- W2972145391 hasPublicationYear "2007" @default.
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