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Pharmaceutical Research

, Volume 22, Issue 8, pp 1294–1304 | Cite as

Electrohydrodynamic Comminution: A Novel Technique for the Aerosolisation of Plasmid DNA

  • Lee A. Davies
  • Kevin Hannavy
  • Neville Davies
  • Alistair Pirrie
  • Ronald A. Coffee
  • Stephen C. Hyde
  • Deborah R. Gill
Research Paper

Purpose

Naked plasmid DNA (pDNA) is a potential gene transfer agent for lung gene therapies but cannot be aerosolised without degradation using conventional nebulisation devices. This study investigated the viability of an alternative nebulisation technique, electrohydrodynamic (EHD) comminution for the aerosol delivery of naked DNA in vivo.

Methods

Naked pDNA was aerosolised using jet and ultrasonic nebulisers, and by EHD comminution. Degradation associated with the aerosolisation process was investigated using gel electrophoresis and by transfection studies in cell culture. Optimised formulations for EHD aerosolisation of pDNA were developed and in vivo deposition and reporter gene expression were investigated in mice.

Results

Unlike conventional nebulisation devices, EHD comminution of plasmids up to 15 kb in size resulted in no detectable pDNA degradation. EHD formulations containing up to 1 mg/ml pDNA were developed and shown to produce monodisperse aerosols suitable for targeted lung delivery in humans. Aerosolisation studies in vivo demonstrated detectable levels of pDNA deposition and measurable luciferase reporter gene expression in the lungs of exposed mice.

Conclusions

This study demonstrates for the first time that respirable aerosols of naked pDNA can be generated without plasmid degradation and that EHD comminution is an appropriate technique for the aerosolisation of delicate gene transfer agents.

Key Words

aerosolisation electrohydrodynamic gene transfer naked DNA 

Abbreviations

CF

cystic fibrosis

DC-Chol/DOPE

3β[N-(N′, N′-dimethylaminoethane)-carbomoyl] cholesterol and dioleoylphosphatidylethanolamine

DMEM

Dulbecco’s modified Eagle medium

EHD

electrohydrodynamic

GSD

geometric standard deviation

GTA

gene transfer agent

OC

open circular

pDNA

plasmid DNA

PCR

polymerase chain reaction

PEI

polyethylenimine

RF

respiratory fraction

SC

supercoiled

VMD

volume median diameter

Notes

Acknowledgments

All research included in this study was sponsored by Electrosols Ltd., Oxford, United Kingdom and by the Medical Research Council, United Kingdom.

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Copyright information

© Springer Science + Business Media, Inc. 2005

Authors and Affiliations

  • Lee A. Davies
    • 1
    • 2
  • Kevin Hannavy
    • 1
  • Neville Davies
    • 2
  • Alistair Pirrie
    • 2
  • Ronald A. Coffee
    • 2
  • Stephen C. Hyde
    • 1
  • Deborah R. Gill
    • 1
  1. 1.GeneMedicine Research Group, Nuffield Department of Clinical Laboratory SciencesUniversity of Oxford, John Radcliffe HospitalOxfordUK
  2. 2.Electrosols Ltd., Magdalen CentreOxford Science ParkOxfordUK

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