title: Use of magnetic nanoparticles and oscillating magnetic field for non-viral gene transfer into mouse cornea.
abstract: BACKGROUND
  Inherited, iatrogenic, and metabolic corneal disease could be potentially treated by supplying a functional gene or changing the expression levels of specific genes. Viral-based gene therapy is efficient, but restricted by evoking immune responses and inflammation. This study aimed to transfect mouse cornea with a non-viral based (oscillating magnetofection) method.
  
  
  METHODS
  Cultured mouse corneas were treated with magnetic nanoparticles (MNP) tethered to CAG promoter and green fluorescent protein (GFP) reporter plasmids exposed to a 1 Hz, 2 Hz, and 4 Hz oscillating magnetic field for 30 min and 60 min and in three DNA:MNP ratios (1:2, 1:1, 2:3). Corneas were cultured for up to 3 days and their green fluorescent channel intensity and number of GFP-positive cells were recorded. Transfection efficiency was estimated as the percentage of GFP-positive cells per total cells in a microscopic field.
  
  
  RESULTS
  Control experiments with absent magnetic exposure showed no GFP-positive cells. The optimum condition was recorded at 3:2 DNA:MNP ratio, 1 Hz magnetic oscillation, and 30 min duration of magnetic exposure (mean GFP-positive endothelial cell count 191·7 [SD 54·5], p=0·009; mean green fluorescent intensity 85·3 [SD 48·5]; and average transfection efficiency 23·3% [range 10·6-30·9]).
  
  
  CONCLUSIONS
  A novel non-viral method of transfecting cornea, magnetofection, is demonstrated and gives proof of principle for its translation into corneal gene therapy.
  
  
  BACKGROUND
  Welsh Clinical Academic Training Scheme.
authors: Cardiff Centre for Vision Sciences, Cardiff University, Cardiff, UK. Electronic address: waisieneng@gmail.com.
authors: Wai
authors: W
authors: Siene Ng
authors: Cardiff Centre for Vision Sciences, Cardiff University, Cardiff, UK.
authors: Katie
authors: K
authors: Binley
authors: School of Dentistry, Cardiff University, Cardiff, UK.
authors: Bing
authors: B
authors: Song
authors: Cardiff Centre for Vision Sciences, Cardiff University, Cardiff, UK.
authors: James E
authors: JE
authors: Morgan