Delivery of plasmid IGF-1 to chondrocytes via cationized gelatin nanoparticles

The objective of the present study was to investigate the use of gelatin and cationized-gelatin nanoparticles for the nonviral delivery of the plasmid DNA encoding for insulin-like growth factor (IGF)-1 to adult canine articular chondrocytes in vitro; plasmid for enhanced green fluorescence protein (EGFP) was used as a marker gene. The spherical cationized gelatin nanoparticles were on average 172 nm in diameter, compared with the often ellipsoid-shaped unmodified (noncationized) gelatin particles that generally appeared to be 10 μm to greater than 20 μm in length. The zeta potential of the positively charged cationized gelatin nanoparticles containing the plasmid was around 20 mV compared with about 2 mV for the unmodified gelatin particles. There was no noticeable fluorescence from the cells treated with the nanoparticles prepared with the original (noncationized) gelatin particles containing the pEGFP. In contrast, numerous cells in the group transfected with the cationized gelatin-pEGFP nanoparticles were found to fluoresce demonstrating the transfection of the cells. There was five-fold elevation in the amount of IGF-1 produced by the cells treated with the cationized gelatin nanoparticles containing the IGF-1 plasmid compared with the unmodified (noncationized) gelatin particles. There was a clear effect of varying the weight ratio of plasmid IGF-1 in the cationized gelatin nanoparticles on the IGF-1 in the medium of cells exposed to the nanoparticles for 5 h. A peak in the amount of released IGF-1 was detected at a gelatin:IGF-1 weight ratio of 250:1.