Preparation of Biodegradable Gelatin Nanospheres with a Narrow Size Distribution for Carrier of Cellular Internalization of Plasmid DNA

Abstract

The objective of this study is to design biodegradable nanospheres of cationized gelatin as a carrier of cellular internalization of plasmid DNA. Ethylenediamine was chemically introduced into the carboxyl groups of gelatin to obtain cationized gelatin. The gelatin solution was filtered through a glass membrane under high pressure and dropped into 2-butanol, acetone or a mixture of the two to form nanospheres of cationized gelatin. The microspheres of cationized gelatin were prepared by the conventional water-in-oil emulsion method. The resulting nano- and microspheres of cationized gelatin were dehydrothermally treated at 160°C for different time periods to allow them to cross-link chemically. The size of nanospheres, prepared by the filtration method and changed by the type of solvents, was 1.86, 0.83 or 0.24 μm. The in vitro degradation of spheres became faster as the time period of dehydrothermal treatment was shorter. The degradation time of spheres in HCl solution linearly increased with an increase in the cross-linking time, irrespective of the sphere size. However, in the collagenase solution, when compared at the similar cross-linking density, the smaller spheres were degraded more slowly than the larger ones. The plasmid DNA incorporated in the nanospheres was released from the nanospheres with their degradation. The nanospheres incorporating plasmid DNA were internalized into cells, and intracellularly degraded with time to release plasmid DNA. The time period of plasmid DNA release was prolonged by increasing the nanosphere degradation time.