RESUMO
We have prepared and characterised injectable adenovirus-loaded polymeric microparticles to be used for in vitro and in vivo gene transfer studies. Microparticles were prepared by the water-in-oil-in-water solvent evaporation method using a novel system where the emulsification process is carried out by the turbulent injection of the phases in the total recirculation one-machine system (TROMS) apparatus. In vitro studies were performed to assess the amount of infectious adenovirus released from the microparticles, showing that these microparticles release higher amounts of infectious adenovirus than microparticles prepared by standard emulsification techniques. We also tested whether sustained release in vivo could overcome the short-lived gene expression profile which is typical of adenovirus delivery into muscle. Intramuscular injection of adenovirus-loaded microparticles in immunocompetent mice showed transgene (beta-galactosidase) expression for at least 7 weeks in two out of four muscles injected with adenovirus-loaded microparticles prepared by TROMS, but not in control muscles injected with purified adenovirus stocks.