PMID: 15367377Sep 16, 2004Paper

In vitro characterization and delivery of chitosan-DNA microparticles into mammalian cells

Journal of Pharmacy & Pharmaceutical Sciences : a Publication of the Canadian Society for Pharmaceutical Sciences, Société Canadienne Des Sciences Pharmaceutiques
Tugce Dastan, Kadir Turan

Abstract

Chitosan has a high potential for transferring DNA molecules into mammalian cells because of its cationic properties. In the present study, we have investigated DNA encapsulation efficiency and loading capacity of chitosan microparticles prepared in different conditions, as well as in vitro DNA release from microparticles, and transfection of different cell lines with chitosan-DNA microparticles, which may be employed in future in vivo studies. Plasmid DNAs were amplified in Escherichia coli DH5alpha and isolated by the alkali SDS-lysis method. Chitosan-DNA microparticles were prepared by the coacervation method by using different concentrations of chitosan and plasmid DNAs. In vitro release experiments were performed in PBS at 37 degrees C and DNA release was monitored spectrophotometrically. Transfection efficiency of chitosan-DNA microparticles into mammalian cells was determined by measuring the beta-galactosidase activity in cell lysates. DNA encapsulation efficiency and loading capacity of microparticles was altered depending on the chitosan and DNA concentrations. Approximately 75-85% of DNA was encapsulated into the chitosan-DNA microparticles. The average size of microparticles was found to be approximately 2 microm. I...Continue Reading

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