title: Cationic lipid nanosystems as carriers for nucleic acids.
abstract: Solid lipid nanoparticles (SLNs) consisting of tristearin or tribehenin, and monoolein aqueous dispersions (MADs) consisting of glyceryl-monoolein have been studied as potential nanocarriers for nucleic acids. The cationic character of nanocarriers was obtained by adding cationic surfactants, such as diisobutylphenoxyethyl-dimethylbenzyl ammonium chloride (DEBDA) or PEG-15 Cocopolyamine (PCPA), to the lipid composition. The products were characterised in terms of size and morphology by Cryo-TEM and PCS. The charge properties were determined by measuring the zeta potential. Our experimental protocol enabled us to obtain homogeneous and stable cationic nanosystems within 3-6 months of production. Assessment of cytotoxicity on HepG2 cells by MTT assays indicated that MAD preparations were less toxic than SLN, and in general PCPA-containing formulations are less cytotoxic than DEBDA-containing ones. The formation of electrostatic complexes with salmon sperm or plasmid DNA, used as model nucleic acids, was evaluated by electrophoresis on agarose gel. The results confirmed that all the formulations studied are able to form the complex. Finally, we investigated the ability of SLN and MAD to deliver DNA into HepG2 cells, and to this purpose we exploited expression plasmids for green fluorescent protein or firefly luciferase. Although with reduced efficiency, the results showed that the produced nanocarriers are able to convey plasmids into cells. The data obtained encourage further study aimed at improving these new formulations and proposing them as novel in vitro transfection reagents with potential application to in vivo delivery of nucleic acids.
authors.affiliation: Department of Life Sciences and Biotechnology, University of Ferrara, Ferrara, Italy. Electronic address: crt@unife.it.
authors.forename: Rita
authors.initials: R
authors.lastname: Cortesi
authors.forename: Matteo
authors.initials: M
authors.lastname: Campioni
authors.forename: Laura
authors.initials: L
authors.lastname: Ravani
authors.forename: Markus
authors.initials: M
authors.lastname: Drechsler
authors.forename: Mirko
authors.initials: M
authors.lastname: Pinotti
authors.forename: Elisabetta
authors.initials: E
authors.lastname: Esposito
mesh_terms: D000818:Animals; D004247:DNA / Q000737:chemistry*; D018014:Gene Transfer Techniques; D056945:Hep G2 Cells; D006801:Humans; D008055:Lipids / Q000737:chemistry*; D008422:Materials Testing; D053758:Nanoparticles / Q000737:chemistry*; D010957:Plasmids / Q000737:chemistry*; D012474:Salmon; D014162:Transfection / Q000379:methods*
chemical_list: D008055:Lipids; D004247:DNA