Solid lipid nanoparticles (SLN) consisting of tristearin or tribehenin, and monoolein aqueous dispersions (MAD) consisting of glyceril-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 characterized 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 from 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 complex 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 the green fluorescent protein or the firefly luciferase. Although with reduced efficiency, the results showed that the produced nanocarriers are able to convey plasmids within 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.

Cationic lipid nanosystems as carriers for nucleic acids

CORTESI, Rita
Primo
;
CAMPIONI, Matteo
Secondo
;
RAVANI, Laura;PINOTTI, Mirko
Penultimo
;
ESPOSITO, Elisabetta
Ultimo
2014

Abstract

Solid lipid nanoparticles (SLN) consisting of tristearin or tribehenin, and monoolein aqueous dispersions (MAD) consisting of glyceril-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 characterized 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 from 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 complex 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 the green fluorescent protein or the firefly luciferase. Although with reduced efficiency, the results showed that the produced nanocarriers are able to convey plasmids within 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.
Cortesi, Rita; Campioni, Matteo; Ravani, Laura; M., Drechsler; Pinotti, Mirko; Esposito, Elisabetta
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11392/1868717
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