Gene therapy is a pharmacological regimen of great interest for the treatment of genetic, neoplastic and infectious diseases. Recently, the use of lipid dispersions as gene delivery systems has attracted wide attention of worldwide formulators due to their potential applications. This study focuses on the preparation and characterisation of nanostructured lipid carrier (NLC) and monoolein aqueous dispersions (MAD) as potential nanocarriers intended to convey nucleic acids. As cationic surfactants, diisobutylphenoxyethyl-dimethylbenzyl ammonium chloride (DEBDA) or PEG-15 Cocopolyamine (PCPA) were employed. In the experimental conditions used, it was possible to obtain homogeneous and stable cationic nanosystems within 3-6 months from production. MAD and NLC were subjected to MTT assay for in vitro cytotoxicity on HepG2 cells indicating that MAD-PCPA are less cytotoxic than MAD-DEBDA. The formation of electrostatic complex with Salmon Sperm DNA, used as model nucleic acid, was evaluated by electrophoresis on agarose gel. The results confirm that all the formulations studied are able to form the complex. Finally, studies were performed in vitro transfection on HepG2 cells using a plasmid encoding GFP and one encoding for luciferase. The encouraging data obtained allow to further study aimed to achieve in vitro transfection agents alternative to those currently available and to identify possible in vivo applications.
Cationic nanostructured lipid carrier (NLC) and monoolein aqueous dispersions (MAD) as potential carriers for nucleic acids
CORTESI, Rita;RAVANI, Laura;PINOTTI, Mirko;MENEGATTI, Enea;ESPOSITO, Elisabetta
2011
Abstract
Gene therapy is a pharmacological regimen of great interest for the treatment of genetic, neoplastic and infectious diseases. Recently, the use of lipid dispersions as gene delivery systems has attracted wide attention of worldwide formulators due to their potential applications. This study focuses on the preparation and characterisation of nanostructured lipid carrier (NLC) and monoolein aqueous dispersions (MAD) as potential nanocarriers intended to convey nucleic acids. As cationic surfactants, diisobutylphenoxyethyl-dimethylbenzyl ammonium chloride (DEBDA) or PEG-15 Cocopolyamine (PCPA) were employed. In the experimental conditions used, it was possible to obtain homogeneous and stable cationic nanosystems within 3-6 months from production. MAD and NLC were subjected to MTT assay for in vitro cytotoxicity on HepG2 cells indicating that MAD-PCPA are less cytotoxic than MAD-DEBDA. The formation of electrostatic complex with Salmon Sperm DNA, used as model nucleic acid, was evaluated by electrophoresis on agarose gel. The results confirm that all the formulations studied are able to form the complex. Finally, studies were performed in vitro transfection on HepG2 cells using a plasmid encoding GFP and one encoding for luciferase. The encouraging data obtained allow to further study aimed to achieve in vitro transfection agents alternative to those currently available and to identify possible in vivo applications.I documenti in SFERA sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
