title: DOTAP/UDCA vesicles: novel approach in oligonucleotide delivery.
abstract: The relatively hydrophilic bile acid, ursodeoxycholic acid (UDCA), was used as an additive to DOTAP cationic liposomes to evaluate the effect on the cellular uptake of an oligonucleotide. Nuclear magnetic resonance studies were applied to estimate the relative amount of incorporated UDCA into the lipidic bilayers. DOTAP or DOTAP-UDCA vesicles (MixVes; DOTAP/UDCA molar ratios 1:0.25, 1:0.5, 1:1, and 1:2) formed complexes with 5'-fluorescein conjugated 29-mer phosphorothioate oligonucleotides (PS-ODNs) and studied using gel electrophoresis. In addition, the complexes were tested after transfection to assess the cellular uptake and the localization of the oligo in a HaCaT cell line by the use of cytofluorimetric and confocal microscopic analysis. DOTAP lipid formulated in the presence of a defined amount of UDCA forms more stable, flexible, and active MixVes. In particular, the MixVes at 1:0.25 and 1:0.5 molar ratios increase and modify the cellular uptake of PS-ODNs if compared with DOTAP liposomes 3 hours after the transfection studies. Moreover, the in vitro data suggest that these new formulations are not toxic.
authors: Department of Pharmaceutical Sciences, University of Modena and Reggio Emilia, Modena, Italy.
authors: Barbara
authors: B
authors: Ruozi
authors: Renata
authors: R
authors: Battini
authors: Monica
authors: M
authors: Montanari
authors: Adele
authors: A
authors: Mucci
authors: Giovanni
authors: G
authors: Tosi
authors: Flavio
authors: F
authors: Forni
authors: Maria Angela
authors: MA
authors: Vandelli
mesh_terms: D016923:Cell Death / Q000187:drug effects; D016503:Drug Delivery Systems / Q000379:methods*; D005229:Fatty Acids, Monounsaturated / Q000494:pharmacology*; D006801:Humans; D008081:Liposomes / Q000494:pharmacology*; D009682:Magnetic Resonance Spectroscopy; D018625:Microscopy, Atomic Force; D018613:Microscopy, Confocal; D009841:Oligonucleotides / Q000008:administration & dosage* / Q000494:pharmacology*; D000644:Quaternary Ammonium Compounds / Q000494:pharmacology*; D014162:Transfection; D014580:Ursodeoxycholic Acid / Q000494:pharmacology*
chemical_list: D005229:Fatty Acids, Monounsaturated; D008081:Liposomes; D009841:Oligonucleotides; D000644:Quaternary Ammonium Compounds; D014580:Ursodeoxycholic Acid; C070046:1,2-dioleoyloxy-3-(trimethylammonium)propane