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A physicochemical characterization of the interaction between DC-Chol/DOPE cationic liposomes and DNA

AuthorsRodríguez-Pulido, Alberto; Ortega, Francisco José; Llorca, Óscar ; Aicart, Emilio; Junquera, Elena
Issue Date2-Oct-2008
PublisherAmerican Chemical Society
CitationJournal of Physical Chemistry B 112(39):12555-12565(2008)
AbstractA 1:1 mixture of the cationic lipid 3β-[N-(N′,N′-dimethylaminoethane)-carbamoyl]cholesterol hydrochloride (DC-Chol) and the zwitterionic lipid, 1,2-dioleoyl-sn-glycero-3-phosphoetanolamine (DOPE), has been used to compact calf-thymus DNA (CT-DNA) in aqueous buffered solution at 298.15 K. The formation process of this lipoplex has been analyzed by means of electrophoretic mobility, cryo-TEM, dynamic light scattering, and fluorescence spectroscopy techniques. The experimental results indicate that DC-Chol/DOPE liposomes are mostly spherical and unilamellar, with a mean diameter of around 99 ± 10 nm and a bilayer with a thickness of 4.5 ± 0.5 nm. In the presence of CT-DNA, DC-Chol/DOPE/CT-DNA lipoplexes are formed by means of a strong entropically driven surface electrostatic interaction, as confirmed by zeta potential and fluorescence results, as a consequence of which DNA is compacted and condensed at the surface of the cationic liposomes. The negative charges of DNA phosphate groups are neutralized by the positive charges of cationic liposomes at the isoneutrality L/D ratio, (L/D) around 4, obtained from electrophoretic, fluorescence, and DLS measurements. The decrease in the fluorescence emission intensity of ethidium bromide, EtBr, initially intercalated between DNA base pairs, as long as the association between the biopolymer and the cationic liposomes takes place has permitted one to confirm its electrostatic character as well as to evaluate the different microenvironments of varying polarity of DNA-double helix, liposomes, and/or lipoplexes. Electronic microscopy reveals a rich scenario of possible nanostructures and morphologies for the lipoplexes, from unilamellar DNA-coated liposomes to multilamellar lipoplexes passing through cluster-like structures and several intermediate morphologies
Description11 páginas, 13 figuras, 2 tablas, 3 gráficos -- PAGS nros. 12555-1565
Publisher version (URL)http://dx.doi.org/10.1021/jp804066t
Appears in Collections:(CIB) Artículos
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