A physicochemical characterization of the interaction between DC-Chol/DOPE cationic liposomes and DNA.
A 1:1 mixture of the cationic lipid 3beta-[ 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) varphi 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.
Rodríguez-Pulido
Alberto
A
Departamento de Química Física I, Facultad de Ciencias Químicas, Universidad Complutense de Madrid, Madrid, Spain.
Ortega
Francisco
F
Llorca
Oscar
O
Aicart
Emilio
E
Junquera
Elena
E
D000818:Animals; D002417:Cattle; D002784:Cholesterol / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism; D004247:DNA / Q000737:chemistry* / Q000378:metabolism; D004586:Electrophoresis; D008051:Lipid Bilayers / Q000737:chemistry / Q000378:metabolism; D008081:Liposomes / Q000737:chemistry* / Q000378:metabolism; D046529:Microscopy, Electron, Transmission; D049329:Nanostructures / Q000737:chemistry; D010714:Phosphatidylethanolamines / Q000737:chemistry* / Q000378:metabolism; D013050:Spectrometry, Fluorescence
C410262:1,2-dioleoyl-glycero-3-phosphatidyl ethanolamine; D008051:Lipid Bilayers; D008081:Liposomes; D010714:Phosphatidylethanolamines; C070648:3-(N-(N',N'-dimethylaminoethane)carbamoyl)cholesterol; D004247:DNA; C427203:calf thymus DNA; D002784:Cholesterol