Abstract
The interaction between calf-thymus DNA, ctDNA, and various single-chained surfactants with different functional groups at the end of hydrophobic tail was studied with the goal of investigating the influence of the functional group nature on surfactant DNA compacting efficiency. The surfactants investigated were dodecyltriethylammonium bromide (DTEABr), triethyl(1-phenoxydodecyl)ammonium bromide (12PhBr), triethyl(2-naphthoxydodecyl)ammonium bromide (12NBr) and 11-(isonicotinoyloxy)-N,N,N-triethyl-1-undecanaminium bromide (11PyBr). Results made evident that the surfactants’ tendencies to self-aggregation is the key factor determining their efficiency to compact the nucleic acid. Subsequently, DOPE/12NBr/pEGFP-C1 lipoplexes, with different cationic surfactant molar fractions (α) and mass ratios (L/D), were prepared and characterized. DOPE is a zwitterionic phospholipid 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine, and the plasmid pEGFP-C1 carries a GFP coding sequence with the necessary regulatory elements for constitutive expression of the gene in human cells. 12NBr was chosen because it was the most efficient DNA compacting agent among the surfactants investigated. Finally, the cytotoxicity and transfection efficiency (TE) of DOPE/12NBr/pDNA lipoplexes, with different compositions, were investigated.
Highlights
Research on the interactions between surfactants and nucleic acids has drawn great attention in the last decades for its significance in biomedical and biotechnological applications, for the possibility of using surfactant/DNA systems for gene delivery [1,2,3,4,5,6,7]
In order to investigate if the formation of the DOPE/12NBr/pDNA lipoplexes was accompanied by a morphological change of the pDNA, circular dichroism (CD) spectra were registered for various α at different L/D values
For lipoplexes with α = 0.2 and L/D = 20 and α = 0.3 and L/D = 16 the hydrodynamic diameter found by TEM were 178 ± 46 nm and 203 ± 55 nm, respectively, in agreement with those estimated by Dynamic Light Scattering (DLS), within experimental errors
Summary
Research on the interactions between surfactants and nucleic acids has drawn great attention in the last decades for its significance in biomedical and biotechnological applications, for the possibility of using surfactant/DNA systems for gene delivery [1,2,3,4,5,6,7]. Cytotoxicity of these lipoplexes in a human bone osteosarcoma epithelial U2OS cell line was studied, and the transfection efficiency (TE) of these DOPE/12NBr/pDNA lipoplexes in the same cell line was investigated These studies are relevant considering that, after a better understanding of the DNA compaction process by the surfactants studied, the search for more effective cationic surfactant nonviral vectors for gene therapy would be a priority research goal, given the number of rare disorders in the population which need medical solutions. Was studied, and the transfection efficiency (TE) of these DOPE/12NBr/pDNA lipoplexes in the same cell line was investigated
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