Influence of phospholipid composition on cationic emulsions/DNA complexes: physicochemical properties, cytotoxicity, and transfection on Hep G2 cells

Abstract

Background Cationic nanoemulsions have been recently considered as potential delivery systems for nucleic acids. This study reports the influence of phospholipids on the properties of cationic nanoemulsions/DNA plasmid complexes. Methods Nanoemulsions composed of medium-chain triglycerides, stearylamine, egg lecithin or isolated phospholipids, ie, DSPC, DOPC, DSPE, or DOPE, glycerol, and water were prepared by spontaneous emulsification. Gene transfer to Hep G2 cells was analyzed using real-time polymerase chain reaction. Results The procedure resulted in monodispersed nanoemulsions with a droplet size and zeta potential of approximately 250 nm and +50 mV, respectively. The complexation of cationic nanoemulsions with DNA plasmid, analyzed by agarose gel retardation assay, was complete when the complex was obtained at a charge ratio of ≥1.0. In these conditions, the complexes were protected from enzymatic degradation by DNase I. The cytotoxicity of the complexes in Hep G2 cells, evaluated by MTT assay, showed that an increasing number of complexes led to progressive toxicity. Higher amounts of reporter DNA were detected for the formulation obtained with the DSPC phospholipid. Complexes containing DSPC and DSPE phospholipids, which have high phase transition temperatures, were less toxic in comparison with the formulations obtained with lecithin, DOPC, and DOPE. Conclusion The results show the effect of the DNA/nanoemulsion complexes composition on the toxicity and transfection results.

Self-assembled hyaluronan nanocapsules for the intracellular delivery of anticancer drugs

“…This was evident in the differences in the cellular toxicities of the two nanocapsules at the highest tested concentration (1 mg/mL). This finding correlates to the presence of the cationic surfactant CTAB in high concentration, which is in agreement with previous reports 39 . In addition, the marked difference in viability between the HA-based nanocapsules and the surfactant solution composed of Tween®80/Solutol/CTAB at 350 μg/mL suggests the beneficial effect of HA surrounding the surfactant layer as well as the correct isolation of the system from the free surfactants 40 .…”

Nasal Administration of Cationic Nanoemulsions as CD73-siRNA Delivery System for Glioblastoma Treatment: a New Therapeutical Approach

“…The main physicochemical characteristics of NE and complexes obtained in this study were in accordance with those reported in previous studies using NE obtained through microfluidization under similar conditions [40,47]. The correlation between the increase in charge ratio and the efficiency of oligonucleotide complexation corroborates data already reported in the literature [48][49][50][51]. In addition, an efficient system for CD73 siRNA in vitro and in vivo delivery was developed.…”

Post-modification of preformed liposomes with novel non-phospholipid poly(ethylene glycol)-conjugated hexadecylcarbamoylmethyl hexadecanoic acid for enhanced circulation persistence in vivo

“…As shown in Figure 3, HDAS-PEG as well as liposomes containing HDAS-PEG did not show any effect on the viability of these two cell lines. In general, cationic lipids and phospholipids with low phase transition temperature are toxic to cells (Fraga et al, 2011; Senior et al, 1991). Liposomes bearing rigid lipid chains provide more stable and less toxic interface between biological system and the formulation.…”