Liposome/DNA systems: correlation between association, hydrophobicity and cell viability
Small unilamellar vesicles associated with plasmid DNA showed maximum association efficiency for a cationic mixture of egg phosphatidylcholine (EPC):1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE):di-1,2-dioleoyl-3-trimethyl ammonium propane (DOTAP) (16:8:1 molar ratio) [65%], followed by neutral lipids EPC:1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine (DMPE):cholesterol (Chol) (2:2:1 molar ratio) [30%], and a polymerized formulation 1,2-bis(10,12-tricosadiynoyl)sn-glycero-3-phosphocholine (DC8,9PC):DMPE:Chol (2:2:1 molar ratio) [11%]. The hydrophobicity factor (HF) for these formulations followed the trend DC8,9PC:DMPE:CHOL < EPC:DMPE:Chol < EPC:DOPE DOTAP, and DNA association did not alter this trend. Results suggest that the higher the HF value, the more fluid the membrane and the higher the efficiency of DNA association. On the other hand, no differences were observed in cell toxicity with lipids up to 1 mg/ml in VERO cells.
KeywordsCell viability Hydrophobicity Liposomes Plasmid DNA Polymeric lipid
We thank Lic. Silvina Mangano, Lic. Ricardo Gargini and Lic. Marcelo Argüelles for their technical assistance in VP7 plasmid DNA obtention and helpful discussions. This work was supported by grants from CIC (Comisión de Investigaciones Científicas), CONICET (Consejo Nacional de Investigaciones Científicas y Técnicas), and the Universidad Nacional de Quilmes (Buenos Aires, Argentina). Silvia del V. Alonso is a Scientific Research Career member of the CONICET, Argentina.
- Bakaltcheva I, Williams WP, Schmitt JM, Hincha DK (1994) The solute permeability of thylakoid membranes is reduced by low concentrations of trehalose as a co-solute. Biochim Biophys Acta 3:38–44Google Scholar
- Borgatti M, Breda L, Cortesi R, Nastruzzi C, Romanelli A, Saviano M, Bianchi N, Mischiati C, Pedone C, Gambari R (2002) Cationic liposomes as delivery systems for double-stranded PNA-DNA chimeras exhibiting decoy activity against NF-κB transcription factors. Biochem Pharmacol 64:609–616PubMedCrossRefGoogle Scholar
- Chiaramoni NS, Speroni L, Taira MC, Alonso Romanowski S (2003) Lipid-DNA formulations: an approach to biodistribution assays. Biocell 27:86–86Google Scholar
- Lelkes PI, Miller IR (1980) Perturbations of membrane structure by optical probes: II. Differential scanning calorimetry of dipalmitoyllecithin and its analogs interacting with Merocyanine 540. J Membr Biol 31:1–15Google Scholar
- R.R.C. New (1990) Ficoll flotation method for DNA. In: Rickwood D (ed) Liposomes: a practical approach. IRL Press, UK, pp 95–96Google Scholar
- Sambrook J, Fritsch EF, Maniatis T (1989) Quantitation of DNA ns RNA. In: Nolan C (ed) Molecular cloning: a laboratory manual, vol 3, 2nd edn. Cold Spring Harbor Laboratory Press, pp E5–E7Google Scholar
- Savva M, Aljaberi A, Feigand J, Beer Stolz D (2005) Correlation of the physicochemical properties of symmetric 1,3-dialkoylamidopropane-based cationic lipids containing single primary and tertiary amine polar head groups with in vitro transfection activity. Colloids Surf B Biointerfaces 43:43–56PubMedCrossRefGoogle Scholar