Novel cationic liposome formulation for the delivery of an oligonucleotide decoy to NF-kappaB into activated macrophages.
ABSTRACT Nuclear factor-kappaB (NF-kappaB) is involved in several pathological processes, such as inflammation. Pro-inflammatory genes expression can be down-regulated by using an oligonucleotide (ODN) decoy to NF-kappaB. Cationic liposomes are largely used to improve ODN uptake into cells, although a higher transfection efficiency and a lower toxicity are required to use them in therapy. In this work, we investigated the potential of a novel liposome formulation, based on the recently synthesised cationic lipid (2,3-didodecyloxypropyl) (2-hydroxyethyl) dimethylammonium bromide (DE), as the delivery system for a double stranded ODN decoy to NF-kappaB. Liposomes composed of DE or DE mixed with 1,2-dioleyl-sn-glycero-3-phosphoethanolamine or cholesterol as helper lipids were complexed with ODN at different +/- charge ratios. In vitro uptake and the effect of ODN, naked or complexed with DE-containing liposomes, were evaluated in lipopolysaccharide-stimulated RAW 264.7 macrophages. The use of helper lipids increased liposome physical stability up to 1 year at 4 degrees C. ODN complexed with DE/cholesterol liposomes, at the +/- charge ratio of 8, showed a limited cytotoxicity and the highest inhibition of nitrite production, inducible nitric oxide synthase protein expression and NF-kappaB/DNA binding activity. Confocal microscopy confirmed a high ODN cell uptake obtained with DE/cholesterol liposomes at the highest +/- charge ratio.
Article: Nano and microtechnologies for the delivery of oligonucleotides with gene silencing properties.[show abstract] [hide abstract]
ABSTRACT: Oligonucleotides (ONs) are synthetic fragments of nucleic acid designed to modulate the expression of target proteins. DNA-based ONs (antisense, antigene, aptamer or decoy) and more recently a new class of RNA-based ONs, the small interfering RNAs (siRNAs), have gained great attention for the treatment of different disease states, such as viral infections, inflammation, diabetes, and cancer. However, the development of therapeutic strategies based on ONs is hampered by their low bioavailability, poor intracellular uptake and rapid degradation in biological fluids. The use of a non-viral carrier can be a powerful tool to overcome these drawbacks. Lipid or polymer-based nanotechnologies can improve biological stability and cellular uptake of ONs, with possibility of tissue and/or cellular targeting. The use of polymeric devices can also produce a prolonged release of the ON, thus reducing the need of frequent administrations. This review summarizes advantages and issues related to the main non-viral vectors used for ON delivery.Molecules 02/2009; 14(8):2801-23. · 2.39 Impact Factor
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ABSTRACT: The development of carbon nanotubes (CNTs) for biomedical and biotechnological applications has gained great promise recently, especially for their interesting use in the delivery of therapeutically active molecules to targeted cells. The interaction between cells and this nanomaterial is a critical feature that is responsible for the pharmacological effect as well as for any eventual toxicity. With respect to the latter aspect, in this manuscript we have evaluated a few parameters that seem to be involved in the cytotoxic profile of CNTs (both Single-walled (SWCNTs) and Multi-walled (MWCNTs) nanotubes), such as their sidewall functionalization, tubes' length, solubility, concentration and purity. Among them, we identified the last as the most crucial factor: we have shown that our ultrapure, totally dispersible, carbon nanotubes not only display lack of toxicity in the range of concentrations normally used (10-150 μg/ml), but they also pave the way for an extensive use of this material for several biomedical purposes.Current Nanoscience 03/2010; 6(2):141-154. · 1.78 Impact Factor