Microencapsulation of nanoparticulate complexes of DNA with cationic lipids and polymers in pectin beads for targeted gene delivery |
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Authors: | Richard D. Harvey Sandrine Bourgeois Peter Pietzonka Laurent Desire Elias Fattal |
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Affiliation: | (1) Centre d’Etudes Pharmaceutiques, CNRS UMR 8612, Université Paris-Sud, 5 Rue Jean Baptiste Clément, 92296 Chatenay-Malabry, France;(2) Present address: Department of Pharmacy, King’s College London, Franklin Wilkins Building, 150 Stamford Street, SE1 9NN London, UK;(3) Present address: Novartis Pharm Ag, Technical Research & Development, CH-4002 Basel, Switzerland;(4) ExonHit Therapeutics SA, 65 Boulevard Massena, 75013 Paris, France |
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Abstract: | Nanoparticulate complexes of plasmid DNA (pDNA) with cationic liposomes/polymer, of approx 200 nm diameter, were encapsulated with a high degree of efficiency within calcium pectinate gel beads. Electron microscopy showed the DNA nanocomplexes to be evenly distributed throughout the gel matrix. Controlled release of pDNA-lipid nanocomplexes was achieved by the action of pectinase enzymes, whereas release of naked and polymer-complexed DNA was found to be more greatly influenced by the swelling behavior of the polysaccharide matrices in buffer alone. Physical degradation of pDNA within pectin beads was found to be accelerated during bead drying, most probably as a result of shear forces generated within the gel matrices by the evaporation of water. Plasmid complexation with cationic liposomes provided a greater degree of protection for the DNA during bead drying than complexation with cationic polymer, and was shown to successfully transfect cultured cells after release from the beads, via the action of pectinase. Observations concerning the physical stability of nanocomplexed pDNA, and its encapsulation within and release from pectin gel beads, are discussed with reference to the electrostatic interactions existing between the various components. |
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Keywords: | Calcium pectinate DNA nanocomplexes cationic liposomes polyethylenimine gene delivery |
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