title: Chitosan nanoparticles for siRNA delivery: optimizing formulation to increase stability and efficiency.
abstract: This study aims at developing chitosan-based nanoparticles suitable for an intravenous administration of small interfering RNA (siRNA) able to achieve (i) high gene silencing without cytotoxicity and (ii) stability in biological media including blood. Therefore, the influence of chitosan/tripolyphosphate ratio, chitosan physicochemical properties, PEGylation of chitosan as well as the addition of an endosomal disrupting agent and a negatively charged polymer was assessed. The gene silencing activity and cytotoxicity were evaluated on B16 melanoma cells expressing luciferase. We monitored the integrity and the size behavior of siRNA nanoparticles in human plasma using fluorescence fluctuation spectroscopy and single particle tracking respectively. The presence of PEGylated chitosan and poly(ethylene imine) was essential for high levels of gene silencing in vitro. Chitosan nanoparticles immediately released siRNA in plasma while the inclusion of hyaluronic acid and high amount of poly(ethylene glycol) in the formulation improved the stability of the particles. The developed formulations of PEGylated chitosan-based nanoparticles that achieve high gene silencing in vitro, low cytotoxicity and high stability in plasma could be promising for intravenous delivery of siRNA.
authors.affiliation: Université Catholique de Louvain, Pharmaceutics and Drug Delivery Group, Louvain Drug Research Institute, 1200 Brussels, Belgium.
authors.forename: H
authors.initials: H
authors.lastname: Ragelle
authors.affiliation: University of Liège, Center for Education and Research on Macromolecules, 4000 Liège, Belgium.
authors.forename: R
authors.initials: R
authors.lastname: Riva
authors.affiliation: Université Catholique de Louvain, Pharmaceutics and Drug Delivery Group, Louvain Drug Research Institute, 1200 Brussels, Belgium.
authors.forename: G
authors.initials: G
authors.lastname: Vandermeulen
authors.affiliation: Ghent University, Laboratory of General Biochemistry and Physical Pharmacy, Faculty of Pharmaceutical Sciences, 9000 Ghent, Belgium.
authors.forename: B
authors.initials: B
authors.lastname: Naeye
authors.affiliation: Université Catholique de Louvain, Molecules, Solids and Reactivity, Institute of Condensed Matter and Nanosciences, 1348 Louvain-la-Neuve, Belgium.
authors.forename: V
authors.initials: V
authors.lastname: Pourcelle
authors.affiliation: Université Catholique de Louvain, Molecules, Solids and Reactivity, Institute of Condensed Matter and Nanosciences, 1348 Louvain-la-Neuve, Belgium.
authors.forename: C S
authors.initials: CS
authors.lastname: Le Duff
authors.affiliation: Université Catholique de Louvain, Institute of Condensed Matter and Nanosciences, Bio & Soft Matter, 1348 Louvain-la-Neuve, Belgium.
authors.forename: C
authors.initials: C
authors.lastname: D'Haese
authors.affiliation: Université Catholique de Louvain, Institute of Condensed Matter and Nanosciences, Bio & Soft Matter, 1348 Louvain-la-Neuve, Belgium.
authors.forename: B
authors.initials: B
authors.lastname: Nysten
authors.affiliation: Ghent University, Laboratory of General Biochemistry and Physical Pharmacy, Faculty of Pharmaceutical Sciences, 9000 Ghent, Belgium; Ghent University, Center for Nano- and Biophotonics, 9000 Ghent, Belgium.
authors.forename: K
authors.initials: K
authors.lastname: Braeckmans
authors.affiliation: Ghent University, Laboratory of General Biochemistry and Physical Pharmacy, Faculty of Pharmaceutical Sciences, 9000 Ghent, Belgium.
authors.forename: S C
authors.initials: SC
authors.lastname: De Smedt
authors.affiliation: University of Liège, Center for Education and Research on Macromolecules, 4000 Liège, Belgium.
authors.forename: C
authors.initials: C
authors.lastname: Jérôme
authors.affiliation: Université Catholique de Louvain, Pharmaceutics and Drug Delivery Group, Louvain Drug Research Institute, 1200 Brussels, Belgium. Electronic address: veronique.preat@uclouvain.be.
authors.forename: V
authors.initials: V
authors.lastname: Préat
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D002626:Chemistry, Pharmaceutical; D048271:Chitosan / Q000008:administration & dosage* / Q000737:chemistry; D020868:Gene Silencing; D049409:Luciferases, Firefly / Q000235:genetics; D051379:Mice; D053758:Nanoparticles / Q000008:administration & dosage* / Q000737:chemistry; D011092:Polyethylene Glycols / Q000737:chemistry; D011094:Polyethyleneimine / Q000737:chemistry; D034741:RNA, Small Interfering / Q000008:administration & dosage* / Q000737:chemistry
chemical_list: D034741:RNA, Small Interfering; D011092:Polyethylene Glycols; D011094:Polyethyleneimine; D048271:Chitosan; D049409:Luciferases, Firefly
keywords: Chitosan; Gene silencing; Nanoparticles; Stability in plasma; siRNA delivery