title: Evaluation of antisense oligonucleotide loaded chitosan nanoparticles; characterization and antisense effect.
abstract: The objective of this study was to investigate the effect of different formulation parameters [i.e. molecular weight and concentration of chitosan, concentration of tripolyphosphate (TPP) and use of alginate] on physico chemical and antisense properties of antisense oligonucleotide (AsODN) loaded chitosan nanoparticles (NPs). Preparation methods of phosphodiester (PO) and phosphorothioate (PS) AsODNs-NPs were also compared. AsODN was designed to target the beta-galactosidase (beta-gal) gene. HeLa cells were used for in vitro transfection studies and beta-gal was assayed spectrophotometrically. AsODN-NPs obtained were in general positively charged with size between 221.4-525.7 nm depending on formulation. Encapsulation efficiency of NPs depended on the type of backbone of the AsODN. PO-AsODN encapsulation into NPs (78-94%) was less efficient than PS encapsulation (91-98%). The pH of the chitosan solution affected AsODN entrapment. PO-NPs exhibited faster AsODN release than NPs containing PS. In general higher beta-gal inhibition was obtained after transfection of AsODN-NPs in cell culture studies. PS-NPs exhibited a higher inhibition effect and the highest (90.71%) inhibition was obtained with formulation PT-2. PS-adsorbed NPs showed an 88% reduction in beta-gal. This study can form the basis for forthcoming in vivo studies related to AsODN carrier systems that will use chitosan.
authors.affiliation: Department of Pharmaceutical Biotechnology, Faculty of Pharmacy, Marmara University, Istanbul, Turkey.
authors.forename: S
authors.initials: S
authors.lastname: Ozbaş-Turan
authors.forename: J
authors.initials: J
authors.lastname: Akbuğa
authors.forename: B
authors.initials: B
authors.lastname: Enneli
mesh_terms: D000464:Alginates / Q000737:chemistry; D002626:Chemistry, Pharmaceutical; D048271:Chitosan; D003692:Delayed-Action Preparations; D004339:Drug Compounding; D005079:Excipients; D006367:HeLa Cells; D006801:Humans; D053758:Nanoparticles; D016376:Oligonucleotides, Antisense / Q000008:administration & dosage / Q000494:pharmacology*; D010316:Particle Size; D011122:Polyphosphates / Q000737:chemistry; D013056:Spectrophotometry, Ultraviolet; D001616:beta-Galactosidase / Q000235:genetics
chemical_list: D000464:Alginates; D003692:Delayed-Action Preparations; D005079:Excipients; D016376:Oligonucleotides, Antisense; D011122:Polyphosphates; D048271:Chitosan; D001616:beta-Galactosidase; C005692:triphosphoric acid