title: Mono-N-carboxymethyl chitosan (MCC) and N-trimethyl chitosan (TMC) nanoparticles for non-invasive vaccine delivery.
abstract: Mucosal application of a vaccine can effectively induce both systemic and mucosal immune responses. In general, mucosal applications of antigens result in poor immune responses. Therefore, adjuvant/delivery systems are required to enhance the immune response. Chitosan is a cationic biopolymer which exerts advantages as a vaccine carrier due to its immune stimulating activity and bioadhesive properties that enhance cellular uptake and permeation as well as antigen protection. Similar effects are also shown by chitosan derivatives. In this study, the nanoparticulate systems were prepared by using differently charged chitosan derivatives, N-trimethyl chitosan (TMC, polycationic), and mono-N-carboxymethyl chitosan (MCC, polyampholytic) for mucosal immunisation. The derivatives were synthesised and characterised in-house. The aqueous dispersions of the derivatives were also prepared for comparison. The cytotoxicity studies (MTT assay) on Chinese hamster ovary (CHO-K1) cell lines showed that cell viability was in the order of MCC, chitosan and TMC. Nanoparticles were prepared using ionic gelation method and loaded with tetanus toxoid (TT). Nanoparticles with high loading efficacy (>90% m/m), particle size within the range of 40-400nm, with a negative surface charge for MCC and positive surface charge for TMC and chitosan were obtained. The structural integrity of the TT in the formulations was confirmed by SDS-PAGE electrophoresis analysis. The effective uptake of the FITC-BSA loaded nanoparticles into the cells was demonstrated by cellular uptake studies using J774A.1 cells. Immune responses induced by the formulations loaded with tetanus toxoid were studied in vivo in Balb/c mice. Enhanced immune responses were obtained with intranasal (i.n.) application of nanoparticle formulations. Chitosan and TMC nanoparticles which have positively charged surfaces induced higher serum IgG titres when compared to those prepared with MCC which are negatively charged and smaller in size. Nanoparticle formulations developed in this study can be used as promising adjuvant/delivery systems for mucosal immunisation.
authors.affiliation: Hacettepe University, Faculty of Pharmacy, Department of Pharmaceutical Technology, Ankara, Turkey.
authors.forename: B
authors.initials: B
authors.lastname: Sayin
authors.forename: S
authors.initials: S
authors.lastname: Somavarapu
authors.forename: X W
authors.initials: XW
authors.lastname: Li
authors.forename: M
authors.initials: M
authors.lastname: Thanou
authors.forename: D
authors.initials: D
authors.lastname: Sesardic
authors.forename: H O
authors.initials: HO
authors.lastname: Alpar
authors.forename: S
authors.initials: S
authors.lastname: Senel
mesh_terms: D000276:Adjuvants, Immunologic / Q000008:administration & dosage* / Q000737:chemistry / Q000633:toxicity; D000281:Administration, Intranasal; D000818:Animals; D016466:CHO Cells; D002470:Cell Survival / Q000187:drug effects; D002626:Chemistry, Pharmaceutical; D048271:Chitosan / Q000008:administration & dosage* / Q000737:chemistry / Q000276:immunology / Q000633:toxicity; D006224:Cricetinae; D003412:Cricetulus; D004337:Drug Carriers; D004339:Drug Compounding; D016650:Fluorescein-5-isothiocyanate / Q000031:analogs & derivatives / Q000378:metabolism; D018928:Immunity, Mucosal / Q000187:drug effects; D007074:Immunoglobulin G / Q000097:blood; D007279:Injections, Subcutaneous; D008264:Macrophages / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D053758:Nanoparticles; D012710:Serum Albumin, Bovine / Q000378:metabolism; D013745:Tetanus Toxoid / Q000008:administration & dosage* / Q000737:chemistry / Q000276:immunology; D014612:Vaccines / Q000008:administration & dosage* / Q000737:chemistry / Q000276:immunology
chemical_list: D000276:Adjuvants, Immunologic; D004337:Drug Carriers; D007074:Immunoglobulin G; C118071:N-trimethyl chitosan chloride; D013745:Tetanus Toxoid; D014612:Vaccines; C039217:fluorescein isothiocyanate bovine serum albumin; C426873:mono-N-carboxymethylchitosan; D012710:Serum Albumin, Bovine; D048271:Chitosan; D016650:Fluorescein-5-isothiocyanate