title: Low molecular weight chitosan nanoparticles as new carriers for nasal vaccine delivery in mice.
abstract: High molecular weight (Mw) chitosan (CS) solutions have already been proposed as vehicles for nasal immunization. The aim of the present work was to investigate the potential utility of low Mw CS in the form of nanoparticles as new long-term nasal vaccine delivery vehicles. For this purpose, CS of low Mws (23 and 38 kDa) was obtained previously by a depolymerization process of the commercially available CS (70 kDa). Tetanus toxoid (TT), used as a model antigen, was entrapped within CS nanoparticles by an ionic cross-linking technique. TT-loaded nanoparticles were first characterized for their size, electrical charge, loading efficiency and in vitro release of antigenically active toxoid. The nanoparticles were then administered intranasally to conscious mice in order to study their feasibility as vaccine carriers. CS nanoparticles were also labeled with FITC-BSA and their interaction with the rat nasal mucosa examined by confocal laser scanning microcopy (CLSM). Irrespective of the CS Mw, the nanoparticles were in the 350 nm size range, and exhibited a positive electrical charge (+40 mV) and associated TT quite efficiently (loading efficiency: 50-60%). In vitro release studies showed an initial burst followed by an extended release of antigenically active toxoid. Following intranasal administration, TT-loaded nanoparticles elicited an increasing and long-lasting humoral immune response (IgG concentrations) as compared to the fluid vaccine. Similarly, the mucosal response (IgA levels) at 6 months post-administration of TT-loaded CS nanoparticles was significantly higher than that obtained for the fluid vaccine. The CLSM images indicated that CS nanoparticles can cross the nasal epithelia and, hence, transport the associated antigen. Interestingly, the ability of these nanoparticles to provide improved access to the associated antigen to the immune system was not significantly affected by the CS Mw. Indeed, high and long-lasting responses could be obtained using low Mw CS molecules. Furthermore, the response was not influenced by the CS dose (70-200 microg), achieving a significant response for a very low CS dose. In conclusion, nanoparticles made of low Mw CS are promising carriers for nasal vaccine delivery.
authors.affiliation: Department of Pharmacy and Pharmaceutical Technology, School of Pharmacy, Santiago de Compostela, Spain.
authors.forename: Ana
authors.initials: A
authors.lastname: Vila
authors.forename: Alejandro
authors.initials: A
authors.lastname: Sánchez
authors.forename: Kevin
authors.initials: K
authors.lastname: Janes
authors.forename: Isabel
authors.initials: I
authors.lastname: Behrens
authors.forename: Thomas
authors.initials: T
authors.lastname: Kissel
authors.forename: José Luis
authors.initials: JL
authors.lastname: Vila Jato
authors.forename: María José
authors.initials: MJ
authors.lastname: Alonso
mesh_terms: D000281:Administration, Intranasal; D000818:Animals; D048271:Chitosan / Q000008:administration & dosage* / Q000138:chemical synthesis* / Q000737:chemistry; D004337:Drug Carriers / Q000008:administration & dosage / Q000737:chemistry; D004353:Drug Evaluation, Preclinical / Q000379:methods; D016650:Fluorescein-5-isothiocyanate / Q000031:analogs & derivatives*; D005544:Forecasting; D007114:Immunization / Q000379:methods; D007074:Immunoglobulin G / Q000737:chemistry / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D018613:Microscopy, Confocal / Q000379:methods; D008970:Molecular Weight; D045333:Nanotubes, Peptide / Q000737:chemistry*; D009297:Nasal Mucosa / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D010316:Particle Size; D014677:Pharmaceutical Vehicles / Q000008:administration & dosage / Q000737:chemistry; D012710:Serum Albumin, Bovine / Q000276:immunology; D013745:Tetanus Toxoid / Q000276:immunology / Q000378:metabolism; D014612:Vaccines / Q000008:administration & dosage* / Q000276:immunology
chemical_list: D004337:Drug Carriers; D007074:Immunoglobulin G; D045333:Nanotubes, Peptide; D014677:Pharmaceutical Vehicles; D013745:Tetanus Toxoid; D014612:Vaccines; C039217:fluorescein isothiocyanate bovine serum albumin; D012710:Serum Albumin, Bovine; D048271:Chitosan; D016650:Fluorescein-5-isothiocyanate