Uptake studies in rat Peyer's patches, cytotoxicity and release studies of alginate coated chitosan nanoparticles for mucosal vaccination.

The design of particulate vaccine delivery systems, particularly for mucosal surfaces, has been a focus of interest in recent years. In this context, we have previously described the development and the characterization of a new nanosized delivery system, consisting of a model antigen adsorbed to chitosan particles and coated with sodium alginate. In the present work the ovalbumin release profiles from these coated nanoparticles in different pH buffers were investigated and compared to those of the uncoated particles. Cytotoxicity of the polymers and nanoparticles was assessed using the MTT assay. Finally, particle uptake studies in rat Peyer's patches were performed. It was demonstrated that the coating of the nanoparticles with sodium alginate not only avoided a burst release observed with uncoated particles but also increased the stability of the particles at pH 6.8 and 7.4 at 37 degrees C. At neutral pH, the release was lower than 5% after 3.5 h incubation in a low ionic strength buffer. For both, chitosan and alginate polymers, and for the nanoparticles, comparable cell viability data close to 100%, were obtained. Additionally, based on confocal laser scanning microscopy observations, it was shown that alginate coated nanoparticles were able to be taken up by rat Peyer's patches, rendering them suitable carriers for intestinal mucosal vaccination.

[1]  J. Sousa,et al.  Comparison of dissolution profiles of Ibuprofen pellets. , 2003, Journal of controlled release : official journal of the Controlled Release Society.

[2]  H. Junginger,et al.  Enhancement of bronchial octreotide absorption by chitosan and N-trimethyl chitosan shows linear in vitro/in vivo correlation. , 2006, Journal of controlled release : official journal of the Controlled Release Society.

[3]  D. O’hagan Microparticles as vaccine delivery systems , 2006 .

[4]  C. Washington,et al.  Drug release from microparticulate systems , 1996 .

[5]  K. Janes,et al.  Chitosan nanoparticles as delivery systems for doxorubicin. , 2001, Journal of controlled release : official journal of the Controlled Release Society.

[6]  M. Amiji,et al.  Stomach-specific anti-H. pylori therapy. I: Preparation and characterization of tetracyline-loaded chitosan microspheres. , 2002, International journal of pharmaceutics.

[7]  Ching-Yu Chang,et al.  Application of gold nanoparticles to microencapsulation of thioridazine. , 2006, Journal of controlled release : official journal of the Controlled Release Society.

[8]  V. Schijns Immunopotentiators in modern vaccines , 2003 .

[9]  Maria José Alonso,et al.  Comparative Uptake Studies of Bioadhesive and Non-Bioadhesive Nanoparticles in Human Intestinal Cell Lines and Rats: The Effect of Mucus on Particle Adsorption and Transport , 2002, Pharmaceutical Research.

[10]  Vincent W. Bramwell,et al.  Stimulation of spleen cells in vitro by nanospheric particles containing antigen. , 2003, Journal of controlled release : official journal of the Controlled Release Society.

[11]  Krishnendu Roy,et al.  Oral gene delivery with chitosan–DNA nanoparticles generates immunologic protection in a murine model of peanut allergy , 1999, Nature Medicine.

[12]  T. Kissel,et al.  Biodegradable nanoparticles for oral delivery of peptides: is there a role for polymers to affect mucosal uptake? , 2000, European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenstechnik e.V.

[13]  A. Vila,et al.  Low molecular weight chitosan nanoparticles as new carriers for nasal vaccine delivery in mice. , 2004, European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenstechnik e.V.

[14]  V. Dodane,et al.  Effect of chitosan on epithelial permeability and structure. , 1999, International journal of pharmaceutics.

[15]  H. Junginger,et al.  Preparation and characterization of protein-loaded N-trimethyl chitosan nanoparticles as nasal delivery system. , 2006, Journal of controlled release : official journal of the Controlled Release Society.

[16]  C. Czerkinsky,et al.  Mucosal immunisation and adjuvants: a brief overview of recent advances and challenges. , 2003, Vaccine.

[17]  S. Harding Mucoadhesive interactions. , 2003, Biochemical Society transactions.

[18]  I. Kwon,et al.  Protective Immunity of Microsphere-Based Mucosal Vaccines against Lethal Intranasal Challenge withStreptococcus pneumoniae , 1999, Infection and Immunity.

[19]  Min Huang,et al.  Uptake of FITC-Chitosan Nanoparticles by A549 Cells , 2002, Pharmaceutical Research.

[20]  John F. Carpenter,et al.  Physical Stability of Proteins in Aqueous Solution: Mechanism and Driving Forces in Nonnative Protein Aggregation , 2003, Pharmaceutical Research.

[21]  G. Jan,et al.  Comparison of different electrophoretic separations of hen egg white proteins. , 2001, Journal of agricultural and food chemistry.

[22]  W. Shalaby Development of oral vaccines to stimulate mucosal and systemic immunity: barriers and novel strategies. , 1995, Clinical immunology and immunopathology.

[23]  Yumin Du,et al.  Effect of molecular structure of chitosan on protein delivery properties of chitosan nanoparticles. , 2003, International journal of pharmaceutics.

[24]  J. Kreuter,et al.  Preparation and characterization of chitosan microspheres as drug carrier for prednisolone sodium phosphate as model for anti-inflammatory drugs , 1996 .

[25]  C. Remuñán-López,et al.  Enhancement of Nasal Absorption of Insulin Using Chitosan Nanoparticles , 1999, Pharmaceutical Research.

[26]  J. Pedraz,et al.  Biocompatibility evaluation of different alginates and alginate-based microcapsules. , 2005, Biomacromolecules.

[27]  C. T. Chen,et al.  Porous chitosan microsphere for controlling the antigen release of Newcastle disease vaccine: preparation of antigen-adsorbed microsphere and in vitro release. , 1999, Biomaterials.

[28]  Min Huang,et al.  Uptake and Cytotoxicity of Chitosan Molecules and Nanoparticles: Effects of Molecular Weight and Degree of Deacetylation , 2004, Pharmaceutical Research.

[29]  H. Tønnesen,et al.  Alginate in Drug Delivery Systems , 2002, Drug development and industrial pharmacy.

[30]  S. Turco,et al.  A simple model based on first order kinetics to explain release of highly water soluble drugs from porous dicalcium phosphate dihydrate matrices , 1995 .

[31]  K. Neoh,et al.  Antibacterial and mechanical properties of bone cement impregnated with chitosan nanoparticles. , 2006, Biomaterials.

[32]  H E Junginger,et al.  In Vivo Uptake of Chitosan Microparticles by Murine Peyer's Patches: Visualization Studies using Confocal Laser Scanning Microscopy and Immunohistochemistry , 2001, Journal of drug targeting.

[33]  S. Benita,et al.  Nanoparticle characterization : a comprehensive physicochemical approach , 1991 .

[34]  H E Junginger,et al.  Chitosan microparticles for oral vaccination: preparation, characterization and preliminary in vivo uptake studies in murine Peyer's patches. , 2001, Biomaterials.

[35]  W. Elmquist,et al.  Measurement of drug release from microcarriers by microdialysis. , 2005, Journal of pharmaceutical sciences.

[36]  H. Junginger,et al.  Chitosan microparticles for mucosal vaccination against diphtheria: oral and nasal efficacy studies in mice. , 2003, Vaccine.

[37]  N Hussain,et al.  Recent advances in the understanding of uptake of microparticulates across the gastrointestinal lymphatics. , 2001, Advanced drug delivery reviews.

[38]  Y. Okahata,et al.  Mechanism of cell transfection with plasmid/chitosan complexes. , 2001, Biochimica et biophysica acta.

[39]  L. Babiuk,et al.  Mucosal immune responses following oral immunization with rotavirus antigens encapsulated in alginate microspheres. , 2002, Journal of controlled release : official journal of the Controlled Release Society.

[40]  H. Junginger,et al.  Preparation of coated nanoparticles for a new mucosal vaccine delivery system. , 2005, International journal of pharmaceutics.

[41]  M. Neutra,et al.  Cholera Toxin Induces Migration of Dendritic Cells from the Subepithelial Dome Region to T- and B-Cell Areas of Peyer's Patches , 2003, Infection and Immunity.

[42]  L. Lim,et al.  Formulation pH modulates the interaction of insulin with chitosan nanoparticles. , 2002, Journal of pharmaceutical sciences.

[43]  A. Bernkop‐Schnürch,et al.  Improvement in the mucoadhesive properties of alginate by the covalent attachment of cysteine. , 2001, Journal of controlled release : official journal of the Controlled Release Society.

[44]  P. de Vos,et al.  Chemistry and biocompatibility of alginate-PLL capsules for immunoprotection of mammalian cells. , 2002, Journal of biomedical materials research.

[45]  C. Czerkinsky,et al.  Mucosal adjuvants based on cholera toxin and E . coli heat-labile enterotoxin , 2006 .

[46]  P. Tengvall,et al.  Blood protein adsorption onto chitosan. , 2002, Biomaterials.

[47]  L. Babiuk,et al.  Multiple intestinal 'loops' provide an in vivo model to analyse multiple mucosal immune responses. , 2001, Journal of immunological methods.

[48]  R. Mebius,et al.  Structure and function of the spleen , 2005, Nature Reviews Immunology.

[49]  I. Kwon,et al.  Novel mucosal immunization with polysaccharide-protein conjugates entrapped in alginate microspheres. , 1998, Journal of controlled release : official journal of the Controlled Release Society.

[50]  W. Domschke,et al.  Influence of Chitosan Microspheres on the Transport of Prednisolone Sodium Phosphate Across HT-29 Cell Monolayers , 2004, Pharmaceutical Research.

[51]  T. Bowersock,et al.  Effects of cellulose derivatives and poly(ethylene oxide)-poly(propylene oxide) tri-block copolymers (Pluronic)surfactants) on the properties of alginate based microspheres and their interactions with phagocytic cells. , 2002, Journal of Controlled Release.