title: Spray dried microspheres based on chitosan: A promising new carrier for intranasal administration of polymeric antigen BLSOmp31 for prevention of ovine brucellosis.
abstract: Previous studies have demonstrated that parenteral immunization with polymeric antigen BLSOmp31 induced a strong immune response and conferred protection against Brucella ovis in rams. This work describes the development of a novel formulation strategy for the delivery of BLSOmp31 in the nasal mucosa. Chitosan microparticles were prepared by spray-drying technology processes and recombinant chimera BLSOmp31 was loaded by passive adsorption onto chitosan microspheres, which were characterized by means of the evaluation of size, zeta potential, morphology, and loading and release rate of BLSOmp31. The mucoadhesive properties of microspheres were evaluated by studying the interaction between microparticles and mucin. The antigen BLSOmp31 integrity was investigated by SDS-PAGE. The yield of production of spray-drying process was 68.95%. Microspheres had a good sphericity, 1-10 μm of particle size and had a positive charge. The loading capacity was found to be 45.19%. The initial fast release of BLSOmp31 from chitosan microparticles was 60%. The BLSOmp31 integrity was not affected by passive adsorption (ionic interaction). The amount of mucin adsorbed on the surface of CMs-BLSOmp31 was lower than on the surface of blank CMs at neutral pH. In vivo studies were carried out in rams. Intranasal immunization induced systemic and local antibodies. In conclusion, the use of BLSOmp31-loaded chitosan spray-drying microspheres offers a promising way for nasal mucosal vaccination in sheep against brucellosis.
authors.affiliation: Laboratorio de Inmunología, Departamento de Sanidad Animal y Medicina Preventiva (SAMP), Centro de Investigación Veterinaria Tandil (CIVETAN-CONICET-CIC), Facultad de Ciencias Veterinarias, Universidad Nacional del Centro de la Provincia de Buenos Aires, Argentina; Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Buenos Aires, Argentina.
authors.forename: Alejandra Graciela
authors.initials: AG
authors.lastname: Díaz
authors.affiliation: Departamento de Farmacia, Facultad Ciencias Químicas, Unidad de Investigación y Desarrollo en Tecnología Farmacéutica (UNITEFA-CONICET), Universidad Nacional de Córdoba, Argentina; Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Buenos Aires, Argentina.
authors.forename: Daniela Alejandra
authors.initials: DA
authors.lastname: Quinteros
authors.affiliation: Departamento de Farmacia, Facultad Ciencias Químicas, Unidad de Investigación y Desarrollo en Tecnología Farmacéutica (UNITEFA-CONICET), Universidad Nacional de Córdoba, Argentina; Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Buenos Aires, Argentina.
authors.forename: Juan Manuel
authors.initials: JM
authors.lastname: Llabot
authors.affiliation: Departamento de Farmacia, Facultad Ciencias Químicas, Unidad de Investigación y Desarrollo en Tecnología Farmacéutica (UNITEFA-CONICET), Universidad Nacional de Córdoba, Argentina; Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Buenos Aires, Argentina.
authors.forename: Santiago Daniel
authors.initials: SD
authors.lastname: Palma
authors.affiliation: Departamento de Farmacia, Facultad Ciencias Químicas, Unidad de Investigación y Desarrollo en Tecnología Farmacéutica (UNITEFA-CONICET), Universidad Nacional de Córdoba, Argentina; Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Buenos Aires, Argentina.
authors.forename: Daniel Alberto
authors.initials: DA
authors.lastname: Allemandi
authors.affiliation: Inmunova S.A., Buenos Aires, Argentina.
authors.forename: Giselle
authors.initials: G
authors.lastname: Ghersi
authors.affiliation: Inmunova S.A., Buenos Aires, Argentina; Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Buenos Aires, Argentina.
authors.forename: Vanesa
authors.initials: V
authors.lastname: Zylberman
authors.affiliation: Inmunova S.A., Buenos Aires, Argentina; Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Buenos Aires, Argentina.
authors.forename: Fernando Alberto
authors.initials: FA
authors.lastname: Goldbaum
authors.affiliation: Laboratorio de Inmunología, Departamento de Sanidad Animal y Medicina Preventiva (SAMP), Centro de Investigación Veterinaria Tandil (CIVETAN-CONICET-CIC), Facultad de Ciencias Veterinarias, Universidad Nacional del Centro de la Provincia de Buenos Aires, Argentina; Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Buenos Aires, Argentina. Electronic address: silmares@vet.unicen.edu.ar.
authors.forename: Silvia Marcela
authors.initials: SM
authors.lastname: Estein
mesh_terms: D000281:Administration, Intranasal; D000327:Adsorption; D000818:Animals; D000906:Antibodies / Q000097:blood; D000941:Antigens / Q000276:immunology*; D001426:Bacterial Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D042061:Brucella ovis / Q000378:metabolism; D002006:Brucellosis / Q000382:microbiology / Q000517:prevention & control* / Q000662:veterinary; D002626:Chemistry, Pharmaceutical; D048271:Chitosan / Q000737:chemistry*; D004337:Drug Carriers / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D018625:Microscopy, Atomic Force; D008863:Microspheres; D009077:Mucins / Q000737:chemistry / Q000378:metabolism; D010316:Particle Size; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000276:immunology; D012756:Sheep; D017550:Spectroscopy, Fourier Transform Infrared
chemical_list: D000906:Antibodies; D000941:Antigens; D001426:Bacterial Proteins; D004337:Drug Carriers; D009077:Mucins; D011993:Recombinant Fusion Proteins; D048271:Chitosan
keywords: BLSOmp31; Chitosan microspheres; Intranasal immunization; Passive adsorption; Spray-drying; Systemic and local antibodies