CONICET | Buscador de Institutos y Recursos Humanos

INTRODUCTIONInsulin (I) is one of the most widely used peptides in the treatment of insulin-dependent patients. In this context alternativeadministration routes (other than parenteral) are being explored, especially those for pediatric uses. Chitosan (CS) is a biodegradable, biocompatible and non-toxic biopolymer obtained from chitin after a chemical deacetylation process. The formation of I-CS nano-complexes could be an alternative in the development of a new generation of therapeutic peptides, specially designed for pulmonary drug delivery.The objective of this work was to generate nanovehicles (NVs) for the bioactive I based on CS as the encapsulant material by applying nanospray technology. Nanoparticles were characterized and their biological activity assayed in culture cells.EXPERIMENTAL METHODSThe nanodehydration equipment used was a Buchi B-90 model (BÜCHI Labortechnik AG).HPLCchromatography was used to evaluate the content of the trapped protein in the NV. In addition, the particle size distribution and the potential ζ were obtained by dynamic light scattering (DLS).The identification of functional groups and interactions in NVs was achieved by FTIR spectroscopy. The biological activity of I-CS was evaluated as the phosphorylation of AKT by western blot in 3T3 fibroblasts.RESULTS AND DISCUSSIONThe method had a yield of 94% for a ratio of 1: 1 between I and CS 0.1% w/ w solutions. The entrapment efficiency resulted equal to 62.27%. The peaks obtained by DLS corresponded to the majority populations were located at 32 and 413 nm. The potential ζ was (62.8 ± 0.7) mV.I-CS NVs collected from the receiver chamber from Calu-3 culture, increased the phosphorylation of AKT at larger times in 3T3 fibroblasts in comparison to non- encapsulated I.CONCLUSIONNVs exhibited particle size values smaller than 100 nm.AKT activation confirms that the structural and functional stability of encapsulated Insulin are mantained after nanodehydration.