Morais, Ana Heloneida de AraujoCosta, Rafael Oliveira de Araújo2019-06-062019-06-062019-03-29COSTA, Rafael Oliveira de Araújo. Identificação da segurança e potencial aplicação clínica de nanopartículas contendo inibidor de tripsina isolado de sementes de tamarindo (Tamarindus indica L.). 2019. 74f. Dissertação (Mestrado em Bioquímica) - Centro de Biociências, Universidade Federal do Rio Grande do Norte, Natal, 2019.https://repositorio.ufrn.br/jspui/handle/123456789/27159Natural molecules with bioactive action, such as the trypsin inhibitor isolated from tamarind (Tamarindus indica L.) (TTI) stands out among the satietogenic and anti-inflammatory agents currently studied. In this perspective, aiming to maintain TTI intact and protected throughout the gastrointestinal tract, encapsulation was performed by nanoprecipitation in purified chitosan and isolated whey protein, and the particles (ECW) were characterized by different techniques. However, for this, the safety needs to be evaluated, aiming at possible clinical applications. Thus, the objective of this study was to evaluate the cytotoxicity, and subacute blood toxicity of the bioactive dose of ECW in Wistar rats fed a high glycemic index diet. TTI was obtained from protein fractionation using ammonium sulfate. The fraction with the highest inhibitory activity against trypsin, fractionation of 30-60%, was isolated by Chromatography of Affinity in Trypsin-Sepharose and used for the synthesis of nanoparticles. ECW was synthesized respecting the ratio of TTI, purified chitosan and whey protein isolated of 1: 2: 2 w/w/w, respectively, being posteriorly characterized by different physical and chemical analyzes. And the interaction between TTI and the encapsulating agents in neutral pH (water) and acidic pH, through several filtration steps in Amicon® 100 K, being monitored antitrypsin activity. In addition, the cytotoxicity of the ECW at different concentrations (0.5, 2.5 and 5 mg / mL) was evaluated by resazurin assay (Caco-2 and CCD-18Co) and blood toxicity (blood count, hepatic and renal function) in male Wistar rats (n = 10) fed a high glycemic index diet and , administered with ECW (12.5 mg / kg) for days 10 days. The ECW presented spherical particles with a smooth surface according to Scanning Electron Microscopy, mean diameter of 118 nm (17.27), a polydispersity index of 0.373 (0.02), surface charge of -38.26 mV (0.15) and efficiency of encapsulation of 95.3% (0.31). Concerning the interaction in water of TTI and the encapsulating agents through the evaluation of the antitrypsin activity, using Amicon® 100 K, it was observed that in water the ECW protected the TTI and kept it entrapped. However, in acidic pH, there was a gradual release of TTI, evidenced by the strong interaction between TTI and the encapsulating agents used in this study. The cytotoxicity assay indicated high cell viability (> 70%) for Caco-2 and CCD18Co when exposed to the ECW evaluated at different concentrations. As for the subacute blood toxicity of the ECW administered in Wistar rats, no toxic effects were also demonstrated according to the results obtained for the biochemical parameters evaluated. In this study, ECW proved to be effective in protecting TTI when dispersed in water and exposed to acid pH. The concentrations of ECW evaluated in cells, and the experimental model did not show signs of toxicity, suggesting the safe application of these particles and, probably, hepatoprotective effect. Finally, ensuring its application in complementary studies to investigate the efficacy in the treatment of obesity and its complications.Acesso AbertoEncapsulaçãoNanopartículaAtividade antitrípticaCCD18- CoCaco-2Ratos WistarmasterThesisCNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICA