Development, biophysical and toxicical characterization of polymeric nanoparticles

PATRICIO BF; PRIETO MJ; FEAS DA; MARTINEZ CS; ALONSO SDV; BISCH PM; WEISSMÜLLER G

Congreso; FESBE 2015 - XXX Reunião Anual, Federação de Sociedades de Biologia Experimental.; 2015

Introduction: The use of nanoparticles as drug carriers promises properties such as controlled, prolonged and sustained release of the active ingredient, reduction of the required dose for therapeutic effect and the toxic effects. In addition, it is well known that in inflamed tissue, the endotelium possess around 250 nm gaps between cells, so particles with size between 4-250 nm will deliver the therapy through the unhealthy endothelium only. However this kind of therapy requires a toxicological study.The use of zebrafish (Danio rerio) to study acute and chronic toxicity of drug and nanoparticles are growing. They are relatively easy to use and give a first idea of the biodistribution of the nanoparticles. Objectives: The aim of this study is to produce polymeric nanoparticles, evaluating the parameters in relation to the size of the nanoparticles.Methods: Nanoparticles were produced by the method of double emulsion using poly-lactic acid (PLA) and poly-vinyl alcohol (PVA) as polymers. During this process, we test changes in the output power (55, 60 and 90W) to obtain smaller nanoparticles. The nanoparticle morphology and dimension of the nanoparticles were obtained by atomic force microscopy (AFM) using PeakForce Tapping Mode (PFQNM)® and light scattering and were analysed by ANOVA nonparametric test using Graphpad Prism®. The toxicity of the nanoparticles were tested on zebrafish WT. It was analyzed neurotoxocity (N=8/group), cardiotoxicity, hepatotoxicity and histological changes (N=5/group) in three concentrantion of each nanoparticles produced. The results were analyzed with ANOVA test using Graphpad Prism®. All experiments were approved by the ethical committee of Universidad Nacional de Quilmes (CE-UNQ 2/2014).Results: The nanoparticles exhibited spherical shapes, when analyzed through AFM topographic images. In the adhesion and elasticity images it was possible to determine a heterogeneous surface of the nanoparticles, probably because of the use of different polymers in the preparation. The average and size range of the nanoparticles were different depending on the output power of sonication. The best output power was 60W with a mean size of 150±57nm with the lower size of 23nm and the higher size of 470nm. Nanoparticles did not present neurotoxicity. Nanoparticles produced with 55W shown a higher hepatotoxicity. Higher concentrations of nanoparticles produced with 60 and 90W induces tachycardia and low concentration of these same nanoparticles induces bradycardia.Conclusion: Nanoparticles of PLA/PVA were successfully manufactured by the double emulsion methodology and this new mode of analysis by AFM allows the evaluation of the nanoparticles physical properties. Also, it was possible to determine optimal output power sonication to produce nanoparticles, which can be used in further studies. With the zebrafish model it was possible to have an idea of the biodistribution of the nanoparticles and the most affected organs. Apoio Financeiro: Capes, Faperj, CNPq