CAMPERI Silvia Andrea
congresos y reuniones científicas
Dermatan Sulfate/Chitosan Polyelectrolyte Complexes loaded with IRW tripeptide inhibit NFkB nuclear translocation through CD44 receptor activation
A. BLACHMAN; F. FUNEZ; A.C. EIGUREN; S. L. SAAVEDRA; S. A. CAMPERI; R. GLISONI; A. SOSNIK; G. C. CALABRESE
Simposio; XVI Simposio Latinoamericano de Polímeros; 2018
INTRODUCTION Cardiovascular disease is the largest single cause of morbidmortality in the world. However, there is still no pharmaceutical treatment that directly targets the blood vessel wall instead of just controlling the risk factors. We reported an easy and reproducible method to produce a novel nanotechnological polyelectrolyte complex of low molecular weight dermatan sulfate (DS) and chitosan (CS) (DS/CS-PEC) that provides a unique combination of specific binding and control of biological features on endothelial cells1 . The aim of the present work was to study this complex as a drug delivery platform of an anti-inflammatory egg white ovotransferrin-derived tripeptide Ile-Arg-Trp (IRW)2 . EXPERIMENTAL METHODS IRW was obtained on solid phase employing Fmoc chemistry in Rink-Amide-MBHA resin. DS/CS-PEC was produced by ionotropic gelification method in the presence of IRW 50 µM, (n= 10). Besides, CS labeled with fluorescein isothiocyanate was employed to obtain FITC-DS/CS-PEC for confocal studies (n =5). Dh, PDI and Z-potential of complexes were determined by DLS. Murine endothelial cell line (H5V) was incubated with DS/CS-PEC or FITC-DS/CS-PEC (10 µg/ml, according to their DS concentration) for 2 hs after bacterial lipopolysaccharide treatment (LPS, 1.5 µg/ml). Hyaluronic acid receptor CD44 was evaluated by semiquantitative reverse transcriptasepolymerase chain reaction analysis, (n=3). Zimography and NFkB cellular distribution studies (Western blot and immunofluorescences), (n=2) were carried out to analyze the modulator effect on the inflammatory response. Statistical analysis was performed with ANOVA and Bonferroni´s test.RESULTS AND DISCUSSION Table 1. Physicochemical propertiesHigher uptake of PECs was observed in activated endothelial cells, accompanied with a significant increse in CD44 expression.After LPS treatment, the activity of MMP-9 was decreased in the presence of PECs/IRW. Moreover, PECs/IRW diminished NFκB nuclear translocation. CONCLUSION A new nanocomposite structure PEC/IRW was obtained, in which DS guarantee specific uptake by endothelial cells; while CS limits lysosomal degradation and support the gradual release of IRW inside the cell. Moreover, PECs/IRW could modulate the endothelial inflammatory response associated with vascular diseases. REFERENCES 1. Rasente RY et al. Carbohydr. Polym. 144, 370, 2016. 2. Majumder K et al. J. Sci. Food Agric.96, 2303-2011. 2015.