GLISONI Romina Julieta
congresos y reuniones científicas
Dermatan Sulfate/Chitosan Polyelectrolyte complex as a new strategy for diagnosis and treatment of the vessel wall
VASTA M.; BARIANDARÁN A.; FUNEZ F.; GLISONI RJ.; SOSNIK A.; CALABRESE G.C.
Caba, Buenos Aires Argentina
Congreso; III International Congress in Translational Medicine; 2016
IMBS Congress Team
Introduction. Cardiovascular disease is the largest single cause of morbid-mortality 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 have produced polyelectrolyte complexes (PECs) by a simple and reproducible polyelectrolyte complexation method between low molecular mass dermatan sulfate (LMMDS) (polyanionic polysaccharide) and chitosan (CS) (polycationic polysaccharide); with a hydrodynamic radius of around 600 nm. PECs were not cytotoxic for a murine heart endothelium-derived cell line (H5V) and fluorescent confocal microscopy showed the specific uptake of fluorescently-labeled PECs (FITC-PECs) by endothelial cells when they were cultured alone or in the presence of macrophages. The aim of the present work was to analyze the molecular characteristic of the binding of PECs to endothelial cells.Methods. FITC-PECs were prepared employing CS labeled with fluorescein isothiocyanate. The size (Dh), size distribution (PDI) and zeta potential (Z-potential) of PECs were determined by dynamic light scattering (DLS). LMMDS and hialuronic acid (HA) cytotoxicity was evaluated by MTT assays. H5V were incubated with FITC-PECs (10 µg/mL, according to their LMMDS concentration) for 30min in the absence or in the presence of 37.5 µg/mL LMMDS or 50 µg/mL of HA. Fluorescence images from a minimum of 10 fields were collected from each sample. Chemical hypoxia was induced by cobalt (250 mM) for 1 h. Hialuronic acid receptor CD44 was evaluated by Westernblot.Results. FITC-PECs, produced by the ionotropic gelification method, exhibited a main size distribution of 644 ± 81 nm (98%) and an average PDI value of 0.398 ± 0.043 (n= 3). H5V cells treated or not with cobalt presented a homogeneous green dotted signal after incubating with FITC-PECs for 30 min. The presence of a high but nontoxic dose of LMMDS, as well as HA blocked the uptake of FITC-PECs. The expression of CD44 was confirmed for both control and cobalt treated cells. Conclusions. Our in vitro results show that (1) CS-LMMDS PECs interacted with normal and injured endothelium; (2) LMMDS of the PECs was associated to the specific binding to endothelial cell and (3) CD44 receptor could be responsible for the specific interaction. CS-LMMS PECs could be a novel strategy for targeting agents to the vessel wall.