Dermatan Sulfate/Chitosan nanomaterials loaded with IRW modulate human endothelial sterile inflammatory response.

BLACHMAN A.; SOLOTOV N. ; EIGUREN AC.; CAMPERI S.; GLISONI RJ.; CALABRESE GC.

Mar del Plata

Congreso; S A I C . S A F E . S A B . SAP 2 0 1 9 Participan AACYTAL . NANOMED-AR . HCS; 2019

SAIC-NANOMED-ar

The present work describes a novel delivery system for the selective targeting of egg-derived anti-inflammatory tripeptide Ile-Arg-Trp (IRW) to modulate human endothelial cells inflammation, in the context of high levels of oxidized triglyceride-rich lipoproteins (VLDLox). IRW is produced by solid phase peptide synthesis. Dermatan sulfate/Chitosan nanoparticles loaded with IRW (8.00% w/w) (DS/CS-IRW) are prepared by ionotropic gelification method and characterized by dynamic light scattering (DLS) and transmission electron microscopy (TEM). VLDLs were isolated from healthy human volunteers by density-gradient ultracentrifugation and oxidized by 5µM CuSO4 for 2 at 37°C. To analyze the selective binding and uptake, human endothelial cells (EA.hy926) and human macrophages were incubated in the presence of FITC-nanoparticles at the biologically active concentration of DS, 10 µg/mL. Endothelial sterile inflammatory response was evaluated by NFκB subcellular distribution (immunofluorescence) and zymography studies. The incorporation of IRW results in a stable nanoparticle dispersion with a single size population of 539 ± 75 nm (n=6). TEM shows that IRW inclusion resulted in compact spherical-like particles. Co-cultures between endothelial cells and macrophages confirm the selectively interaction of fluorescent DS/CS-IRW with EA.hy926. After incubating endothelial cells with 100 µg protein/mL of VLDLox for 24h, NFκB is localized both at the cytoplasmic and nuclear compartment. Nevertheless, the transcriptional factor is restricted to the cytoplasm in the presence of IRW-loaded nanoparticles. NFκB subcellular distribution was correlated with endothelial inflammatory response through the evaluation of the effect of IRW-loaded nanoparticles on matrix metalloproteinases activity-9. Zymographic analysis reveal a significant decrease in MMP-9 activity after DS/CS-IRW treatment. We report here on the capability of these IRW-loaded complexes to modulate endothelial inflammatory response by as simple and potentially scalable nanotechnology platform.