Fig. 5
Analysis of the effect of nanoparticles on the viability of mouse primary macrophages and human PBMCs. Mouse bone marrow derived macrophages and human PBMCs were mock treated (ctrl) or incubated with four types of nanoparticles (NPR-P, NPR-PG, NPR-PT, NPR-PTG) at four concentrations (25, 50, 100 and 200 μg/mL) for 24 h as indicated in experimental section. (a) Analysis of nanoparticles entry in macrophages using confocal microscopy. A representative experiment 100 μg/mL of NPR-PTG and 200 μg/mL of NPR-PT is shown. (b) Detection of phosphatydylserine translocation (AnnexinV) and cell membrane permeabilisation (7AAD) in macrophages by flow citometry. (c). Analysis of nanoparticles entry in PBMCs using confocal microscopy. A representative experiment 100 μg/mL of NPR-PTG and 200 μg/mL of NPR-PT is shown. (d). Analysis of ΔΨm loss (DIOC6), (e) detection of superoxide anion generation and (f) detection of phosphatydylserine translocation (AnnexinV) and cell membrane permeabilisation (7AAD) in PBMCs by flow citometry. Data represent mean values ± SD from three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001. As positive control (C+) staurosporine 1 μM for macrophages and cladribine 5 μM for PBMCs was used. Scale bar: 30 μm