Fig. 2

NP behavior: drug release and NP uptake. A Thermal behavior of blank NPs and 6BIGOE-loaded NPs prepared by emulsion-diffusion-evaporation (EDE), Cyrene, or PEG 400 method analyzed by DSC. Two heating–cooling cycles at 10 °C/min were applied for run 1 (left) and run 2 (right), shown for the range between 0 to 190 °C. B Cumulative in vitro drug release from 6BIGOE-loaded NPs prepared by EDE, Cyrene, or PEG 400 method in PBS / Tween 80 (1% (m/m)) pH 7.4, up to 168 h. 6BIGOE was quantified by UV/Vis spectroscopy at 520 nm, data are means ± SD of three independent NP batches. C Simplified workflow depicting analysis of fluorescence-labeled NP uptake by human monocytes. D, E Uptake of fluorescein-labeled 6BIGOE-loaded NPs in human monocytes was analyzed using microscopy and flow cytometry. Monocytes (0.8 × 10⁶/mL (D), 1 × 10⁶/mL (E)) were incubated for the indicated times with 6BIGOE-loaded NPs prepared by EDE, Cyrene, or PEG 400 method. D Fluorescence microscopy of monocytes visualizing uptake of fluorescein-labeled 6BIGOE-loaded NPs by monocytes after 3 h. Individual fluorescence channel for FITC (upper panel) and overlay with DAPI nuclear stain (lower panel) are shown. Results presented for one single cell are representative for approx. 100 individual cells analyzed in three independent experiments. E Representative histograms (upper panel) display uptake of fluorescein-labeled 6BIGOE-loaded NPs by CD14⁺ monocytes (PBS 0.1%, negative control). Data are presented as mean fluorescent intensity for 30 min, 3 h or 6 h (lower panel); n = 3 separate donors, one-way ANOVA for multiple comparisons with Dunnett’s correction