Fig. 3

Assessment of cytotoxic potential and biocompatibility of blank and 6BIGOE-loaded NPs. A Cell viability of human monocytes was evaluated by MTT assay. Cells were treated for 24 or 48 h with vehicle (DMSO 0.5% (v/v) for 6BIGOE or PBS 0.1% (v/v) for 6BIGOE-loaded NPs), 6BIGOE, 6BIGOE-loaded NPs prepared by emulsion-diffusion-evaporation (EDE), Cyrene, or PEG 400 method at the indicated 6BIGOE concentrations, or 1 µM staurosporine (STSP, positive control), at 37 °C, and MTT assay was performed. Values are means + SEM; expressed as percentage of control (vehicle = 100%); n = 4 separate donors. Statistical analysis was performed applying repeated-measurement one-way ANOVA with Geisser-Greenhouse correction and Dunnett’s multiple comparisons test, testing treatments against vehicle (0.1% DMSO for 6BIGOE and STSP or 0.1% PBS for NPs). Data was log-transformed prior to analysis for 6BIGOE-loaded NPs at 24 h. (B) Biocompatibility assessment of the blank and 6BIGOE-loaded NPs in an ex ovo shell-less hens egg test. The clustergram illustrates toxic effects of injected (2 µL) 6BIGOE-loaded NPs prepared by EDE, Cyrene, or PEG 400 method. A concentration of 3 µM 6BIGOE calculated on the 6BIGOE drug load was used. Deionized water was used as solvent control, 0.9% NaCl served as negative control and branched PEI was applied as positive control. The columns quantify the time-dependent effect, whereas the rows show the different test samples. The number of affected egg’s correlates to the intensity of the color. Data were collected in two independent experiments with a total number of 10 eggs per NP sample