Fig. 4.

6BIGOE and 6BIGOE-loaded NPs modulate LPS-induced cytokine release from monocytes. A LPS-induced TLR4 stimulation results in modulation of pro- and anti-inflammatory cytokine formation via GSK-3 activity and associated downstream signaling pathways in human monocytes. B, C Human monocytes (10⁶/mL) were pretreated for 3 h with vehicle (0.1% DMSO for 6BIGOE or equivalent amount of blank NP dispersions), 6BIGOE or 6BIGOE-loaded NPs prepared by EDE, Cyrene, or PEG 400 method in a concentration-dependent manner (0.003 to 1 µM) as indicated, and subsequently stimulated with 100 ng/ml LPS for 18 h. (B) Release of TNF-α, IL-β, and IL-6 was determined by ELISA. Values are means + SEM; expressed as percentage of control (vehicle = 100%), n = 4 separate experiments. Statistical analysis was conducted on log-transformed data using one-way ANOVA with Holm-Šídák correction for multiple comparisons of control group (DMSO 0.1%) with 6BIGOE treatments. Unpaired, two-sided Student’s t-test was applied for 6BIGOE-loaded NP treatments and their respective NP blank controls. C Release of IL-10 was determined by ELISA. Values are means + SEM; expressed as percentage of control (vehicle = 100%). n = 4 separate donors. Statistical analysis was conducted applying one-way ANOVA with Dunnett’s correction for multiple comparisons of 6BIGOE at indicated concentrations with control (DMSO 0.1%), and unpaired, two-sided Student’s t-test for comparison of different 6BIGOE-NP treatments with their respective controls (NP blanks)