Figure 5

Size and shape dictates HA particle induced modulation of cytokine production following injection. Female C57BL/6 mice (n = 3) aged 6–8 weeks were injected intraperitoneally with PBS vehicle, S_0.1 (1 mg/mouse), N₅ (1 mg/mouse), R₂₀ (1 mg/mouse), S₂₀ (1 mg/mouse) or S₁₀₀ (1 mg/mouse) HA particles, and 24 h later, peritoneal lavage cells were collected. (A,B) PECs were stained to analyse cell recruitment by flow cytometry. Cells were characterized using a combination of markers for peritoneal resident macrophages (CD11b^hi F4/80^hi), mast cells (SiglecF⁻ cKit⁺) monocytes (CD11b⁺ SiglecF⁻ F4/80⁻ Gr1^int SSC^low), neutrophils (CD11b⁺ SiglecF⁻ F4/80⁻ Gr1^hi) and eosinophils (CD11b⁺ SiglecF⁺ Gr1⁻ SSC^hi). Results are representative of two independent experiments. (A) Representative plots (B) Data are expressed as number of single/live cells. (*p < 0.05, **p < 0.01, ***p < 0.001). (C) PECs were restimulated with complete RPMI 1640 medium or heat-killed Escherichia coli (10 bacteria/cell) for 24 h. Supernatants were assessed for IL-10 and TNF-α secretion by ELISA. Results are mean cytokine concentrations (±SEM) for triplicate samples (vs PBS ***p < 0.001).