Figure 1: Intraperitoneal inoculation of ITO-NPs in mice induces peritonitis through NLRP3 inflammasome. | Scientific Reports

Figure 1: Intraperitoneal inoculation of ITO-NPs in mice induces peritonitis through NLRP3 inflammasome.

From: Endocytosis of indium-tin-oxide nanoparticles by macrophages provokes pyroptosis requiring NLRP3-ASC-Caspase1 axis that can be prevented by mesenchymal stem cells

Figure 1

(a) Schematic diagram of the experimental procedures used in vivo and ex vivo in Balb/c mice to examine the role of ITO-NPs in inducing NLRP3 inflammasome-dependent peritonitis. (b) Flow cytometer analysis of peritoneal cells of control Balb/c mice (black dots), (c) mice receiving ITO-NPs (red dots) and (d) both NLRP3 inflammasome inhibitor glybenclamide (Gly) and ITO-NPs (green dots). (b) Dot-plots show peritoneal cells of control mice stained for CD11b, Gr-1 and Ly-6G; about 20% of the CD45+ cells were peritoneal macrophages phenotypically characterized as positive for CD11b and negative for Gr-1 and Ly-6G. (c) 6-h post-inoculation i.p. of ITO-NPs in Balb/c mice, CD11b+Gr-1+ increases, to reach about 60% Ly-6G positive (neutrophils) and 13% Ly-6G negative (inflamed-monocytes). (d) Balb/c mice receiving i.p. Gly and ITO-NPs show reduced number of CD11b+Ly-6G+ neutrophils. (b–d) One representative experiment out of 4 is shown. (e) Offset histogram overlays representing cell surface density of CD11b antigens with mean fluorescence intensity (MFI x103) on CD45+ peritoneal cells from control mice (black), ITO-NP–inoculated mice (red), Gly– and ITO-NP–receiving mice (green). (f) Absolute number of cells phenotypically characterized as CD11b+Gr-1+, CD11b+Gr-1+Ly-6G+ (neutrophils) and CD11b+Gr-1+Ly-6G− (inflamed-monocytes) among CD45+ peritoneal cells from control mice and mice receiving ITO-NPs or both Gly and ITO-NPs. (g) IL-1β detected in peritoneal fluid from control mice, mice receiving ITO-NPs and both Gly and ITO-NPs. (h) Release of IL-1β, TNF-α and of LDH in mice peritoneal macrophages stimulated ex vivo with phorbol 12-myristate 13-acetate (PMA) with or without 500 μg/mL of ITO-NP exposure for 16 h. (b–h), n = 4 ± SEM, *p < 0.05. Schematic diagram abbreviations, MΦ = macrophages, Neu = neutrophils, Mo = monocytes, BMCs = bone marrow cells, BMDMs = bone marrow-derived macrophages, PMΦ = peritoneal macrophages.

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