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. Author manuscript; available in PMC: 2013 Mar 23.
Published in final edited form as: Nat Commun. 2012 Mar 27;3:755. doi: 10.1038/ncomms1754

Figure 1. NLRP3 inflammasome activation by phagocytosed WD particles.

Figure 1

(a) Transmission electron micrograph of nanometre and micrometre size WD recovered from the periprosthetic tissue of patients undergoing joint revision surgery. Bar corresponds to 1 μm. (b) ultrastructural analysis of micrometre size WD particles (arrows) phagocytosed by myeloid mononuclear cells present in periprosthetic tissue. Bar corresponds to 1 μm. (c) ultrastructural analysis of micrometre size WD particles (arrows) next to a blood vessel in periprosthetic tissue. Bar corresponds to 1 μm. (d) ultrastructural analysis of nanometre and micrometre size WD particles (arrows) phagocytosed by a myeloid mononuclear cells present in periprosthetic tissue. Bar corresponds to 500 nm. (e) FTIR spectroscopic analysis of pre-implant UHMWPE and commercially available UHMWPE particles. The analysis was carried out between 4,000 and 500 cm2 wave number. (f) Two-dimensional electron micrograph cross-section and (g,h) 3D models derived from 200-nm–section tomograms of an untreated dendritic cell. (g) The late endosome multivesicular body presents a definitive spherical shape and well-defined limiting membrane. (h) The model is rotated 25 ° relative to (g). (i) 2D electron micrograph cross-section of a micrometre size WD particle phagocytosed by a cultured dendritic cell, and (j,k) 3D tomographic reconstruction of the same (j) WD particle inside the endosomal compartment. (k) The model is rotated 25 ° relative to (j). (l,m,n) Western blotting for (l) cathepsins B, (m) cathepsin S, and (n) cyclophilin (loading control), from control (Ctr) and WD-particles-treated dendritic cells. Bar graphs represent average and s.e.m. of three independent western blots, P < 0.001 and P < 0.05 values, respectively, ANOVA, Tukey (o) ELIsA for IL-1β secretion from dendritic cells (control, NLRP3, ASC and caspase-1 knockout mice) untreated or treated for 12 h with WD particles (100 μg ml–1), PGN (a TLR2 agonist, at 10 μg ml–1), or WD and PGN combined. The average and s.e.m. of three experiments is shown; asterisk indicates a statistically significant (P < 0.001, ANOVA, Tukey) decrease in IL-1 production in NLRP3, ASC and caspase 1 knockout mice as compared with C57Bl/6 controls.