Figure 3.

Wound-conditioned medium activates the inflammasome in both human and mouse Mp in vitro. A and B: Blood-derived Mp from healthy human volunteers either left nonstimulated (N) or stimulated with chronic wound–conditioned medium (CM) along with vehicle (V), the caspase-1 inhibitor YVAD (Y) (20 μmol/L), or the inflammasome inhibitor glyburide (G) (200 μmol/L) and release of IL-1β and IL-18 measured by ELISA. C and D: Representative flow cytograms of active caspase-1 in human blood–derived Mp, Mp either left nonstimulated or stimulated with chronic wound–conditioned medium. E and F: Bone marrow–derived Mp from wild-type (C57BL/6) mice either left nonstimulated or stimulated with day 10 db/db wound-conditioned medium along with vehicle or the same inhibitors as used for human Mp and release of IL-1β and IL-18 measured by ELISA. G and H: Bone marrow–derived Mp from wild-type (WT), caspase-1 (Casp1), and NLRP-3 knockout (ko) mice left nonstimulated (Non), stimulated with TNF-α and IFN-γ (20 ng/mL each) (CA), or stimulated with day 10 db/db wound-conditioned medium (CM). For all graphs, bars = mean ± SD, n = 5–6 in two separate experiments. Data compared between groups using ANOVA on ranks. *Mean value significantly different from that for nonstimulated controls; **mean value significantly different from that for wound-conditioned medium + vehicle-treated cells; #mean value significantly different from that for nonstimulated controls; ##mean value significantly different from that for wound-conditioned medium–treated wild-type Mp; P < 0.05. FSC, forward scatter.