Skip to main content
. Author manuscript; available in PMC: 2020 Nov 1.
Published in final edited form as: Arterioscler Thromb Vasc Biol. 2019 Sep 5;39(11):2353–2366. doi: 10.1161/ATVBAHA.119.312754

Figure 1. Cutaneous Wound Healing is Impaired Following Recovery from Sepsis and Wound Macrophages Demonstrate Decreased Inflammatory Cytokine Expression.

Figure 1.

A: Wounds were created by 4-mm punch biopsy on the backs of post-sepsis or Sham mice 4 weeks after recovery. The change in wound area was recorded daily with Image J software until complete healing was observed. Representative photographs of the wounds of post-sepsis and sham mice are shown on days 0 and 4 are shown (n=20/group, repeated 2×). *p<0.05 by Mann-Whitney U test. B: Wounds were harvested on day 3, paraffin embedded and sectioned. 5 µM sections were stained with hematoxylin and eosin. Percent re-epithelialization was calculated by measuring distance traveled by epithelial tongues on both sides of wound divided by total distance for full re-epithelialization. Representative images are shown in 2× magnification (n=5/group). Black bar over tissue represents overall distance of the wound, arrowheads identify epithelial tongue ingrowth, scale bar equals 300 μm. *p<0.05 by Student t test with Welch’s correction. C: Wound myeloid cells CD11b+[CD3−CD19−Ly6G−] were isolated from post-sepsis and control mice. Gene expression of Il1b, Nos2, Il23, Il12, and Arginase 1 was quantified using qPCR (n=5/group). **p<0.01 and ***p<0.001 by Student t test with Welch’s correction. D: Post-sepsis and control wound cell isolates were processed for intracellular flow cytometry. The gating strategy used for intracellular flow cytometry selecting live, lineage−, Ly6G−, CD11b+ cells is shown. Flow cytometry quantification of IL1β+ cells (% of Live, Lineage−, Ly6G−, CD11b+, Ly6CHi Cells) in wounds (n = 10/group). *p<0.05 by Student t test with Welch’s correction. E: Levels of IL1β in supernatant of wound macrophages from DIO and control mice following treatment with LPS for 24 hours measured by ELISA (n = 3/group). **p<0.01 by Student t test with Welch’s correction. FMO, fluorescence minus one; FSC, forward scatter; FSC-A, forward scatter area; FSC-H, forward scatter height; SSC, side scatter.