Skip to main content
. Author manuscript; available in PMC: 2019 May 1.
Published in final edited form as: Arterioscler Thromb Vasc Biol. 2018 Mar 1;38(5):1102–1114. doi: 10.1161/ATVBAHA.118.310703

Figure 6. Timed treatment of diabetic mice with anti-MCP-1 antibody post-injury restores normal healing.

Figure 6

(A) The ligand for the blood monocyte CCR2 receptor, MCP-1, was examined in wound macrophages isolated by magnetic cell sorting CD11b+[CD3−CD19−NK1.1−Ly6G−] from DIO and control mice on day 5. Protein levels of MCP-1 as determined by ELISA in control and DIO wound cells on day 5 post-injury. (*P < 0.05; n= 5 mice/group; data is representative of 2 independent experiments). (B) Control and DIO wounds were harvested on day 6 and single cell suspensions were processed for flow cytometry. Ly6CHi[Live,Ly6G−,CD11b+] monocyte/macrophage counts (as gated in Figure 1) are shown. (*P < 0.05; n= 5 mice/group; repeated 1X). (C) DIO mice were wounded with a 4 mm punch biopsy and wound healing was monitored daily using an 8mp iPad camera, internal scale, and NIH ImageJ software. At day 3 post-injury, mice underwent intra-peritoneal injection with either normal rabbit serum (NRS) or purified rabbit anti-mouse MCP-1 antibody (anti-MCP-1) and wound healing was monitored until wound closure. Wound curves were generated as percent of initial wound area. (*P < 0.05, **P < 0.01; n = 10 wounds per group). (D) Control mice were wounded with a 4 mm punch biopsy and wound healing was monitored daily using an 8mp iPad camera, internal scale, and NIH ImageJ software. At day 3 post-injury, mice underwent intra-peritoneal injection with either normal rabbit serum (NRS) or purified rabbit anti-mouse MCP-1 antibody (anti-MCP-1) and wound healing was monitored until wound closure. Wound curves were generated as percent of initial wound area. (n = 10 wounds per group). (E/F) DIO mice were wounded with a 4 mm punch biopsy and on day 3 post-injury, mice underwent intra-peritoneal injection with either normal rabbit serum (NRS) or purified rabbit anti-mouse MCP-1 antibody (anti-MCP-1). Wounds were harvested on day 5 and analyzed by flow cytometry. Gating was identical to that shown in Figure 2. DIO Ly6CHi[Lin−Ly6G−CD11b+] and DIO Ly6CLo[Lin−Ly6G−CD11b+] wound cells from NRS and anti-MCP-1 injected. (*P < 0.05; n =25 mice; tissues of 2 wounds per mouse were pooled for a single biological replicate. Data is representative of 2 independent experiments.) All data are expressed as mean +/− the standard error of the mean (SEM).