Figure 4.

CSO is a potent inducer of mϕ^heal macrophage polarization while attenuating M1 phenotype in chronic diabetic wounds. (A) Wound mϕ were harvested from PVA sponges coated with CSO (300 mg) implanted subcutaneously in db/db mice on day 7 post-implantations. Total RNA was isolated and mRNA expression of Arginase-1, CD206, YM1 and VEGF was measured using RTPCR. CSO, solid bars; control, blank bars. (B) Total protein isolated from day 7 wound mϕ of db/db mice, treated with CSO in vivo, was subjected to capillary electrophoresis immunoassay to measure expression of Arginase 1 protein. (C) Arginase activity of in vivo CSO treated (300 mg) day 7 wound mϕ of db/db mice were measured by Arginase activity assay kit (Colorimetric). (D) mRNA expression of CD74 was measured in day 7 wound mϕ of db/db mice treated with CSO (300 mg). (E) PMA-induced superoxide anion production in day 3 wound mϕ of db/db mice treated in vivo with CSO (300 mg) was measured. For all figure parts, data are expressed as mean ± SEM (n = 3–6); *p < 0.05 compared to wound mϕ treated with equal amounts of white petrolatum (control). (F) Immunocytochemistry (ICC) images of Arginase-1 (red) and NOS2 (red) protein expression in d7 wound mϕ from db/db animals. Day 7 wound mϕ were harvested from PVA sponges coated with CSO (300 mg) implanted subcutaneously. Counter staining was performed using DAPI (nuclear, blue). Scale bar, 20 μm.