(A) (Left) Animals were first injected i.v. with either OVA-NP or DT-OVA-NP (2.5 μl/g) and, 1d later, the wound healing assay began. Two days after wounding, animals were treated with another similar dose of OVA-NP (control) or DT-OVA-NP. Then, mice were sacrificed 3d later (mid-stage repair) and their ears were collected to analyze skin macrophages. (Right) Bar histogram showing the percentage of MΦs out of total CD45+ cells in the ears of OVA-NP- or DT-OVA-NP-treated animals. Data are mean ± SEM (n = 4–7). Statistical significance was assessed by unpaired two-tailed Student’s t-test (**p-value < 0.01). (B) Analysis of mid-stage repair (5d after wounding). The plot illustrates the decrease of the wound area over time expressed as percentage of the initial wound. Data are mean SEM, n = 32 wounds/group. Statistical significance was assessed by two-way ANOVA analysis with Sidak’s post-test (**p-value < 0.01, ****p-value < 0.0001) (C) Histological analysis of wounds at mid-stage of repair from OVA-NP- and DT-OVA-NP-treated animals. Hematoxylin-eosin staining of representative samples is shown. (Hp ep: hyperproliferative epithelium, ep tg: epithelial tongue, G: granulation tissue, d: dermis, p: panniculus carnosus, a: adipose tissue, ml: muscle layer). (D) (Left) Representative immunofluorescence staining of vessels (CD31 staining) and macrophages (Mac-3 staining) in tissue sections of wounds at mid-stage (5d) from OVA-NP- or DT-OVA-NP-treated animals. (Right) Quantification of CD31 and Mac-3 stained area within the granulation tissue is shown. Data are mean ± SD (n = 4–6). Statistical significance was assessed by unpaired two-tailed Student’s t-test (**p<0.01, ***p<0.005).