Fig. 8

H&E and immunostaining of inflamed paw sections collected at end-point from male and female rats treated with CXB-NE or DF-NE. Top Panel: Cross-section of normal male rat paw (A) and normal female rat paw, (B) showing plantar and dorsal aspects (Detail, H&E, 4X). The location of metatarsals (curved arrows) and the site of CFA injection into the plantar aspect (yellow curved arrow) are shown. Immunofluorescence staining shows nuclei (DAPI, blue) macrophages (CD68 staining, green) and NE (purple-pink). Middle Panel: Cross-section of male rat paw, (C) and female rat paw (D) following CFA injection and DF-NE treatment. Note the significant immune infiltration in the plantar zone (shown in white boxed insets) in both specimens, with edema most pronounced in the male paw sample. Immunofluorescence staining reveals co-localization of DFNE (purple-pink) macrophages (CD68 staining, green) following CFA injection. Note the zones of significant macrophage infiltration in both samples and the correlation of DFNE distribution with histopathologic presentation. The localization of DF-NE with macrophage infiltration is most pronounced in the deep dermal and fascial planes of foot. (4X and 20X). Lower Panel: Cross-section of male rat paw E) and female rat paw, F) following CFA injection and CUR-NE treatment. There is a marked reduction in immune infiltration in the plantar zones (shown in white boxed insets) in both specimens with edema scant or absent in paw samples. Immunofluorescence staining reveals co-localization of CUR-NE (purple-pink) with macrophages (CD68 staining, green) in inflammatory zones following CFA injection. Note that the intensity and distribution of macrophage infiltration in both samples are markedly reduced/different from DF-NE samples and correlate well with histopathologic presentation on H&E (4X and 20X). Scale bar: 100 \(\:\mu\:\)M.