Fig. 4

Histologic and suction cutometer analysis of healed wounds. A H + E staining of healed specimens revealed DFO treated irradiated wounds resembled nonirradiated wounds more closely in regards to wound thickness (mean 244.6 vs. 284.3 μm, respectively; *p < 0.05) rather than much thinner vehicle (111.7; ***p < 0.001) and IR untreated wounds (116.5; ***p < 0.001). Representative H + E images of healed wounds of each treatment condition taken at 10 × magnification shown (left). B On TC staining, DFO wounds also had similar collagen density (mean 7.79 × 10⁶) to nonirradiated wounds (8.69 × 10⁶; *p < 0.05) after healing, both of which had significantly higher density than vehicle (3.31 × 10⁶; ***p < 0.001) and IR untreated wounds (3.24 × 10⁶; ***p < 0.001). Representative TC images of healed wounds of each treatment condition taken at 10 × magnification shown (left). C Representative appearance of collagen in Picro images taken at 40 × magnification (left). UMAP representation of collagen ultrastructure in wounds in each treatment condition (right). An approximation of the clustering pattern beneath plotted points highlights DFO (green) clustering more closely to nonirradiated wounds (clear) than vehicle (red) or IR untreated wounds (blue). D Elasticity measured as skin displacement upon application of fixed negative pressure indicated healed DFO wounds had similar elasticity to nonirradiated wounds (mean 0.695 vs. 0.675 mm; p > 0.05). Healed vehicle (0.479;***p < 0.001) and IR untreated (0.465;***p < 0.001) wounds were found to be more stiff. Black scale bar = 100 μm. White scale bar = 25 μm. DFO: deferoxamine; H + E: Hematoxylin and Eosin; IR: irradiation; mm: millimeter; ns: not significant; Picro: Picrosirius Red; POD: post-operative day; TC: Masson’s Trichrome; UMAP: Uniform Manifold Approximation and Projection