Fibroblast proliferation and ECM changes during wound healing

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Hanneke N. Monsuur, Mireille A. Boink, Ester M

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Fibroblast proliferation and ECM changes during wound healing Fibroblast proliferation and ECM changes during wound healing A, BExperimental design of wound healing time course analysis. (A) Wound healing time course. Skin was collected at the indicated time points. (B) Quantification strategy is schematically illustrated. Red boxes upper dermis; black boxes lower dermis; solid lines inside and dashed lines outside the wound site.C, DFibroblast activation during wound repair. Immunofluorescence staining for α‐sma (red) of PDGFRαH2BEGFP (green) wound sites at indicated postwounding (PW) times (C) and quantification of α‐sma fluorescence intensity in pixels per area inside the upper and lower wound bed (n = 4 PW0, PW2, PW7, PW21; n = 3 PW10, PW14; n = 2 PW4 biological replicates) (D).E–HChanges in fibroblast density, proliferation and collagen deposition during wound repair. (E) Immunostaining for Ki67 (red) of PDGFRαH2BEGFP (green) back skin sections at indicated wound healing time points. (F) Quantification of fibroblast proliferation in the upper and lower dermis inside the wound bed over time (n = 4 PW0, PW2, PW21; n = 3 PW4, PW10; n = 2 PW7, PW14 biological replicates). (G) Quantification of fibroblast density during wound repair inside and outside the wound bed (n = 4 PW0, PW2, PW21; n = 3 PW4, PW10; n = 2 PW7, PW14 biological replicates). (H) Polarised light images of Picrosirius red stained back skin section shown as binary image at indicated time points after wounding.IQuantification of proliferating (Ki67‐positive) keratinocytes and fibroblasts during wound repair over time (n = 4 PW0, PW2, PW21; n = 3 PW4, PW10; n = 2 PW7, PW14 biological replicates).Data information: Nuclei were labelled with DAPI (blue) (C, E). Scale bars, 100 μm (C, E). WB, wound bed. Error bars represent standard deviation of the biological replicates. Emanuel Rognoni et al. Mol Syst Biol 2018;14:e8174 © as stated in the article, figure or figure legend