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Fig. 3 | Biological Research

Fig. 3

From: Maintenance of chronicity signatures in fibroblasts isolated from recessive dystrophic epidermolysis bullosa chronic wound dressings under culture conditions

Fig. 3

Functional and molecular markers of fibrosis in RDEB-CW fibroblasts. (a) Representative images of contracted collagen discs after 24 h of in vitro culture. The size decreased as the contractile capability of the fibroblasts increased. Bar: 1 cm. (b) Quantification of contractile assay based on collagen lattices (n = 3). We carried out a statistical analysis to compare the disc contraction between the different experimental groups at 0, 24, 48, and 72 h. Asterisks (*) indicate significant differences versus NHF. Hashtags (#) indicate significant differences versus the RDEB group (p < 0.05, One-way ANOVA with Tukey post-hoc). (c-d) Western blot analysis of protein lysates derived from fibroblast cultures to detect the fibrotic markers α-SMA, C1 and YKL-40. (e-g) Densitometric analysis of western blots displayed in (c-d). Asterisks indicate significant differences (n = 3, one-way ANOVA with Tukey post-hoc). (h) IIF analysis of Collagen I (green) and α-SMA (red). Nuclei were co-stained with DAPI (blue). Bar: 100 μm. (i) Quantification of α-SMA+ cells, expressed as percentage. Fibroblasts were considered as positives for α-SMA specifically when this protein was localized to stress fibers (n = 3). Asterisks indicate significant differences (p < 0.05, One-way ANOVA with Tukey post-hoc). All data are expressed as mean ± SD.

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