Figure 5.

Time-course experiment: histomorphometric analysis of lung and skin fibrosis progression following bleomycin administration. Representative MT- and Picrosirius-stained lung sections (×20 magnification; scale bar: 100 µm) from saline controls and BLM OA+PUMP (15 + 100)-treated mice at different time-points (28, 35, and 42 days post-OA) are shown in A–A’ and B–B’, respectively. Ashcroft scores represented as scatter plots with mean ± SD for each time-point and treatment condition (saline and BLM OA+PUMP 15 + 100; n = 5 mice/group/time-point) are shown in C. Ashcroft scores frequency distribution of different severity groups, classified as mild (0–3), moderate (=4), and severe (≥5), is expressed as fractional (%) representation and plotted as mean ± SD in D. Collagen content quantification performed on Picrosirius sections is expressed as % tissue area of the whole slide area (mean ± SD; n = 5 mice/group/time-point) and presented as a scatter plot in E. Representative MT-stained skin sections (×20 magnification; scale bar: 100 µm) from saline controls and BLM-OA + PUMP (15 + 100)-treated mice at the indicated time-points after OA-BLM administration (28, 35, and 42 days) are shown in F and F’, respectively. Dermal thickness (G) and hypodermal thickness (H) variations in saline controls and BLM OA+PUMP (15 + 100)-treated mice at the indicated time-points (n = 5 mice/group/time-point); data (µm) are given as mean ± SD. For C and E statistical significance was evaluated by one-way ANOVA with Tukey’s test for multiple comparisons (*P < 0.05, **P < 0.01, ***P < 0.001 vs. saline), whereas for D, G, and H, two-way ANOVA with Tukey’s test for multiple comparisons was used (**P < 0.01, ***P < 0.001 vs. saline). BLM, bleomycin; MT, Masson’s trichrome; OA, oropharyngeal aspiration.