Table 1.
Animal studies overview.
Fig 1.
Fibrosis features used for morphometric detection.
Digitalized slides of Masson’s tri-chrome stained lung paraffin sections. Collagen is stained in blue and cell nuclei in red. A: alveolar thickening by collagen deposition. B: fibrotic masses composed of collagen, fibroblasts, and other components.
Fig 2.
Masson’s trichrome stained lung sections from study 3, Bleomycin (A, C, E, F) and Saline (B, D, F, H) animals, A and B at low magnification, scale bar 2.5mm E and F alveolar structure at high magnification, scale bar 50μm. Upper right square: zoom in one alveola. C and D: low scale detection model; false colour image indicates areas of dense fibrosis (green), alveolar tissue (blue), bronchi (red), background (yellow). G and H: high magnification detection; false colour image indicates collagen (blue), lung tissue (red), background (yellow). Upper right square: zoom in one alveola collagen detection.
Fig 3.
Inter study reproducibility of the quantification method.
Automated image analysis was applied to five independent studies, performed over 2 years (3 independent studies for each time point: D14 and D28). A: percentage of fibrotic masses per lung (excluding bronchi for normalization), B: percentage of alveolar collagen deposition. Mean with +/- SEM, Mann-Whitney statistical test. Each dot represents an individual animal and color represents independent studies. Study 2: red, study 3: purple, study 4: dark blue, study 5: green, and study 6: light blue.
Fig 4.
Quantitative analysis of anti-fibrotic compound effects.
Lungs from Wistar rats treated with bleomycin in combination with pirfenidone (0,50%) and BIBF-1120 (50mg/kg) were collected at Day 14 and a Masson’s trichrome staining was performed for fibrosis parameter assessment using automated image analysis. A: percentage of fibrotic masses, B: percentage of alveolar collagen. Mean with +/- SEM, Mann-Whitney statistical test. Each dot represents an individual animal and color represents independent studies. Study 2 in red, and study 6 in blue.