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Whole Slide Quantitative Image Analysis Can Demonstrate Bleomycin Induced Dose‐Dependent Increases in Fibrotic Areas Within Histologically Stained Rat Lung Tissue
Author(s) -
Young Alan,
McElroy Mary,
Naylor Stuart,
Anderson Mark,
Bigley Alison,
McGregor Kieran,
Sherry Lorcan
Publication year - 2017
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.31.1_supplement.656.6
Subject(s) - lesion , parenchyma , bleomycin , lung , saline , pathology , histology , medicine , nuclear medicine , surgery , chemotherapy
Rationale The accurate classification and quantification of specific morphological and cellular components within the bleomycin (Bleo)‐induced rodent lung tissue model is challenging. This study aimed to employ histology in combination with whole slide digital image analysis to quantify the extent of lesion formation and content within lung tissue from Bleo treated rats. Methods Male SD rats, n=5 per group (Gr), were treated with either saline (Gr1) each day for 7 days or 1.5mg/Kg Bleo (Gr2) and 3.0mg/Kg Bleo (Gr3) on one dose occasion or 0.5mg/Kg Bleo (Gr4) and 1.0mg/Kg Bleo (Gr5) each day for 7 days. On day 22, lungs were fixed, paraffin embedded, sectioned and histologically stained using Picrosirius Red. Whole slide images of lung tissue were captured and analysis algorithms developed by OracleBio (using Definiens Tissue Studio® software) to define specific regions of interest (ROI) per section including Parenchyma and Lesion, excluding basal collagen. Collagen content and cell density can then be quantified within the Lesion and Parenchyma ROI, but have not been presented in this abstract. The % Fibrotic Lesion area was calculated relative to the combined Lesion and Parenchyma areas only. Results The mean % Lesion area in Gr1 was 1.1% (95% CI: 0.5 to 2.7). This value was attributed as artefact which was classified during the objective application of the analysis algorithm and was excluded from further analysis. There was a clear trend showing an increase in % Lesion area as the Bleo dose increased from 0.5 to 3.0mg/Kg. The mean % Lesion area in Gr2 was 7.0% (95% CI: 4.5 to 9.6), Gr3 was 9.1% (95% CI: 1.7 to 16.5), Gr4 was 10.1% (95% CI: 6.5 to 13.7) and Gr5 was 16.2% (95% CI: 9.4 to 23.1). A significant difference was observed between Gr2 and Gr5 (p = 0.0124). Conclusion Quantitative whole slide image analysis can be utilised to accurately classify therapeutically relevant regions of interest. This may provide a new tool to assess and better interpret changes in morphological and cellular tissue parameters within histologically stained lung tissue sections from pre‐clinical bleomycin fibrosis models. Support or Funding Information Charles River Laboratories and OracleBio