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Preclinical Laboratory Services

Bleomycin-Induced Fibrosis

Validated Model of Idiopathic Pulmonary Fibrosis/Lung Injury


Bleomycin-induced Lung Injury/Pulmonary Fibrosis (IPF)

Bleomycin is a chemotherapeutic agent used to treat cancers such as Hodgkins lymphoma. One of the side effects is pulmonary toxicity, which can be life threatening in approximately 10% of patients.  

 The mechanism of bleomycin-induced lung injury includes oxidative damage via oxidant-mediated DNA breaks, causing inflammatory reactions in the lungs.

Bleomycin has also been used to induce lung injury in rodents for basic research into pulmonary fibrosis for over a decade. At MD Biosciences, we have refined the oral aspiration administration to allow for more even distribution of disease throughout both right and left lungs, making bleomycin-induced injury in mice a reliable model for research. This is a great model for studying idiopathic pulmonary fibrosis (IPF).

Available Bleomycin Study Readouts:

  • Immunophenotyping of bronchoalveolar lavage fluid (BALF):
    •   Quantitation of T Cells, B Cells, Macrophage, Neutrophils, Eosinophils, Fibroblasts, etc
  • Cytokine Expression Levels (ELISA/Multiplex) in:
    • BALF
    • Lung Tissue Homogenate
    • Plasma/Serum
  • Histology of lung:
    • H&E for morphologic analysis, cellular infiltration, etc (Figure 1)
    • Masson's Trichrome staining for collagen (Figure 2)
    • IHC for specific markers of interest such as CD3, CD11b, etc (Figures 3 & 4)
  • Lung tissue biomarkers
  • Blood biomarkers (protein or nucleic acid)
  • Lung tissue cellularity

Figure 1. Longitudinal Mice Lung Sections (H&E Staining):


A-C (Naïve Group).  A. Subgross image demonstrating normal histologic lung tissue. B. Medium magnification of normal parenchyma with clear airways (asterisks). Figure C. High magnification of normal parenchyma with thin, normal septae and optically-empty alveolar spaces (arrowheads) 

D-F (Bleomycin-Treated Group). D. Subgross image showing cellular consolidation in all lobes (darker areas). E. Medium magnification of affected areas demonstrating obliteration of the alveolar spaces by fibrosis (arrows) surrounding the airways (asterisks). F. High magnification of affected areas with obliteration of alveolar spaces accompanied by inflammation and regenerative hyperplasia of pneumocyte type II (foci marked with arrows, airways marked with asterisks).

G-I (Dexamethasone Group). G. Subgross image showing dark areas of consolidation in all lobes. H. Medium magnification of boxed area in G showing variable fibrosis surrounding airways (asterisks). I. High magnification of boxed area in H showing regions with variable fibrosis demonstrating loss of the normal “spongy” appearance of the lungs, inflammatory infiltration (dark dots) and regnerative hyperplasia (arrow).


Figure 2. Longitudinal Mice Lung Tissue (Masson's Trichrome Staining of Collagen):


A-C (Naïve Group): A. Subgross image showing no significant changes over timecourse of study demonstrating normal lung histology.  B. Medium magnification of normal parenchyma.  C. High magnification of normal parenchyma showing only a small amount of collagen in the perivascular interstitium (arrow), with normal width of alveolar septae (arrowheads). 

D-G (Bleomycin Group): D. Subgross image showing dark areas of cellular consolidation in all lobes (~65%, outlined in blue). E. Medium magnification of affected areas (arrowheads) characterized by obliteration of alveolar spaces due to fibrosis. F. High magnification of the more severely affected areas, with fibrosis appearing to be progress from the center of the lung outwards toward the pleura. G. High magnification of a focus with ongoing fibroplasia, including collections of fibroblasts and collagen (blue material) from within and efface alveolar spaces (asterisks).

H-K (Dexamethasone Group): H. Subgross image showing dark areas of cellular consolidation in all lobes (~35%, outlined in blue). I. Medium magnification of regions with fibrosis of variable severity. J. High magnification of region where alveolar architecture has been completely replaced by fibrous tissue (blue). There is also multifocal mild regenerative hyperplasia (proliferation of pneumocyte type II, arrows). K. High magnification of peripheral field of the lung which is mostly normal, except for early subpleural fibrosis at the edge (arrowheads). 


Figure 3. Lung Sections from Belomycin-Induced Fibrosis Model Stained For Macorphage (IHC Staining with CD11b Antibody):


A. Medium magnification of interstitial perivascular infiltrates showing staining for CD11b-positive cells (macrophage-specific marker). B. High magnification of interstitial perivascular infiltrates of CD11b-positive cells.


Figure 4. Lung Sections From Bleomycin-Induced Fibrosis Model Stained for Lymphocytes (IHC Staining with CD3 Antibody):


A. Rare CD3+ cells (arrows, non-specific staining in epithelium in upper left corner of image). Figure B. Subpleural infiltrate which includes CD3+ lymphocytes 


Species Availability:


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