Fibrosis, Lung, Bleomycin-Induced
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DOCUMENTS/REFERENCES
BIOLOGICAL INFORMATION
BACKGROUND INFORMATION
Pulmonary fibrosis is characterized by alveolar epithelial cell injury, areas of type II cell hyperplasia, accumulation of fibroblasts and myofibroblasts, and the deposition of extracellular matrix proteins. Intratracheal instillation (IT) of bleomycin is a widely used experimental model for lung fibrosis and assessment of anti-fibrotic agents.
ORGANISM
Mouse
ASSAY INFORMATION
ASSAY TYPE
In Vivo
TESTING INFORMATION
PROCEDURE SUMMARY
Male C57BL/6 mice weighing 24 ± 2 g are used. The animals are divided into groups of 10 each. The animals are anesthetized with isoflurane and a single dose of bleomycin at 1.5 IU/kg (dissolved in 50 μl of saline) is administered intratracheally with a PennCentury intrapulmonary aerosolizer on Day 1. Sham control receives the same volume of saline intratracheally. Dosing of vehicle and test articles starts immediately post bleomycin administration once daily through Day 21. Body weight is recorded daily during the study. The animals are sacrificed on Day 22 and the lungs are 0.5 mL of phosphate-buffered saline (PBS) is administered twice through a tracheal cannula after which about 0.6 mL of bronchoalveolar lavage fluid (BALF) is obtained. Total cell counts and differential cell counts in BALF are determined by an automatic hematology analyzer. TGF-β1 in BALF is measured by ELISA (optional). Lung homogenate samples are collected for measurement of hydroxyproline levels (whole lungs) (optional). Lung tissues are formalin-fixed, paraffin-embedded, and stained by hematoxylin and eosin (H&E) or Masson’s trichrome staining for histopathological examination (optional). ANOVA and Dunnett’s test are used to ascertain the difference between vehicle control and treated mice. Significance is set at p<0.05 level.
CRITERIA FOR SIGNIFICANCE
Significant difference is determined at P < 0.05
TURNAROUND TIME
STANDARD
60 Days
CLINICAL RELEVANCE
THERAPEUTIC AREA
Pulmonology
ADDITIONAL INFORMATION
BRAND
PDS
TESTING LOCATION
Taiwan - Taipei
OTHER INFORMATION
We will readily accommodate client-specified alterations. These studies are performed at our AAALAC accredited BSL-2 laboratory in Taipei, Taiwan. All aspects of this work are performed in general accordance with the Guide for the Care and Use of laboratory animals (National Academy Press, Washington, DC, 2011). The study protocol was approved by the Pharmacology Discovery Services IACUC and is performed with the oversight of veterinarians to assure the humane treatment of laboratory animals.
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