The intratracheal instillation of bleomycin in mice induces early damage to alveolar epithelial cells and development of inflammation followed by fibrotic tissue changes and represents the most widely used model of pulmonary fibrosis to investigate human IPF.
Histopathology is the gold standard for assessing lung fibrosis in rodents, however it precludes repeated and longitudinal measurements of disease progression and does not provide information on spatial and temporal distribution of tissue damage.
Here we investigated the use of the Micro-CT technique to allow the evaluation of disease onset and progression at different time-points in the mouse bleomycin model of lung fibrosis. Micro-CT was throughout coupled with histological analysis for the validation of the imaging results.
In bleomycin-instilled and control mice, airways and lung morphology changes were assessed and reconstructed at baseline, 7, 14 and 21 days post-treatment based on Micro-CT images. Ashcroft score, percentage of collagen content and percentage of alveolar air area were detected on lung slides processed by histology and subsequently compared with Micro-CT parameters.
Extent (%) of fibrosis measured by Micro-CT correlated with Ashcroft score, the percentage of collagen content and the percentage of alveolar air area (r 2 = 0.91; 0.77; 0.94, respectively). Distal airway radius also correlated with the Ashcroft score, the collagen content and alveolar air area percentage (r 2 = 0.89; 0.78; 0.98, respectively).
Micro-CT data were in good agreement with histological read-outs as micro-CT was able to quantify effectively and non-invasively disease progression longitudinally and to reduce the variability and number of animals used to assess the damage. This suggests that this technique is a powerful tool for understanding experimental pulmonary fibrosis and that its use could translate into a more efficient drug discovery process, also helping to fill the gap between preclinical setting and clinical practice.
Balikian JP, Jochelson MS, Bauer KA, Skarkin AT, Garnick MB, Canellos GP, et al. Pulmonary complications of chemotherapy regimens containing bleomycin. AJR Am J Roentgenol. 1982;139(3):455–61.
Bargon G. Drug-induced changes in the lungs. [in german]. Rontgenblatter. 1984;37(7):261–5. PubMed
Antije M, Carlos R-LJ, Lingqiao W, Jack G, Martin K. Models of pulmonary fibrosis. Drug Discov Today Dis Model. 2006;3:243–9. CrossRef
N. R. C. Institute of Laboratory Animal Resources Commission on Life Sciences. Guide for the care and use of laboratory animals. Washington D.C.: National Academy Press; 1996.
Jin GY, Bok SM, Han YM, Chung MJ, Yoon KH, Kim SR, et al. Effectiveness of rosiglitazone on bleomycin-induced lung fibrosis: assessed by micro-computed tomography and pathologic scores. Eur J Radiol. 2012;81(8):1901–6.
Lee HJ, Goo JM, Kim NR, Kim MA, Chung DH, Son KR, et al. Semiquantitative measurement of murine bleomycin-induced lung fibrosis in in vivo and postmortem conditions using microcomputed tomography: correlation with pathologic scores--initial results. Invest Radiol. 2008;43(6):453–60.
Vande Velde G, Poelmans J, De Langhe E, Hillen A, Vanoirbeek J, Himmelreich U, et al. Longitudinal micro-CT provides biomarkers of lung disease that can be used to assess the effect of therapy in preclinical mouse models, and reveal compensatory changes in lung volume. Dis Model Mech. 2016;9(1):91–8.
Vos W, De Backer J, Poli G, De Volder A, Ghys L, Van Holsbeke C, et al. Novel functional imaging of changes in small airways of patients treated with extrafine beclomethasone/formoterol. Respiration. 2013;86(5):393–401.
De Backer J, Vos W, Vinchurkar S, Van Holsbeke C, Poli G, Claes R, et al. The effects of extrafine beclometasone/formoterol (BDP/F) on lung function, dyspnea, hyperinflation, and airway geometry in COPD patients: novel insight using functional respiratory imaging. J Aerosol Med Pulm Drug Deliv. 2015;28(2):88–99.
Schepens T, Cammu G, Maes S, Desmedt B, Vos W, and Deseure K. Functional respiratory imaging after neostigmine- or sugammadex-enhanced recovery from neuromuscular blockade in the anaesthetised rat: a randomised controlled pilot study. Braz J Anesthesiol (English Edition). 2016. available online http://dx.doi.org/10.1016/j.bjane.2015.11.004.
Van Holsbeke CS, Leemans G, Vos WG, De Backer JW, Vinchurkar SC, Geldof M, et al. Functional respiratory imaging as a tool to personalize respiratory treatment in patients with unilateral diaphragmatic paralysis. Respir Care. 2014;59(9):e127–31.
De Backer JW, Vos WG, Burnell P, Verhulst SL, Salmon P, De Clerck N, et al. Study of the variability in upper and lower airway morphology in Sprague–Dawley rats using modern micro-CT scan-based segmentation techniques. Anat Rec (Hoboken). 2009;292(5):720–7.
Ashcroft T, Simpson JM, Timbrell V. Simple method of estimating severity of pulmonary fibrosis on a numerical scale. J Clin Pathol. 1988;41(4):467–70.
Hübner RH, Gitter W, El Mokhtari NE, Mathiak M, Both M, Bolte H, et al. Standardized quantification of pulmonary fibrosis in histological samples. Biotechniques. 2008;44(4):507–11. 514–7.
Organ L, Bacci B, Koumoundouros E, Barcham G, Milne M, Kimpton W, et al. Structural and functional correlations in a large animal model of bleomycin-induced pulmonary fibrosis. BMC Pulm Med. 2015;15:81.
King TE, Pardo A, Selman M. Idiopathic pulmonary fibrosis. Lancet. 2011;378(9807):1949–61.
De Langhe E, Vande Velde G, Hostens J, Himmelreich U, Nemery B, Luyten FP, et al. Quantification of lung fibrosis and emphysema in mice using automated micro-computed tomography. PLoS One. 2012;7(8):e43123.
Ask K, Labiris R, Farkas L, Moeller A, Froese A, Farncombe T, et al. Comparison between conventional and “clinical” assessment of experimental lung fibrosis. J Transl Med. 2008;6:16.
Stellari FF, Ruscitti F, Pompilio D, Ravanetti F, Tebaldi G, Macchi F, et al. Heterologous Matrix Metalloproteinase Gene Promoter Activity Allows In Vivo Real-time Imaging of Bleomycin-Induced Lung Fibrosis in Transiently Transgenized Mice. Front Immunol. 2017;8:199.
Peng R, Sridhar S, Tyagi G, Phillips JE, Garrido R, Harris P, et al. Bleomycin induces molecular changes directly relevant to idiopathic pulmonary fibrosis: a model for “active” disease. PLoS One. 2013;8(4):e59348.
Pammolli F, Magazzini L, Riccaboni M. The productivity crisis in pharmaceutical R & D. Nat Rev Drug Discov. 2011;10(6):428–38.
Cook D, Brown D, Alexander R, March R, Morgan P, Satterthwaite G, et al. Lessons learned from the fate of AstraZeneca’s drug pipeline: a five-dimensional framework. Nat Rev Drug Discov. 2014;13(6):419–31.
Paul SM, Mytelka DS, Dunwiddie CT, Persinger CC, Munos BH, Lindborg SR, et al. How to improve R & D productivity: the pharmaceutical industry’s grand challenge. Nat Rev Drug Discov. 2010;9(3):203–14.
- Longitudinal assessment of bleomycin-induced lung fibrosis by Micro-CT correlates with histological evaluation in mice
Paula van Heijningen
Cedric Van Holsbeke
Franco Fabio Stellari
- BioMed Central
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