Characterization of transient and progressive pulmonary fibrosis by spatially correlated phase contrast microCT, classical histopathology and atomic force microscopy
Computers in Biology and Medicine. 2024-02-01; 169: 107947
DOI: 10.1016/j.compbiomed.2024.107947
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D’Amico L(1), Svetlove A(2), Longo E(3), Meyer R(4), Senigagliesi B(5), Saccomano G(6), Nolte P(7), Wagner WL(8), Wielpütz MO(8), Leitz DHW(9), Duerr J(9), Mall MA(9), Casalis L(3), Köster S(10), Alves F(11), Tromba G(3), Dullin C(12).
Author information:
(1)University of Trieste, Department of Physics, Via Alfonso Valerio 2, Trieste,
34127, Italy; Elettra Sincrotrone Trieste S.C.p.A., s.s. 14 km 163, 500 in Area
Science Park, Basovizza, 34149, Italy.
(2)Translational Molecular Imaging, Max-Plank-Institute for Multidisciplinary
Sciences, Hermann-Rein-Straße 3, Göttingen, 37075, Germany; Cluster of
Excellence “Multiscale Bioimaging: from Molecular Machines to Networks of
Excitable Cells” (MBExC), Robert-Koch-Str. 40, Göttingen, 37075, Germany.
(3)Elettra Sincrotrone Trieste S.C.p.A., s.s. 14 km 163, 500 in Area Science
Park, Basovizza, 34149, Italy.
(4)Institute for X-ray Physics, University of Göttingen, Friedrich-Hund-Platz 1,
Göttingen, 37077, Germany.
(5)Interdisciplinary Institute for Neuroscience, University of Bordeaux-UMR 5297
and CNRS, 146 Rue Léo Saignat, Bordeaux, 33000, France.
(6)Elettra Sincrotrone Trieste S.C.p.A., s.s. 14 km 163, 500 in Area Science
Park, Basovizza, 34149, Italy; University of Trieste, Department of Architecture
and Engineering, Via Alfonso Valerio 6/1, Trieste, 34127, Italy.
(7)Faculty of Engineering and Health, University of Applied Sciences and Arts,
Göttingen, 37085, Germany; Institute for Diagnostic and Interventional
Radiology, University Medical Center, Göttingen, 37075, Germany.
(8)Diagnostic and Interventional Radiology, University Hospital Heidelberg,
Heidelberg, Germany.
(9)Department of Pediatric Respiratory Medicine, Immunology and Critical Care
Medicine, Charite – University Hospital Berlin, Berlin, 13353, Germany; German
Center for Lung Research (DZL), associated partner site, Berlin, Germany; Berlin
Institute of Health at Charité – Universitätsmedizin Berlin, Berlin, 10117,
Germany.
(10)Cluster of Excellence “Multiscale Bioimaging: from Molecular Machines to
Networks of Excitable Cells” (MBExC), Robert-Koch-Str. 40, Göttingen, 37075,
Germany; Institute for X-ray Physics, University of Göttingen,
Friedrich-Hund-Platz 1, Göttingen, 37077, Germany.
(11)Translational Molecular Imaging, Max-Plank-Institute for Multidisciplinary
Sciences, Hermann-Rein-Straße 3, Göttingen, 37075, Germany; Cluster of
Excellence “Multiscale Bioimaging: from Molecular Machines to Networks of
Excitable Cells” (MBExC), Robert-Koch-Str. 40, Göttingen, 37075, Germany;
Institute for Diagnostic and Interventional Radiology, University Medical
Center, Göttingen, 37075, Germany; Department for Haematology and Medical
Oncology, University Medical Center, Göttingen, 37075, Germany.
(12)Elettra Sincrotrone Trieste S.C.p.A., s.s. 14 km 163, 500 in Area Science
Park, Basovizza, 34149, Italy; Translational Molecular Imaging,
Max-Plank-Institute for Multidisciplinary Sciences, Hermann-Rein-Straße 3,
Göttingen, 37075, Germany; Institute for Diagnostic and Interventional
Radiology, University Medical Center, Göttingen, 37075, Germany; Diagnostic and
Interventional Radiology, University Hospital Heidelberg, Heidelberg, Germany.
Electronic address: .
Pulmonary fibrosis (PF) is a severe and progressive condition in which the lung
becomes scarred over time resulting in pulmonary function impairment. Classical
histopathology remains an important tool for micro-structural tissue assessment
in the diagnosis of PF. A novel workflow based on spatial correlated
propagation-based phase-contrast micro computed tomography (PBI-microCT), atomic
force microscopy (AFM) and histopathology was developed and applied to two
different preclinical mouse models of PF – the commonly used and well
characterized Bleomycin-induced PF and a novel mouse model for progressive PF
caused by conditional Nedd4-2 KO. The aim was to integrate structural and
mechanical features from hallmarks of fibrotic lung tissue remodeling.
PBI-microCT was used to assess structural alteration in whole fixed and paraffin
embedded lungs, allowing for identification of fibrotic foci within the 3D
context of the entire organ and facilitating targeted microtome sectioning of
planes of interest for subsequent histopathology. Subsequently, these sections
of interest were subjected to AFM to assess changes in the local tissue
stiffness of previously identified structures of interest. 3D whole organ
analysis showed clear morphological differences in 3D tissue porosity between
transient and progressive PF and control lungs. By integrating the results
obtained from targeted AFM analysis, it was possible to discriminate between the
Bleomycin model and the novel conditional Nedd4-2 KO model using agglomerative
cluster analysis. As our workflow for 3D spatial correlation of PBI, targeted
histopathology and subsequent AFM is tailored around the standard procedure of
formalin-fixed paraffin-embedded (FFPE) tissue specimens, it may be a powerful
tool for the comprehensive tissue assessment beyond the scope of PF and
preclinical research.
Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.
DOI: 10.1016/j.compbiomed.2024.107947
PMID: 38211385
Conflict of interest statement: Declaration of competing interest Conflict of
interest statement for the manuscript: None Declared