Characterization of transient and progressive pulmonary fibrosis by spatially correlated phase contrast microCT, classical histopathology and atomic force microscopy

L. D' Amico et al., Computers in Biology and Medicine 169, 107947 (2024); doi: 10.1016/j.compbiomed.2024.107947
Graphical illustration of the presented pipeline. Whole FFPE mouse lungs were scanned with PBI at the SYRMEP beamline of the Italian synchrotron, resulting in a 3D representation of the entire specimen (first row) which allowed screening and identification of planes of interest. Subsequently, the specimens were sectioned. Adjacent sections were used for classical histology (second row) and AFM (third row). The results were then fused using elastic registration which resulted in spatial integration of the information into a single data set.
ABSTRACT

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.

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Characterization of transient and progressive pulmonary fibrosis by spatially correlated phase contrast microCT, classical histopathology and atomic force microscopy
D'Amico L., SvetloveA., Longo E., Meyer R., SenigagliesiB., SaccomanoG., Nolte P., Wagner W.L., WielpützM.O., LeitzD.H.W., DuerrJ., Mall M.A., CasalisL., KösterS., Alves F., Tromba G., DullinC.
Computers in Biology and Medicine, Vol. 169, 107947 (2024)doi: 10.1016/j.compbiomed.2024.107947
Ultima modifica il Giovedì, 04 Aprile 2024 18:16