University of Oulu

Pirhonen J, Arola J, Sädevirta S, Luukkonen P, Karppinen S-M, Pihlajaniemi T, et al. (2016) Continuous Grading of Early Fibrosis in NAFLD Using Label-Free Imaging: A Proof-of-Concept Study. PLoS ONE 11(1): e0147804.

Continuous grading of early fibrosis in NAFLD using label-free imaging : a proof-of-concept study

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Author: Pirhonen, Juho1,2; Arola, Johanna3,4; Sädevirta, Sanja5,2;
Organizations: 1Departments of Anatomy, Faculty of Medicine, University of Helsinki, Helsinki, Finland
2Minerva Foundation Institute for Medical Research, Helsinki, Finland
3Department of Pathology, Faculty of Medicine, University of Helsinki, Helsinki, Finland
4Department of Pathology, HUSLAB, Helsinki, Finland
5Department of Medicine, Faculty of Medicine, University of Helsinki, Helsinki, Finland
6Faculty of Biochemistry and Molecular Medicine, Oulu Center for Cell-Matrix Research, Biocenter Oulu, Oulu, Finland
Format: article
Version: published version
Access: open
Online Access: PDF Full Text (PDF, 14.3 MB)
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Language: English
Published: Public Library of Science, 2016
Publish Date: 2019-10-24


Background and Aims: Early detection of fibrosis is important in identifying individuals at risk for advanced liver disease in non-alcoholic fatty liver disease (NAFLD). We tested whether second-harmonic generation (SHG) and coherent anti-Stokes Raman scattering (CARS) microscopy, detecting fibrillar collagen and fat in a label-free manner, might allow automated and sensitive quantification of early fibrosis in NAFLD.

Methods: We analyzed 32 surgical biopsies from patients covering histological fibrosis stages 0–4, using multimodal label-free microscopy. Native samples were visualized by SHG and CARS imaging for detecting fibrillar collagen and fat. Furthermore, we developed a method for quantitative assessment of early fibrosis using automated analysis of SHG signals.

Results: We found that the SHG mean signal intensity correlated well with fibrosis stage and the mean CARS signal intensity with liver fat. Little overlap in SHG signal intensities between fibrosis stages 0 and 1 was observed. A specific fibrillar SHG signal was detected in the liver parenchyma outside portal areas in all samples histologically classified as having no fibrosis. This signal correlated with immunohistochemical location of fibrillar collagens I and III.

Conclusions: This study demonstrates that label-free SHG imaging detects fibrillar collagen deposition in NAFLD more sensitively than routine histological staging and enables observer-independent quantification of early fibrosis in NAFLD with continuous grading.

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Series: PLoS one
ISSN: 1932-6203
ISSN-E: 1932-6203
ISSN-L: 1932-6203
Volume: 11
Issue: 1
Article number: e0147804
DOI: 10.1371/journal.pone.0147804
Type of Publication: A1 Journal article – refereed
Field of Science: 3121 General medicine, internal medicine and other clinical medicine
Funding: This work was financially supported by the Academy of Finland (EI grants 131489, 263841, 272130; TP grant 251314; S-MK grant 259872, HYJ), European Union/European Federation of Pharmaceutical Industries and Associations Innovative Medicines Initiative Joint Undertaking (European Medical Information Framework EMIF grant no. 115372, HYJ), the Sigrid Juselius (EI, TP, HYJ), Special governmental subsidy for health sciences research (EVO, Helsinki University Central Hospital grant, HYJ) and the Novo Nordisk Foundation (HYJ). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Academy of Finland Grant Number: 251314
Detailed Information: 251314 (Academy of Finland Funding decision)
259872 (Academy of Finland Funding decision)
Copyright information: © 2016 Pirhonen et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.