Effect of liver biopsy size on MASLD fibrosis assessment by second-harmonic generation/two-photon excitation fluorescence microscopy

Background & Aims: Fibrosis in metabolic dysfunction-associated steatotic liver disease (MASLD) is a prognostic indicator and clinical trial efficacy endpoint. Second-harmonic generation/two-photon excitation fluorescence (SHG/TPEF) microscopy images unstained tissue sections and, when integ...

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Main Authors: Daniel T. Field, Yayun Ren, Kutbuddin Akbary, Elaine Chng, Dean Tai, Nikolai V. Naoumov, David E. Kleiner, Jonathan A. Fallowfield, Timothy J. Kendall, Arun J. Sanyal
Format: Article
Language:English
Published: Elsevier 2025-08-01
Series:JHEP Reports
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Online Access:http://www.sciencedirect.com/science/article/pii/S2589555925001272
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author Daniel T. Field
Yayun Ren
Kutbuddin Akbary
Elaine Chng
Dean Tai
Nikolai V. Naoumov
David E. Kleiner
Jonathan A. Fallowfield
Timothy J. Kendall
Arun J. Sanyal
author_facet Daniel T. Field
Yayun Ren
Kutbuddin Akbary
Elaine Chng
Dean Tai
Nikolai V. Naoumov
David E. Kleiner
Jonathan A. Fallowfield
Timothy J. Kendall
Arun J. Sanyal
author_sort Daniel T. Field
collection DOAJ
description Background & Aims: Fibrosis in metabolic dysfunction-associated steatotic liver disease (MASLD) is a prognostic indicator and clinical trial efficacy endpoint. Second-harmonic generation/two-photon excitation fluorescence (SHG/TPEF) microscopy images unstained tissue sections and, when integrated with artificial intelligence models, generates a continuous fibrosis value (qFibrosis) and ordinal qFibrosis stage. The impact of biopsy size and location on the accuracy of these approaches has not been assessed in MASLD, leaving quality assurance procedures undefined. Methods: One unstained section each from 100 hepatectomy/explant MASLD cases, 20 of each pathologist-assigned Non-alcoholic Steatohepatitis Clinical Research Network (NASH-CRN) fibrosis stage (F0–F4), were used to create virtual core biopsies by cropping regions from within the whole parent section. Regions varied in length (5–30 mm) with a fixed width of 0.9 mm, width (0.5–1.3 mm) with a fixed length of 15 mm, or position within the whole parent section. SHG/TPEF was used, and the qFibrosis continuous value and stage of the virtual core biopsies were determined for comparison with those of the whole parent section. Results: The qFibrosis continuous value and stage correlated strongly with pathologist-assigned NASH-CRN stage (rs = 0.92). Increasing the length and width of virtual biopsies increased the correlation between the qFibrosis continuous value and the agreement with the qFibrosis stage of the whole parent section, stabilising between 20–26 mm in length and 0.9 mm in width. The position within the tissue did not influence qFibrosis metrics. Conclusions: Longer (>20 mm) and wider (>0.9 mm) biopsies provide more accurate fibrosis assessment using SHG/TPEF. Biopsy position and orientation do not influence accuracy. Impact and implications: Fibrosis assessment is an important prognostic indicator and clinical trial endpoint in metabolic dysfunction-associated steatotic liver disease, but liver biopsy sampling variation quality assurance has not been investigated for second-harmonic generation/two-photon excitation fluorescence (SHG/TPEF) microscopy quantification of fibrosis. Longer (>20 mm) and wider (>0.9 mm) biopsies allow for more accurate digital assessment of fibrosis. Clinical trials should incorporate suitable protocols to verify biopsy sizes that optimise digital fibrosis assessment using SHG/TPEF.
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spelling doaj-art-97e3e05f2f7f4940be3c3fd2d477723f2025-08-20T03:28:13ZengElsevierJHEP Reports2589-55592025-08-017810144910.1016/j.jhepr.2025.101449Effect of liver biopsy size on MASLD fibrosis assessment by second-harmonic generation/two-photon excitation fluorescence microscopyDaniel T. Field0Yayun Ren1Kutbuddin Akbary2Elaine Chng3Dean Tai4Nikolai V. Naoumov5David E. Kleiner6Jonathan A. Fallowfield7Timothy J. Kendall8Arun J. Sanyal9Edinburgh Pathology, University of Edinburgh, Edinburgh, UKHistoindex Pte. Ltd., SingaporeHistoindex Pte. Ltd., SingaporeHistoindex Pte. Ltd., SingaporeHistoindex Pte. Ltd., SingaporeDivision of Medicine, University College London, London, UKLaboratory of Pathology, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MA, USACentre for Inflammation Research, Institute for Regeneration and Repair, University of Edinburgh, Edinburgh, UKEdinburgh Pathology, University of Edinburgh, Edinburgh, UK; Centre for Inflammation Research, Institute for Regeneration and Repair, University of Edinburgh, Edinburgh, UK; Corresponding author. Address: Centre for Inflammation Research, Institute for Regeneration and Repair, University of Edinburgh, 5 Little France Drive, Edinburgh, EH16 4UU, UK. Tel.: +44-0131-2427007.Stravitz-Sanyal Institute of Liver Disease and Metabolic Health, Virginia Commonwealth University School of Medicine, Richmond, VA, USABackground & Aims: Fibrosis in metabolic dysfunction-associated steatotic liver disease (MASLD) is a prognostic indicator and clinical trial efficacy endpoint. Second-harmonic generation/two-photon excitation fluorescence (SHG/TPEF) microscopy images unstained tissue sections and, when integrated with artificial intelligence models, generates a continuous fibrosis value (qFibrosis) and ordinal qFibrosis stage. The impact of biopsy size and location on the accuracy of these approaches has not been assessed in MASLD, leaving quality assurance procedures undefined. Methods: One unstained section each from 100 hepatectomy/explant MASLD cases, 20 of each pathologist-assigned Non-alcoholic Steatohepatitis Clinical Research Network (NASH-CRN) fibrosis stage (F0–F4), were used to create virtual core biopsies by cropping regions from within the whole parent section. Regions varied in length (5–30 mm) with a fixed width of 0.9 mm, width (0.5–1.3 mm) with a fixed length of 15 mm, or position within the whole parent section. SHG/TPEF was used, and the qFibrosis continuous value and stage of the virtual core biopsies were determined for comparison with those of the whole parent section. Results: The qFibrosis continuous value and stage correlated strongly with pathologist-assigned NASH-CRN stage (rs = 0.92). Increasing the length and width of virtual biopsies increased the correlation between the qFibrosis continuous value and the agreement with the qFibrosis stage of the whole parent section, stabilising between 20–26 mm in length and 0.9 mm in width. The position within the tissue did not influence qFibrosis metrics. Conclusions: Longer (>20 mm) and wider (>0.9 mm) biopsies provide more accurate fibrosis assessment using SHG/TPEF. Biopsy position and orientation do not influence accuracy. Impact and implications: Fibrosis assessment is an important prognostic indicator and clinical trial endpoint in metabolic dysfunction-associated steatotic liver disease, but liver biopsy sampling variation quality assurance has not been investigated for second-harmonic generation/two-photon excitation fluorescence (SHG/TPEF) microscopy quantification of fibrosis. Longer (>20 mm) and wider (>0.9 mm) biopsies allow for more accurate digital assessment of fibrosis. Clinical trials should incorporate suitable protocols to verify biopsy sizes that optimise digital fibrosis assessment using SHG/TPEF.http://www.sciencedirect.com/science/article/pii/S2589555925001272HumansMetabolic dysfunction-associated steatotic liver diseaseNon-alcoholic fatty liver diseaseLiver cirrhosisBiopsyArtificial intelligence
spellingShingle Daniel T. Field
Yayun Ren
Kutbuddin Akbary
Elaine Chng
Dean Tai
Nikolai V. Naoumov
David E. Kleiner
Jonathan A. Fallowfield
Timothy J. Kendall
Arun J. Sanyal
Effect of liver biopsy size on MASLD fibrosis assessment by second-harmonic generation/two-photon excitation fluorescence microscopy
JHEP Reports
Humans
Metabolic dysfunction-associated steatotic liver disease
Non-alcoholic fatty liver disease
Liver cirrhosis
Biopsy
Artificial intelligence
title Effect of liver biopsy size on MASLD fibrosis assessment by second-harmonic generation/two-photon excitation fluorescence microscopy
title_full Effect of liver biopsy size on MASLD fibrosis assessment by second-harmonic generation/two-photon excitation fluorescence microscopy
title_fullStr Effect of liver biopsy size on MASLD fibrosis assessment by second-harmonic generation/two-photon excitation fluorescence microscopy
title_full_unstemmed Effect of liver biopsy size on MASLD fibrosis assessment by second-harmonic generation/two-photon excitation fluorescence microscopy
title_short Effect of liver biopsy size on MASLD fibrosis assessment by second-harmonic generation/two-photon excitation fluorescence microscopy
title_sort effect of liver biopsy size on masld fibrosis assessment by second harmonic generation two photon excitation fluorescence microscopy
topic Humans
Metabolic dysfunction-associated steatotic liver disease
Non-alcoholic fatty liver disease
Liver cirrhosis
Biopsy
Artificial intelligence
url http://www.sciencedirect.com/science/article/pii/S2589555925001272
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