Ren Yayun, Chang Jason P E, Tai Dean, Filozof Claudia M, Kleiner David E, Sanyal Arun J
HistoIndex Pte Ltd, Singapore.
Department of Gastroenterology and Hepatology, Singapore General Hospital, Singapore.
Clin Gastroenterol Hepatol. 2025 Jul 30. doi: 10.1016/j.cgh.2025.06.042.
BACKGROUND & AIMS: Fibrosis stage is a key determinant of outcomes in metabolic dysfunction-associated steatohepatitis (MASH). Assessment of fibrosis change is a critical component of the assessment of therapeutic efficacy of interventions. Conventional assessment of fibrosis stage is limited by sampling and inter- and intra-observer variability. Second harmonic generation/2-photon excitation fluorescence (SHG/TPEF) imaging provides an alternate way to assess fibrosis from unstained histologic sections. The aim of this study is to establish repeatability and reproducibility of SHG/TPEF imaging in metabolic dysfunction-associated steatohepatitis histologic samples.
The SHG/TPEF images were acquired by 3 Genesis machines for the included samples. Each sample was scanned 3 times by each machine at different time points. qFibrosis stage (qFS) and qFibrosis continuous value (qFC) were calculated by an artificial intelligence-based algorithm that was previously correlated with the Nonalcoholic Steatohepatitis Clinical Research Network fibrosis staging system.
The overall intra-/inter-system weighted kappas of qFS were 0.880 (95% confidence interval [CI], 0.844-0.915) and 0.850 (95% CI, 0.811-0.889), respectively. Using the pairwise agreement method, the intra-/inter-system agreements of qFS were 89.16% (95% CI, 0.856-0.921) and 86.46% (95% CI, 0.829-0.890), respectively. For qFC, the intra-/inter-system agreements were 87.53% (95% CI, 0.837-0.911) and 78.05% (95% CI, 0.737-0.824), respectively. Furthermore, the Bland-Altman method was used to evaluate the intra-/inter-system agreements of qFC, with 93.77% (95% CI, 0.911-0.960) and 93.50% (95% CI, 0.908-0.959), respectively.
The qFibrosis system has good repeatability and reproducibility for fibrosis staging. It can provide an accurate reference for pathologists to stage liver fibrosis more objectively.
纤维化阶段是代谢功能障碍相关脂肪性肝炎（MASH）预后的关键决定因素。纤维化变化的评估是干预治疗效果评估的重要组成部分。传统的纤维化阶段评估受抽样以及观察者间和观察者内变异性的限制。二次谐波产生/双光子激发荧光（SHG/TPEF）成像提供了一种从未经染色的组织学切片评估纤维化的替代方法。本研究的目的是确定SHG/TPEF成像在代谢功能障碍相关脂肪性肝炎组织学样本中的可重复性和再现性。
使用3台Genesis机器对纳入的样本采集SHG/TPEF图像。每台机器在不同时间点对每个样本扫描3次。通过一种基于人工智能的算法计算定量纤维化阶段（qFS）和定量纤维化连续值（qFC），该算法先前已与非酒精性脂肪性肝炎临床研究网络纤维化分期系统相关联。
qFS的总体系统内/系统间加权卡帕值分别为0.880（95%置信区间[CI]，0.844 - 0.915）和0.850（95%CI，0.811 - 0.889）。采用配对一致性方法，qFS的系统内/系统间一致性分别为89.16%（95%CI，0.856 - 0.921）和86.46%（95%CI，0.829 - 0.890）。对于qFC，系统内/系统间一致性分别为87.53%（95%CI，0.837 - 0.911）和78.05%（95%CI，0.737 - 0.824）。此外，采用Bland - Altman方法评估qFC的系统内/系统间一致性，分别为93.77%（95%CI，0.911 - 0.960）和93.50%（95%CI，0.908 - 0.959）。
定量纤维化系统在纤维化分期方面具有良好的可重复性和再现性。它可为病理学家更客观地进行肝纤维化分期提供准确参考。
