Abstract
The evolution of cirrhosis is marked by quantitative and qualitative modifications of the fibrosis tissue and an increasing risk of complications such as hepatocellular carcinoma (HCC). Our purpose was to identify by FTIR imaging the spectral characteristics of hepatic fibrosis in cirrhotic patients with and without HCC. FTIR images were collected at projected pixel sizes of 25 and 2.7 μm from paraffinized hepatic tissues of five patients with uncomplicated cirrhosis and five cirrhotic patients with HCC and analyzed by k-means clustering. When compared to the adjacent histological section, the spectral clusters corresponding to hepatic fibrosis and regeneration nodules were easily identified. The fibrosis area estimated by FTIR imaging was correlated to that evaluated by digital image analysis of histological sections and was higher in patients with HCC compared to those without complications. Qualitative differences were also observed when fibrosis areas were specifically targeted at higher resolution. The partition in two clusters of the fibrosis tissue highlighted subtle differences in the spectral characteristics of the two groups of patients. These data show that the quantitative and qualitative changes of fibrosis tissue occurring during the course of cirrhosis are detectable by FTIR imaging, suggesting the possibility of subclassifying cirrhosis into different steps of severity.
Highlights
Cirrhosis is defined as an advanced stage of diffuse liver fibrosis in response to a chronic injury causing lobular disorganization with formation of fibrous septa and nodules of morphologically and Molecules 2020, 25, 4092; doi:10.3390/molecules25184092 www.mdpi.com/journal/moleculesMolecules 2020, 25, 4092 functionally abnormal hepatocytes [1]
Analysis of Spectral Images of Whole Hepatic Samples Acquired with a Projected Pixel Size of 25 μm Histological and spectral images of two hepatic samples from patients with uncomplicated cirrhosis and cirrhosis complicated by hepatocellular carcinoma (HCC) are presented as examples in Figures 1 and 2
(A) Histological section stained with Masson’s trichrome stain showing fibrosis in green. (B) Digital image analysis with fibrosis area in red. (C–F) K-means clustering of Fourier transform infrared (FTIR) spectral images of an adjacent
Summary
Cirrhosis is defined as an advanced stage of diffuse liver fibrosis in response to a chronic injury causing lobular disorganization with formation of fibrous septa and nodules of morphologically and Molecules 2020, 25, 4092; doi:10.3390/molecules25184092 www.mdpi.com/journal/moleculesMolecules 2020, 25, 4092 functionally abnormal hepatocytes [1]. A histological scoring system, the Laennec system [5,6], has been proposed to subclassify cirrhosis into three categories of severity according to the width of fibrous septa and size of nodules. Using this system, it has been shown that increasing severity of cirrhosis was tightly correlated with both the clinical stage of cirrhosis and the grade of portal hypertension [7]. The incidence of hepatic decompensation and HCC significantly correlated with the histological subclassification of cirrhosis [9] This scoring system is semi-quantitative and not routinely used
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