Study of liver in HBV-related hepatocellular carcinoma: Stereology shows quantitative differences in liver structure

Abstract

Hepatocellular carcinoma is one of the main consequences of liver chronic disease. Hepatocellular carcinoma-related changes may be seen in patients with chronic hepatitis B. The aim of the current study was to quantitate liver tissue elements by stereological technique in patients with hepatitis B-related cancer and compare the results with control and only hepatitis B group. Needle liver biopsies from 40 patients with only chronic hepatitis B infection, from 41 patients with only early hepatocellular carcinoma, from 40 patients with early hepatitis B-related cancer and 30 healthy subjects (control group) were analyzed by stereological method using systematic uniform random sampling method. Haematoxylin and eosin stained sections were used. The numerical density of hepatocytes, hepatocyte volume, numerical density of Kupffer cells, volume density of the connective tissue in the portal space, and volume density of the connective tissue were assessed. Quantitative analysis of liver samples indicated that there were statistically significant differences in the numerical density of hepatocytes, hepatocyte volume, numerical density of Kupffer cells, volume density of the connective tissue in the portal space, and volume density of the connective tissue between control and hepatitis B-related cancer and hepatitis B groups. Quantitative, stereological technique is simple and reliable for evaluating HCC in chronic hepatitis B. It is useful for assessing the liver tissue parameters. Stereology is recommended for the diagnosis of people prone to cancer in patients with chronic hepatitis B.

Figure 1.

A light microscopic image from a selected liver section after superimposing a point-counting grid on it. The image shows a section of liver from the control healthy group.

Figure 2.

Stereological estimation of the number of hepatocytes by the Cavalieri-physical disector combination method. The image shows a section of liver from the control healthy group. In the counting frame, all the profiles trapped completely inside the frame and the profiles hit by the inclusion edge (green lines) must be counted (shown with red star). However, any profile hitting the exclusion edges and their extensions (shown in red) must be excluded from counting.

Figure 3.

Stereological estimation of the number of Kupffer cells by the Cavalieri-physical disector combination method. The image shows a section of liver from the control healthy group. In the counting frame, all the profiles trapped completely inside the frame and the profiles hit by the inclusion edge (green lines) must be counted (shown with red star). However, any profile hitting the exclusion edges and their extensions (shown in red) must be excluded from counting.

Figure 4.

Light-microscopic images of Mayer’s Hematoxylin-stained optical sections of liver from the control healthy subjects (A), only chronic HBV infection (B), only early hepatocellular carcinoma (C), and early HBV-related HCC (D) groups depicting details of the liver microstructure. B) Chronic hepatitis B pattern with significant lobular inflammation, ballooned hepatocytes; this biopsy shows severe inflammation with necrosis in numerous area and loss of architecture. C) Feature of low grade nodule showing a slight increase in cellularity, compared with the control tissue without architectural atypia (A). D) Growth patterns of hepatocellular carcinoma with HBV infection, trabecular pattern with pseudoglandular growth pattern and giant cell formation are obvious. H&E: 400x.

Figure 5.

Numerical density of hepatocytes (n×10³) / mm, in control (C), only chronic HBV (HBV), only early hepatocellular carcinoma (HCC) and early HBV-related HCC (HBV+HCC). Coefficient of error for point counting is less than 0.05. ^aP<0.001 compared to control group; ^bP<0.05 compared to HBV group; ^cP<0.05 compared to HCC group; ^dP<0.05 compared to control group.

Figure 6.

Volume of hepatocytes (μm), in control (C), only chronic HBV (HBV), only early hepatocellular carcinoma (HCC) and early HBV-related HCC (HBV+HCC). Coefficient of error for point counting is less than 0.05. ^aP<0.001 compared to control group; ^bP<0.05 compared to HBV group; ^cP<0.05 compared to HCC group; ^dP<0.05 compared to control group.

Figure 7.

Numerical density of Kupffer cells (n×10³) / mm, in control (C), only chronic HBV (HBV), only early hepatocellular carcinoma (HCC) and early HBV-related HCC (HBV+HCC). Coefficient of error for point counting is less than 0.05. ^aP<0.001 compared to control group; ^bP<0.05 compared to HBV group; ^cP<0.05 compared to HCC group; ^dP<0.05 compared to control group.

Figure 8.

Volume density of the connective tissue in the portal space, in control (C), only chronic HBV (HBV), only early hepatocellular carcinoma (HCC) and early HBV-related HCC (HBV+HCC). Coefficient of error for point counting is less than 0.05. ^aP<0.001 compared to control group; ^bP<0.05 compared to HBV group; ^cP<0.05 compared to HCC group; ^dP<0.05 compared to control group.

Figure 9.

Volume density of the connective tissue, in control (C), only chronic HBV (HBV), only early hepatocellular carcinoma (HCC) and early HBV-related HCC (HBV+HCC). Coefficient of error for point counting is less than 0.05. ^aP<0.001 compared to control group; ^bP<0.05 compared to HBV group; ^cP<0.05 compared to HCC group; ^dP<0.05 compared to control group.