Steatosis In Liver Biopsies Research Articles

Histopathological Changes in Liver of Patients With Chronic Pancreatitis Requiring Surgical Intervention and Their Clinical Significance.

The histopathological changes in the liver and their clinical implication in chronic pancreatitis (CP) have not been studied well. We analyzed the incidence, risk factors, and long-term outcomes of these changes in CP. Chronic pancreatitis patients who underwent surgery with intraoperative liver biopsy from 2012 to 2018 formed the study group. Based on liver histopathology, 3 groups were formed: normal liver, group NL; fatty liver, group FL; and inflammation/fibrosis, group FS. The risk factors and long-term outcomes, including mortality, were evaluated. Among 73 patients, 39 (53.4%) had idiopathic, and 34 (46.6%) had alcoholic CP. The median age was 32 years, 52 (71.2%) were males and comprised NL, n = 40 (55%); FL, n = 22 (30%); and FS, n = 11 (15%). The preoperative risk factors were comparable among NL and FL groups. Overall 14 of 73 patients (19.2%) (NL, 5 of 40; FL, 5 of 22; FS, 4 of 11 [P = 0.82]) had died at median follow-up of 36 months (range, 25-85 months). The main causes of mortality were tuberculosis and severe malnutrition secondary to pancreatic insufficiency. The mortality is higher in patients with inflammation/fibrosis or steatosis in liver biopsy, and such patients need monitoring for progression of liver disease and pancreatic insufficiency.

Noninvasive Estimation of Hepatic Steatosis by Controlled Attenuation Parameter in Living Donor Liver Transplant.

Estimation of liver fat among living donor candidates is necessary before living donor liver transplant. This study aimed to investigate the usefulness of the controlled attenuation parameter compared with liver biopsy for pretransplant estimation of hepatic steatosis in living liver donors. In this retrospective study, we included all individuals who underwent transient elastography with controlled attenuation parameter and ultrasonography-guided liver biopsy as a part of donor evaluations before living donor liver transplant. Clinical and laboratory data of living donor candidates were reviewed and collected. Of 49 donor candidates included in this study, 21 (42.9%) had different degrees of hepatic macrosteatosis. Of the 21 donor candidates who had hepatic steatosis in liver biopsy, 13 individuals were diagnosed to have steatosis in transient elastography. Of the 28 donor candidates without hepatic steatosis in liver biopsy, 26 individuals showed no steatosis in transient elastography (odds ratio: 21.12; 95% CI, 3.91- 114.08; P < .001). Controlled attenuation parameter was useful in discriminating presence (P = .001) and grade of hepatic steatosis (P = .009) compared with liver biopsy with good sensitivity and specificity. The controlled attenuation parameter is a noninvasive method for detection of hepatic steatosis in living donor candidates and can be used as an adjunct to liver biopsy for screening of living donor candidates before liver transplant.

Multiscale and Multimodal Optical Imaging of the Ultrastructure of Human Liver Biopsies.

The liver as the largest organ in the human body is composed of a complex macroscopic and microscopic architecture that supports its indispensable function to maintain physiological homeostasis. Optical imaging of the human liver is particularly challenging because of the need to cover length scales across 7 orders of magnitude (from the centimeter scale to the nanometer scale) in order to fully assess the ultrastructure of the entire organ down to the subcellular scale and probe its physiological function. This task becomes even more challenging the deeper within the organ one hopes to image, because of the strong absorption and scattering of visible light by the liver. Here, we demonstrate how optical imaging methods utilizing highly specific fluorescent labels, as well as label-free optical methods can seamlessly cover this entire size range in excised, fixed human liver tissue and we exemplify this by reconstructing the biliary tree in three-dimensional space. Imaging of tissue beyond approximately 0.5 mm length requires optical clearing of the human liver. We present the successful use of optical projection tomography and light-sheet fluorescence microscopy to derive information about the liver architecture on the millimeter scale. The intermediate size range is covered using label-free structural and chemically sensitive methods, such as second harmonic generation and coherent anti-Stokes Raman scattering microscopy. Laser-scanning confocal microscopy extends the resolution to the nanoscale, allowing us to ultimately image individual liver sinusoidal endothelial cells and their fenestrations by super-resolution structured illumination microscopy. This allowed us to visualize the human hepatobiliary system in 3D down to the cellular level, which indicates that reticular biliary networks communicate with portal bile ducts via single or a few ductuli. Non-linear optical microscopy enabled us to identify fibrotic regions extending from the portal field to the parenchyma, along with microvesicular steatosis in liver biopsies from an older patient. Lastly, super-resolution microscopy allowed us to visualize and determine the size distribution of fenestrations in human liver sinusoidal endothelial cells for the first time under aqueous conditions. Thus, this proof-of-concept study allows us to demonstrate, how, in combination, these techniques open up a new chapter in liver biopsy analysis.

The correlation between fatty liver disease and serum and histological viral parameters in patients with chronic hepatitis B

Objective To investigate the correlation between non-alcoholic fatty liver disease and serum and histological viral parameters in patients with chronic hepatitis B (CHB). Methods Clinical and laboratory data from patients with CHB who received liver biopsy from 2009 to 2015 were collected. Patients were divided into steatosis and non-steatosis groups based on the presence of steatosis in liver biopsies. Propensity score matching (PSM) was conducted to adjust the confounding bias including age, sex, body mass index (BMI), total cholesterol (TC) and triglyceride (TG). Correlation of liver fatty and viral parameters was compared between steatosis and non-steatosis groups. Student t test, χ2 test, rank sum test and Pearson correlation test were employed to analyze the data. Results A total of 874 patients with a mean age of (37.0±10.1) years were enrolled in the study, with 690 males and 184 females, and 270 (30.9%) patients were diagnosed with steatosis by liver biopsy. Age, gender, BMI, TC and TG were significantly different between the two groups before PSM (all P 0.05). Serum hepatitis B virus (HBV) DNA, hepatitis B surface antigen (HBsAg) level, proportion of hepatitis B e antigen (HBeAg) positivity, HBsAg and hepatitis B core antigen (HBcAg) immunohistological staining in liver tissue were not significantly different between steatosis and non-steatosis groups after PSM (all P>0.05). Patients in steatosis group were stratified into two groups according to the degree of steatosis confirmed by liver biopsies: mild steatosis group (F1) and medium to severe steatosis group (F2-F4). The serum alanine aminotransferase (ALT), HBV DNA, HBsAg level, proportion of HBeAg positivity, immunohistological HBsAg and HBcAg staining in liver tissue between those two groups showed no differences (all P>0.05). The mean rank of liver inflammation and fibrosis in F1 group were 129.9 and 128.2, respectively, which were both significantly higher than those in F2-F4 group (105.9 and 108.5, respectively; both P<0.05). Steatosis was negatively correlated with either inflammatory grade (r=-0.183, P=0.005) or fibrosis stage (r=-0.150, P =0.020). Conclusions There is no correlation between serum viral factors and hepatic steatosis. Hepatic steatosis is not associated with the expressions of HBsAg and HBcAg in liver tissue. The severity of steatosis is negatively correlated with both liver inflammation and fibrosis. Key words: Hepatitis B, chronic; Fatty liver; Liver biopsy; Propensity score matching

Factors associated with steatosis in liver biopsies of individuals with chronic hepatitis C infection in southern Brazil

Background and study aimsInfection by the hepatitis C virus (HCV) is associated with various metabolic disorders that are collectively referred to as dysmetabolic syndrome associated with HCV. Hepatic steatosis is a common finding in chronic HCV infection and has been reported in 30–70% of patients. Here, we determine the prevalence of steatosis in patients with HCV, identify the characteristics associated with the presence of steatosis in liver biopsies and assess the association between steatosis and the severity of liver disease. Patients and methodsThis analytic cross-sectional study evaluated HCV carriers (adults) at the Gastroenterology and Hepatology Outpatient Clinic of a public university hospital between July 2013 and June 2014 using retrospective data collection. The patients were divided into two groups according to the presence or absence of steatosis in their liver biopsies. The groups were compared for the presence of risk factors for steatosis and clinical, laboratory, virological and histological characteristics. ResultsOne hundred and four patients aged 49.5±9.3years were included in the study; 56.0% of the patients were men. Steatosis was observed in 65.4% of the liver biopsies. When comparing individuals with and without steatosis, patients with steatosis exhibited a higher proportion of non-1 genotype (43.9 vs. 20.7%; p=0.034), higher median triglyceride levels (101.0 vs. 75.0; p=0.034), ferritin levels (333.0 vs. 193.5; p=0.025) and gamma glutamyl transferase levels (2.92 xULN vs. 1.87; p=0.030). Multivariate analysis demonstrated that triglyceride levels were independently associated with the presence of steatosis (OR=1.016; 95% CI 1.002–1.031; p=0.026). ConclusionsHepatic steatosis was observed in 65.4% of individuals with HCV. We observed that elevated triglyceride levels were associated independently with the presence of hepatic steatosis; we did not demonstrate an association between hepatic steatosis and histological severity.

Liver ultrasound scanning in the detection of hepatic steatosis and fibrosis in NASH patients

Abstract Background Nonalcoholic fatty liver disease (NAFLD) has become the most prevalent cause of liver disease in western countries. The development of nonalcoholic steatohepatitis (NASH) and fibrosis identifies the risk group with an increased incidence of liver-related deaths. Aim The aim of the present study is to investigate how accurately liver ultrasound (US) can contribute toward the prediction of the severity of liver damage in NAFLD, and to determine whether it may be an easily available, inexpensive, noninterventional, widely used screening method. Methods Fifty-four obese patients with variable degrees of BMI were recruited in the present study. Assessment of full medical history, anthropometric measurements, biochemical studies, abdominal US, liver biopsy for histological examination, and determination of the NAFLD activity score (NAS) score were carried out on all patients to identify NASH patients . Liver steatosis was evaluated using liver US, and graded according to a semiquantitative scale from 1 to 4. Liver histological examination was carried out to identify patients with NASH, borderline NASH, or non-NASH according to the NAS score. Results According to the NAS score, patients were divided into non-NASH patients (eight patients), borderline NASH patients (24 patients), NASH patients (20 patients), and patients with NASH and fibrosis (two patients). Alanine aminotransferase and γ-glutamyl transpeptidase were significantly higher in NASH patients. Correlating the grading of hepatic steatosis by liver US and NAS score, grade 1 was found in 37.5% of patients with non-NASH, 33.3% of patients with borderline NASH, and only in 5% of patients with NASH, whereas grade 4 steatosis was found in 20% of NASH patients and 4.2% of patients with borderline NASH; none of the non-NASH patients were diagnosed with grade 4 hepatic steatosis. The sensitivity of liver US in detecting grades of steatosis in liver biopsy was 61% in grade 1, 25% in grade 2, and 75% in grade 3. There was a direct correlation between grading of steatosis in the histological examination and the presence of NASH, P less than 0.000. Conclusion Liver US is not only sensitive in the detection of hepatic steatosis, but also in the prediction of the presence of NASH; therefore, it can be used as a simple, noninvasive, low-cost method for the screening of NAFLD and for the early identification of patients in need of aggressive intervention.

A critical analysis of three quantitative methods of assessment of hepatic steatosis in liver biopsies

The issue of adequately quantitatively evaluating hepatic steatosis is still unresolved. Therefore, we compared three methods of quantitative assessment. Two groups of mice (n = 10 each) were fed standard chow (10% fat, SC group) or a high-fat diet (60% fat, HF group) for 16 weeks, and hepatic triglyceride (HT) and liver tissue were then studied. Paraplast-embedded tissues stained by hematoxylin and eosin (H-E) were compared to frozen sections stained by Oil Red-O (ORO). In addition, the volume density of steatosis (Vv[steatosis, liver]) was measured by point counting (P-C, sections H-E or ORO) or by image analysis (I-A, sections ORO). HT was significantly higher in the HF group (104% greater, P = 0.0004) than in the SC group. With P-C and H-E, Vv[steatosis, liver] was 4.80 ± 0.90% in the SC group and 33.50 ± 3.17% in the HF group (600% greater, P < 0.0001). With P-C and ORO, Vv[steatosis, liver] was 4.86 ± 0.89% in the SC group and 25.21 ± 1.27% in the HF group (420% greater, P < 0.0001). With I-A and ORO, Vv[steatosis, liver] was 4.17 ± 0.85% in the SC group and 23.35 ± 1.58% in the HF group (460% greater, P < 0.0001). Correlations between Vv[steatosis, liver] and HT were strong and significant in all methods. In conclusion, all methods were appropriate and reproducible. In P-C and H-E, there is a slight overestimation of steatosis in the HF animals in comparison to frozen sections and ORO; in frozen sections, differences between P-C and I-A are insignificant.

Significance of Microvesicular Steatosis in Nonalcoholic Fatty Liver Disease (NAFLD)

Purpose: NAFLD is associated with accumulation of excess fat in hepatocytes visualized as macrovesicular steatosis. Some patients with NAFLD also have nonzonal aggregates of microvesicular steatosis but its significance is unknown. Diffuse microvesicular steatosis is associated with mitochondrial injury; a factor that plays an important role in the disease progression. We evaluated the frequency of microvesicular steatosis and assessed its association with histologic markers of severity in patients with NAFLD. Methods: All adult patients' biopsies in the NAFLD Database Study or pre-treatment PIVENS Trial in the NASH Clinical Research Network in the US were evaluated in a cross sectional study. Liver biopsies were reviewed centrally by the Pathology subcommittee. Microvesicular steatosis is defined as discrete, nonzonal patches of “foamy” hepatocytes with central nucleus and too numerous to count small droplets of fat. The relationship between microvesicular steatosis and different NAFLD histological features was tested by multiple logistic regression analysis after controlling for age, gender, race, BMI and diabetes. Results: 1022 liver biopsy samples were studied. 102 (10%) had documented microvesicular steatosis. Patients with and without microvesicular steatosis were comparable in age (50.2±12.5 vs. 49.5±11.9 years, p=0.56), female gender (66% vs. 63%, p=0.59), BMI (36.1±7.6 vs. 34.5±6.4 kg/m2, p=0.07) and diabetes (25% vs. 30%, p=0.36). Microvesciular steatosis was associated with higher grades of steatosis and ballooning, presence of Mallory bodies and advanced fibrosis. NAFLD patients with NASH had significantly higher microvesicular steatosis than those without NASH. Conclusion: The presence of microvesicular steatosis in liver biopsies of NAFLD was associated with greater overall degrees of activity and fibrosis. These observations suggest that microvesicular steatosis may play an important role in mediating cell injury and disease progression in NAFLD.Table: Table. Relationship between microvesicular steatosis and different histological features of NASH from multiple logistic regression analysis

Diagnosis and Quantitation of Fibrosis

Hepatic fibrosis is the final common pathway for many different liver insults. Originally considered to be irreversible, hepatic fibrosis is now known to be a dynamic process with a significant potential for resolution. The diagnosis and quantitation of fibrosis have traditionally relied on liver biopsy. However, there are a number of drawbacks including the invasive nature of the procedure, sampling error, and interobserver variability. This article reviews the current role of liver biopsy in the assessment of hepatic fibrosis and discusses the role of the newer noninvasive methods including serum markers and radiologic tests.

Impact of hepatic steatosis on viral kinetics and treatment outcome during antiviral treatment of chronic HCV infection

Liver steatosis is highly prevalent in chronic hepatitis C virus (HCV) infection, especially in patients infected with genotype 3 virus, but its significance for the outcome of antiviral treatment is not fully understood. We have monitored steatosis in liver biopsies from 231 patients with chronic HCV infection who received pegylated recombinant interferon-alpha and ribavirin in a phase III study (DITTO trial). The degree of steatosis, along with relevant metabolic parameters, was correlated with the early disappearance of virus and with the final outcome of treatment. Our data suggest that the presence of steatosis impairs the early reduction of viral load during treatment in patients infected with HCV genotype 3 and non-3. Steatosis negatively affected the final outcome of treatment mainly in patients infected with HCV genotype non-3 virus. Based on these findings, we propose that interventions aiming at reducing hepatic steatosis prior to the onset of antiviral therapy may be of benefit to patients infected with HCV of the non-3 genotypes. Patients infected with genotype 3, on the other hand, should be offered early antiviral treatment.

Semiquantitative evaluation overestimates the degree of steatosis in liver biopsies: a comparison to stereological point counting

The degree of steatosis in liver biopsies is usually assessed by a morphological semiquantitative approach in which the histopathologist uses a four-graded scale: 0–3 or none, slight, moderate and severe. Scores 1–3 are considered to correspond to fat deposition in <33, 33–66 and >66% of the hepatocytes. There is a considerable inter- and intra-individual variation in such scoring methods and a more standardized and quantitative approach is preferable. In the present study, we compare the semiquantitative technique with the stereological point counting method in the assessment of hepatic steatosis. A total of 75 archived liver needle biopsies were used. They were selected according to the original routine diagnosis of slight, moderate or severe steatosis. In all, 10 randomly selected images from each biopsy were digitized into a computer, a point grid lattice was superimposed and the number of hits on fat globules was counted. A pathologist scored the specimens in a four-graded scale as described above. The mean liver biopsy area (volume) with fat in hepatocytes was 2.2% for grade 1, 9.2% for grade 2 and 23.1% for grade 3. The kappa value for the semiquantitative estimates was 0.71 for the unweigthed kappa and 0.87 for weighted kappa. The intraclass correlation coefficient (ICC) was 0.99 for images counted twice and 0.95 when two sets of images were captured from the same biopsy. These ICCs indicate excellent agreement and above that of the semiquantitative estimates. In conclusion, the area/volume of fat content of the hepatocytes is greatly overemphasized in semiquantitative estimation. Furthermore, the point counting technique has a better reproducibility than visual evaluation and should be preferred in estimates of liver steatosis in scientific studies and in clinical contexts when the amount of steatosis is important for treatment and prognosis, such as liver transplantation.

Quantitative assessment of fibrosis and steatosis in liver biopsies from patients with chronic hepatitis C

Backgrounds—Hepatic fibrosis is one of the main consequences of liver disease. Both fibrosis and steatosis may be seen in some patients with chronic hepatitis C and alcoholic liver disease (ALD)....