Introduction

This paired biopsy study aimed to investigate the predictive value of various non-invasive scores for identifying the temporal evolution of hepatic fibrosis in patients with non-alcoholic fatty liver disease (NAFLD).

Aims & Methods

A total of 69 patients with NAFLD who had undergone two paired liver biopsies (mean interval: 21.3 ± 9.7 months) were examined. Fibrosis progression and regression were defined as an increase or decrease of at least one stage in fibrosis between the initial and second biopsy. Fibrosis-4 index (FIB-4), NAFLD Fibrosis Score (NFS), Agile 3+ score, Agile 4 score, and FibroScan-AST (FAST) score were calculated at the time of the initial biopsy.

Results

Upon comparing the results of paired biopsies, 45% of the participants (n = 31) showed no change in fibrosis, 26% (n = 18) experienced fibrosis progression, and 29% (n = 20) had evidence of fibrosis regression. In multivariable logistic regression analysis, the only independent predictive factor for progressive fibrosis was the FAST score. The odds ratio for progressive fibrosis was found to increase by 19% (95% confidence interval: 8–38%, p < 0.05) with every unit increase in FAST score at the time of the initial biopsy. No independent predictors for fibrosis regression were identified.

Conclusion

Patients with higher baseline FAST scores were more likely to experience fibrosis progression independent of potential confounders. Consequently, FAST could serve not only as a diagnostic tool for screening fibrosis in NAFLD, but also as a prognostic indicator.

Introduction

The part of NAFLD children has a progressive sub-phenotype known as nonalcoholic steatohepatitis (NASH), characterized by the ability to develop advanced fibrosis, cirrhosis, and end-stage liver disease [1]. Given the high prevalence of NAFLD and invasiveness of the liver biopsy, there is an essential need to find safe non-invasive modalities to determine liver fibrosis in children [2].

Aims & Methods

To investigate serum fibrosis markers in obese children with non-alcoholic fatty liver disease (NAFLD) depending on the presence of liver fibrosis.
94 children aged 9 to 17 years (average age 12.15 ± 2.51 years) were included in the study. According to the transient elastography data (FibroScan®502touch, Echosence, France) and body mass index, children were divided into four groups: group 1 - 27 obese NAFLD children with fibrosis ≥F1, group 2 - 35 obese NAFLD children without fibrosis, group 3 - 18 obese children without NAFLD and fibrosis. Group 4 (control) consisted of 14 children with normal weight without NAFLD and liver fibrosis. The serum levels of vascular endothelial growth factor (VEGF) (Wuhan Fine Biotech Co Ltd, China), transforming growth factor-beta 1 (TGF-β1) (IBL International, Germany), cytokeratin-18 (CK-18) (IDL Biotech AB, Sweden) were determined by enzyme-linked immunosorbent assay (ELISA). Serum content of hydroxyproline free (HPf) and hydroxyproline protein-bound (HPp/b) according to Osadchuk and glycosaminoglycans (GAG) according to Rimington were evaluated.

Results

In 1 group children, a significant increase in the median level of CK-18 (in 2.2 times, p<0.05), VEGF (in 2.4 times, p<0.05), and TGF-β1 (in 2.7 times, p<0.05) was found compared to 4 group children. Moreover, CK-18 level was significantly higher in 1 group of patients compared to 2 group (in 1.4 times, p<0.05) and 3 group (in 1.8 times, p<0.05). In all group children, an increase in the serum HPp/b content was found: in 1 group - in 1.2 times (p<0.01), in 2 group - in 1.2 times (p<0.05), in 3 group - in 1.3 times (p<0.05) compared to 4 group. The changes in the content of hydroxyproline led to an increase in the HPp/b/HPf ratio in all groups: 1.4 times (p<0.01) in 1 and 2 groups, in 1.3 times (p<0.01) in 3 group. Also, in 1 group of children median GAG level was increased by 1.3 times (p<0.001).
In children with NAFLD HPf content negatively correlated with CAP (r= - 0.408; p<0.01), CK-18 (r=-0.469; p<0.01), the ratio HPp/b/HPf positively correlated with CAP (r=0.307; p<0.05). CK-18 positively correlated with fibrosis degree (according to METAVIR) (r=0.468; p=0.008) and liver steatosis degree (r=0.357; p=0.048). A positive correlation was also found between VEGF levels and the degree of liver fibrosis (r=0.372, p=0.036).

Conclusion

Serum levels of VEGF, TGF-β1, HPp/b, and GAG were significantly elevated in children with non-alcoholic fatty liver fibrosis. Assessment of these parameters may be useful for early diagnosis of liver fibrosis in children with NAFLD.

References

1. Schwimmer JB, Johnson JS, Angeles JE, et al. Microbiome Signatures Associated With Steatohepatitis and Moderate to Severe Fibrosis in Children With Nonalcoholic Fatty Liver Disease. Gastroenterology. 2019;157(4):1109-1122. doi:10.1053/j.gastro.2019.06.028.
2. Alkhouri N. Putting it all together: Noninvasive diagnosis of fibrosis in nonalcoholic fatty liver disease in adults and children. Clin Liver Dis (Hoboken). 2017;9(6):134-137. Published 2017 Jun 29. doi:10.1002/cld.636

Introduction

It has been suggested that gut transition time (GTT) and gut microbiome could have a possible role in non-alcoholic fatty liver disease (NAFLD) development.

Aims & Methods

Our study aims to compare the GTT between healthy volunteers and NAFLD patients, regarding their microbiome composition.
Method: A total of 34 healthy volunteers and 34 NAFLD patients, without previous history of intestinal resection, inflammatory bowel disease, radiotherapy, or course of antibiotic, probiotic, and immunosuppressive therapy 2 weeks prior, were asked to provide a fresh stool sample. The gut transition time was measured by the timing of the „blue poop“ appearance after ingestion of two chocolate muffins with 1.8g of blue dye gel (dr Oetker). The microbiome composition was determined using a 16s rRNA sequencing technique with a focus on V3-V4 hypervariable region.

Results

The two groups had similar age and sex distribution. The group of NAFLD patients had significantly longer GTT when compared to healthy volunteers (36h vs 41h, p=0.0137), with a lower Bristol Stool chart score (2.32 vs 2.76, p=0.037). The main difference between gut microbiome composition in the two groups was a greater abundance of bacterial genera Faecalibacterium (0.0162 vs 0.0245, p=0.043) and bacterial family Lachnospiraceae (0.279 vs 0.331, p=0.036) in the group of healthy volunteers. NAFLD patients showed a higher abundance of Intestinibacter genera (0.00582 vs 0.003421, p=0.041). The correlation substudy showed a positive correlation between gut transition time and abundance of Methanobacteriaceae (Rho=0.231, p=0.03) and Clostridiaceae (Rho=0.197, p=0.01) bacterial families.

Conclusion

Prolonged gut transition time paralleled by gut microbiome composition disturbance might play an important role in NAFLD pathogenesis.

Introduction

Nonalcoholic fatty liver disease (NAFLD) is the most common cause of chronic liver disease worldwide, with a prevalence of 25% in the adult population. NAFLD encompasses a wide spectrum of histological conditions, from simple steatosis to nonalcoholic steatohepatitis (NASH), fibrosis and cirrhosis. Inflammation is one of the principal hallmarks of NAFLD. Polyclonal free light chains (FLCs) λ and κ reflect B-cell activation and high serum levels of FLCs are observed in a variety of inflammatory conditions.

Aims & Methods

The aim of this study is to evaluate the potential role of FLCs as biomarkers of inflammation and fibrosis in NAFLD/NASH patients. 254 patients with metabolic liver disease were consecutively enrolled in the study. 89 patients had simple steatosis, 88 had NASH, 77 had liver cirrhosis (45 with clinically significant portal hypertension [CSPH]). Diagnosis of NASH was histologically assessed. 101 patients had compensated advanced chronic liver disease (cACLD) based on histology (F>3) or elastography (liver stiffness measurement>15 kPa). Medical and pharmacological anamnesis, anthropometric measurements and laboratory tests (included λ and κ FLCs) were obtained for all patients.

Results

The concentration of total FLCs levels in patients with cirrhosis were higher compared with patients with NAFLD/NASH (p<0.01), while no significant difference in λ and κ FLCs levels were observed between NASH and NAFLD patients. Among patients who underwent biopsy, λFLCs tended to increase gradually with lobular inflammation severity, and patients with advanced fibrosis (F3-4) had a significantly higher FLCs compared with patients with mild-to-moderate fibrosis (F<3; p<0.05). Patients with clinically significant portal hypertension (CSPH) showed higher levels of FLCs compared to patients without CSPH (p<0.01). At multivariate logistic analysis both kFLCs and λFLCs are significantly associated with diagnosis of CSPH and cACLD, independently from age, sex, diabetes, dyslipidemia and obesity. Diagnostic performance of cACLD and CSPH were high for both kFLCs (AUC 0.79 and 0.87 respectively) and λFLCs (AUC 0.81 and 0.89 respectively).

Conclusion

This study highlights the role of adaptive immune system in the pathogenesis of NAFLD. FLCs may be considered useful biomarkers of advanced liver disease (cACLD and CSPH) in patients with NAFLD.

Introduction

Liver stiffness measurements has become a "gold standard" for liver fibrosis assessment. New techniques based on the same physical principles have become available, but little is known about their comparative efficacy, especially in subjects with non-alcoholic fatty liver disease.

Aims & Methods

Aim: to compare the data of liver stiffness measurement with the use of FibroScan 502 (EchoSens, France) and iLivTouch FT200 (Wuxi Hisky Medical Technologies Co., China) devices.
Method:
Subjects with non-alcoholic fatty liver disease (per EASL guidelines) who gave informed consent were eligible for the study (NCT05224037 at clinicaltrials.gov). All participants were examined on the same day with the use of both, iLivTouch FT200 (Wuxi Hisky Medical Technologies Co., China) (data are marked as LT) with a standard sensor, and Fibroscan 502 (EchoSens, France) with M or XL sensors depending on the patient's characteristics (FS). We compared the efficacy of examination and analyzed the results obtained with both devices depending on the stage of liver fibrosis. The null hypothesis was the absence of differences between the data obtained using both devices.

Results

The data of 105 people (58 women, mean age per group 52.8±12.6 years, BMI: 32.3±9.48 kg/m²) were available. Success rate for FS was 68.7 ± 29% compared to LT: 86.9±19%; p<0.001. In 5 patients, it was impossible to conduct examination with FS, while it was possible to obtained at least 10 measurements using LT (in all cases BMI was >35 kg/m²). In 1 case, it was not possible to obtain sufficient number of measurements with LT, although FS gave successful result.
There was significant direct correlation between the data of liver stiffness measurements obtained with both devices: Spearman rank R=0.56. However, comparison revealed significant difference in values: 9.6±6.6 kPa with LT vs 6.9±4.5 kPa using FS, p<0.0001.
In patients with NAFLD with advanced stages of liver fibrosis (F2+, n=29) the values of FS and LT were similar: 11.8±5.9 kPa for FS vs 13.9±7.9 kPa for LT, p=0.2. However, the values of liver stiffness differed significantly when subjects had milder stages (F0-1, n=71,) of liver fibrosis: 4.9±1.0 kPa for FS vs 7.4±3.0 kPa, p<0.001. The agreement in liver fibrosis stage assessment (using cut-off values provided by the manufacturers) were as follows: for F0 - 56.9%, for F1-42.9%, F2-43.8%, F3-66.7%, and F4-70% of cases.

Conclusion

The results of liver stiffness obtained with Fibroscan and iLivTouch correlate closely. However, the agreement between studied techniques in subjects with non-alcoholic fatty liver disease is low.To make the measurements obtained with iLivTouch more accurate it may be necessary to develop new cut-off values that corresponds with different stages of liver fibrosis.
This work was performed with financial support of Ministry of Science and Higher Education of Russia, research grant No. FGMF-2022–0005