Introduction

Chronic liver disease (CLD) is a disease of the liver that involves progressive destruction and regeneration of the liver parenchyma, leading to fibrosis and cirrhosis. Although liver fibrosis and cirrhosis have been considered irreversible, it has recently been suggested that even advanced fibrosis and cirrhosis may be reversible. The use of platelet-rich plasma (PRP) in tissue regeneration is a developing area of research for clinicians and specialists. Platelets contain proteins required for hemostasis as well as numerous growth factors. They have a prominent role in suppressing the progression of liver fibrosis in vitro and in vivo. Elevating growth factors induced by platelet transfusion can enhance liver function in patients with CLD and cirrhosis. However, PRP could increase the risk of rejection or immune response in allogenic transfusion; therefore, searching for effective therapeutic strategies is much needed.

Aims & Methods

This study evaluated the possible regenerative effect of purified platelet-derived growth factors on an experimentally induced model of liver cirrhosis. Purified Platelet Derived Growth Factors were prepared using a patented method (Gooris & Fahmy, 2019). Twenty-eight mice were included and categorized into four groups. The 1^st group acted as a normal control group. The 2^nd group acted as a Purified Platelet Derived Growth Factors control group and was injected subcutaneously with Purified Platelet Derived Growth Factors only twice weekly for eight weeks. The 3^rd group acted as a pathological control group and was injected intraperitoneally with thioacetamide (TAA) three times a week for eight weeks. At the same time, the 4^th group was injected with both Purified Platelet Derived Growth Factors and TAA for eight weeks. After eight weeks, animals were euthanized, and liver specimens and blood samples were collected for pathological and biochemical assessment.

Results

Our results showed that Purified Platelet Derived Growth Factors administration could improve the hepatic status of the mice, as cirrhotic livers were regenerated back into normal-appearing parenchyma when compared to the pathological control group. In addition, the levels of ALT, AST, and total bilirubin decreased significantly (P<0.001), and the Relative expression of hepatic Alpha-SMA was significantly down-regulated (P<0.001) after Purified Platelet Derived Growth Factors administration.

Conclusion

Purified Platelet Derived Growth Factors can be used as a new therapeutic modality for liver cirrhosis.

References

• Gooris T and Fahmy HM (2019): A method for preparing a growth factors containing platelet releasate. United States patent application US 16/462,452. Oct 24.

Introduction

Alcoholic liver cirrhosis (ALC) is a complex condition marked by numerous complications, often leading to a poor prognosis and high mortality rates^​. This study evaluates the clinical characteristics, causes of hospitalization, and factors associated with prolonged hospital stay and in-hospital mortality among patients with alcoholic liver cirrhosis.

Aims & Methods

A retrospective study was conducted on 357 patients diagnosed with alcoholic liver cirrhosis in a tertiary gastroenterology department. The mean age was 57.8 ± 9.8 years, and 85% were male. Data on causes of hospital admission, length of stay, and clinical complications were analyzed. Associations with prolonged hospitalization (defined as >3 days) and in-hospital mortality were assessed using univariate and multivariate statistical analyses.

Results

The overall in-hospital mortality rate was 21.5%. The most common causes of admission included urinary tract infections (57.7%), hepato-renal syndrome (28.2%), digestive hemorrhage (22.6%), acute alcoholic hepatitis (19.3%), spontaneous bacterial peritonitis (8.4%), and pulmonary infections (5.6%). The median hospital stay was 6.6 days.
Variables significantly associated with prolonged hospitalization in univariate analysis included ascites (p<0.0001), spontaneous bacterial peritonitis (p=0.01), hepatic encephalopathy (p<0.0001), urinary tract infections (p<0.0001), and hepato-renal syndrome (p=0.09). In multivariate analysis, only urinary tract infection was independently associated with prolonged hospital stay.
Regarding in-hospital mortality, significant associations in univariate analysis were found with spontaneous bacterial peritonitis (p<0.0001), acute alcoholic hepatitis (p<0.0001), digestive hemorrhage (p<0.0001), and hepato-renal syndrome (p<0.0001). In multivariate analysis, only hepato-renal syndrome remained independently associated with mortality (p<0.0001).

Conclusion

Urinary tract infection is an independent predictor of prolonged hospitalization in patients with alcoholic liver cirrhosis, while hepato-renal syndrome is the only independent predictor of in-hospital mortality. These findings highlight the importance of early recognition and management of infections and renal complications in this high-risk population.

Introduction

Hepatocyte sheet (HS) transplantation is expected to be an effective therapeutic strategy as an alternative to liver transplantation. We have developed a unique HS formation technique and have conducted study on the improvement of liver function (Miyamoto et al., Regen Ther. 2021). However, although HS has shown improvement in liver function for a short period of time, long-term functional improvement has not been achieved. This may be due to the lack of multidimensional structures in existing hepatocyte sheets, although the liver maintains various functions by possessing unique structures such as sinusoids and biliary ducts. In fact, the lack of biliary structures in the HS may induce liver damage due to bile stasis. (Fujii et al., J Tissue EngRegen Med, 2018). In other words, the formation of bile drainage mechanism is considered an important issue to maintain long-term liver function. In parallel, we have established a biliary drainage system with a luminal structure in vitro using chemically induced hepatic progenitor cells (CLiPs) (Huang et al., Biotechnol Bioeng. 2021).

Aims & Methods

In this study, we attempted to develop a new novel Hepato-Biliary Sheet (HBS) based on our biliary drainage system and evaluated its structure and hepatic function.Mouse embryonic fibroblasts (MEFs) and GFP-positive rat-derived CLiPs were sequentially seeded in temperature-responsive culture dishes and cultured for 2 weeks to induce the biliary drainage system. HBS was formed by seeding and culturing primary mature rat hepatocytes. The HBS was then implanted on the liver surface of a mouse model of liver failure, which was created by administration of Retrorsine and 70% hepatectomy, and its structure and function were evaluated in vivo.

Results

First, immunofluorescence image analysis of the prepared HBS revealed a structure consisting of CLiP-derived bile ducts (GFP-positive) and mature hepatocytes (Alb-positive, Glutamine-positive). To evaluate the incorporation of bile into the HBS, a fluorescent reagent (Cell Tracker Orange) as a marker substance was added to culture medium and bile excretion via hepatocytes bile duct cells was observed 24 hours later. Compared to HS prepared from mature rat hepatocytes alone, drug metabolism (CYP3A4 activity) and albumin production were significantly improved. CYP3A4 activities in HBS and HS were 139.3±59.9 and 75.4±47.1 (LUM/hr) (p<0.0001) and albumin productions were 1.5±0.5×104 and 1.1±0.1 x104 (ng /day) (p=0.005), respectively. Next, when the HBS was transplanted to the liver surface in non-liver failure mouse, it was confirmed that a bile duct-like structure (CK7-positive) was maintained and that mature hepatocytes (ALB-positive) were adjacent to the bile duct-like structure, and that vascular components (CD34-positive) derived from mice were infiltrated, confirming long-term cell viability, at least 1 month. Finally, to evaluate the effect of transplantation in a mouse model of liver failure, rat albumin concentration in blood was measured, and the HBS transplantation group showed higher albumin concentration than the HS group at 7 days (9.5×10^5±8.2×10^5 ng/mL vs 5.8×10^5±6.1×10^4ng/mL, P<0.05) and 28 days (1.1×10^6±9.2×10^5 ng/mL vs 3.2×10^5±3.2×10^5 ng/mL, P<0.05),respectively.

Conclusion

We have observed formations of hepatocyte and bile duct structure in HBS which is confirmed to have biliary drainage and drug metabolizing activities. HBS are expected to improve liver function, and to play a valuable role in the treatment of liver diseases through regenerative medicine.

References

Miyamoto D, Sakai Y, Huang Y, et al. Functional changes of cocultured hepatocyte sheets subjected to continuous liver regeneration stimulation in cDNA-uPA/SCID mouse: Differences in transplantation sites. Regen Ther. 2021;18:7-11.
Fujii M, Yamanouchi K, Sakai Y, et al. In vivo construction of liver tissue by implantation of a hepatic non-parenchymal/adipose-derived stem cell sheet. J Tissue Eng Regen Med. 2018;12:287-295.
Huang Y, Sakai Y, Hara T, et al. Bioengineering of a CLiP‐derived tubular biliary‐duct‐like structure for bile transport in vitro. Biotechnol Bioeng. 2021;118:2572-2584

Disclosure

This study was supported in part by the research grant from Eisai Co., Ltd.

Introduction

Liver cirrhosis is a late stage of liver fibrosis characterized by progressive deterioration of liver function, leading to potentially life-threatening complications. Recent studies focusing on bacterial alterations of gut microbiota have shown their association with liver disease pathogenesis, decompensation, and hospitalization rates. Recently, there has been increasing recognition of the potential role of gut fungal changes in liver cirrhosis, however, further investigation in this field is warranted.

Aims & Methods

The aim of this study was to determine gut fungi composition changes in patients with liver cirrhosis and to assess whether these changes are related to liver disease severity. For the analysis were used DNA samples, extracted from the liver cirrhosis patients’ and healthy controls’ stool samples. Internal transcribed spacer (ITS) was amplified and sequenced using MiSeq Illumina sequencing machine. Bioinformatical and statistical analysis were performed to reveal the fungal profile of each study group.

Results

Forty-five patients with liver cirrhosis and 45 healthy controls were included in the study. No significant differences in age or gender were observed between the cirrhosis and control groups. Cirrhosis patients showed significantly reduced fungal richness compared to the control cohort. However, no difference was observed in the Shannon alpha diversity index. No significant alterations in fungal compositions were noted concerning gender and etiology within the cirrhosis group. Patients who had Child-Pugh B and C liver cirrhosis had a significant shift in stool fungi composition compared to those in the Child-Pugh A group. Patients with Child C liver cirrhosis had a significantly increased fungi abundance compared with the Child A group. In accordance with the MELD score, patients with score ≥15, had significantly increased richness and fungal diversity. Furthermore, an elevated abundance of the Candida genus was observed in liver cirrhosis patients group, while Saccharomyces genus was increased in the control group.

Conclusion

Patients with liver cirrhosis had reduced richness in stool fungal composition and an increased abundance of the Candida genus when compared with healthy controls. Moreover, patients with worse liver function had significant shifts in fecal fungal composition, suggesting that fungal alterations may be associated with cirrhosis progression.

Introduction

This study aims to investigate the causal estimates of the correlation between liver cirrhosis and colorectal cancer (CRC) and assess the mediation effect of modifiable risk factors direct to clinical treatments.

Aims & Methods

We performed a two-step mendelian randomization (MR) analysis using publicly accessible summary-level data on liver cirrhosis and modifiable risk factors from databases except for the FinnGen, as well as CRC from the FinnGen. Single-nucleotide polymorphisms (SNPs) significantly related to exposures were selected as instrumental variables. A random-effects inverse variance weighted (IVW) method was used in the primary analysis and sensitivity analyses were complemented to ensure the reliability of causal associations. Gene set enrichment analysis (GSEA) was performed to explore the potential biological mechanisms of the causal associations.

Results

In the IVW analysis, liver cirrhosis was causally associated with CRC without adjusting for confounding factors (OR = 1.193, 95%CI = 1.008, 1.411, p =0.040). Multivariate MR analysis showed that body mass index, whole body fat mass, waist to hip ratio (WHR) and alcohol consumption were independent risk factors of CRC (p values < 0.05). The estimate of the mediator, WHR, is 0.013 and the standard error is 0.006. WHR explained 7.45% of the total effect of cirrhosis on CRC risk. GSEA analysis identified two intersection genes which worked both in liver cirrhosis and CRC tissues, namely SERPINE1 and ARKND1. The potential biological mechanisms might include epithelial–mesenchymal transition, coagulation, inflammatory response, and etc.

Table 1 The mediator effect of modified risk factors on the causal association between liver cirrhosis and colorectal cancer.

Conclusion

Genetically predicted liver cirrhosis had potentially risk causal effect on CRC with the mediation by WHR. SERPINE1 and ARKND1 might be candidate targets for preventing CRC in patients with liver cirrhosis.