Characterization of Pro‐apoptotic Effect of Liver Failure Plasma on Primary Human Hepatocytes and Its Modulation by Molecular Adsorbent Recirculation System Therapy

Abstract

Plasma from patients with liver failure may contain toxic molecules that cause hepatocyte apoptosis and worsen liver disease, suggesting that removal of pro-apoptotic factors is an appropriate therapeutic strategy. We investigated the apoptosis of human hepatocytes induced by plasma from patients with both acute and acute-on-chronic liver disease, and the effect of molecular adsorbent dialysis (molecular adsorbent recirculation system [MARS] dialysis) on this. Apoptotic effects of acute and acute-on-chronic liver failure plasmas from 46 patients were assessed on cultured primary human hepatocytes using terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) labeling and caspase 3 activation. In 11 patients undergoing MARS dialysis, the pro-apoptotic effect of their plasma was analyzed before and after therapy. Acute liver failure plasma induced more apoptosis than normal plasma (within 4-6 h of culture, a 2.5-fold increase by TUNEL labeling, 1.8-fold by caspase 3 activation), via a pathway involving caspase 8, suggesting involvement of the death-receptor pathway. However, not all acute liver failure plasmas were significantly more pro-apoptotic than normal plasma. Plasma from patients with acutely decompensated chronic liver disease induced apoptosis at the same rate as normal plasma. MARS dialysis improved biochemical parameters indicating effective removal of albumin-bound molecules, but the apoptotic effects of the plasma were unchanged. Thus, plasma of patients with acute liver failure, compared to normal plasma, induced increased apoptosis of primary human hepatocytes by a caspase-8- and caspase-3-dependent pathway. The apoptosis induced in the presence of liver failure plasma was not reduced by MARS dialysis.

HepaRG-Progenitor Cell Derived Hepatocytes Cultured in Bioartificial Livers Are Protected from Healthy- and Acute Liver Failure-Plasma Induced Toxicity

“…The applied 3D culture platform provides a flexible culture surface, which may be essential for maintaining cell-cell contact. Loss of adhesion is one of the first events in plasma-induced toxicity, and anchorage proteins, such as E-cadherin and the beta1-integrin receptor are known to protect against hepatocyte dedifferentiation and apoptosis [28][29][30][31]. In addition, matrix rigidity is known to limit hepatocyte differentiation, in part through transcription factor HNF4A [32].…”

“…Due to the scarcity of human ALF plasma there was no opportunity to assess the effect of plasma from ALF patients. In this study we used a model of paracetamol-induced liver failure which is commonly associated with mitotoxicity and disruption of cell tight junctions [33], although also ALF-plasma of other origins has been reported to impair mitochondrial activity to varying degrees [31,34]. However, the detrimental effects of ALF-plasma are likely to vary between etiologies, patients and clinical status, implicating the necessity of close monitoring of biocomponent functionality during therapy.…”

Artificial Organs 2007: A Year in Review

“…Rebecca Saich et al. (55) of the Royal Free & University College Medical School, Hampstead Campus (London, UK) investigated the apoptosis of human hepatocytes induced by plasma from patients with both acute and acute‐on‐chronic liver disease, and the effect of molecular adsorbent dialysis (MARS dialysis) on this. Apoptotic effects of plasmas from 46 patients were assessed on cultured primary human hepatocytes.…”

Structure and biocompatibility of poly(vinyl alcohol)-based and agarose-based monolithic composites with embedded divinylbenzene-styrene polymeric particles

“…Apart from extracorporeal blood purification, the removal of toxic metabolites from human or animal plasma is challenging in view of production of cell growth media. In particular, removal of toxic metabolites related to liver failure or pro-inflammatory cytokine tumor necrosis factor is a need for the studies performed with cultured hepatocytes (Saich et al 2007; Jones and Czaja 1998). The above production processes and research applications are required for adsorbent devices with low resistance to the flow of plasma combined with effective adsorption of various toxins.…”