Characterization of Chemically Induced Hepatotoxicity in Collagen Sandwiches of Rat Hepatocytes

Farkas, Dora; Tannenbaum, Steven R.

doi:10.1093/toxsci/kfi145

Cited by 34 publications

(15 citation statements)

References 34 publications

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“…In these conditions, the expression of liver specific genes, such as transporters, nuclear receptor and CYPs was stable over the whole culture duration. However, in this setting the addition of a low percentage of serum as well as the presence of EGF in the cell culture medium did not improve the cells status, despite some published evidences showing enhancement of long-term maintenance and survival of rat hepatocytes (Farkas and Tannenbaum, 2005). (Suppl.…”

Section: Discussionmentioning

confidence: 61%

Determination of liver specific toxicities in rat hepatocytes by high content imaging during 2-week multiple treatment

Germano

Uteng

Philippe

et al. 2015

Toxicology in Vitro

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DILI is a major safety issue during drug development and one of the leading causes for market withdrawal. Despite many efforts made in the past, the prediction of DILI using in vitro models remains very unreliable. In the present study, the well-established hepatocyte Collagen I-Matrigel™ sandwich culture was used, mimicking chronic drug treatment after multiple incubations for 14 days. Ten drugs associated with different types of specific preclinical and clinical liver injury were evaluated at non-cytotoxic concentrations. Mrp2-mediated transport, intracellular accumulation of neutral lipids and phospholipids were selected as functional endpoints by using Cellomics™ Arrayscan® technology and assessed at five timepoints (day 1, 3, 7, 10, 14). Liver specific functional impairments after drug treatment were enhanced over time and could be monitored by HCI already after few days and before cytotoxicity. Phospholipidosis-inducing drugs Chlorpromazine and Amiodarone displayed the same response as in vivo. Cyclosporin A, Chlorpromazine, and Troglitazone inhibited Mrp2-mediated biliary transport, correlating with in vivo findings. Steatosis remained difficult to be reproduced under the current in vitro testing conditions, resulting into false negative and positive responses. The present results suggest that the repeated long-term treatment of rat hepatocytes in the Collagen I-Matrigel™ sandwich configuration might be a suitable tool for safety profiling of the potential to induce phospholipidosis and impair Mrp2-mediated transport processes, but not to predict steatosis.

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Section: Discussionmentioning

confidence: 61%

Determination of liver specific toxicities in rat hepatocytes by high content imaging during 2-week multiple treatment

Germano

Uteng

Philippe

et al. 2015

Toxicology in Vitro

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“…Its viscosity (0.84–0.95cP at 20°C) is slightly lower than that of H 2 O (1.00cP at 20°C) even though its density is much higher. CCl 4 has also been used as co-solvent in cell culture studies [23]. …”

Section: Methodsmentioning

confidence: 99%

“…Also unlikely is the negative effect of the selected solvents on the structural integrity of collagen as they are often used as co-solvents in biological studies [19, 20, 23–26]. To further verify this, a pilot study was performed by treating demineralized bone samples (N=2) in each of the selected solvents for three days at ambient temperature and then having them tested in tension.…”

Section: Methodsmentioning

confidence: 99%

Water residing in small ultrastructural spaces plays a critical role in the mechanical behavior of bone

Samuel

Sinha

Zhao

et al. 2014

Bone

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Water may affect the mechanical behavior of bone by interacting with the mineral and organic phases through two major pathways: i.e. hydrogen bonding and polar interactions. In this study, dehydrated bone was soaked in several solvents (i.e. water, heavy water (D2O), ethylene glycol (EG), dimethylformamide (DMF), and carbon tetrachloride(CCl4)) that are chemically harmless to bone and different in polarity, hydrogen bonding capability and molecular size. The objective was to examine how replacing the original matrix water with the solvents would affect the mechanical behavior of bone. The mechanical properties of bone specimens soaked in these solvents were measured in tension in a progressive loading scheme. In addition, bone specimens without any treatments were tested as the baseline control whereas the dehydrated bone specimens served as the negative control. The experimental results indicated that 22.3±5.17vol% of original matrix water in bone could be replaced by CCl4, 71.8±3.77vol% by DMF, 85.5±5.15vol% by EG, and nearly 100% by D2O and H2O, respectively. CCl4 soaked specimens showed similar mechanical properties with the dehydrated ones. Despite of great differences in replacing water, only slight differences were observed in the mechanical behavior of EG and DMF soaked specimens compared with dehydrated bone samples. In contrast, D2O preserved the mechanical properties of bone comparable to water. The results of this study suggest that a limited portion of water (<15vol% of the original matrix water) plays a pivotal role in the mechanical behavior of bone and it most likely resides in small matrix spaces, into which the solvent molecules larger than 4.0Å cannot infiltrate.

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“…The idea of direct cell–cell and cell–matrix interactions promoting survival was suggested and subsequently demonstrated by reports that maintenance of differentiated phenotype in hepatocytes was enhanced by sandwiching mono-layers of cells between layers of extracellular matrix [37,38,74–78]. Studies of various tumor models also long since demonstrated that maintenance of differentiated phenotype was enhanced when cells form spheroids or multi-layer cell aggregates [79,80].…”

Section: Evolving Trends In Liver Tissue Engineering In Vitromentioning

confidence: 99%

Liver tissue engineering in the evaluation of drug safety

Dash

Inman

Hoffmaster

et al. 2009

Expert Opinion on Drug Metabolism & Toxicology

144

130

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Assessment of drug–liver interactions is an integral part of predicting the safety profile of new drugs. Existing model systems range from in vitro cell culture models to FDA-mandated animal tests. Data from these models often fail, however, to predict human liver toxicity, resulting in costly failures of clinical trials. In vitro screens based on cultured hepatocytes are now commonly used in early stages of development, but many toxic responses in vivo seem to be mediated by a complex interplay among several different cell types. We discuss some of the evolving trends in liver cell culture systems applied to drug safety assessment and describe an experimental model that captures complex liver physiology through incorporation of heterotypic cell–cell interactions, 3D architecture and perfused flow. We demonstrate how heterotypic interactions in this system can be manipulated to recreate an inflammatory environment and apply the model to test compounds that potentially exhibit idiosyncratic drug toxicity. Finally, we provide a perspective on how the range of existing and emerging in vitro liver culture approaches, from simple to complex, might serve needs across the range of stages in drug discovery and development, including applications in molecular therapeutics.

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Characterization of Chemically Induced Hepatotoxicity in Collagen Sandwiches of Rat Hepatocytes

Cited by 34 publications

References 34 publications

Determination of liver specific toxicities in rat hepatocytes by high content imaging during 2-week multiple treatment

Determination of liver specific toxicities in rat hepatocytes by high content imaging during 2-week multiple treatment

Water residing in small ultrastructural spaces plays a critical role in the mechanical behavior of bone

Liver tissue engineering in the evaluation of drug safety

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