Understanding the biokinetics of ibuprofen after single and repeated treatments in rat and human in vitro liver cell systems

Truisi, Germaine L.; Consiglio, Emma Di; Parmentier, Céline; Savary, Camille; Pomponio, Giuliana; Bois, Frédéric; Lauer, Birthe; Jossé, Rozenn; Hewitt, Philip; Mueller, Stefan O.; Richert, Lysiane; Guillouzo, André; Testai, Emanuela

doi:10.1016/j.toxlet.2015.01.006

Cited by 29 publications

(26 citation statements)

References 54 publications

(7 reference statements)

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“…The Relay method (Di and Obach, 2015), which involves transferring the supernatant from hepatocyte incubations to freshly thawed hepatocytes at the end of the incubation time, is used to prolong the total incubation time up to 20h depending how many transfer cycles are performed. However, results from other studies, demonstrating that for the clearance identification and for in vitro in vivo extrapolation purposes it is necessary to know the time course of the concentration in the cells relevant for toxicity and not only the concentration-time course in the supernatant (Truisi et al 2015;Mielke et al, 2017) indicates that the "relay method" has some limitations depending on the intended application for which it is used. The "relay method" should be considered as a model to qualitatively evaluate low clearance compounds while for quantitative measurements other cellular-based models may be more suitable.…”

Section: Accepted Manuscriptmentioning

confidence: 99%

Establishing a systematic framework to characterise in vitro methods for human hepatic metabolic clearance

Gouliarmou¹,

Lostia²,

Coecke³

et al. 2018

Toxicology in Vitro

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Hepatic metabolic clearance is one of the most important factors driving the overall kinetics of chemicals including substances used in various product categories such as pesticides, biocides, pharmaceuticals, and cosmetics. A large number of in vitro systems from purified isozymes and subcellular organelles to hepatocytes in simple cultures and in complex scaffold setups are available for measuring hepatic metabolic clearance for different applications. However, there is currently no approach for systematically characterising and comparing these in vitro methods in terms of their design, applicability and performance. To address this, existing knowledge in the field of in vitro human hepatic metabolic clearance methods was gathered and analysed in order to establish a framework to systematically characterise methods based on a set of relevant components. An analogous framework would be also applicable for non-human in vitro systems. The components are associated with the biological test systems used (e.g. subcellular or cells), the in vitro method (e.g. number of cells, test item solubility), related analytical techniques, data interpretation methods (based on substrate depletion/metabolite formation), and performance assessments (precision and accuracy of clearance measurements). To facilitate the regulatory acceptance of this class of methods, it is intended that the framework provide the basis of harmonisation work within the OECD.

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Section: Accepted Manuscriptmentioning

confidence: 99%

Establishing a systematic framework to characterise in vitro methods for human hepatic metabolic clearance

Gouliarmou¹,

Lostia²,

Coecke³

et al. 2018

Toxicology in Vitro

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“…Despite this, many in vitro studies are performed between 24 and 48 h, therefore exposing the cells to a range of acute doses not comparable to therapeutic concentrations (Germano et al 2014). Repeated-dose administration is a more relevant scenario for therapeutics, being usually evaluated in in vivo studies which yield information on general characteristics of the toxicity, delayed responses and/or cumulative effects (Truisi et al 2015). To mimic long-term repeated exposures in vitro, cell models which retain hepatic phenotype for a sufficiently long time should be used.…”

Section: Discussionmentioning

confidence: 99%

Long-term and mechanistic evaluation of drug-induced liver injury in Upcyte human hepatocytes

et al. 2018

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Drug-induced liver injury (DILI) constitutes one of the most frequent reasons of restricted-use warnings as well as withdrawals of drugs in postmarketing and poses an important concern for the pharmaceutical industry. The current hepatic in vivo and in vitro models for DILI detection have shown clear limitations, mainly for studies of long-term hepatotoxicity. For this reason, we here evaluated the potential of using Upcytes human hepatocytes (UHH) for repeated-dose long-term exposure to drugs. The UHH were incubated with 15 toxic and non-toxic compounds for up to 21 days using a repeated-dose approach, and, in addition to conventional examination of effects on viability, the mechanisms implicated in cell toxicity were also assessed by means of high-content screening. The UHH maintained the expression and activity levels of drug-metabolizing enzymes for up to 21 days of culture and became more sensitive to the toxic compounds after extended exposures, showing inter-donor differences which would reflect variability among the population. The assay also allowed to detect the main mechanisms implicated in the toxicity of each drug as well as identifying special susceptibilities depending on the donor. UHH can be used for a long-term repeated detection of DILI at clinically relevant concentrations and also offers key mechanistic features of drug-induced hepatotoxicity. This system is therefore a promising tool in preclinical testing of human relevance that could help to reduce and/or replace animal testing for drug adverse effects.

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“…Primary human hepatocytes (PHHs) are the gold standard with respect to in vitro metabolic activity; thus, it useful to comparatively scrutinize the activity of the cells discussed herein with PHH cultures. EROD activity values for fresh PHHs are between 0.14 and 0.96 pmol/min/mg protein, 24 hours post seeding (Alexandre et al, ; Truisi et al, ); values for cryopreserved PHH cultures vary between 1.68 and 6.73 pmol/min/mg protein 96 hours post‐thawing (Roymans et al, ). PHH CYP 3A4 activity levels, measured by testosterone 6β‐hydroxylation, are between 26.6 and 67.4 pmol/min/mg protein 96 hours post‐thawing (Roymans et al, ); one study noted 55.0 pmol/min/mg protein for freshly cultured cells (Lübberstedt et al, ).…”

Section: Discussionmentioning

confidence: 99%

The development and prevalidation of an in vitro mutagenicity assay based on MutaMouse primary hepatocytes, Part I: Isolation, structural, genetic, and biochemical characterization

Cox

Zwart

Luijten

et al. 2018

Environ and Mol Mutagen

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To develop an improved in vitro mammalian cell gene mutation assay, it is imperative to address the known deficiencies associated with existing assays. Primary hepatocytes isolated from the MutaMouse are ideal for an in vitro gene mutation assay due to their metabolic competence, their “normal” karyotype (i.e., neither transformed nor immortalized), and the presence of the MutaMouse transgene for rapid and reliable mutation scoring. The cells were extensively characterized to confirm their utility. Freshly isolated cells were found to have a hepatocyte‐like morphology, predominantly consisting of binucleated cells. These cells maintain hepatocyte‐specific markers for up to 3 days in culture. Analyses revealed a normal murine hepatocyte karyotype with a modal ploidy number of 4 n . Fluorescence in situ hybridization analysis confirmed the presence of the lambda shuttle vector on chromosome 3. The doubling time was determined to be 22.5 ± 3.3 h. Gene expression and enzymatic activity of key Phase I and Phase II metabolic enzymes were maintained for at least 8 and 24 h in culture, respectively. Exposure to β‐naphthoflavone led to approximately 900‐ and 9‐fold increases in Cyp1a1 and Cyp1a2 gene expression, respectively, and approximately twofold induction in cytochrome P450 (CYP) 1A1/1A2 activity. Exposure to phenobarbital resulted in an approximately twofold increase in CYP 2B6 enzyme activity. Following this characterization, it is evident that MutaMouse primary hepatocytes have considerable promise for in vitro mutagenicity assessment. The performance of these cells in an in vitro gene mutation assay is assessed in Part II. Environ. Mol. Mutagen. 60:331–347, 2019. © 2018 The Authors. Environmental and Molecular Mutagenesis published by Wiley Periodicals, Inc. on behalf of Environmental Mutagen Society.

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Understanding the biokinetics of ibuprofen after single and repeated treatments in rat and human in vitro liver cell systems

Cited by 29 publications

References 54 publications

Establishing a systematic framework to characterise in vitro methods for human hepatic metabolic clearance

Establishing a systematic framework to characterise in vitro methods for human hepatic metabolic clearance

Long-term and mechanistic evaluation of drug-induced liver injury in Upcyte human hepatocytes

The development and prevalidation of an in vitro mutagenicity assay based on MutaMouse primary hepatocytes, Part I: Isolation, structural, genetic, and biochemical characterization

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