title: Understanding the biokinetics of ibuprofen after single and repeated treatments in rat and human in vitro liver cell systems.
abstract: Common in vitro toxicity testing often neglects the fate and intracellular concentration of tested compounds, potentially limiting the predictability of in vitro results for in vivo extrapolation. We used in vitro long-term cultures of primary rat (PRH) and human hepatocytes (PHH) and HepaRG cells to characterise and model the biokinetic profile of ibuprofen (IBU) after single and daily repeated exposure (14 days) to two concentrations. A cross-model comparison was carried out at 100μM, roughly corresponding to the human therapeutic plasma concentration. Our results showed that IBU uptake was rapid and a dynamic equilibrium was reached within 1 or 2 days. All three cell systems efficiently metabolised IBU. In terms of species-differences, our data mirrored known in vivo results. Although no bioaccumulation was observed, IBU intracellular concentration was higher in PRH due to a 10-fold lower metabolic clearance compared to the human-derived cells. In HepaRG cells, IBU metabolism increased over time, but was not related to the treatment. In PHH, a low CYP2C9 activity, the major IBU-metabolising CYP, led to an increased cytotoxicity. A high inter-individual variability was seen in PHH, whereas HepaRG cells and PRH were more reproducible models. Although the concentrations of IBU in PRH over time differed from the concentrations found in human cells under similar exposure conditions.
authors.affiliation: Non-Clinical Safety, Merck Serono Research, Merck KGaA, Darmstadt, Germany; Institut für Angewandte Biowissenschaften, Karlsruhe Institute of Technology (KIT), Karlsruhe, Germany. Electronic address: Germaine.Truisi@web.de.
authors.forename: Germaine L
authors.initials: GL
authors.lastname: Truisi
authors.affiliation: Mechanism of Toxicity Unit, Environment and Primary Prevention Department, Istituto Superiore di Sanità, Rome, Italy. Electronic address: emma.diconsiglio@iss.it.
authors.forename: Emma Di
authors.initials: ED
authors.lastname: Consiglio
authors.affiliation: KaLy-Cell, 20A Rue du Général Leclerc, Plobsheim, France. Electronic address: c.parmentier@kaly-cell.com.
authors.forename: Céline
authors.initials: C
authors.lastname: Parmentier
authors.affiliation: Institut National de la Santé et de la Recherche Médicale (Inserm), UMR991, Université de Rennes 1, Rennes, France. Electronic address: camille.savary@univ-rennes1.fr.
authors.forename: Camille C
authors.initials: CC
authors.lastname: Savary
authors.affiliation: Mechanism of Toxicity Unit, Environment and Primary Prevention Department, Istituto Superiore di Sanità, Rome, Italy. Electronic address: giuliana.pomponio@gmail.com.
authors.forename: Giuliana
authors.initials: G
authors.lastname: Pomponio
authors.affiliation: Institut National de L'Environnement Industriel et des Risques, DRC/VIVA/METO, Verneuil en Halatte, France. Electronic address: frederic.bois@utc.fr.
authors.forename: Frederic
authors.initials: F
authors.lastname: Bois
authors.affiliation: Non-Clinical Safety, Merck Serono Research, Merck KGaA, Darmstadt, Germany. Electronic address: lauerbi@web.de.
authors.forename: Birthe
authors.initials: B
authors.lastname: Lauer
authors.affiliation: Institut National de la Santé et de la Recherche Médicale (Inserm), UMR991, Université de Rennes 1, Rennes, France. Electronic address: rozenn.josse@gmail.com.
authors.forename: Rozenn
authors.initials: R
authors.lastname: Jossé
authors.affiliation: Non-Clinical Safety, Merck Serono Research, Merck KGaA, Darmstadt, Germany. Electronic address: Philip.Hewitt@merckgroup.com.
authors.forename: Philip G
authors.initials: PG
authors.lastname: Hewitt
authors.affiliation: Non-Clinical Safety, Merck Serono Research, Merck KGaA, Darmstadt, Germany; Institut für Angewandte Biowissenschaften, Karlsruhe Institute of Technology (KIT), Karlsruhe, Germany. Electronic address: stefan.o.mueller@t-online.de.
authors.forename: Stefan O
authors.initials: SO
authors.lastname: Mueller
authors.affiliation: KaLy-Cell, 20A Rue du Général Leclerc, Plobsheim, France; Laboratoire de Toxicologie Cellulaire, EA4267, Université de Franche-Comté, Besançon, France. Electronic address: l.richert@kaly-cell.com.
authors.forename: Lysiane
authors.initials: L
authors.lastname: Richert
authors.affiliation: Institut National de la Santé et de la Recherche Médicale (Inserm), UMR991, Université de Rennes 1, Rennes, France. Electronic address: andre.guillouzo@univ-rennes1.fr.
authors.forename: André
authors.initials: A
authors.lastname: Guillouzo
authors.affiliation: Mechanism of Toxicity Unit, Environment and Primary Prevention Department, Istituto Superiore di Sanità, Rome, Italy. Electronic address: emanuela.testai@iss.it.
authors.forename: Emanuela
authors.initials: E
authors.lastname: Testai
mesh_terms: D000818:Animals; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000737:chemistry / Q000493:pharmacokinetics* / Q000633:toxicity; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D022781:Hepatocytes / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D007052:Ibuprofen / Q000737:chemistry / Q000493:pharmacokinetics* / Q000633:toxicity; D008099:Liver / Q000166:cytology / Q000378:metabolism*; D008297:Male; D015233:Models, Statistical; D061251:Primary Cell Culture; D051381:Rats; D017208:Rats, Wistar
chemical_list: D000894:Anti-Inflammatory Agents, Non-Steroidal; D003577:Cytochrome P-450 Enzyme System; D007052:Ibuprofen
keywords: Biokinetics; HepaRG cells; Ibuprofen; In vitro; Primary human hepatocytes; Primary rat hepatocytes