{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,24]],"date-time":"2026-09-24T17:20:15Z","timestamp":1790270415511,"version":"4.1.0"},"reference-count":29,"publisher":"Oxford University Press (OUP)","issue":"1","license":[{"start":{"date-parts":[[2024,11,7]],"date-time":"2024-11-07T00:00:00Z","timestamp":1730937600000},"content-version":"vor","delay-in-days":24,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100000265","name":"Medical Research Council","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100000265","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Centre for Drug Safety Science","award":["MR\/L006758\/1"],"award-info":[{"award-number":["MR\/L006758\/1"]}]},{"name":"TransQST Consortium"},{"name":"Innovative Medicines Initiative 2 Joint Undertaking","award":["116030"],"award-info":[{"award-number":["116030"]}]},{"name":"European Union\u2019s Horizon 2020"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2025,1,1]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Acetaminophen is commonly used as a reference hepatotoxin to demonstrate that in vitro human liver platforms can emulate features of clinical drug-induced liver injury. However, the induction of substantial cell death in these models typically requires acetaminophen concentrations (\u223c10\u2009mM) far higher than blood concentrations of the drug associated with clinical hepatotoxicity (\u223c1 mM). Using the cytochrome P450 inhibitor 1-aminobenzotriazole, we show that acetaminophen toxicity in cultured human, mouse, and rat hepatocytes is not dependent on N-acetyl-p-benzoquinonimine formation, unlike the in vivo setting. This finding highlights the limitation of using acetaminophen as a reference hepatotoxin for the evaluation of in vitro liver models. Hence, we make recommendations on the selection of reference hepatotoxins for this purpose.<\/jats:p>","DOI":"10.1093\/toxsci\/kfae133","type":"journal-article","created":{"date-parts":[[2024,10,2]],"date-time":"2024-10-02T15:18:10Z","timestamp":1727882290000},"page":"35-40","source":"Crossref","is-referenced-by-count":8,"title":["Limitations of acetaminophen as a reference hepatotoxin for the evaluation of in vitro liver models"],"prefix":"10.1093","volume":"203","author":[{"given":"Lucia A","family":"Livoti","sequence":"first","affiliation":[{"name":"Department of Pharmacology & Therapeutics, Institute of Systems, Molecular & Integrative Biology, University of Liverpool , Liverpool, L69 3GE,","place":["United Kingdom"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rowena","family":"Sison-Young","sequence":"additional","affiliation":[{"name":"Department of Pharmacology & Therapeutics, Institute of Systems, Molecular & Integrative Biology, University of Liverpool , Liverpool, L69 3GE,","place":["United Kingdom"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4971-2346","authenticated-orcid":false,"given":"Dennis","family":"Reddyhoff","sequence":"additional","affiliation":[{"name":"Certara Predictive Technologies , Sheffield, S1 2BJ,","place":["United Kingdom"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ciar\u00e1n P","family":"Fisher","sequence":"additional","affiliation":[{"name":"Certara Predictive Technologies , Sheffield, S1 2BJ,","place":["United Kingdom"]},{"name":"GSK Research and Development Ltd , Stevenage, SG1 2NY,","place":["United Kingdom"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Iain","family":"Gardner","sequence":"additional","affiliation":[{"name":"Certara Predictive Technologies , Sheffield, S1 2BJ,","place":["United Kingdom"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rafael","family":"Diaz-Nieto","sequence":"additional","affiliation":[{"name":"Liverpool University Hospitals NHS Foundation Trust , Liverpool, L7 8YE,","place":["United Kingdom"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Christopher E","family":"Goldring","sequence":"additional","affiliation":[{"name":"Department of Pharmacology & Therapeutics, Institute of Systems, Molecular & Integrative Biology, University of Liverpool , Liverpool, L69 3GE,","place":["United Kingdom"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4101-1913","authenticated-orcid":false,"given":"Ian M","family":"Copple","sequence":"additional","affiliation":[{"name":"Department of Pharmacology & Therapeutics, Institute of Systems, Molecular & Integrative Biology, University of Liverpool , Liverpool, L69 3GE,","place":["United Kingdom"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2024,10,14]]},"reference":[{"key":"2024122307504207700_kfae133-B1","doi-asserted-by":"crossref","first-page":"1251","DOI":"10.1016\/0006-2952(93)90277-4","article-title":"Zonation of acetaminophen metabolism and cytochrome P450 2e1-mediated toxicity studied in isolated periportal and perivenous hepatocytes","volume":"45","author":"Anundi","year":"1993","journal-title":"Biochem Pharmacol"},{"key":"2024122307504207700_kfae133-B2","doi-asserted-by":"crossref","first-page":"215","DOI":"10.1039\/C9LC00768G","article-title":"Liver microphysiological systems development guidelines for safety risk assessment in the pharmaceutical industry","volume":"20","author":"Baudy","year":"2020","journal-title":"Lab Chip"},{"key":"2024122307504207700_kfae133-B3","doi-asserted-by":"crossref","first-page":"1501","DOI":"10.1007\/s11010-023-04814-z","article-title":"Dose-dependent effects of acetaminophen and ibuprofen on mitochondrial respiration of human platelets","volume":"479","author":"Be\u0163iu","year":"2024","journal-title":"Mol Cell Biochem"},{"key":"2024122307504207700_kfae133-B4","doi-asserted-by":"crossref","first-page":"2099","DOI":"10.1016\/0024-3205(74)90092-7","article-title":"Species differences in hepatic glutathione depletion, covalent binding and hepatic necrosis after acetaminophen","volume":"14","author":"Davis","year":"1974","journal-title":"Life Sci"},{"key":"2024122307504207700_kfae133-B5","doi-asserted-by":"crossref","first-page":"2979","DOI":"10.1007\/s00204-016-1845-1","article-title":"Evidence-based selection of training compounds for use in the mechanism-based integrated prediction of drug-induced liver injury in man","volume":"90","author":"Dragovic","year":"2016","journal-title":"Arch Toxicol"},{"key":"2024122307504207700_kfae133-B6","doi-asserted-by":"crossref","first-page":"935","DOI":"10.1016\/j.jhep.2021.06.021","article-title":"Advanced preclinical models for evaluation of drug-induced liver injury\u2014Consensus Statement by the European Drug-Induced Liver Injury Network [PRO-EURO-DILI-NET]","volume":"75","author":"Fernandez-Checa","year":"2021","journal-title":"J Hepatol"},{"key":"2024122307504207700_kfae133-B7","doi-asserted-by":"crossref","first-page":"42","DOI":"10.1016\/j.tiv.2018.12.017","article-title":"VIVD: virtual in vitro distribution model for the mechanistic prediction of intracellular concentrations of chemicals in in vitro toxicity assays","volume":"58","author":"Fisher","year":"2019","journal-title":"Toxicol In Vitro"},{"key":"2024122307504207700_kfae133-B8","doi-asserted-by":"crossref","first-page":"16084","DOI":"10.1038\/s41598-017-16491-2","article-title":"Real-time in vivo imaging reveals localised NRF2 stress responses associated with direct and metabolism-dependent drug toxicity","volume":"7","author":"Forootan","year":"2017","journal-title":"Sci Rep"},{"key":"2024122307504207700_kfae133-B9","doi-asserted-by":"crossref","first-page":"330","DOI":"10.1016\/0041-008X(83)90020-0","article-title":"Hamster hepatocytes in culture as a model for acetaminophen toxicity studies with inhibitors of drug metabolism","volume":"71","author":"Harman","year":"1983","journal-title":"Toxicol Appl Pharmacol"},{"key":"2024122307504207700_kfae133-B10","doi-asserted-by":"crossref","first-page":"439","DOI":"10.1007\/s00204-016-1694-y","article-title":"Mechanistic evaluation of primary human hepatocyte culture using global proteomic analysis reveals a selective dedifferentiation profile","volume":"91","author":"Heslop","year":"2017","journal-title":"Arch Toxicol"},{"key":"2024122307504207700_kfae133-B11","doi-asserted-by":"crossref","first-page":"961","DOI":"10.1016\/j.tiv.2008.02.001","article-title":"Interspecies differences in acetaminophen sensitivity of human, rat, and mouse primary hepatocytes","volume":"22","author":"Jemnitz","year":"2008","journal-title":"Toxicol In Vitro"},{"key":"2024122307504207700_kfae133-B12","doi-asserted-by":"crossref","first-page":"732","DOI":"10.1016\/j.tiv.2015.02.011","article-title":"The utility of HEPG2 cells to identify direct mitochondrial dysfunction in the absence of cell death","volume":"29","author":"Kamalian","year":"2015","journal-title":"Toxicol In Vitro"},{"key":"2024122307504207700_kfae133-B13","doi-asserted-by":"crossref","first-page":"575","DOI":"10.1016\/j.cld.2013.07.001","article-title":"Drug-induced acute liver failure","volume":"17","author":"Lee","year":"2013","journal-title":"Clin Liver Dis"},{"key":"2024122307504207700_kfae133-B14","doi-asserted-by":"crossref","first-page":"185","DOI":"10.1007\/978-1-59745-019-5_13","article-title":"Isolation and culture of adult mouse hepatocytes","volume":"633","author":"Li","year":"2010","journal-title":"Methods Mol Biol"},{"key":"2024122307504207700_kfae133-B15","volume-title":"Acetaminophen. Livertox\u2014clinical and research information on drug-induced liver injury","author":"LiverTox","year":"2012"},{"key":"2024122307504207700_kfae133-B16","doi-asserted-by":"crossref","first-page":"230","DOI":"10.1124\/jpet.115.223537","article-title":"A cytochrome P450-independent mechanism of acetaminophen-induced injury in cultured mouse hepatocytes","volume":"354","author":"Miyakawa","year":"2015","journal-title":"J Pharmacol Exp Ther"},{"key":"2024122307504207700_kfae133-B17","doi-asserted-by":"crossref","first-page":"21","DOI":"10.1016\/j.toxlet.2020.12.005","article-title":"Mitochondrial protein adduct and superoxide generation are prerequisites for early activation of c-Jun n-terminal kinase within the cytosol after an acetaminophen overdose in mice","volume":"338","author":"Nguyen","year":"2021","journal-title":"Toxicol Lett"},{"key":"2024122307504207700_kfae133-B18","doi-asserted-by":"crossref","first-page":"221","DOI":"10.1055\/s-0039-1679919","article-title":"Acetaminophen hepatotoxicity","volume":"39","author":"Ramachandran","year":"2019","journal-title":"Semin Liver Dis"},{"key":"2024122307504207700_kfae133-B19","doi-asserted-by":"crossref","first-page":"207","DOI":"10.1016\/j.toxlet.2016.06.1127","article-title":"Cytotoxicity evaluation using cryopreserved primary human hepatocytes in various culture formats","volume":"258","author":"Richert","year":"2016","journal-title":"Toxicol Lett"},{"key":"2024122307504207700_kfae133-B20","doi-asserted-by":"crossref","first-page":"112","DOI":"10.1093\/toxsci\/kfad085","article-title":"A systems approach reveals species differences in hepatic stress response capacity","volume":"196","author":"Russomanno","year":"2023","journal-title":"Toxicol Sci"},{"key":"2024122307504207700_kfae133-B21","doi-asserted-by":"publisher","first-page":"630","DOI":"10.1016\/j.jhep.2024.04.026","article-title":"Control compounds for preclinical drug-induced liver injury assessment: consensus-driven systematic review by the PROEURODILI network","volume":"81","author":"Segovia-Zafra","year":"2024","journal-title":"J Hepatol"},{"key":"2024122307504207700_kfae133-B22","first-page":"3917","article-title":"Isolation and primary culture of rat hepatic cells","volume":"64","author":"Shen","year":"2012","journal-title":"J Vis Exp"},{"key":"2024122307504207700_kfae133-B23","doi-asserted-by":"crossref","first-page":"523","DOI":"10.1111\/j.1533-2500.2012.00556.x","article-title":"Plasma and cerebrospinal fluid pharmacokinetic parameters after single-dose administration of intravenous, oral, or rectal acetaminophen","volume":"12","author":"Singla","year":"2012","journal-title":"Pain Pract"},{"key":"2024122307504207700_kfae133-B24","doi-asserted-by":"crossref","first-page":"1385","DOI":"10.1007\/s00204-016-1745-4","article-title":"A multicenter assessment of single-cell models aligned to standard measures of cell health for prediction of acute hepatotoxicity","volume":"91","author":"Sison-Young","year":"2017","journal-title":"Arch Toxicol"},{"key":"2024122307504207700_kfae133-B25","doi-asserted-by":"crossref","first-page":"869","DOI":"10.1016\/S0140-6736(19)31894-X","article-title":"Acute liver failure","volume":"394","author":"Stravitz","year":"2019","journal-title":"Lancet"},{"key":"2024122307504207700_kfae133-B26","doi-asserted-by":"crossref","first-page":"415","DOI":"10.1007\/BF00755791","article-title":"A microtiter plate assay for total glutathione and glutathione disulfide contents in cultured\/isolated cells: performance study of a new miniaturized protocol","volume":"10","author":"Vandeputte","year":"1994","journal-title":"Cell Biol Toxicol"},{"key":"2024122307504207700_kfae133-B27","doi-asserted-by":"crossref","first-page":"131","DOI":"10.1038\/s41573-019-0048-x","article-title":"Managing the challenge of drug-induced liver injury: a roadmap for the development and deployment of preclinical predictive models","volume":"19","author":"Weaver","year":"2020","journal-title":"Nat Rev Drug Discov"},{"key":"2024122307504207700_kfae133-B28","doi-asserted-by":"crossref","first-page":"266","DOI":"10.1016\/j.taap.2014.05.010","article-title":"Mechanisms of acetaminophen-induced cell death in primary human hepatocytes","volume":"279","author":"Xie","year":"2014","journal-title":"Toxicol Appl Pharmacol"},{"key":"2024122307504207700_kfae133-B29","doi-asserted-by":"crossref","first-page":"325","DOI":"10.1093\/toxsci\/kfs283","article-title":"Purinergic receptor antagonist A438079 protects against acetaminophen-induced liver injury by inhibiting P450 isoenzymes, not by inflammasome activation","volume":"131","author":"Xie","year":"2013","journal-title":"Toxicol Sci"}],"container-title":["Toxicological Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/academic.oup.com\/toxsci\/advance-article-pdf\/doi\/10.1093\/toxsci\/kfae133\/60481692\/kfae133.pdf","content-type":"application\/pdf","content-version":"am","intended-application":"syndication"},{"URL":"https:\/\/academic.oup.com\/toxsci\/article-pdf\/203\/1\/35\/60481692\/kfae133.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/academic.oup.com\/toxsci\/article-pdf\/203\/1\/35\/60481692\/kfae133.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,12,23]],"date-time":"2024-12-23T02:51:47Z","timestamp":1734922307000},"score":1,"resource":{"primary":{"URL":"https:\/\/academic.oup.com\/toxsci\/article\/203\/1\/35\/7885107"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,10,14]]},"references-count":29,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2024,10,14]]},"published-print":{"date-parts":[[2025,1,1]]}},"URL":"https:\/\/doi.org\/10.1093\/toxsci\/kfae133","relation":{},"ISSN":["1096-6080","1096-0929"],"issn-type":[{"value":"1096-6080","type":"print"},{"value":"1096-0929","type":"electronic"}],"subject":[],"published-other":{"date-parts":[[2025,1]]},"published":{"date-parts":[[2024,10,14]]}}}