{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,28]],"date-time":"2026-04-28T03:47:33Z","timestamp":1777348053373,"version":"3.51.4"},"reference-count":38,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,4,28]],"date-time":"2026-04-28T00:00:00Z","timestamp":1777334400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Toxicol."],"abstract":"<jats:p>\n                    Drug-induced liver injury (DILI) is a major cause of drug development failure and market withdrawal. Despite the use of animal and human-derived preclinical models, none reliably predict human hepatotoxicity. Conventional animal studies, based on group-level averages, overlook inter-individual variability critical to idiosyncratic DILI. Although several alternatives to animal testing are available, implementation of alternative methodologies into safety evaluations is very slow and to date, no standalone validated alternative models to assess systemic toxicity exist. To address this, we developed a novel serum-educated rat liver spheroid model that captures individual metabolic diversity, allowing DILI to be assessed both\n                    <jats:italic>in vitro<\/jats:italic>\n                    and\n                    <jats:italic>in vivo<\/jats:italic>\n                    within the same animal. Building on this approach, the present study aimed to validate the model\u2019s ability to detect DILI caused by well-characterized hepatotoxic drugs, including diclofenac and bosentan, in rat models. Rats were treated orally with diclofenac or bosentan for 28 days. Blood was collected pre-dosing to generate individualized spheroids containing rat hepatocytes, stellate cells, and macrophages. These spheroids were exposed to each drug across a ten-point concentration range for 3\u00a0days and cell viability was quantified using the CellTiter-Glo ATP assay. Clinical chemistry analyses of ALT, AST, ALP, albumin, and bilirubin were performed at\n                    <jats:italic>in vivo<\/jats:italic>\n                    study termination on day 29.\n                    <jats:italic>In vitro<\/jats:italic>\n                    data were analyzed using PredictCan-MIND to derive DILI severity scores and correlate\n                    <jats:italic>in vitro<\/jats:italic>\n                    cytotoxicity with\n                    <jats:italic>in vivo<\/jats:italic>\n                    biomarkers.\n                    <jats:italic>In vivo<\/jats:italic>\n                    , diclofenac caused no significant liver enzyme elevations, although some rats, exclusively females, displayed subtle toxicity revealing sex-dependent susceptibility. Corresponding\n                    <jats:italic>in vitro<\/jats:italic>\n                    spheroids confirmed hepatocellular injury in these individuals. Bosentan produced a mild cholestatic response\n                    <jats:italic>in vivo<\/jats:italic>\n                    without consistent enzyme elevation, while\n                    <jats:italic>in vitro<\/jats:italic>\n                    analysis showed clear hepatotoxicity in 8 of 10 rats. Notably,\n                    <jats:italic>in vitro<\/jats:italic>\n                    DILI severity correlated strongly with\n                    <jats:italic>in vivo<\/jats:italic>\n                    ALP levels, consistent with bosentan\u2019s known cholestatic mechanism. The serum-educated rat liver spheroid model captures inter-individual and sex-related differences in hepatotoxicity and demonstrates translational concordance with\n                    <jats:italic>in vivo<\/jats:italic>\n                    cholestatic markers. This approach improves DILI prediction, aligns with the 3Rs principle, and supports a potential 50%\u201370% reduction in animal use for preclinical liver toxicity testing.\n                  <\/jats:p>","DOI":"10.3389\/ftox.2026.1784229","type":"journal-article","created":{"date-parts":[[2026,4,28]],"date-time":"2026-04-28T03:20:00Z","timestamp":1777346400000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":0,"title":["A new in vitro rat liver platform capturing biological diversity for drug-induced liver injury assessment and reduction of animal use in drug discovery and development"],"prefix":"10.3389","volume":"8","author":[{"given":"Pralhad","family":"Wangikar","sequence":"first","affiliation":[{"name":"PRADO Pvt Ltd","place":["Pune, India"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sara","family":"Cherradi","sequence":"additional","affiliation":[{"name":"PredictCan Biotechnologies, Biop\u00f4le Eurom\u00e9decine, Cap Sigma","place":["Grabels, France"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Pradhnya","family":"Choudhari","sequence":"additional","affiliation":[{"name":"PRADO Pvt Ltd","place":["Pune, India"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Aditi","family":"Wangikar","sequence":"additional","affiliation":[{"name":"PRADO Pvt Ltd","place":["Pune, India"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Vaibhav","family":"Madiwal","sequence":"additional","affiliation":[{"name":"PRADO Pvt Ltd","place":["Pune, India"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Salom\u00e9","family":"Roux","sequence":"additional","affiliation":[{"name":"PredictCan Biotechnologies, Biop\u00f4le Eurom\u00e9decine, Cap Sigma","place":["Grabels, France"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nisha","family":"Banerjee","sequence":"additional","affiliation":[{"name":"PRADO Pvt Ltd","place":["Pune, India"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Pranav","family":"Panzade","sequence":"additional","affiliation":[{"name":"PRADO Pvt Ltd","place":["Pune, India"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Pallavi","family":"Gangarde","sequence":"additional","affiliation":[{"name":"PRADO Pvt Ltd","place":["Pune, India"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nithish","family":"Kannan","sequence":"additional","affiliation":[{"name":"PRADO Pvt Ltd","place":["Pune, India"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hong Tuan","family":"Duong","sequence":"additional","affiliation":[{"name":"PredictCan Biotechnologies, Biop\u00f4le Eurom\u00e9decine, Cap Sigma","place":["Grabels, France"]}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1965","published-online":{"date-parts":[[2026,4,28]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"85","DOI":"10.1186\/gm489","article-title":"Emerging hepatotoxicity biomarkers and their potential to improve understanding and management of drug-induced liver injury","volume":"5","author":"Aubrecht","year":"2013","journal-title":"Genome Med."},{"key":"B2","doi-asserted-by":"publisher","first-page":"715","DOI":"10.1016\/j.therap.2018.02.004","article-title":"Safety data and withdrawal of hepatotoxic drugs","volume":"76","author":"Babai","year":"2021","journal-title":"Therapie"},{"key":"B3","doi-asserted-by":"publisher","first-page":"655","DOI":"10.1093\/toxsci\/kfx289","article-title":"Comparison of hepatic 2D sandwich cultures and 3D spheroids for long-term toxicity applications: a multicenter study","volume":"162","author":"Bell","year":"2018","journal-title":"Toxicol. 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