20.500.12556/DKUM-97609 A probabilistic and context-dependent cell culture modelling framework for regulatory toxicology The persistent gap between preclinical findings and clinical outcomes highlights the limitations of current in vitro models in regulatory toxicology. While New Approach Methodologies (NAMs) promise mechanistic insight, reduced reliance on animal testing, and enhanced human relevance, their translational accuracy remains constrained by oversimplified assumptions. This manuscript identifies foundational aspects of human physiology—degeneracy, interconnected pathways, variability among individuals, biological rhythms, and context dependency—that are often underrepresented in existing systems. I propose reframing biological effects as probabilistic outcomes rather than deterministic events, recognizing that cells and organisms operate through overlapping networks where redundancy, variability, and timing shape the likelihood of specific responses. Building on this framework, I outline a probabilistic and context-dependent cell culture model that integrates viability, functional fidelity, pathway mapping, temporal resolution, and Bayesian inference. Translational relevance is further strengthened by anchoring in vitro measurements to clinically meaningful benchmarks, incorporating patient perspectives, and aligning with regulatory oversight. Although challenges remain—including replicating cell–cell communication, multi-organ synchronization, and harmonizing probabilistic outputs with deterministic regulatory frameworks—embedding these principles into NAMs offers a pathway to overcome the translational bottleneck. By embracing complexity rather than reducing it, NAMs can evolve into tools that are not only mechanistically informative but also predictive, acceptable, and implementable in real-world milieu, ultimately advancing safer and more ethical toxicological assessment. context dependency new approach methodologies (NAMs) organ-based in vitro model probabilistic modeling translational research translational toxicology true true true Frontiers Angleški jezik Angleški jezik Znanstveno delo 2026-03-25 03:13:52 2026-03-27 10:05:05 2026-03-28 03:17:21 0000-00-00 00:00:00 2026 0 0 str. 1-15 marec 2026, št. članka 1765753 Letn. 8 0000-00-00 Zaloznikova Objavljeno NiDoloceno 2026-03-03 2026-03-03 2026-03-24 60 272874499 10.3389/ftox.2026.1765753 2673-3080 RAZ_Madorran_Eneko_2026.pdf RAZ_Madorran_Eneko_2026.pdf 1 A043A2387438076CB90F09E6575CA132 55122233fd56c125471b2ef4156483368f699af30dc3439b2ebc2b3ad29daf21 c5281169-29bb-11f1-9c37-00155d0001a1 https://dk.um.si/Dokument.php?lang=slv&id=204855 https://doi.org/10.3389/ftox.2026.1765753 0 e661460d-29bb-11f1-9c37-00155d0001a1 https://dk.um.si/Dokument.php?lang=slv&id=204856 https://www.frontiersin.org/journals/toxicology/articles/10.3389/ftox.2026.1765753/full 1 45495964-27f0-11f1-bbad-00155d0001a1 https://dk.um.si/Dokument.php?lang=slv&id=204767 Medicinska fakulteta 0 0 0