In Vitro New Approach Methodologies for Toxicity Assessment of Emerging Food Contaminants in Indonesia: A Proposed Strategy for BPOM Regulatory Integration.

The accelerating global diversification of food chemicals-encompassing mycotoxins, heavy metals, organophosphate pesticide residues and the increasingly prevalent burden of microplastics and nanoplastics (MNPs)-creates an urgent demand for rapid, human-relevant and ethically acceptable toxicity testing platforms. Conventional animal-based bioassays are insufficient to characterise the thousands of substances awaiting safety evaluation, a deficit particularly acute in tropical food-producing regions including Indonesia where environmental contamination profiles and regulatory infrastructure differ markedly from high-income countries. New approach methodologies (NAMs), comprising validated in vitro cell and organoid systems, high-throughput screening (HTS), multiomics toxicogenomics, in silico modelling and adverse outcome pathway (AOP) frameworks, represent a paradigm shift in food safety toxicology. This review critically appraises current evidence on in vitro NAMs for emerging food contaminants, with emphasis on (i) 2D cell line and 3D organoid/organ-on-chip models for hepatotoxicity, enterotoxicity and nephrotoxicity; (ii) mechanistic pathways-oxidative stress, mitochondrial dysfunction, genotoxicity and intestinal barrier disruption-as interrogated through standardised in vitro endpoints; (iii) AOP frameworks as regulatory bridges between molecular initiating events and adverse outcomes; (iv) high-throughput transcriptomics (HTTr) and Cell Painting integration for mechanism-informed hazard prioritisation; and (v) in vitro-to-in vivo extrapolation (IVIVE) via physiologically based toxicokinetic (PBTK) modelling. Critical gaps in chronic-exposure data, mixture toxicity assessment and BPOM regulatory acceptance are discussed. A novel evidence-based NAM validation roadmap tailored to the Southeast Asian regulatory context is proposed.