On this page, you will find an overview of RIVM publications related to alternatives to animal testing and safety assessment of chemicals and pharmaceuticals.

Publications 2025

Title Authors Journal
Mechanisms of developmental neurotoxicity mediated by perturbed thyroid hormone homeostasis in the brain: an adverse outcome pathway network Dierichs, N.T.O.M.; Piersma, A.H.; Peeters, R.P.; Visser, W.E.; Meiman, M.E.; Hessel, E.V.S. Crit Rev Toxicol 2024; epub ahead of print
The European Partnership PARC’s Role in Actively Promoting the Uptake of New Approach Methodologies and Next-Generation Risk Assessment into Regulatory Risk Assessment Practice Herzler, M.; Luijten, M.; Marx-Stoelting, P.; Rivière, G. Curr Opinion Toxicology 2025; 42:100517
Developing a global education hub for animal-free innovation Janssens, M.R.E.; Salvatori, D.; Hogervorst, J.; Nonis, C.; Bailey J.; Bajramovic, J.; Burgers, A.; Caloni, F.; Kienhuis, A.; et al.  ALTEX 2025: accepted manuscript
Mapping physiology: A systems biology approach for the development of alternative methods in toxicology Staumont, B.; Ladeira, L.; Gamba, A.; Heusinkveld, H.J.; Piersma, A.; Fritsche, E.; Masereeuw, R.; Vanhaecke, T.; Teunis, M.; Luechtefeld, T.H.; Hartung, T.; Jover, J.; Vinken, M.; Geris, L. ALTEX 2025: accepted manuscript

Publications 2024

Title Authors Journal
Animal-free safety assessment of chemicals: an innovation system perspective Hoogstraten, M.J.; Vriend, J.; Leeuw, V.C.de; Negro, S.O.; Moors, E.H.M.; Kienhuis, A.S.; Hoekman, J. Arch Toxicol 2024: online ahead of print
Proposal for a qualification system for New Approach Methodologies (NAMs) in the food and feed sector: example of implementation for nanomaterial risk assessment Haase, A.; Barroso, J.; Bremer-Hoffman, S.; Fessard, V.; Gutleb, A.C.; Mast, J.; McVey, E.; Mertens, B.; Oomen, A.G.; et al. EFSA supporting publication 2024: 21(9):EN-9008
Review of New Approach Methodologies for Application in Risk Assessment of Nanoparticles in the Food and Feed Sector: Status and Challenges Usmani, S.M.; Bremer-Hoffmann, S.; Cheyns, K.; Cubadda, F.; Dumit, V.I.; Escher, S.E.; Fessard, V.; Gutleb, A.C.; Leger, T.; Liu, Y.C.; Mast, J.; McVey, E.; Mertens, B.; Montalvo, D.; Oomen, A.G.; et al. EFSA supporting publication 2024: 21(9):EN-8826
The zebrafish embryo as a model for chemically-induced steatosis: A case study with three pesticides Heusinkveld, H.J.; Zwart, E.P.; de Haan, A.; Braeuning, A.; Alarcan, J.; van der Ven L.T.M. Toxicology 2024; 508:153927
Utilization of human stem cells to examine neurotoxic impacts on differentiation de Leeuw, V.C.; Hessel, E.V.S. Advances Neurotoxicol 2024: online ahead of print
Chemical respiratory sensitization - Current status of mechanistic understanding, knowledge gaps and possible identification methods of sensitizers Hargitai, R.; Parrakova, L.; Szatmari, T.; Monfort-Lanzas, P.; Galbiati, V.; Audouze K.; Jornod, F.; Staal, Y.C.M.; Burla, S.; Chary, A.; Gutleb, A.C.; Lumniczky, K.; Vandebriel, R.J.; Gostner, J.M. Front Toxicol 2024; 5
Evaluation of rat and rabbit embryofetal development studies with pharmaceuticals: the added value of a second species Roos, P.; Angasta, C.; Piersma, A.H.; van Meer, P.J.K.; Theunissen, P.T. Crit Rev Toxicol 2024; online ahead of print
How to optimize communication about animal and animal-free research methods Homberg, J.R.; Piersma, A.H.; Krul, C.A.M.; Genzel, L.; Kienhuis, A.; ter Gast, E.; Wolvekamp, M. Front Vet Sci 2024; 11
New approach methods to assess developmental and adult neurotoxicity for regulatory use: a PARC work package 5 project Tal, T.; Myhre, O.; Fritsche, E.; Ruegg, J.; Craenen, K.; Aiello-Holden, K.; Hessel, E.; Heusinkveld, H.J.; et al. Front Toxicol; 2024;6

How neurobehavior and brain development in alternative whole-organism models can contribute to prediction of developmental neurotoxicity

 Collins, E-M.S.; Hessel, E.V.S.; Hughes, S. NeuroToxicology 2024; 102:48-57

New approach methodologies to enhance human health risk assessment of immunotoxic properties of chemicals — a PARC (Partnership for the Assessment of Risk from Chemicals) project

 Snapkow, I.; Smith, N.M.; Arnesdotter, E.; Beekmann, K.; Blanc, E.B.; Braeuning, A.; Staal, Y.C.M.; Vandebriel, R. J.; et al. Front Toxicol 2024; 6

A roadmap towards a human-centric safety assessment of advanced therapy medicinal products

 Chien, H.T.; Leeuw, V.C. de; Esterik, J.C.J. van; Russel, F.G.M.; Kienhuis, A.S.; Theunissen, P.T.; Meer, P. van Regul Toxicol Pharmacol 2024; 150:105631

New Approach Methodologies (NAMs) for ad hoc human health risk assessment of food and non-food products - Proceedings of a workshop

 Wit, L.de; Hendriks, H.; Engelen, J. van; Heusinkveld, H.; Kienhuis, A.; Rorije, E.; Woutersen, M.;  Zee, van der M.; Jeurissen, S. Regul Toxicol Pharmacol 2024; 105615

In Vitro Assessment of Translocation and Toxicological Effects of Nicotine and Ethyl Maltol from e-Cigarettes Using Air–Liquid Interface-Cultured Bronchial Epithelial Cells

 Staal, Y.C.M.; Gremmer, E.; Duijm, G.; Duistermaat, E.; Fokkens, P.; Lensen, D.; Hodemakers, H.M.; Maas, L.; Remels, A.; Talhout, R. Applied in Vitro Toxicol 2024; 10(1)
Beyond Encapsulation: Exploring Macrophage-Fibroblast Crosstalk in Implant-Induced Fibrosis Krishna Sudarsanam, P.; Alsema, E.C.; Beijer, N.R.M.; Kootenm T. van; Boer, J. de Tissue Engineering Part B:  Reviews 2023; online ahead of print
The role of trust in the use of artificial intelligence for chemical risk assessment Wassenaar, P.N.H.; Minnema, J.; Vriend, J.; Peijnenburg, W.J.G.M.; Pennings, J.L.A.; Kienhuis, A. Regul Toxicol Pharmacol 2024; 148:105589
Report of the First ONTOX Stakeholder Network Meeting: Digging Under the Surface of ONTOX Together With the Stakeholders  Diemar, M.G.; Vinken, M.; Roggen, E.L.  Alternatives to Laboratory animals 2024; 52(2):117-31
Title Authors Journal
An integrated approach to testing and assessment (IATA) to support grouping and read-across of nanomaterials in aquatic systems Cross, R.K., Spurgeon, D., Svendsen, C., Lahive, E., Little, S., von der Kammer, F., Loosli, F., Matzke, M., Fernandex, T.F., Stone, V., Peijnenburg, W.J.G.M., Bleeker, A.J. Nano Today 2024; 54:102065
An adverse outcome pathway for chemical-induced Parkinson’s disease: Calcium is key Meerman, J.J., Legler, J., Piersma, A.H., Westerink, R.H.S., Heusinkveld, H.J. NeuroToxicology  2023: 99: 226-243
The Importance of Variations in In VitroDosimetry to Support Risk Assessment of Inhaled Toxicants Staal, Y.C.M., Geraets, L., Rothen-Rutishauer, B., Clift, M.J.D., Braakhuis, H., Kienhuis, A.S., Bos, P.J.M. ALTEX 2023; 41(1), 91-103
New approach methodologies to facilitate and improve the hazard assessment of non-genotoxic carcinogens—a PARC project  Audebert, M., Assmann, A.S., Azqueta, A., Babica, P. Benfenati, E., Bortoli, S. Heusinkveld, H.J., Luijten, M., et al. Front Toxicol 2023; 5
New approach methodologies in human regulatory toxicology – Not if, but how and when!  Schmeisser, S., Miccoli, A., Bergen, M. von, Berggren, E., Braeuning, A., Busch, W., Luijten, M.,  et al. Environment Int 2023; 178:108082
Variability of in vivo potency assays of whole-cell pertussis, inactivated polio, and meningococcal B vaccines Walstijn, C. van, Verweij, S., Care, R., Rigsby, P., Clapper, E-B., Markey, K., Vandebriel, R.J., Stickings, P. Hoefnagel, M.H.N. Vaccine 2023; 41(38):5603-5613
Retinoic acid signaling pathway perturbation impacts mesodermal-tissue development in the zebrafish embryo: Biomarker candidate identification using transcriptomics  Samrani, L.M.M., Dumont, F., Hallmark, N., Bars, R., Tinwell, H., Pallardy, M., Piersma, A.H. Regulatory Toxicology and Pharmacology 2023; 142:105438
Transferability and reproducibility of exposed air-liquid interface co-culture lung models Braakhuis, H.M., Gremmer, E.R., Bannuscher, A. Drasler, B., Keshavan, S., Rothen-Rutishauser, B., Birk, B., Verlohner, A. Vandebriel, R.J., et al. NanoImpact 2023; 31:100466
An adverse outcome pathway network for chemically-induced oxidative stress leading to (non-)genotoxic carcinogenesis Veltman, C.H.J., Pennings, J.L.A., Water, B. van de, Luijten, M. Chem Res Toxicol 2023; online ahead of print

The State of the Art and Challenges of In Vitro Methods for Human Hazard Assessment of Nanomaterials in the Context of Safe-by-Design

 Ruijter, N., Soeteman-Hernandez, L.G., Carriere, M., Boyles, M., McLean, P., Catalan, J., et al. Nanomaterials 2023; 13(3): 472

Prolonged differentiation of neuron-astrocyte co-cultures results in emergence of dopaminergic neurons

 de Leeuw, V.C., van Oostrom, C.T.M., Zwart, E.P., Heusinkveld H.J., Hessel, E.V.S. Int J Mol Sci 2023; 24(4): 3608

Innovating human chemical hazard and risk assessment through a holistic approach

 Piersma, A.H., Heusinkveld, H., Hessel, E., Kienhuis, A. Current Opinion in Toxicology 2023; 33: 100386
Neurodegeneration in a regulatory context: The need for speed Meerman, J. J., Wolterink, G., Hessel, E. V. S., de Jong, E., & Heusinkveld, H. J. Current Opinion in Toxicology 2023; 33: 100383
Application of AOPs to assist regulatory assessment of chemical risks – Case studies, needs and recommendations Bajard, L., Adamovsky, O., Audouze, K., Baken, K., Barouki, R., Beltman, J. B., Luijten, et al. Environmental Research 2023, 217:114650
Dynamic regulation of gene expression and morphogenesis in the zebrafish embryo test after exposure to all-trans retinoic acid.  Samrani, L. M. M., Pennings, J. L. A., Hallmark, N., Bars, R., Tinwell, H., Pallardy, M., & Piersma, A. H.  Reproductive Toxicology 2023: 115:8-16
Towards achieving a modern science-based paradigm for agrochemical carcinogenicity assesshttps://doi.org/10.1016/j.yrtph.2022.105301ment. Hilton, G. M., Corvi, R., Luijten, M., Mehta, J., & Wolf, D. C. Regulatory Toxicology and Pharmacology 2023; 137:105301
Title Authors Journal

Smoking-associated exposure of human primary bronchial epithelial cells to aldehydes: impact on molecular mechanisms controlling mitochondrial content and function 

 Tulen, C.B.M., Duistermaat, E., Cremers, J.W.J.M., Klerx, W.N.M., Fokkens, P.H.B., Weibolt, N., Kloosterboer, N., Dentener, M.A., Gremmer, E.R., Jessen, P.J.J., Koene, E.J.C., Maas, L., Opperhuizen, A., van Schooten, F.J., Staal, Y.C.M., Remels, A.H.V. Cells 2022; 11(21): 3481
The Current Status and Work of Three Rs Centres and Platforms in Europe Neuhaus, W., Reininger-Gutmann, B., Rinner, B., Plasenzotti, R., Wilflingseder, D., De Kock, J., Vanhaecke, T., Rogiers, V., Jírová, D., Kejlova, K., Knudsen, L., Nielsen, R., Kleuser, B., Kral, V., Thöne-Reineke, C., Hartung, T., Pallocca, G., Rovida, C., Leist, M., & Spielmann, H. Alternatives to laboratory animals : ATLA 2022: 50(6):381-413
Applicability of generic PBK modelling in chemical hazard assessment: A case study with IndusChemFate. Fragki, S., Piersma, A. H., Westerhout, J., Kienhuis, A., Kramer, N. I., & Zeilmaker, M. J.   Regulatory Toxicology and Pharmacology 2022; 136:105267
Smoking-Associated Exposure of Human Primary Bronchial Epithelial Cells to Aldehydes: Impact on Molecular Mechanisms Controlling Mitochondrial Content and Function Tulen, C. B. M., Duistermaat, E., Cremers, J. W. J. M., Klerx, W. N. M., Fokkens, P. H. B., Weibolt, N., Kloosterboer, N., Dentener, M. A., Gremmer, E. R., Jessen, P. J. J., Koene, E. J. C., Maas, L., Opperhuizen, A., van Schooten, F. J., Staal, Y. C. M., & Remels, A. H. V. Cells 2022; 11(21):3481
Analyses of Transcriptomics Cell Signalling for Pre-Screening Applications in the Integrated Approach for Testing and Assessment of Non-Genotoxic Carcinogens.  Oku, Y., Madia, F., Lau, P., Paparella, M., McGovern, T., Luijten, M., & Jacobs, M. N.   International Journal of Molecular Sciences 2022; 23(21):12718
Neuronal differentiation pathways and compound-induced developmental neurotoxicity in the human neural progenitor cell test (hNPT) revealed by RNA-seq. de Leeuw, V. C., van Oostrom, C. T. M., Wackers, P. F. K., Pennings, J. L. A., Hodemaekers, H. M., Piersma, A. H., & Hessel, E. V. S. Chemosphere 2022: 304:135298
Development of a cell line-based in vitro assay for assessment of Diphtheria, Tetanus and acellular Pertussis (DTaP)-induced inflammasome activation Vandebriel, R. J., Stalpers, C. A. L., Vermeulen, J. P., Remkes, M., Schmelter, M., Broere, F., & Hoefnagel, M. H. N. Vaccine 2022; 40(39):5601-7
Prioritization of chemicals in food for risk assessment by integrating exposure estimates and new approach methodologies: A next generation risk assessment case study Luijten, M., Sprong, R. C., Rorije, E., & van der Ven, L. T. M. Frontiers in Toxicology 2022: 4
Transcriptome and proteome analysis of innate immune responses to inactivated Leptospira and bivalent Leptospira vaccines in canine 030-D cells.  Novak, A., Pennings, J. L. A., van der Maas, L., Meiring, H. D., Ludwig, I., Verkoeijen, S., Rutten, V., Broere, F., & Sloots, A.  Scientific Reports 2022; 12(1):13418
Next-generation risk assessment of chemicals – Rolling out a human-centric testing strategy to drive 3R implementation: The RISK-HUNT3R project perspective. Pallocca, G., Moné, M. J., Kamp, H., Luijten, M., van de Water, B., & Leist, M.  ALTEX - Alternatives to animal experimentation 2022; 39(3):419-26
Identification of proteome markers for drug-induced liver injury in zebrafish embryos. Driessen, M., van der Plas - Duivesteijn, S., Kienhuis, A. S., van den Brandhof, E.-J., Roodbergen, M., van de Water, B., Spaink, H. P., Palmblad, M., van der Ven, L. T. M., & Pennings, J. L. A. Toxicology 2022: 477:153262
Development of a Roadmap for Action on New Approach Methodologies in Risk Assessment. Escher, S. E., Partosch, F., Konzok, S., Jennings, P., Luijten, M., Kienhuis, A., de Leeuw, V., Reuss, R., Lindemann, K.-M., & Bennekou, S. H. EFSA Supporting Publications 2022; 19(6):7341E
Dose Addition in the Induction of Craniofacial Malformations in Zebrafish Embryos Exposed to a Complex Mixture of Food-Relevant Chemicals with Dissimilar Modes of Action. Van Der Ven Leo, T. M., Van Ommeren, P., Zwart Edwin, P., Gremmer Eric, R., Hodemaekers Hennie, M., Heusinkveld Harm, J., van Klaveren Jacob, D., & Rorije, E. Environmental Health Perspectives 2022; 130(4):047003
Potential of concentration-response data to broaden regulatory application of in vitro test guidelines. Jacobs, M. N., Ezendam, J., Hakkert, B., & Oelgeschlaeger, M. ALTEX - Alternatives to animal experimentation 2022; 39(2):315-21
The Rise of Three Rs Centres and Platforms in Europe Neuhaus, W., Reininger-Gutmann, B., Rinner, B., Plasenzotti, R., Wilflingseder, D., , Kienhuis, A., et al. Alternatives to laboratory animals : ATLA 2022; 50(2):90-120
AOP report: Development of an adverse outcome pathway for oxidative DNA damage leading to mutations and chromosomal aberrations Cho, E., Allemang, A., Audebert, M., Chauhan, V., Dertinger, S., Hendriks, G., Luijten, M., Marchetti, F., Minocherhomji, S., Pfuhler, S., Roberts, D. J., Trenz, K., & Yauk, C. L. Environmental and Molecular Mutagenesis 2022; 63(3):118-34
Pluripotent stem cell assays: Modalities and applications for predictive developmental toxicity Piersma, A. H., Baker, N. C., Daston, G. P., Flick, B., Fujiwara, M., Knudsen, T. B., Spielmann, H., Suzuki, N., Tsaioun, K., & Kojima, H.  Current Research in Toxicology 2022: 3:100074
Regulation of Clostridium tetani Neurotoxin Expression by Culture Conditions.  Pennings, J. L. A., Abachin, E., Esson, R., Hodemaekers, H., Francotte, A., Claude, J.-B., Vanhee, C., Uhlrich, S., & Vandebriel, R. J. Toxins 2022; 14(1)
Integrating in vitro chemical transplacental passage into a generic PBK model: A QIVIVE approach Fragki, S., Hoogenveen, R., van Oostrom, C., Schwillens, P., Piersma, A. H., & Zeilmaker, M. J. Toxicology 2022; 465:153060
Neuromodulatory and neurotoxic effects of e-cigarette vapor using a realistic exposure method Staal, Y. C. M., Li, Y., Gerber, L.-S., Fokkens, P., Cremers, H., Cassee, F. R., Talhout, R., Westerink, R. H. S., & Heusinkveld, H. J. Inhalation Toxicology 2022:1-10
Title Authors Journal
Integrating 3Rs approaches in WHO guidelines for the batch release testing of biologicals. Lilley, E., Isbrucker, R., Ragan, I., & Holmes, A. Biologicals 2021; 74:24-7
Exploring Neurobehaviour in Zebrafish Embryos as a Screening Model for Addictiveness of Substances Havermans, A., Zwart, E. P., Cremers, H. W. J. M., van Schijndel, M. D. M., Constant, R. S., Mešković, M., et al. Toxics 2021; 9(10:250
Developmental Neurotoxicity of Environmentally Relevant Pharmaceuticals and Mixtures Thereof in a Zebrafish Embryo Behavioural Test Atzei, A., Jense, I., Zwart, E. P., Legradi, J., Venhuis, B. J., van der Ven, L. T. M., Heusinkveld, H. J., Hessel, E. V. S. International Journal of Environmental Research and Public Health 2021; 18(13):6717
Safer chemicals using less animals: kick-off of the European ONTOX project Vinken, M., Benfenati, E., Busquet, F., Castell, J., Clevert, D. A., de Kok, , et al. Toxicology 2021; 458:152486
Optimization of an air-liquid interface in vitro cell co-culture model to estimate the hazard of aerosol exposures He, R. W., Braakhuis, H. M., Vandebriel, R. J., Staal, Y. C. M., Gremmer, E. R., Fokkens, , et al. Journal of aerosol science 2021; 153:105703
An ontology for developmental processes and toxicities of neural tube closure. Heusinkveld, H. J., Staal, Y. C. M., Baker, N. C., Daston, G., Knudsen, T. B., & Piersma, A. H. Reproductive Toxicology 2021; 99:160-7
Title Authors Journal
An efficient neuron-astrocyte differentiation protocol from human embryonic stem cell-derived neural progenitors to assess chemical-induced developmental neurotoxicity. de Leeuw, V. C., van Oostrom, C. T. M., Westerink, R. H. S., Piersma, A. H., Heusinkveld, H. J., & Hessel, E. V. S. Reproductive Toxicology 2020; 98:107-16
The effects of aliphatic alcohols and related acid metabolites in zebrafish embryos - correlations with rat developmental toxicity and with effects in advanced life stages in fish van der Ven, L. T. M., Schoonen, W. G., Groot, R. M., den Ouden, F., Heusinkveld, H. J., Zwart, E. P., Hodemaekers, H. M., Rorije, E., de Knecht, J.
Toxicology and Applied Pharmacology 2020; 407:115249
Distinguishing mode of action of compounds inducing craniofacial malformations in zebrafish embryos to support dose-response modeling in combined exposures. Heusinkveld, H. J., Schoonen, W. G., Hodemaekers, H. M., Nugraha, A., Sirks, J. J., Veenma, V., Sujan, C., Pennings, J. L. A., Wackers, P. F., Palazzolo, L., Eberini, I., Rorije, E., & van der Ven, L. T. M. Reproductive Toxicology 2020; 96:114-27
An Air-liquid Interface Bronchial Epithelial Model for Realistic, Repeated Inhalation Exposure to Airborne Particles for Toxicity Testing Braakhuis, H. M., He, R., Vandebriel, R. J., Gremmer, E. R., Zwart, E., Vermeulen, J. P., Fokkens, P., Boere, J., Gosens, I., & Cassee, F. R.  Journal of visualized experiments : JoVE 2020; 
The virtual human in chemical safety assessment Piersma, A. H., van Benthem, J., Ezendam, J., Staal, Y. C. M., Kienhuis, A. S. Current Opinion in Toxicology 2019; 15:26-32
Design and validation of an ontology-driven animal-free testing strategy for developmental neurotoxicity testing. Toxicology and applied pharmacology Hessel, E. V. S., Staal, Y. C. M., & Piersma, A. H.
Toxicology and Applied Pharmacology 2018; 354:136-52
Rethinking developmental toxicity testing: Evolution or revolution? Scialli, A. R., Daston, G., Chen, C., Coder, P. S., Euling, S. Y., Foreman, J., Hoberman, A. M., Hui, J., Knudsen, T., Makris, S. L., Morford, L., Piersma, A. H., Stanislaus, D., & Thompson, K. E. Birth defects research 2018; 119(10):480-850