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Cited by 7Animal testing and alternativesEnvironmental Toxicology and EcotoxicologyCarcinogens and Genotoxicity Assessment

Gaining confidence in in vitro toxicity data with measured exposure concentrations

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Henneberger, Luise, Bardehly, Sophia, König, Maria, Escher, Beate I.·2025·NAM Journal
DOI 10.1016/j.namjnl.2025.100048PMC PMC13289270
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Abstract

Cell-based in vitro bioassays are essential New Approach Methodologies (NAMs) that allow the bioactivity screening for thousands of chemicals in a time- and cost-efficient manner. One major obstacle for the widespread acceptance of in vitro bioassays is the undefined exposure in these tests. The present study aimed to complement in vitro testing of cytotoxicity as well as specific endpoints, such as the activation of oxidative stress response, with measured exposure concentrations as additional readout. Freely dissolved concentrations ( C free ) of 24 test chemicals in four different cell-based in vitro bioassays were measured in 96-well plate format at the time point of dosing and after 24 h of exposure in the assay plates for all tested concentrations of the chemicals. Freely dissolved fractions ( f free ) in the medium decreased with increasing hydrophobicity. Hydrophilic chemicals like atenolol had f free of >90 %, while the most hydrophobic test chemical chrysene had an f free of only 1.4 % at the beginning of the test and 0.7 % after 24 h of exposure to the cells. For 12 of the 24 chemicals C free decreased >20 % during the bioassays. Depending on the chemical, the loss may be due to sorption to well plate plastic, cellular metabolism or abiotic degradation (e.g., hydrolysis). The simultaneous measurement of toxicity and exposure allows the identification of chemicals with unstable exposure concentrations and/or low bioavailability and provides a quantitative measure of bioactivity that can directly be applied for in vitro -to- in vivo extrapolation (IVIVE).

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Sorted by citations

Authors
Henneberger, L., Bardehly, S., König, M., & Escher, B. I.
Affiliations
Helmholtz Centre for Environmental Research (DE)
Corresponding author
Henneberger, Luise
ORCID
Venue
NAM Journal
Publisher
Elsevier BV
Published
2025
Volume
1
Pages
100048
ISSN
3050-6204 (ISSN-L)
DOI
10.1016/j.namjnl.2025.100048
PMC
PMC13289270
OpenAlex
W4414149340
Cited by
7
References
1
Citations by year
2026: 7
FWCI
8.35
Type
journal article
Language
en
Open access
Licence
Locations
  • NAM journal. (publishedVersion, cc-by-nc)
  • PubMed (publishedVersion)
  • PubMed Central (publishedVersion, cc-by-nc)
Funders
Helmholtz Association
Topics
  • Health Sciences › Veterinary › Small Animals › Animal testing and alternatives
  • Physical Sciences › Environmental Science › Health, Toxicology and Mutagenesis › Environmental Toxicology and Ecotoxicology
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › Carcinogens and Genotoxicity Assessment
Keywords
freely dissolved concentration, bioavailability, in vitro-in vivo extrapolation, new approach methodologies, cytotoxicity, oxidative stress response, hydrophobicity, chemical stability, chrysene, atenolol

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