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Humanising nanotoxicology: replacement of animal-derived products in the application of integrated approaches to testing and assessment of nanomaterial inhalation hazard

Fraser, Roma, Campbell, Keira, Pokorski, Pawel, MacKinnon, Eve, McAllister, Katie, Neves, Karla B., Murphy, Fiona·2025·Frontiers in Bioengineering and Biotechnology
DOI 10.3389/fbioe.2025.1526808PMC PMC11861073
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Abstract

Over the past decade, the development of nanomaterials (NMs) has surged, highlighting their potential benefits across multiple industries. However, concerns regarding human and environmental exposure remain significant. Traditional in vivo models for safety assessments are increasingly viewed as unfeasible and unethical due to the diverse forms and biological effects of NMs. This has prompted the design of Novel Approach Methods (NAMs) to streamline risk assessment and predict human hazards without relying on animal testing. A critical aspect of advancing NAMs is the urgent need to replace animal-derived products in assay protocols. Incorporating human or synthetic alternatives can significantly reduce the ethical burden of animal use while enhancing the relevance of toxicity testing. This study evaluates the impact of removing animal-derived products from standard acellular and in vitro assays recommended in a published Integrated Approaches to Testing and Assessment (IATA) for inhaled NMs. We specifically assessed the effects of replacing fetal bovine serum with human platelet lysate in acellular reactivity tests and in vitro toxicity testing using a panel of well-characterized NMs. Significant differences in acellular NM reactivity and dramatic changes in A549 cell growth rates and responses to NMs were observed under different media conditions. Our findings demonstrate that variations in experimental setup can fundamentally impact NM hazard assessment, influencing the interpretation of results within specific assays and across tiered testing strategies. Further investigation is needed to support a shift toward more ethical toxicity testing that does not rely on animal-derived materials.

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  • Replacing major components, such as serum, ECM preparations or metabolic activation systems, may alter assay biology.

    “Moreover, replacing major components, such as serum, ECM preparations or metabolic activation systems may alter assay biology. Changes in baseline phenotype, sensitivity, or readouts, have been reported even when technical adaptation is successful (Perez-Diaz et al., 2023; Reichstein et al., 2023; Fraser et al., 2025).”

    From Challenges and solutions in transitioning to animal-free standards: a comprehensive analysis of components in human cell-based developmental neurotoxicity assays (2026) · cites this paper as Fraser et al., 2025

    The Discussion reports that replacing fetal bovine serum with human platelet lysate changed cell growth and phenotype and could affect nanotoxicity assay results (section 4; page not provided). This supports the point about serum, but the passages do not establish the same effect for ECM preparations or metabolic activation systems. (LLM, conf=0.86)

    Experimental — accuracy not yet measured

  • The increasing availability of recombinant and chemically defined reagents provides opportunities to reduce reliance on animal-derived materials and to improve the definition of cell culture systems used in toxicological testing.

    “The increasing availability of recombinant and chemically defined reagents provides opportunities to reduce reliance on animal-derived materials and to improve the definition of cell culture systems used in toxicological testing (Nesterenko et al., 2020; Cassotta et al., 2022; Fraser et al., 2025).”

    From Challenges and solutions in transitioning to animal-free standards: a comprehensive analysis of components in human cell-based developmental neurotoxicity assays (2026) · cites this paper as Fraser et al., 2025

  • Changes in baseline phenotype, sensitivity, or readouts have been reported even when technical adaptation is successful.

    “Moreover, replacing major components, such as serum, ECM preparations or metabolic activation systems may alter assay biology. Changes in baseline phenotype, sensitivity, or readouts, have been reported even when technical adaptation is successful (Perez-Diaz et al., 2023; Reichstein et al., 2023; Fraser et al., 2025).”

    From Challenges and solutions in transitioning to animal-free standards: a comprehensive analysis of components in human cell-based developmental neurotoxicity assays (2026) · cites this paper as Fraser et al., 2025

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Cited by

Sorted by citations

Authors
Fraser, R., Campbell, K., Pokorski, P., MacKinnon, E., McAllister, K., Neves, K. B., & Murphy, F.
Venue
Frontiers in Bioengineering and Biotechnology
Publisher
Frontiers Media SA
Published
2025-02-12
Volume
13
ISSN
2296-4185
DOI
10.3389/fbioe.2025.1526808
PMC
PMC11861073
Cited by
4
References
1
Licence
creativecommons.org/licenses/by/4.0/ (vor)
Funders
Animal Free Research United Kingdom

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