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Toxic or not toxic? The specifications of the standard ISO 10993-5 are not explicit enough to yield comparable results in the cytotoxicity assessment of an identical medical device

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Gruber, Sarah, Nickel, Angela·2023·Frontiers in Medical Technology
DOI 10.3389/fmedt.2023.1195529PMC PMC10304299

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Abstract

Background: Medical device manufacturers are obliged to prove the biocompatibility of their products when they come into contact with the human body. The requirements for the biological evaluation of medical devices are specified by the international standard series ISO 10993. Part five of this series describes the performance of in vitro cytotoxicity tests. This test evaluates the effects of medical device use on cell health. The existence of the specific standard suggests that the tests will produce reliable and comparable results. However, the ISO 10993-5 offers wide latitude in the test specifications. In the past, we noticed inconsistencies of the results from different laboratories. Objective: To determine if the specifications of the standard ISO 10993-5 are explicit to ensure the comparability of test results and, if not, identify potential influencing factors. Methods: An interlaboratory comparison was conducted for the in vitro cytotoxicity test according to ISO 10993-5. Fifty-two international laboratories evaluated the cytotoxicity for two unknown samples. One was polyethylene (PE) tubing, which is expected to be non-cytotoxic and the other was polyvinyl chloride (PVC) tubing, for which a cytotoxic potential was presumed. All laboratories were asked to perform an elution test with predefined extraction specifications. The other test parameters were freely chosen by the laboratories according to the guidelines set by the standard. Results: To our surprise only 58 percent of the participating laboratories identified the cytotoxic potential of both materials as expected. Particularly for PVC a considerable variation of the results between the laboratories was observed [mean = 43 ± 30 (SD), min = 0, max = 100]. We showed that ten percent serum supplementation to the extraction medium, as well as longer incubation of the cells with the extract, greatly increased the test sensitivity for PVC. Conclusion: The results clearly show that the specifications set by the ISO 10993-5 are not explicit enough to obtain comparable results for an identical medical device. To set requirements that ensure reliable cytotoxicity assessments, further research will be necessary to identify the best test conditions for specific materials and/or devices and the standard needs to be revised accordingly.

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  • In HaCaT keratinocyte test systems, cell viability over 70-80% at 24 h should be considered nontoxic.

    “In the HaCaT keratinocyte test systems, results with cell viability of over 70-80% at 24 h should be considered nontoxic [30 - 32].”

    From Formulation development, physical stability, and preliminary safety screening of cosmetic emulsions containing banana peel extract (2026) · cites this paper as [32]

    The Methods (Data analysis; page unavailable) define cytotoxicity as cell viability reduced by more than 30% relative to untreated cells, supporting a nontoxic classification above 70%. The Results (Influence of cell culture setup on test sensitivity; page unavailable) say ISO 10993-5 specifies at least 24 hours of incubation. The paper does not establish a 70–80% threshold specifically for HaCaT keratinocytes at 24 hours. (LLM, conf=0.85)

    Experimental — accuracy not yet measured

  • Interlaboratory studies show substantial variability in IC50 and related cytotoxicity metrics, frequently in the range of approximately 20% to >40%, depending on system and endpoint.

    “Interlaboratory studies further show substantial variability in IC50 and related cytotoxicity metrics, frequently in the range of approximately 20% to >40%, depending on system and endpoint (Tani et al., 1999; Gruber and Nickel, 2023).”

    From Cytotoxicity across OECD methods is not a single endpoint: context-dependent roles, variability and implications for decision-making (2026) · cites this paper as Gruber and Nickel, 2023

    The Results section (‘Outcome of the interlaboratory comparison: highly variable cytotoxicity test results for PVC’; page unavailable) reports substantial variation: only 62% of laboratories classified PVC as cytotoxic, and 42% did not classify at least one material as expected. It does not report interlaboratory variability in IC50 or establish the claimed approximately 20% to >40% range for IC50 and related metrics across systems and endpoints; its percentages describe classification outcomes. (LLM, conf=0.87)

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Authors
Gruber, S., & Nickel, A.
Affiliations
Johner Institut (Germany) (DE)
Corresponding author
Gruber, Sarah
Venue
Frontiers in Medical Technology
Publisher
Frontiers Media SA
Published
2023-06-14
Volume
5
Pages
1195529-1195529
ISSN
2673-3129 (ISSN-L)
DOI
10.3389/fmedt.2023.1195529
PMC
PMC10304299
OpenAlex
W4380684791
Cited by
200
References
0
Citations by year
2026: 113 · 2025: 63 · 2024: 21 · 2023: 3
FWCI
44.90
Type
journal article
Language
en
Open access
Licence
Locations
  • Frontiers in Medical Technology (publishedVersion, cc-by)
  • medRxiv (acceptedVersion, cc-by-nd)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Topics
  • Health Sciences › Health Professions › Health Information Management › Healthcare Regulation
  • Health Sciences › Veterinary › Small Animals › Animal testing and alternatives
  • Health Sciences › Medicine › Surgery › Orthopaedic implants and arthroplasty
Keywords
ISO 10993-5, cytotoxicity assay, medical device safety, inter-laboratory variability, test result comparability, PVC pipes, polyethylene pipes, elution test, serum supplementation, test sensitivity

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