Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2024 Jan:123:108513.
doi: 10.1016/j.reprotox.2023.108513. Epub 2023 Nov 26.

Validation of a zebrafish developmental defects assay as a qualified alternative test for its regulatory use following the ICH S5(R3) guideline

A M J Weiner  1 , I Irijalba  2 , M P Gallego  2 , I Ibarburu  2 , L Sainz  3 , F Goñi-de-Cerio  3 , C Quevedo  2 , A Muriana  4
Affiliations

Validation of a zebrafish developmental defects assay as a qualified alternative test for its regulatory use following the ICH S5(R3) guideline

A M J Weiner et al. Reprod Toxicol. 2024 Jan.

Abstract

Zebrafish is a popular toxicology model and provides an ethically acceptable small-scale analysis system with the complexity of a complete organism. Our goal is to further validate this model for its regulatory use for reproductive and developmental defects by testing the compounds indicated in the "Guideline on detection of reproductive and developmental toxicity for human pharmaceuticals" (ICH S5(R3) guideline.) To determine the embryotoxic and developmental risk of the 32 reference compounds listed in the ICH S5(R3) guideline, the presence of morphological alterations in zebrafish embryos was analyzed at two different stages to calculateLC50 and EC50 values for each stage. Teratogenic Indexes were established as the ratio between LC50 and EC50 critical for the proper compound classification as teratogenic when it is ≥ 2. A total of three biological replicates have been conducted to study the reproducibility of the assay. The chemicals' concentration in the medium and internally in the zebrafish embryos was evaluated. In this study, the 3 negative compounds were properly categorized while 23 compounds out of the 29 reference ones (sensitivity of 79.31%) were classified as teratogenic in zebrafish. The 6 that had false-negative results were classified 4 as inconclusive, 1 as not toxic, and 1 compound resulted toxic for zebrafish embryos under testing conditions. After the bioavailability experiments, some of the obtained inconclusive results were refined. The developmental defects assay in zebrafish gives an accuracy of 89.66%, sensitivity of 88.46%, specificity and repeatability of 100% compared to mammals; therefore, this is a well-integrated strategy using New Alternative Methods, to minimize the use of animals in developmental toxicity studies.

Keywords: Bench Marking Concentration (BMC); Bioavailability; Developmental defects; Embryotoxicity; ICH S5 (R3) Guideline; New alternative method (NAM); Zebrafish.

PubMed Disclaimer

Conflict of interest statement

Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Arantza Muriana reports equipment, drugs, or supplies and writing assistance were provided by GAIKER. Arantza Muriana reports a relationship with GAIKER that includes: consulting or advisory. The author is co-founder and R&D Director of BBD BioPhenix SLU, and all the coauthors except Sainz L and De Goñi F are nowadays employed or have been employed previously by BBD BioPhenix SLU, a CRO specialized in the use of zebrafish for efficacy, toxicity and ecotoxicity assays, including offering teratogenicity, reprotoxicity, neurotoxicity, or endocrine disruption assays for Pharma Industry, Cosmetic Industry, or Agrochemical Industry, including the assays described in the paper. Sainz L and De Goñi F work in GAIKER, a company that BBD BioPhenix SLU outsources to develop bioavailability assays for their clients, in this case, they offered their services to analyze the compounds inside the zebrafish for this paper.

Similar articles

See all similar articles

Cited by

LinkOut - more resources