A methodological framework for assessing foetal skeletal and visceral development in teratological investigations

Ujházy, Eduard; Farkaš, Michal; Brucknerová, Ingrid; Mach, Mojmír; Dubovický, Michal

doi:10.2478/intox-2021-0014

Abstract

This paper outlines the methodical approaches essential for the accurate and reproducible evaluation of foetal skeletal and visceral abnormalities in teratology studies. Teratological investigations are critical for assessing the potential of drugs, chemicals, or environmental agents to induce developmental toxicity. The established methodology involves a systematic two-part process. Firstly, visceral examinations use techniques like microdissection and serial sectioning to identify gross and microscopic changes in internal organs. Secondly, skeletal evaluations utilize staining procedures, such as Alizarin red S and Alcian blue, to visualize and analyze the ossification and structural integrity of the foetal skeleton. These standardized procedures are vital for the reliable detection and characterization of malformations and variations. A rigorous, methodical approach ensures the scientific validity of the data, which is fundamental for effective risk assessment and regulatory decision-making concerning human and environmental health.

References

Burdan F, Szumito J, Dudka J, Klepacz R, Blaszszak M, Solecky M, Korobowitz A, Chatas A, Klepacki J, Palczak M, Žuchnik-Wrona A, Hadata-Kiš, Urbanowicz Z, Wójtowicz Z. (2005). Morphological studies in modern teratological investigations. Folia Morphol 64(1): 1–8.
Search in Google Scholar Back to article
Carson F, Hladik Ch. (2009). Histotechnology: A self-instructional text (3rd ed.). ASCP Press, ISBN 978-0-89189-581-7.
Search in Google Scholar Back to article
Critchell KF. (2013). Fetal soft tissue examination by serial sectioning – Methodology. In Teratogenicity testing. Methods and Protocols. Part 1. (Barow PC ed.) pp. 233–253, Springer Protocols. Human Press.
Search in Google Scholar Back to article
Estevan C, Pamics D, Vilanova E, Sogorb MA. (2017). Chapter 9 – OECD Guidelines for In Vivo Testing of Reproductive Toxicity, in Reproductive and Developmental Toxicology (Second Edition) (Gupta RC ed) pp. 163–178, Academic Press, London.
Search in Google Scholar Back to article
Chahoud I, Buschmann J, Clark R, Druga A, Falke H, Faqi A, Hansen E, Heinrich-Hirsch B, Hellwig J, Lingk W, Parkinson M, Paumgartten FJ, Pfeil R, Platzek T, Scialli AR, Seed J, Stahlmann R, Ulbrich B, Wu X, Yasuda M, Younes M, Solecki R. (1999). Classification terms in developmental toxicology: need for harmonisation. Report of the Second Workshop on the Terminology in Developmental Toxicology Berlin, 27–28 August 1998. Reprod Toxicol 13: 77–82.
Search in Google Scholar Back to article
Lorke D. (1977) Evaluation of skeleton, in Methods in prenatal toxicology (Neu-bert D, Merker HJ, Kwasigroch G eds) pp. 145–152, Thieme, Stuttgart.
Search in Google Scholar Back to article
Ovchinnikov D. (2009). Alcian blue/alizarin red staining of cartilage and bone in mouse. Cold Spring Harb Protoc 2009(3): pdb.prot5170.
Search in Google Scholar Back to article
Peters PWJ. (1977). Double staining of fetal skeletons for cartilage and bone, in Methods in prenatal toxicology (Neubert D, Merker J, Kwasigroch TE eds) pp. 153–154, George Thieme Publisher, Stuttgart.
Search in Google Scholar Back to article
Puchtler H, Meloan SN, Terry MS. (1969). On the History and Mechanism of Alizarin Red S Stains for Calcium. J Histochem Cytochem 17(2): 110–124.
Search in Google Scholar Back to article
Reynaud L, Monrozier AJ. (2013). Skeletal examination by alizarin staining – Methodology, in Teratogenicity testing. Methods and Protocols. Part 1. (Barow PC ed.) pp. 201–221, Springer Protocols. Human Press.
Search in Google Scholar Back to article
Solecki R, Burgin H, Buschmann J, Clark R, Duverger M, Fialkowski O, Guittin P, Hazelden KP, Hellwig J, Hoffmann E, Hofmann T, Hubel U, Khalil S, Lingk W, Mantovani A, Moxon M, Muller S, Parkinson M, Paul M, Paumgartten F, Pfeil R, Platzek T, Rauch-Ernst M, Scheevelenbos A, Seed J, Talsness CE, Yasuda M, Younes M, Chahoud I. (2001). Harmonisation of rat fetal skeletal terminology and classification. Report of the Third Workshop on the Terminology in Developmental Toxicology. Berlin, 14–16 September 2000. Re-prod Toxicol 15: 713–721.
Search in Google Scholar Back to article
Solecki R, Bergmann B, Burgin H, Buschmann J, Clark R, Druga A, Van Duijnhoven EA, Duverger M, Edwards J, Freudenberger H, Guittin P, Hakaite P, Heinrich-Hirsch B, Hellwig J, Hofmann T, Hubel U, Khalil S, Klaus A, Kudicke S, Lingk W, Meredith T, Moxon M, Muller S, Paul M, Paumgartten F, Rohrdanz E, Pfeil R, Rauch-Ernst M, Seed J, Spezia F, Vickers C, Woelffel B, Chahoud I. (2003). Harmonization of rat fetal external and visceral terminology and classification. Report of the Fourth Workshop on the Terminology in Developmental Toxicology, Berlin, 18–20 April 2002. Reprod Toxicol 17: 625–637.
Search in Google Scholar Back to article
Staples RE. (1974). Detection of visceral alterations in mammalian fetuses. Teratology 9: A37–A38.
Search in Google Scholar Back to article
Tyll WR, Marr CM. (2006). Developmental toxicity testing – Methodology, in Developmental and reproductive toxicology. Second edition. (Hood RD, ed) pp. 425–487, CRC Taylor & Francis Group, Boca Raton, London, New York.
Search in Google Scholar Back to article
Ujházy, E, Preinerová M, Jozefík M. (1979). Effects of cyclophosphamide on the prenatal development of Swiss strain mice. Neoplasma 26(5): 529–537.
Search in Google Scholar Back to article
Wilson JG. (1973). Collection and interpretation of results. In: Environment and birth defects (Lee DHK, Hewson W, Okun D, eds) pp. 181–188, Academic Press, New York.
Search in Google Scholar Back to article
Wise LD, Beck SL, Beltrame D, Beyer BK, Chahoud I, Clark RL, Clark R, Druga AM, Feuston MH, Guittin P, Henwood SM, Kimmel CA, Lindstrom P, Palmer AK, Petrere JA, Solomon HM, Yasuda M, York RG. (1997). Terminology of developmental abnormalities in common laboratory mammals (version 1). Teratology 55: 249–292.
Search in Google Scholar Back to article

A methodological framework for assessing foetal skeletal and visceral development in teratological investigations

Abstract

Paradigm

My account