Skip to main content
SpringerLink
Log in

Skin Equivalent Models: Protocols for In Vitro Reconstruction for Dermal Toxicity Evaluation

  • Protocol
  • First Online:
Toxicity Assessment

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2240))

Abstract

This chapter presents the protocols for developing of skin equivalents (SE) and reconstructed human epidermis (RHE) models for dermal toxicity evaluation as an alternative method to animal use in research. It provides a detailed protocol for the in vitro reconstruction of human skin from primary keratinocytes, melanocytes, and fibroblasts obtained from foreskin biopsies, including the procedures for reconstruction of a stratified epidermis on a polyester membrane. SE and RHE developed through these methods have been proven suitable not only for dermal toxicity studies, but also for investigating of pathological conditions in the skin, such as diabetes and invasion of melanoma.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution

Subscribe and save

Springer+ Basic
$34.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or eBook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 99.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 129.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 179.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Similar content being viewed by others

References

  1. Worth A, Barroso J, Bremer S et al (2014) Alternative methods for regulatory toxicology—a state-of-the-art review. JRC Sci Policy Rep 26797. https://doi.org/10.2788/11111

  2. Mathur AK, Khanna SK (2002) Dermal toxicity due to industrial chemicals. Skin Pharmacol Physiol 15:147–153. https://doi.org/10.1159/000063543

    Article  CAS  Google Scholar 

  3. Welss T, Basketter DA, Schröder KR (2004) In vitro skin irritation: facts and future. State of the art review of mechanisms and models. Toxicol Vitr 18:231–243. https://doi.org/10.1016/j.tiv.2003.09.009

    Article  CAS  Google Scholar 

  4. Muhammad F, Riviere JE (2007) Dermal toxicity. Vet. Toxicol:263–276

    Google Scholar 

  5. Park Y-H, Kim JN, Jeong SH et al (2010) Assessment of dermal toxicity of nanosilica using cultured keratinocytes, a human skin equivalent model and an in vivo model. Toxicology 267:178–181. https://doi.org/10.1016/j.tox.2009.10.011

    Article  CAS  PubMed  Google Scholar 

  6. Roguet R, Cohen C, Leclaire J et al (2000) Use of a standardized reconstructed epidermis kit to assess in vitro the tolerance and the efficacy of cosmetics. Int J Cosmet Sci 22:409–419

    Article  CAS  Google Scholar 

  7. Abd E, Yousef SA, Pastore MN et al (2016) Skin models for the testing of transdermal drugs. Clin Pharmacol 8:163–176. https://doi.org/10.2147/CPAA.S64788

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Kim K, Park H, Lim K-M (2015) Phototoxicity: its mechanism and animal alternative test methods. Toxicol Res 31:97–104. https://doi.org/10.5487/TR.2015.31.2.097

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Netzlaff F, Lehr C-M, Wertz PW, Schaefer UF (2005) The human epidermis models EpiSkin®, SkinEthic® and EpiDerm®: an evaluation of morphology and their suitability for testing phototoxicity, irritancy, corrosivity, and substance transport. Eur J Pharm Biopharm 60:167–178. https://doi.org/10.1016/j.ejpb.2005.03.004

    Article  CAS  PubMed  Google Scholar 

  10. Ho J, Heisler E, Weimans S et al Phototoxicity in vitro: Investigation of photoreactions in the skin using the reconstructed epidermis. Epidermal Skin Test 1000 1000:1000

    Google Scholar 

  11. Wills JW, Hondow N, Thomas AD et al (2016) Genetic toxicity assessment of engineered nanoparticles using a 3D in vitro skin model (EpiDerm™). Part Fibre Toxicol 13:50. https://doi.org/10.1186/s12989-016-0161-5

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. do Nascimento Pedrosa T, Catarino CM, Pennacchi PC et al (2017) A new reconstructed human epidermis for in vitro skin irritation testing. Toxicol In Vitro 42:31–37. https://doi.org/10.1016/j.tiv.2017.03.010

    Article  CAS  Google Scholar 

  13. Brohem CA, da Silva Cardeal LB, Tiago M et al (2011) Artificial skin in perspective: concepts and applications. Pigment Cell Melanoma Res 24:35–50. https://doi.org/10.1111/j.1755-148X.2010.00786.x

    Article  PubMed  Google Scholar 

  14. De Vuyst E, Charlier C, Giltaire S et al (2014) Reconstruction of normal and pathological human epidermis on polycarbonate filter. Methods Mol Biol 1195:191–201. https://doi.org/10.1007/7651_2013_40

    Article  PubMed  Google Scholar 

  15. Poumay Y, Dupont F, Marcoux S et al (2004) A simple reconstructed human epidermis: preparation of the culture model and utilization in in vitro studies. Arch Dermatol Res 296:203–211. https://doi.org/10.1007/s00403-004-0507-y

    Article  CAS  PubMed  Google Scholar 

  16. Groeber F, Schober L, Schmid FF et al (2016) Catch-up validation study of an in vitro skin irritation test method based on an open source reconstructed epidermis (phase II). Toxicol Vitr 36:254–261. https://doi.org/10.1016/j.tiv.2016.07.008

    Article  CAS  Google Scholar 

  17. De Wever B, Goldberg A, Eskes C et al (2015) “Open source”–based engineered human tissue models: a new gold standard for nonanimal testing through openness, transparency, and collaboration, promoted by the ALEXANDRA Association. Appl Vitr Toxicol 1:5–9. https://doi.org/10.1089/aivt.2014.0011

    Article  Google Scholar 

  18. Pennacchi PC, de Almeida MES, Gomes OLA et al (2015) Glycated reconstructed human skin as a platform to study the pathogenesis of skin aging. Tissue Eng Part A 21:2417–2425. https://doi.org/10.1089/ten.tea.2015.0009

    Article  CAS  PubMed  Google Scholar 

  19. Brohem CA, Massaro RR, Tiago M et al (2012) Proteasome inhibition and ROS generation by 4-nerolidylcatechol induces melanoma cell death. Pigment Cell Melanoma Res 25:354–369. https://doi.org/10.1111/j.1755-148X.2012.00992.x

    Article  CAS  PubMed  Google Scholar 

  20. Faiao-Flores F, Alves-Fernandes DK, Pennacchi PC et al (2016) Targeting the hedgehog transcription factors GLI1 and GLI2 restores sensitivity to vemurafenib-resistant human melanoma cells. Oncogene 36(13):1849–1861. https://doi.org/10.1038/onc.2016.348

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. da Silva Cardeal LB, Brohem CA, Correa TCS et al (2006) Higher expression and activity of metalloproteinases in human cervical carcinoma cell lines is associated with HPV presence. Biochem Cell Biol 84:713–719. https://doi.org/10.1139/o06-084

    Article  PubMed  Google Scholar 

Download references

Acknowledgments

The authors thank the Histological Techniques Sector of ICB-USP for the technical support. Financial support was provided by FAPESP (Grants# 2013/12682-9, 2011/07441-7, 2011/14327-6, 2011/22812-1, 2013/00735-0, 2014/24400-0), CAPES, CNPq- INCT-if, and RENAMA.

Author information

Authors and Affiliations

  1. Skin Biology and Melanoma Laboratory, School of Pharmaceutical Sciences, University of São Paulo, Butantã, SP, Brazil

    Tatiana do Nascimento Pedrosa, Carolina Motter Catarino, Paula Comune Pennacchi, Silvia Berlanga de Moraes Barros & Silvya Stuchi Maria-Engler

Authors
  1. Tatiana do Nascimento Pedrosa
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Carolina Motter Catarino
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Paula Comune Pennacchi
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Silvia Berlanga de Moraes Barros
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Silvya Stuchi Maria-Engler
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Silvya Stuchi Maria-Engler .

Editor information

Editors and Affiliations

  1. MitoLab, Department of Life Sciences, Center for Neurosciences and Cell Biology of the University of Coimbra, Coimbra, Portugal

    Carlos Manuel Marques Palmeira

  2. School of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, Ribeirão Preto, São Paulo, Brazil

    Danielle Palma de Oliveira

  3. Department of Chemistry, Faculty of Philosophy, Sciences and Letters of Ribeirão Preto, University of São Paulo, Ribeirão Preto, São Paulo, Brazil

    Daniel Junqueira Dorta

Rights and permissions

Reprints and permissions

Copyright information

© 2021 Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

do Nascimento Pedrosa, T., Catarino, C.M., Pennacchi, P.C., de Moraes Barros, S.B., Maria-Engler, S.S. (2021). Skin Equivalent Models: Protocols for In Vitro Reconstruction for Dermal Toxicity Evaluation. In: Palmeira, C.M.M., de Oliveira, D.P., Dorta, D.J. (eds) Toxicity Assessment. Methods in Molecular Biology, vol 2240. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1091-6_3

Download citation

  • DOI: https://doi.org/10.1007/978-1-0716-1091-6_3

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1090-9

  • Online ISBN: 978-1-0716-1091-6

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation