4.5 Article

Immortalized keratinocytes cells generates an effective model of Epidermal Human Equivalent for irritation and corrosion tests

Journal

TOXICOLOGY IN VITRO
Volume 71, Issue -, Pages -

Publisher

PERGAMON-ELSEVIER SCIENCE LTD
DOI: 10.1016/j.tiv.2020.105069

Keywords

Epidermal human exposure; Irritation; Corrosion; OECD 439; OECD 431; HaCaT; MTT and HPLC validation method

Categories

Funding

  1. Coordination for the Improvement of Higher Education Personnel -Brazil (CAPES) [001]
  2. Sao Paulo Research Foundation (FAPESP) [2016/15660-4, 2017/04926-6, 2019/14527-7]

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The researchers developed a skin model constructed using immortalized cells, which showed good predictive potential in chemical assessments and compliance with regulatory guidelines for safety evaluation.
Three-dimensional skin models, also named 3D skin models, human skin equivalents (HSEs), or Human Epidermal Equivalents (HEEs), have been increasingly used for chemical assessments in terms of efficacy and safety. Considering this, we developed an HEE model using immortalized HaCaT cells, aiming to overcome the limitation of primary tissue source. Our 3D model (HaCaT-HEE) exhibited important markers of cell differentiation (CK10, CK14, involucrin, and filaggrin), although the stratum corneum was shown to be modest. Besides, the model showed a good prediction potential considering membrane permeability, sensitivity, specificity, and accuracy in distinguishing irritant and corrosive effects after exposure to selected chemicals recommended by the OECD protocols. We also validated the formazan determination for the MTT method using High-Performance Liquid Chromatography (HPLC). For that, we considered carry over, linearity, reproducibility/robustness, accuracy, precision, selectivity, and matrix effect, according to the Food and Drug Administration (FDA) guideline. Based on our results, we can conclude that our model has an acceptable predictive value for the safety evaluation of compounds after skin exposure, with the great advantage of being constructed using immortalized cells.

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