A potential in vitro epidermal equivalent assay to determine sensitizer potency

Toxicol In Vitro. 2011 Feb;25(1):347-57. doi: 10.1016/j.tiv.2010.10.008. Epub 2010 Oct 19.

Abstract

Most in vitro assays aim to distinguish sensitizers from non-sensitizers. Few aim to classify sensitizers according to potency. Here, we describe a potential method for classifying sensitizers according to their irritant potency with the aid of in house epidermal equivalents (EE). Sixteen sensitizers were applied topically in a dose response to EE for 24h. The EE-EC(50) value (effective chemical concentration required to reduce cell viability by 50%) and the EE-IL-1α(10)(×) value (chemical concentration which increases IL-1α secretion by 10-fold) were calculated. From 16 sensitizers, EE-EC(50) and/or EE-IL-1α(10×) values were obtained from 12 skin sensitizers. EE-EC(50) and IL-1α(10×) values decreased in proportion to increasing sensitizer potency. The in vitro assay correlated with existing in vivo mouse and human sensitization data (LLNA, HRIPT), and showed low intra- and inter-experimental variability. Additionally DNCB and resorcinol were correctly assessed as extreme and moderate sensitizers using commercial EE (EST1000™ and RHE™). In conclusion, our data supports the view that irritancy may in part be a factor determining sensitizer potency. Since this assay does not distinguish sensitizers from non-sensitizers, its potential application is in a tiered strategy, where Tier 1 identifies sensitizers which may then tested in Tier 2, this assay, which determines sensitizer potency.

Publication types

  • Comparative Study
  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Allergens / classification
  • Allergens / toxicity*
  • Animal Testing Alternatives
  • Cell Survival / drug effects
  • Cells, Cultured
  • Dermatitis, Allergic Contact / prevention & control
  • Dinitrochlorobenzene / toxicity
  • Epidermis / drug effects*
  • Epidermis / immunology*
  • Epidermis / metabolism
  • Foreskin / cytology
  • Humans
  • Infant, Newborn
  • Inhibitory Concentration 50
  • Interleukin-1alpha / metabolism
  • Irritants / classification
  • Irritants / toxicity*
  • Keratinocytes / drug effects
  • Keratinocytes / immunology
  • Keratinocytes / metabolism
  • Male
  • Osmolar Concentration
  • Reproducibility of Results
  • Resorcinols / toxicity
  • Technology Transfer
  • Toxicity Tests* / methods

Substances

  • Allergens
  • Dinitrochlorobenzene
  • IL1A protein, human
  • Interleukin-1alpha
  • Irritants
  • Resorcinols
  • resorcinol