An investigation into the effect of freezing conditions on the barrier function of reconstructed human epidermis using Raman spectroscopy and percutaneous permeation

LE STUDIUM Multidisciplinary Journal, 2020, 4, 79-88

Yuri Dancik 1,2,*, Hichem Kichou 2, Christophe Eklouh-Molinier 2, Martin Soucé 2 , Emilie Munnier 2, Igor Chourpa 2 and Franck Bonnier 2,* 
1 Le STUDIUM Institute of Advanced Studies, 1 rue Dupanloup, 45000 Orléans, France
2 Faculté de Pharmacie, Université de Tours, 31 Avenue Monge, EA 6295 NanoMédicaments et NanoSondes, 37200 Tours, France
*Authors to whom correspondence should be addressed.


Reconstructed human epidermis (RHE) is an emerging skin model in pharmaceutical, toxicological and cosmetic sciences, yielding scientific and ethical advantages. RHEs remain costly, however, due to consumables and time required for their culture and a short shelf-life. Storing, i.e., freezing RHE could help reduce costs but little is known on the effects of freezing on the barrier function of RHE. We studied such effects using commercial EpiSkin™ RHE stored at −20, −80 and −150 °C for 1 and 10 weeks. We acquired intrinsic Raman spectra in the stratum corneum (SC) of the RHEs as well as spectra obtained following topical application of resorcinol in an aqueous solution. In parallel, we quantified the effects of freezing on the permeation kinetics of resorcinol from time-dependent permeation experiments. Principal component analyses discriminated the intrinsic SC spectra and the spectra of resorcinol-containing RHEs, in each case on the basis of the freezing conditions. Permeation of resorcinol through the frozen RHE increased 3- to 6-fold compared to fresh RHE, with the strongest effect obtained from freezing at −20 °C for 10 weeks. Due to the extensive optimization and standardization of EpiSkin™ RHE, the effects observed in our work may be expected to be more pronounced with other RHEs.


Reconstructed human epidermis, storage conditions, Raman spectroscopy, skin barrier, permeation, resorcinol
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Le STUDIUM Multidisciplinary Journal