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