Confocal Raman Spectroscopy Coupled With in Vitro Permeation Testing to Study the Effects of Formalin Fixation on the Skin Barrier Function of Reconstructed Human Epidermis

Authors

Hichem Kichou, UPR CNRS 4301 CBM, département NMNS « NanoMédicaments et NanoSondes », Université de Tours, 31 avenue Monge, 37200 Tours.
Franck Bonnier, LVMH Recherche, 185 Av. de Verdun, 45800, Saint-Jean-de-Braye, France.
Amanda C. Caritá, FOCAS Research Institute, TU Dublin, City Campus, Camden Row, Dublin 8, D08 CKP1, Ireland
Hugh Byrne, FOCAS Research Institute, TU Dublin, City Campus, Camden Row, Dublin 8, D08 CKP1, IrelandFollow
Igor Choupra, UPR CNRS 4301 CBM, département NMNS « NanoMédicaments et NanoSondes », Université de Tours, 31 avenue Monge, 37200 Tours.
Emilie Munnier, UPR CNRS 4301 CBM, département NMNS « NanoMédicaments et NanoSondes », Université de Tours, 31 avenue Monge, 37200 Tours.

Author ORCID Identifier

0000-0002-1735-8610

Document Type

Article

Disciplines

Atomic, Molecular and Chemical Physics, Analytical chemistry, Biochemical research methods, Biophysics, Biomaterials, Biomaterials

Publication Details

https://www.sciencedirect.com/science/article/pii/S1386142524007832

https://doi.org/10.1016/j.saa.2024.124617

Abstract

Confocal Raman Spectroscopy is recognised as a potent tool for molecular characterisation of biological specimens. There is a growing demand for In Vitro Permeation Tests (IVPT) in the pharmaceutical and cosmetic areas, increasingly conducted using Reconstructed Human Epidermis (RHE) skin models. In this study, chemical fixation of RHE in 10% Neutral Buffered Formalin for 24 hours has been examined for storing RHE samples at 4°C for up to 21 days. Confocal Raman Spectroscopy, combined with Principal Components Analysis, revealed the molecular-level effects of fixation, notably in protein and lipid conformation within the stratum corneum and viable epidermis. IVPT by means of high-performance liquid chromatography, using caffeine as a model compound, showed minimal impact of formalin fixation on the cumulative amount, flux, and permeability coefficient after 12 hours. While the biochemical architecture is altered, the function of the model as a barrier to maintain rate-limiting diffusion of active molecules within skin layers remains intact. This study opens avenues for enhanced flexibility and utility in skin model research, promising insights into mitigating the limited shelf life of RHE models by preserving performance in fixed samples for up to 21 days.

DOI

10.1016/j.saa.2024.124617

Recommended Citation

Kichou, Hichem; Bonnier, Franck; Caritá, Amanda C.; Byrne, Hugh; Choupra, Igor; and Munnier, Emilie, "Confocal Raman Spectroscopy Coupled With in Vitro Permeation Testing to Study the Effects of Formalin Fixation on the Skin Barrier Function of Reconstructed Human Epidermis" (2024). Articles. 5.
https://arrow.tudublin.ie/focasart/5

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