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Introduction: The fundamental function of the epidermis is to provide an inside-out barrier to water loss and an outside-in barrier to penetration of external irritants. Transepidermal water loss (TEWL) has been extensively used as a method of estimating the skin barrier quality, typically without any consideration of directionality. The validity of TEWL as an estimate of skin permeability to external substances has been controversial in vitro and in vivo. The aim of this work was to assess the relationship between TEWL and the penetration of a topically applied external marker (caffeine) in healthy skin in vivo before and following a challenge to the barrier., Methods: The skin barrier was challenged by application of aqueous solutions of mild cleanser products under occlusion for 3 h on the forearms of nine human participants. Skin barrier quality was evaluated before and after the challenge by measuring the TEWL rate and the permeated amount of topically applied caffeine using in vivo confocal Raman microspectroscopy., Results: No skin irritation was observed following the skin barrier challenge. TEWL rates and the caffeine penetrated amount in the stratum corneum after the challenge were not correlated. A weak correlation was observed when the changes were corrected to water-only treatment. TEWL values can be influenced by environmental conditions as well as the skin temperature and water content., Conclusions: Measuring TEWL rates is not always representative of the outside-in barrier. TEWL may be useful in differentiating large changes in skin barrier function (e.g., between healthy and compromised skin) but is less sensitive to small variations following topical application of mild cleansers., (© 2023 The Authors. Skin Research and Technology published by John Wiley & Sons Ltd.)

Background: Monitoring the transcutaneous permeation of exogenous molecules using conventional techniques generally requires long pre-analytical preparation or labelling of samples. However, Raman spectroscopy is a label-free and non-destructive method which provides spatial distribution of tracked actives in skin. The aim of our study was to prove the interest of Raman imaging coupled with multivariate curve resolution alternating least square (MCR-ALS) analysis in monitoring retinol penetration into frozen and living human skin., Materials and Methods: After topical treatment of skin samples by free or encapsulated retinol, thin cross sections were analysed by Raman imaging (up to 100 µm depth). Mann-Whitney test was used to identify retinol spectroscopic markers in skin. MCR-ALS was used to estimate retinol contribution in Raman spectral images. Heat maps were constructed to compare the distribution of free and encapsulated retinol in skin models., Results: We identified the bands at 1158, 1196 and 1591 cm -1 as specific features for monitoring retinol in skin. Moreover, our MCR-ALS results showed an improvement of retinol penetration (up to 30 µm depth) with the encapsulated form as well as storage reservoir formation in stratum corneum, for each skin model. Finally, greater retinol penetration into living skin was observed., Conclusion: This study shows a proof of concept for the evaluation of retinol penetration in skin using Raman imaging coupled with MCR-ALS. This concept needs to be validated on more subjects to include inter-individual variability but also other factors affecting skin permeation (age, sex, pH, etc). Our study can be extended to other actives., (© 2021 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Non-enzymatic glycation is a post-translational modification of long-lived matrix proteins such as type I collagen. It occurs during aging and leads to the formation of advanced glycation end-products (AGEs). AGE accumulation is associated with severe complications in chronic and age-related diseases. The assessment of modifications induced by this (patho)physiological process represents an interest in biology and medicine for a better patient care. The objective of our work was to position the interest of Raman spectroscopy in the quantification of collagen glycation. Two types of in vitro glycation were used by incubating collagen samples, at different durations, with ribose or glyoxylic acid; these reducing agents acting on the chemical specificity of the glycation reaction. Glycation efficiency was evaluated by the liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) quantification of carboxymethyllysine (CML) and pentosidine, which are among the most studied AGEs. Raman data were processed by PCA coupled to validity indices and Lasso regression as multivariate analysis tools. Regression models were constructed by considering the LC-MS/MS results as reference values. A marked variability was observed within the Raman datasets making difficult the identification of spectral differences between control and ribose-treated collagen samples. By taking advantage of the chemical specificity of the glyoxylic acid treatment leading to CML formation, on one hand, and the feature selection included in the Lasso algorithm, on the other hand, Raman markers associated with glycation were identified. The assigned vibrations corresponded to modifications of side chains of collagen. In addition, a threshold of CML concentration was determined as quantitative indicator of the applicability of Raman spectroscopy for potential patient follow-up purposes. Although lacking in sensitivity to directly detect AGEs in collagen, Raman spectroscopy allows to highlight the molecular modifications of collagen induced by glycation., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

The variable configuration of Raman spectroscopic platforms is one of the major obstacles in establishing Raman spectroscopy as a valuable physicochemical method within real-world scenarios such as clinical diagnostics. For such real world applications like diagnostic classification, the models should ideally be usable to predict data from different setups. Whether it is done by training a rugged model with data from many setups or by a primary-replica strategy where models are developed on a 'primary' setup and the test data are generated on 'replicate' setups, this is only possible if the Raman spectra from different setups are consistent, reproducible, and comparable. However, Raman spectra can be highly sensitive to the measurement conditions, and they change from setup to setup even if the same samples are measured. Although increasingly recognized as an issue, the dependence of the Raman spectra on the instrumental configuration is far from being fully understood and great effort is needed to address the resulting spectral variations and to correct for them. To make the severity of the situation clear, we present a round robin experiment investigating the comparability of 35 Raman spectroscopic devices with different configurations in 15 institutes within seven European countries from the COST (European Cooperation in Science and Technology) action Raman4clinics. The experiment was developed in a fashion that allows various instrumental configurations ranging from highly confocal setups to fibre-optic based systems with different excitation wavelengths. We illustrate the spectral variations caused by the instrumental configurations from the perspectives of peak shifts, intensity variations, peak widths, and noise levels. We conclude this contribution with recommendations that may help to improve the inter-laboratory studies.

Raman spectroscopy is a candidate technique for diagnosis applications in medicine due to its high molecular specificity. Optimizing the pre-treatment applied for Raman data is important for exploiting Raman signals and ensuring their relevance in medical diagnosis. One of the crucial steps in data pre-processing, normalization, can affect significantly the result interpretation. To select the appropriate normalization method, a strategy based on validity indices (VI) is proposed in this study. VI are based on measuring the quality of data partitioning without involving a full sequence of supervised classification. The approach was tested on Raman data acquired from control and in vitro glycated proteins (albumin and collagen). Protein glycation is a process involved in the molecular ageing of tissues that leads to the formation of products altering the functional and structural properties of proteins. Different methods of normalization were applied on the data sets: integrated intensity of the phenylalanine band, integrated intensity of the amide I band, standard normal variate (SNV), multiplicative signal correction (MSC), and extended multiplicative signal correction (EMSC) that performs simultaneously baseline correction and normalization. Following normalization, principal component analysis (PCA) was applied and VI were calculated from PCA scores resulting from each of the normalization methods mentioned. Based on VI quantitative values, our experiments permit to illustrate the effect of normalization on the data separability of control and glycated samples, and to determine the most appropriate normalization and simultaneously the most discriminant principal components to exploit vibrational information associated with glycation-induced modifications. In parallel, principal component analysis - linear discriminant analysis (PCA-LDA) was carried out for positioning the interest of VI in regard to a common chain of data processing.

Correction for 'Towards normalization selection of Raman data in the context of protein glycation: application of validity indices to PCA processed spectra' by Fatima Alsamad et al., Analyst, 2020, DOI: 10.1039/c9an02155h.