Highlighting the hygroscopic capacities of apiogalacturonans.

To meet the needs of dehydrated skin, molecules with a high hygroscopic potential are necessary to hydrate it effectively and durably. In this context, we were interested in pectins, and more precisely in apiogalacturonans (AGA), a singular one that is currently only found in a few species of aquatic plants. As key structures in water regulation of these aquatic plants and thanks to their molecular composition and conformations, we hypothesized that they could have beneficial role for skin hydration. Spirodela polyrhiza is a duckweed known to be naturally rich in AGA. The aim of this study was to investigate the hygroscopic potential of AGA. Firstly, AGA models were built based on structural information obtained from previous experimental studies. Molecular dynamics (MD) simulations were performed, and the hygroscopic potential was predicted in silico by analyzing the frequency of interaction of water molecules with each AGA residue. Quantification of interactions identified the presence of 23 water molecules on average in contact with each residue of AGA. Secondly, the hygroscopic properties were investigated directly in vivo. Indeed, the water capture in the skin was measured in vivo by Raman microspectroscopy thanks to the deuterated water (D20) tracking. Investigations revealed that AGA significantly capture and retain more water in the epidermis and deeper than a placebo control. Not only do these original natural molecules interact with water molecules, but they capture and retain them efficiently in the skin.

Utilizing deep learning for dermal matrix quality assessment on in vivo line-field confocal optical coherence tomography images.

BACKGROUND: Line-field confocal optical coherence tomography (LC-OCT) is an imaging technique providing non-invasive "optical biopsies" with an isotropic spatial resolution of ∼1 µm and deep penetration until the dermis. Analysis of obtained images is classically performed by experts, thus requiring long and fastidious training and giving operator-dependent results. In this study, the objective was to develop a new automated method to score the quality of the dermal matrix precisely, quickly, and directly from in vivo LC-OCT images. Once validated, this new automated method was applied to assess photo-aging-related changes in the quality of the dermal matrix. MATERIALS AND METHODS: LC-OCT measurements were conducted on the face of 57 healthy Caucasian volunteers. The quality of the dermal matrix was scored by experts trained to evaluate the fibers' state according to four grades. In parallel, these images were used to develop the deep learning model by adapting a MobileNetv3-Small architecture. Once validated, this model was applied to the study of dermal matrix changes on a panel of 36 healthy Caucasian females, divided into three groups according to their age and photo-exposition. RESULTS: The deep learning model was trained and tested on a set of 15 993 images. Calculated on the test data set, the accuracy score was 0.83. As expected, when applied to different volunteer groups, the model shows greater and deeper alteration of the dermal matrix for old and photoexposed subjects. CONCLUSIONS: In conclusion, we have developed a new method that automatically scores the quality of the dermal matrix on in vivo LC-OCT images. This accurate model could be used for further investigations, both in the dermatological and cosmetic fields.

Investigation of the molecular signature of greying hair shafts.

OBJECTIVE: Hair greying (i.e. canitie) is a physiological process occurring with the loss of melanin production and deposition within the hair shafts. Many studies reported the oxidation as the main biological process underlying this defect of pigmentation. Even though the overall appearance and biomechanical properties of hairs are reported to be altered with greying, there is a lack of information about molecular modifications occurring in grey hair shafts. The aim of this study was thus to investigate the molecular signature and associated changes occurring in greying hair shafts by confocal Raman microspectroscopy. METHODS: This study was conducted on pigmented, intermediate (i.e. grey) and unpigmented hairs taken from 29 volunteers. Confocal Raman microspectroscopy measurements were acquired directly on hair shafts. RESULTS: Automatic classification of Raman spectra revealed 5 groups displaying significant differences. Hence, the analysis of the molecular signature highlighted the existence of 3 sub-groups within grey hair: light, medium and dark intermediate. Among molecular markers altered in the course of greying, this study identified for the first time a gradual modification of lipid conformation (trans/gauche ratio) and protein secondary structure (α-helix/ß-sheet ratio), referring respectively to an alteration of barrier function and biomechanical properties of greying hair. CONCLUSION: This study thus reports for the first time a highly specific molecular signature as well as molecular modifications within grey hair shaft.

OBJECTIF: Le grisonnement du cheveu (i.e. canitie) est un processus physiologique correspondant à l'altération de la production et du dépôt des pigments de mélanine au sein de la tige pilaire. De nombreuses études identifient l'oxydation en tant que principal phénomène à l'origine de ce défaut de pigmentation. L'apparence globale et les propriétés biomécaniques des cheveux grisonnants sont également rapportées comme étant altérées. Cependant, il existe un manque d'information concernant les modifications moléculaires ayant lieu dans la tige pilaire grisonnante. Le but de cette étude était donc d'investiguer par microspectroscopie confocale Raman la signature moléculaire de la tige pilaire grisonnante ainsi que les changements biologiques associés. MÉTHODES: Cette étude a été réalisée sur des cheveux pigmentés, intermédiaires (i.e. gris) et non pigmentés, prélevés sur 29 volontaires. Les mesures par microspectroscopie Raman confocale ont directement été acquises sur la tige pilaire. RÉSULTATS: Une classification automatique des spectres Raman a permis de révéler 5 groupes présentant des différences significatives. Ainsi, l'analyse de la signature moléculaire spectrale identifie 3 sous-groupes au sein des cheveux gris : intermédiaires clairs, moyens et foncés. Parmi les marqueurs moléculaires altérés au cours du grisonnement, cette étude identifie pour la première fois une modification graduelle de la conformation des lipides (ratio trans /gauche) et de la structure secondaire des protéines (ratio hélice α/feuillets ß). Ces marqueurs correspondent respectivement à l'altération de la fonction barrière et des propriétés biomécaniques des cheveux gris. CONCLUSION: Cette étude met en évidence pour la première fois une signature moléculaire extrêmement précise ainsi que des modifications moléculaires en lien avec le grisonnement de la tige pilaire.

Development and characterization of a 3D in vitro model mimicking acneic skin.

Acne is an inflammatory skin disease of the pilosebaceous unit, involving four essential factors: hyperseborrhoea combined to a modification of sebum composition, colonization by Cutibacterium (C.) acnes, hyperkeratinization and secreted inflammation. Understanding and mimicking compromised skin is essential to further develop appropriate therapeutic solutions. This study aimed to develop new in vitro 3D models mimicking acneic skin, by combining two main factors involved in the physiopathology, namely, altered sebum composition and C. acnes invasion. Normal human keratinocytes were first used to generate reconstructed human epidermis (RHE) that were then left untreated (control) or treated topically with a combination of both peroxidized squalene and C. acnes cultures. Once validated, this model considered relevant to mimic acneic skin, was further improved by using different phylotypes of C. acnes strains specifically isolated from healthy and acneic patients. While both phylotypes IB and II did not significantly alter RHE, C. acnes IA1 strains induce major acneic skin hallmarks such as hyperkeratinization, secreted inflammation and altered barrier function. Interestingly, these results are obtained independently of the origin of IA1 phylotypes (acneic vs. healthy patient), thus suggesting a role of the ecosystem in controlling C. acnes virulence in healthy skin. In conclusion, by combining two major factors involved in the physiopathology of acne, we (1) succeeded to design in vitro 3D models mimicking this skin disorder and (2) highlighted how C. acnes phylotypes can have an impact on epidermal physiology. These relevant models will be suitable for the substantiation of therapeutic molecules dedicated to acne treatment.

Autophagy in human keratinocytes: an early step of the differentiation?

Studies have established that autophagy constitutes an efficient process to recycle cellular components and certain proteins. The phenomenon was demonstrated primarily in response to nutrient starvation, and there are increasing evidences that it is implied in differentiation. Keratinocyte differentiation was going along an activation of lysosomal enzymes and organelle clearance, and terminal steps are sometimes described as a specialized form of cell death leading to corneocytes. We examined whether initiation of the process in human keratinocyte HaCaT involves autophagy. The KSFM™ culture medium was substituted by M199, which contains a low glucose concentration but a high calcium level (known to induce differentiation). Metabolic stress reduced enhanced cell number in G(1) phase, without apoptotic features (ΔΨmt and membrane integrity are unchanged). Morphological changes were associated with a lower integrin ß1 expression and modifications of protein levels involved in keratinocyte differentiation (involucrin, keratin K10 and ΔNp63α). Whereas autophagic signalling was supported by SIRT1 and pAMPK (T172) increase according to time kinetic, which led to the disappearance of mTOR phosphorylated on S2448 residue. The significant Bcl-X(L) level reduction with stress promoted autophagy, by the release of Beclin-1, whereas ATG5-ATG12 and LC3-II that are involved in autophagosome formation were enhanced significantly. Then, the level of lysosomal protein cathepsin B rose to execute autophagy. Kinetic studies established that autophagy would constitute an early signalling process required for keratinocyte commitment in differentiation pathway.