Ultraviolet transmission microscopy for the imaging of topical sunscreen emulsions.

OBJECTIVE: Microscopy is widely used during the development and testing of topical formulations; however, it often lacks the ability to be chemically specific with regard to what is being imaged. This article describes how moving outside of the visible light region and into different parts of the ultraviolet (UV) spectrum enables differently UV absorbing components in topical emulsions to be directly visualized using optical transmission microscopy. METHODS: Optical transmission microscopy of different sunscreen emulsions was carried out using a custom-built microscope, imaging in the UVB (313 nm), UVA (365 nm) and visible light (546 nm) and with different magnifications. RESULTS: By using light of different wavelengths, direct visualization of different UV absorbing ingredients within the product emulsion using optical transmission microscopy has been performed and the locations of the UV absorbing actives in the formulations imaged. CONCLUSIONS: Microscopy has long been a valuable tool for the skin researcher, providing structural information about the products and how they perform. By moving outside of the spectral region of visible light and into the UV, it has been possible for the first time to directly image different SPF ingredients within topical formulations using optical microscopy.

OBJECTIF: La microscopie est largement utilisée dans le processus de développement et d'analyse des formulations topiques; cependant, elle ne parvient pas souvent à être chimiquement spécifique en ce qui concerne l'objet de l'imagerie. Le présent article décrit comment le déplacement au-delà de la zone de lumière visible et dans différentes parties du spectre ultraviolet (UV) permet aux ingrédients qui absorbent différemment les UV dans les émulsions topiques, d'être directement visualisés grâce à la microscopie à transmission optique. MÉTHODES: La microscopie à transmission optique de différentes émulsions de crème solaire a été réalisée à l'aide d'un microscope sur mesure, de l'imagerie dans les UVB (313 nm), les UVA (365 nm) et la lumière visible (546 nm) et à différents grossissements. RÉSULTATS: En utilisant la lumière de différentes longueurs d'onde, une visualisation directe des différents ingrédients absorbant les UV dans l'émulsion du produit grâce à la microscopie à transmission optique a été réalisée et les emplacements des substances actives absorbant les UV dans les formulations ont fait l'objet d'imagerie. CONCLUSIONS: La microscopie a été depuis longtemps un outil précieux pour les spécialistes de la peau, en fournissant des informations structurelles sur les produits et leurs performances. En se déplaçant au-delà de la région spectrale de la lumière visible, dans les UV, il a été possible pour la première fois d'obtenir directement une imagerie des différents ingrédients du facteur de protection solaire dans des formulations topiques à l'aide de la microscopie optique.

Blue Light and the Skin.

In photodermatology, UV radiation is the component of the solar system that has attracted the most interest as it represents the greatest risk of skin damage from solar exposure. Efficient protection strategies have therefore been developed to protect skin against powerful solar radiation. Recently, there has been increasing evidence to suggest that less energetic radiation, such as visible light and infrared radiation, might also influence skin physiology. Yet, it remains unclear, regarding risk assessment, whether visible light irradiation induces positive or negative effects in skin and when appropriate protection is needed. This review focuses primarily on blue light as part of the visible spectrum and sets out current mechanistic understanding of the benefits and risks of blue-light exposure to skin. Furthermore, it discusses phototherapies and potential strategies for protecting against detrimental effects of blue light such as hyperpigmentation and premature skin aging.

Effect of regioisomers of hydroxystearic acids as peroxisomal proliferator-activated receptor agonists to boost the anti-ageing potential of retinoids.

INTRODUCTION: We report on the in vitro and ex vivo effects of chiral (R)-10-hydroxystearic acid (10-HSA) compared with other mono-hydroxystearic acid regioisomers and stearic acid (SA) together with its benefit when combined with retinol. METHODS: Following treatment with hydroxystearic acids peroxisomal proliferator-activated receptor alpha (PPARα) activity was determined in a luciferase reporter gene assay, collagen type I was assessed in primary human dermal fibroblasts by immunohistochemistry, modification of the intracellular fibroblast collagen proteome was studied by mass-spectrometry-based proteomics and collagen type III was assessed by immunohistochemistry on human ex vivo skin. RESULTS: 10-HSA was the most effective PPARα agonist (15.7× induction; p < 0.001), followed by 9-HSA (10.1× induction) and then 12-HSA (4.9× induction) with 17-HSA (1.7× induction) being similar to the effects of stearic acid (1.8× induction). Collagen type I levels were increased in primary human fibroblasts by 2.12× and 1.56× for 10-HSA and 9-HSA, respectively, in vitro with the10-HSA being significant (p < 0.05), whereas 12-HSA and SA had no statistical effect over the untreated control. 10-HSA and 12-HSA modified the intracellular fibroblast collagen proteome slightly with significant increases in collagen alpha-1 (VI) and alpha-3 (VI) proteins but only 10-HSA increased levels of collagen alpha-2 (V), alpha-1 (III), alpha-1 (I) and alpha-2 (I) (all p < 0.05) with the increases being significantly different between 10-HSA and 12-HSA for collagen alpha-1 (I), collagen-3 (VI) and collagen alpha-2 (I) (p < 0.01). Collagen type III in ex vivo skin was increased +47% (p < 0.05) by 0.05% (1.7 mM) retinol, +70% (p < 0.01) by 0.01% (0.33 mM) 10-HSA and the combination increased levels by +240% (p < 0.01 for either ingredient). CONCLUSION: Chiral (R)-10-HSA has been shown to be superior to 9, 12 and 17-HSA as a PPARα agonist. Moreover, 10-HSA stimulated collagen synthesis in monolayer fibroblast culture as assessed by proteomics and immunohistochemically. Furthermore, we also show the synergistic effects of 10-HSA with retinol on collagen III synthesis in skin explants. These results further highlight the efficacy of 10-HSA as a cosmetically acceptable PPARα agonist and anti-ageing ingredient.

INTRODUCTION: Nous rapportons les effets in vitro et ex vivo de l'acide chiral (R)-10-hydroxystéarique (10-HSA) par rapport à d'autres régioisomères d'acide mono-hydroxystéarique et à l'acide stéarique (SA) ainsi que ses avantages lorsqu'il est associé au rétinol. MÉTHODES: Après un traitement avec des acides hydroxystéariques, l'activité du récepteur alpha activé par les proliférateurs peroxysomaux (PPARα) a été déterminée dans un test du gène rapporteur de la luciférase, le collagène de type I a été évalué dans les fibroblastes dermiques humains primaires par immunohistochimie, la modification du protéome du collagène des fibroblastes intracellulaires a été étudiée par spectrométrie de masse. La protéomique et le collagène de type III ont été évalués par immunohistochimie sur la peau humaine ex vivo. RÉSULTATS: la 10-HSA était l'agoniste PPARα le plus efficace (induction 15,7X ; p<0,001), suivi de la 9-HSA (induction 10,1X) puis de la 12-HSA (induction 4,9X) avec la 17-HSA (induction 1,7X) étant similaire aux effets de l'acide stéarique (induction 1,8X). Les niveaux de collagène de type I ont été augmentés dans les fibroblastes humains primaires de 2,12X et 1,56X pour la 10-HSA et la 9-HSA respectivement in vitro, la 10-HSA étant significative (p<0,05) : alors que la 12-HSA et la SA n'ont eu aucun effet statistique sur le témoin non traité. La 10-HSA et la 12-HSA ont légèrement modifié le protéome du collagène des fibroblastes intracellulaires avec des augmentations significatives des protéines de collagène alpha-1 (VI) et alpha-3 (VI), mais seule la 10-HSA a augmenté les niveaux de collagène alpha-2 (V), alpha -1 (III), alpha-1 (I) et alpha-2 (I) (tous p<0,05) avec des augmentations significativement différentes entre 10-HSA et 12-HSA pour le collagène alpha-1 (I), le collagène- 3 (VI) et Collagène alpha-2 (I) (p<0,01). Le collagène de type III dans la peau ex vivo a augmenté de +47 % (p<0,05) de 0,05 % (1,7 mM) de rétinol, de +70 % (p<0,01) de 0,01 % (0,33 mM) de 10-HSA et la combinaison a augmenté les niveaux de +240 % (p<0,01 pour chaque ingrédient). CONCLUSION: La chiral (R)-10-HSA s'est avérée supérieure à 9, 12 et 17-HSA en tant qu'agoniste de PPARα. De plus, la 10-HSA a stimulé la synthèse de collagène dans la culture de fibroblastes monocouche telle qu'évaluée par protéomique et immunohistochimique. De plus, nous montrons également les effets synergiques de la 10-HSA avec le rétinol sur la synthèse du collagène III dans les explants de peau. Ces résultats soulignent en outre l'efficacité de la 10-HSA en tant qu'agoniste de PPARα et ingrédient anti-âge cosmétiquement acceptable.

Mitochondrial and glycolytic activity of UV-irradiated human keratinocytes and its stimulation by a Saccharomyces cerevisiae autolysate.

Cutaneous aging is correlated with mitochondrial dysfunction and a concomitant decline in energy metabolism that can be accelerated by extrinsic factors such as UV radiation (UVR). In this study we compared cellular bioenergetics of normal and UV-irradiated primary human epidermal keratinocytes. Moreover, we investigated the influence of a Saccharomyces cerevisiae autolysate (SCA) on stressed keratinocytes to regain cellular homeostasis. Cellular metabolism was assessed by extracellular flux analysis which measures oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) as well as by ATP quantification. The expression level of ten mitochondria related genes in normal and UVR-stimulated (60mJ/cm(2) UVB) keratinocytes was quantified by real-time PCR and the impact of SCA addition was determined. Sublethal UV stress increased mitochondrial dysfunction in keratinocytes which resulted in reduced viability, uncoupled oxidative phosphorylation, and down-regulated mitochondrial gene expression. Particularly, gene expression of SHDA, UPC2, BID, and ATP5A1 was reduced about twofold within 4h. Treatment of keratinocytes with SCA shifted cellular metabolism towards a more energetic status by increasing the respiratory rate and glycolysis. SCA also stimulated cellular ATP production after short (4h) and prolonged (22h) incubations and induced the expression of genes related to mitochondrial function towards normal expression levels upon UV irradiation. The decreased respiratory capacity of UV-irradiated keratinocytes was partially compensated by the addition of SCA which enhanced glycolytic activity and thereby increased cellular resistance to environmental stress.

Towards a DNA-like duplex without hydrogen bonds.

The inverse quadrupolar moments of the phenyl and pentafluorophenyl residues in the base pair P-F5 promotes strong intramolecular stacking interactions in DNA duplexes. The more natural base pairs are replaced by this novel pair the higher the thermodynamic stability of the resulting duplex if they are arranged in an alternating fashion.