Background/aims: The study of changes in skin structure with age is becoming all the more important with the increase in life. The atrophy that occurs during aging is accompanied by more profound changes, with a loss of organization within the elastic collagen network and alterations in the basal elements. The aim of this study is to present a method to determine the mechanical properties of total human skin in vivo compared with dermal equivalents (DEs) using indentation and static friction tests. Methods: A new bio-tribometer working at a low contact pressure for the characterization the mechanical properties of the skin has been developed. This device, based on indentation and static friction tests, also allows to characterize the skin in vivo and reconstructed DEs in a wide range of light contact forces, stress and strain.
Most of adult women exhibit cellulite on the hips, buttock and thighs. Although extracellular matrix and lymphatic system disorders can increase its appearance, cellulite basically results from an excessive fat storage in the adipose tissue which exerts considerable pressure on the surrounding skin tissue and creates a dimpled irregular appearance. Caffeine, the most widely used anti-cellulite ingredient, favours fat break-down by inhibiting the phosphodiesterase enzyme and encouraging a high intracellular level of cAMP. A series of studies has shown that spermine and spermidine, two ubiquitous polyamines, encouraged fat storage and slowed fat break-down in the adipose tissue. Besides, it was shown that heparan sulfate glycosaminoglycans had a strong affinity for polyamines. To design a new cosmetic ingredient with anti-cellulite properties, we used molecular modelling to screen several ingredients with a structure similar to that of heparan sulfate glycosaminoglycans. This way, we identified sulfo-carrabiose as a potent molecule for trapping spermine and spermidine. These virtual results were first confirmed in tubo where sulfo-carrabiose was shown to dose-dependently inactivate spermine and spermidine. In vitro, adipocytes cultured with sulfo-carrabiose exhibited a significant reduction of lipogenesis and a significant increase of lipolysis. When sulfo-carrabiose was incorporated in a cosmetic formula, significant improvements were observed in thigh circumference, with better results than those obtained with caffeine after 28 days of use. Furthermore, a combination of caffeine and sulfo-carrabiose led to results significantly better than those obtained with caffeine alone. As measured by fringe projection, thigh volume was also significantly reduced after sulfo-carrabiose treatment. Finally, the appearance of cellulite assessed by clinical evaluation was also significantly reduced within 28 days.
Proopiomelanocortin is a precursor peptide that gives rise to several neuropeptides including adrenocorticotrophic hormone (ACTH) and β-endorphin. POMC-derived peptides have been shown to be synthesized in human epidermis where they modulate numerous skin functions. Because we previously observed that melanocortin receptor-2 and μ-opioid receptor 1, the respective receptors for ACTH and β-endorphin decreased with ageing in human epidermis, we have selected an active ingredient (INCI name: Achillea millefolium extract) able to upregulate receptor expressions. The aim of the present work was first to evaluate the effect of A. millefolium extract on the expression pattern of various epidermal differentiation markers ex vivo in normal human skin biopsies using quantitative image analysis and second to evaluate its capacity to rejuvenate the appearance of skin surface in vivo. Results show an improved expression profile of cytokeratin 10, transglutaminase-1 and filaggrin in cultured skin biopsies as well as an increased epidermal thickness. In vivo, a 2-month treatment with A. millefolium extract at 2% significantly improved the appearance of wrinkles and pores compared with placebo. Results were also directionally better than those of glycolic acid that was chosen as reference resurfacing molecule.
Lysyl oxidase-like (LOXL) is an extracellular enzyme that catalyses the cross-linking between microfibrils and tropoelastin (TE), thereby ensuring elastic fibre functionality. With ageing, LOXL expression decreases, thus participating in the loss of skin elasticity. In a previous study, we showed that a dill seed extract [INCI name: Peucedanum graveolens (Dill) extract] could increase LOXL expression in cultured dermal fibroblasts. Besides, we showed a good correlation between the measurements of skin elasticity obtained in vitro and in vivo using a fully automated bio-tribometer designed to measure the biomechanical properties of soft and complex materials like skin. The aim of this study was to evaluate the ability of the dill extract to improve skin elasticity in vitro and in vivo using different models. Using the bio-tribometer, we first showed that the lateral elasticity of dermis equivalents (DEs) treated with the dill extract at 1% was significantly increased by +29% (P < 0.01) when compared to untreated DEs. In vivo, skin firmness and elastic recovery measured using cutometry methods were also significantly improved compared to placebo in volunteers treated for 56 days with a formula containing 1% of dill extract. Moreover, the clinical evaluation evidenced significant improvements in 'skin elasticity' compared to placebo. A majority of subjects treated with the dill extract also noted significant improvements in skin elasticity, firmness and slackness of the jaw line. Finally, mean wrinkle area and length were also significantly reduced compared to placebo after 84 days as measured using silicone replicas taken from the crow's feet. In summary, this study showed that the dill extract could improve elasticity of DEs in vitro as well as skin biomechanical properties and appearance in vivo. It also highlights the relevance of using the bio-tribometer as an exploratory tool for the measurement of skin elasticity in vitro.
Tissue engineered skin usually consist of a multi-layered visco-elastic material composed of a fibrillar matrix and cells. The complete mechanical characterization of these tissues has not yet been accomplished. The purpose of this study was to develop a multiscale approach to perform this characterization in order to link the development process of a cultured skin to the mechanical properties. As a proof-of-concept, tissue engineered skin samples were characterized at different stages of manufacturing (acellular matrix, reconstructed dermis and reconstructed skin) for two different aging models (using cells from an 18- and a 61-year-old man). To assess structural variations, bi-photonic confocal microscopy was used. To characterize mechanical properties at a macroscopic scale, a light-load micro-mechanical device that performs indentation and relaxation tests was designed. Finally, images of the internal network of the samples under stretching were acquired by combining confocal microscopy with a tensile device. Mechanical properties at microscopic scale were assessed. Results revealed that adding cells during manufacturing induced structural changes, which provided higher elastic modulus and viscosity. Moreover, senescence models exhibited lower elastic modulus and viscosity. This multiscale approach was efficient to characterize and compare skin equivalent samples and permitted the first experimental assessment of the Poisson's ratio for such tissues.
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