ABSTRACT
Research on new ingredients that can prevent excessive melanin production in the skin while considering efficacy, safety but also environmental impact is of great importance to significantly improve the profile of existing actives on the market and avoid undesirable side effects. Here, the discovery of an innovative technology for the management of hyperpigmentation is described. High-throughput screening tests on a wide chemical diversity of molecules and in silico predictive methodologies were essential to design an original thiopyridinone backbone and select 2-mercaptonicotinoyl glycine (2-MNG) as exhibiting the most favorable balance between the impact on water footprint, skin penetration potential and performance. The effectiveness of 2-MNG was confirmed by topical application on pigmented reconstructed epidermis and human skin explants. In addition, experiments have shown that unlike most melanogenesis inhibitors on the market, this molecule is not a tyrosinase inhibitor. 2-MNG binds to certain melanin precursors, preventing their integration into growing melanin and leading to inhibition of eumelanin and pheomelanin synthesis, without compromising the integrity of melanocytes.
Subject(s)
Glycine , Melanins , Melanocytes , Humans , Melanocytes/drug effects , Melanocytes/metabolism , Melanins/biosynthesis , Melanins/metabolism , Glycine/analogs & derivatives , Glycine/pharmacology , Glycine/chemistry , Monophenol Monooxygenase/metabolism , Monophenol Monooxygenase/antagonists & inhibitors , MelanogenesisABSTRACT
INTRODUCTION: Hyperpigmentation disorders are very frequent, affect the quality of life and may become a psychological burden for afflicted patients. Many anti-pigmenting or depigmenting agents are available with various efficacy and almost no comparative data. 2-mercaptonicotinoyl glycine (2-MNG) was recently proposed as a viable candidate showing safe and effective results on hyperpigmentation control in vitro and in vivo. OBJECTIVES: A Bayesian network meta-analysis (BNMA) was conducted to map and rank the anti-pigmenting and depigmenting efficacy of 2-MNG 0.5% on UV daylight (UVDL)-induced pigmentation together with 13 other reference molecules. A comparison in the kinetics of 2-MNG 0.5% was also performed. METHODOLOGY: Fourteen studies were conducted, for each, on 15-30 women of skin phototype III in Shanghai, China and Paris, France. The products were applied on mini zone, in randomized and blinded protocol, on the back, 5 days a week during 6 weeks, at a dose of 4 mg/cm2. During the second week, volunteers were exposed under to varying minimum erythemal dose of UVDL during 4 consecutive days-adapted to obtain a similar induction of skin pigmentation regardless of the population. Assessments were performed instrumentally using Chromameter®. Ascorbic acid 7% was used as a positive control for all experiments. A Bayesian network meta-analysis was then established to map and follow the kinetics of 2-MNG 0.5% performance with 13 reference molecules (glutathione 2%, kojic acid 1%, hydroquinone 4%, ascorbyl glucoside 2%, niacinamide 4%, etc.). RESULTS: 2-MNG 0.5% dominated the ranking at all time points with a significant high probability of strong efficacy against UVDL-induced pigmentation. Ascorbic acid 7% ranks second after 4 days of irradiations (D12) whereas hydroquinone 4% ranks second 1 month after irradiations (D40). In the kinetics, 2-MNG at 0.5% was effective as from the end of irradiations (D12) to the study endpoint (D40). This suggested an immediate and persistent efficacy across all timepoints evaluated. CONCLUSION: The BNMA revealed a rapid and lasting efficacy of 2-MNG 0.5% on the anti-pigmenting and depigmenting phases of the clinical protocol. 2-MNG 0.5% ranked first, with immediate and lasting effect compared to 13 other references. This study is the first allowing comparison between reference anti-pigmenting and depigmenting agents and will help clinicians for proposing the most effective approach for their patients.
Subject(s)
Bayes Theorem , Hyperpigmentation , Ultraviolet Rays , Humans , Ultraviolet Rays/adverse effects , Hyperpigmentation/prevention & control , Network Meta-Analysis , Skin Pigmentation/drug effects , Skin Pigmentation/radiation effects , FemaleABSTRACT
Characterizing melanins in situ and determining their 3D z-epidermal distribution is paramount for understanding physiological/pathological processes of melanin neosynthesis, transfer, degradation or modulation with external UV exposure or cosmetic/pharmaceutical products. Multiphoton fluorescence intensity- and lifetime-based approaches have been shown to afford melanin detection, but how can one quantify melanin in vivo in 3D from multiphoton fluorescence lifetime (FLIM) data, especially since FLIM imaging requires long image acquisition times not compatible with 3D imaging in a clinical setup? We propose an approach combining (i) multiphoton FLIM, (ii) fast image acquisition times, and (iii) a melanin detection method called Pseudo-FLIM, based on slope analysis of autofluorescence intensity decays from temporally binned data. We compare Pseudo-FLIM to FLIM bi-exponential and phasor analyses of synthetic melanin, melanocytes/keratinocytes coculture and in vivo human skin. Using parameters of global 3D epidermal melanin density and z-epidermal distribution profile, we provide first insights into the in vivo knowledge of 3D melanin modulations with constitutive pigmentation versus ethnicity, with seasonality over 1 year and with topical application of retinoic acid or retinol on human skin. Applications of Pseudo-FLIM based melanin detection encompass physiological, pathological, or environmental factors-induced pigmentation modulations up to whitening, anti-photoaging, or photoprotection products evaluation.
Subject(s)
Epidermis/metabolism , Imaging, Three-Dimensional , Melanins/metabolism , Melanocytes/metabolism , Microscopy, Fluorescence, Multiphoton , Skin Pigmentation , Administration, Cutaneous , Adolescent , Adult , Aged , Cells, Cultured , Coculture Techniques , Dermatologic Agents/administration & dosage , Epidermis/drug effects , Female , Humans , Melanocytes/drug effects , Middle Aged , Predictive Value of Tests , Skin Pigmentation/drug effects , Time Factors , Treatment Outcome , Tretinoin/administration & dosage , Vitamin A/administration & dosage , Young AdultABSTRACT
Background: Repeated nonextreme sun exposures induce skin pigmentation by increasing melanin production and by oxidizing preexisting melanin and melanin precursors. This leads to skin disorders and skin color heterogeneity such as hyperpigmented spots. Objective: We assessed 31 randomized, controlled clinical trials to determine the potential of vitamin C to limit ultraviolet (UV) daylight-induced pigmentation, considering dose response and different skin type populations (Caucasian and Chinese). Materials and Methods: Thirty-one intraindividual, randomized, controlled clinical trials involving Caucasian and Chinese subjects (15-35 healthy male or female volunteers per study, 741 total volunteers) 18 to 50 years of age with Phototype III and individual typology angle (ITA) value between 28 and 49 degrees were analyzed. The 31 studies assessed the potential of vitamin C (formulated with the copolymer Styrène-Anhydride Maléique [SMA]) to decrease pigmentation induced by UV daylight exposure. Results were combined using a Bayesian meta-analysis to provide probabilistic evidence of the effects of vitamin C by dose and population. Results: Vitamin C was effective in reducing pigmentation induced by UV daylight-simulated expositions (4 days at 0.75 Individual Minimal Erythemal Dose [MEDi]) in a dose-dependent manner. During the depigmentation phase, no additive value was provided by the vitamin C, suggesting that the lightening properties described in the literature for vitamin C correspond to an antipigmenting quality rather than a depigmenting effect. Conclusion: Vitamin C is a valuable and safe dermocosmetic antipigmenting compound with a strong effect at 10% possibly useful in preventing signs of photoaging.
ABSTRACT
The melanosome pattern was characterized systematically in keratinocytes in situ in highly, moderately, and lightly pigmented human skin, classified according to the individual typological angle, a colorimetric measure of skin color phenotype. Electron microscopy of skin samples showed qualitatively and quantitatively that in highly pigmented skin, although melanosomes are mostly isolated and distributed throughout the entire epidermis, clusters are also observed in the basal layer. In moderately and lightly pigmented skin, melanosomes are concentrated in the first layer of the epidermis, isolated-but for most of them, grouped as clusters of melanocores delimited by a single membrane. Electron tomography resolving intracellular three-dimensional organization of organelles showed that clustered melanocores depict contacts with other cellular compartments, such as endoplasmic reticulum and mitochondria. Additionally, immunogold labelling showed that clusters of melanocores do not correspond to autophagosomes or melanophagosomes but that they present, similarly to melanosomes in melanocytes, features of nonacidic, nondegradative organelles. Overall, these observations suggest that melanocore clusters do not correspond to autophagic organelles but represent reservoirs or protective structures for melanosome integrity and function. These results open avenues for understanding the basis of skin pigmentation in different skin color phenotypes.
Subject(s)
Keratinocytes/ultrastructure , Melanosomes/ultrastructure , Organelles/ultrastructure , Skin Pigmentation , Adult , Autophagosomes/ultrastructure , Epidermis/ultrastructure , Female , Humans , Microscopy, ElectronABSTRACT
With aging, epidermal homeostasis and barrier function are disrupted. In a previous study, we analyzed the transcriptomic response of young skin epidermis after stratum corneum removal, and obtained a global kinetic view of the molecular processes involved in barrier function recovery. In the present study, the same analysis was performed in aged skin in order to better understand the defects which occur with aging. Thirty healthy male volunteers (67 ± 4 years old) were involved. Tape-strippings were carried out on the inner face of one forearm, the other unstripped forearm serving as control. At 2, 6, 18, 30 and 72 h after stripping, TEWL measurements were taken, and epidermis samples were collected. Total RNA was extracted and analyzed using DermArray(®) cDNA microarrays. The results highlighted that barrier function recovery and overall kinetics of gene expression were delayed following stripping in aged skin. Indeed, the TEWL measurements showed that barrier recovery in the young group appeared to be dramatically significant during the overall kinetics, while there were no significant evolution in the aged group until 30 h. Moreover, gene expression analysis revealed that the number of modulated genes following tape stripping increased as a function of time and reached a peak at 6 h after tape stripping in young skin, while it was at 30 h in aged skin, showing that cellular activity linked to the repair process may be engaged earlier in young epidermis than in aged epidermis. A total of 370 genes were modulated in the young group. In the aged group, 382 genes were modulated, whose 184 were also modulated in the young group. Only eight genes that were modulated in both groups were significantly differently modulated. The characterization of these genes into 15 functional families helped to draw a scenario for the aging process affecting epidermal repair capacity.
Subject(s)
Epidermis/physiology , Gene Expression/physiology , Skin Aging/physiology , Wound Healing/genetics , Age Factors , Aged , Gene Expression Profiling , Healthy Volunteers , Humans , Male , Middle AgedABSTRACT
To study human skin pigmentation in a physiological in vitro model, we developed a pigmented reconstructed skin reproducing the three-dimensional architecture of the melanocyte environment and the interactions of melanocyte with its cellular partners, keratinocytes, and fibroblasts. Co-seeding melanocytes and keratinocytes onto a fibroblast-populated collagen matrix led to a correct integration of melanocytes within the epidermal basal layer, but melanocytes remained amelanotic even after supplementation with promelanogenic factors. Interestingly, normalization of keratinocyte differentiation using keratinocyte growth factor instead of epidermal growth factor finally allowed an active pigmentary system to develop, as shown by the expression of key melanogenic markers, the production, and transfer of melanosome-containing melanin into keratinocytes. Various degrees of constitutive pigmentation were reproduced using melanocytes from different skin phenotypes. Furthermore, induction of pigmentation was achieved by treatment with known propigmenting molecules, αMSH and forskolin, thus demonstrating the functionality of the pigmentary system. This pigmented full-thickness skin model therefore represents a highly relevant tool to study the role of cell-cell, cell-matrix, and mesenchymal-epithelial interactions in the control of skin pigmentation.
Subject(s)
Colforsin/pharmacology , Fibroblast Growth Factor 7/physiology , Melanocytes/metabolism , Skin Pigmentation , Skin/cytology , alpha-MSH/pharmacology , Animals , Cells, Cultured , Colorimetry , Humans , Mice , Microscopy, Electron, Transmission , Skin/metabolismABSTRACT
The stratum corneum (SC) is a superficial skin compartment that protects the body from the outside environment. Any disturbance of this function induces cascading steps of molecular and cellular repair in the whole epidermis. The aim of this study was to investigate epidermal gene expression following SC removal by tape stripping. Twenty-nine healthy male volunteers were included (27 +/- 4 years old). Tape stripping was processed on one inner forearm, the other unstripped forearm served as a control. Epidermis samples were collected at 2, 6, 19, 30 and 72 h after tape stripping. Trans-epidermal water loss measurements were performed at each step to monitor barrier restoration. Total RNA was extracted from collected epidermis samples and analysed by using DermArray cDNA microarrays. Among 4000 genes under investigation, we found that the expression of 370 genes varied significantly at least once during the time following stripping. Using an original clustering method, the modulated genes were gathered into eight groups. A functional characterization of the clusters enabled us to get a dynamic and global view of the main molecular processes taking place during epidermal recovery.
Subject(s)
Epidermis/injuries , Epidermis/metabolism , Gene Expression , Wound Healing/genetics , Adult , Databases, Genetic , Gene Expression Profiling , Genomics , Humans , Kinetics , Male , Multigene Family , Oligonucleotide Array Sequence Analysis , Polymerase Chain Reaction , Young AdultABSTRACT
We previously identified constitutive Smad signaling in human melanoma cells despite resistance to transforming growth factor-beta (TGF-beta) control of cell proliferation. This led us to investigate the effect of inhibitory Smad7 overexpression on melanoma cell behavior. Using the highly metastatic cell line, 1205-Lu, we thus generated melanoma cell clones constitutively expressing Smad7, and their mock-transfected counterparts. Stable expression of Smad7 resulted in an inhibition of constitutive Smad2/3 phosphorylation, and in a reduced TGF-beta response of Smad3/Smad4-driven gene transactivation, as measured using transfected Smad3/4-specific reporter gene constructs. Smad7 overexpression, however, did not alter their proliferative capacity and resistance to TGF-beta-driven growth inhibition. On the other hand, expression of Smad7 efficiently reduced the capacity of human melanoma cells to invade Matrigel in Boyden migration chambers, while not affecting their motility and adhesion to collagen and laminin. Gelatin zymography identified reduced MMP-2 and MMP-9 secretion by Smad7-expressing melanoma cells as compared with their control counterparts. Smad7-expressing melanoma cells exhibited a dramatically reduced capacity to form colonies under anchorage-independent culture conditions, and, when injected subcutaneously into nude mice, were largely delayed in their ability to form tumors. These results suggest that TGF-beta production by melanoma cells not only affects the tumor environment but also directly contributes to tumor cell aggressiveness through autocrine activation of Smad signaling.
Subject(s)
Melanoma/pathology , Skin Neoplasms/pathology , Smad7 Protein/biosynthesis , Animals , Cell Adhesion , Cell Movement , Gene Expression Profiling , Humans , Matrix Metalloproteinase 9/metabolism , Melanoma/metabolism , Mice , Mice, Nude , Neoplasm Invasiveness , Neoplasm Metastasis , Signal Transduction , Skin Neoplasms/metabolism , Smad7 Protein/physiology , Transforming Growth Factor beta , Tumor Cells, CulturedABSTRACT
By its numerous properties and importance in cheese technology (production of colour, flavour, bacteriocins and resistance to salt) Brevibacterium linens is a major cheese ripening bacteria. However, the genetic approach of such biological functions has been hindered, up to now, by the lack of tools necessary to realise genetic modifications in this species. Our objective was to demonstrate that it is possible to genetically modify several strains exhibiting interesting technological properties, especially the production of sulphur compounds. We worked with a phenotypically and genetically diverse collection of 11 strains. We genetically transformed several Brevi. linens with acceptable rates with plasmids classically used to transform lactic acid bacteria and other Gram+ bacteria. These results open up new prospects to investigate the most interesting Brevi. linens metabolic pathways both at the biochemical and genetic level.