Identification of New Biological Pathways Involved in Skin Aging From the Analysis of French Women Genome-Wide Data.

Skin aging is an ineluctable process leading to the progressive loss of tissue integrity and is characterized by various outcomes such as wrinkling and sagging. Researchers have identified impacting environmental factors (sun exposure, smoking, etc.) and several molecular mechanisms leading to skin aging. We have previously performed genome-wide association studies (GWAS) in 502 very-well characterized French women, looking for associations with four major outcomes of skin aging, namely, photoaging, solar lentigines, wrinkling, and sagging, and this has led to new insights into the molecular mechanisms of skin aging. Since individual SNP associations in GWAS explain only a small fraction of the genetic impact in complex polygenic phenotypes, we have made the integration of these genotypes into the reference Kegg biological pathways and looked for associations by the gene set enrichment analysis (GSEA) approach. 106 pathways were tested for association with the four outcomes of skin aging. This biological pathway analysis revealed new relevant pathways and genes, some likely specific of skin aging such as the WNT7B and PRKCA genes in the "melanogenesis" pathway and some likely involved in global aging such as the DDB1 gene in the "nucleotide excision repair" pathway, not picked up in the previously published GWAS. Overall, our results suggest that the four outcomes of skin aging possess specific molecular mechanisms such as the "proteasome" and "mTOR signaling pathway" but may also share common molecular mechanisms such as "nucleotide excision repair."

Extracellular Vesicles in Human Skin: Cross-Talk from Senescent Fibroblasts to Keratinocytes by miRNAs.

Extracellular vesicles (EVs) and their miRNA cargo are intercellular communicators transmitting their pleiotropic messages between different cell types, tissues, and body fluids. Recently, they have been reported to contribute to skin homeostasis and were identified as members of the senescence-associated secretory phenotype of human dermal fibroblasts. However, the role of EV-miRNAs in paracrine signaling during skin aging is yet unclear. Here we provide evidence for the existence of small EVs in the human skin and dermal interstitial fluid using dermal open flow microperfusion and show that EVs and miRNAs are transferred from dermal fibroblasts to epidermal keratinocytes in 2D cell culture and in human skin equivalents. We further show that the transient presence of senescent fibroblast derived small EVs accelerates scratch closure of epidermal keratinocytes, whereas long-term incubation impairs keratinocyte differentiation in vitro. Finally, we identify vesicular miR-23a-3p, highly secreted by senescent fibroblasts, as one contributor of the EV-mediated effect on keratinocytes in in vitro wound healing assays. To summarize, our findings support the current view that EVs and their miRNA cargo are members of the senescence-associated secretory phenotype and, thus, regulators of human skin homeostasis during aging.

Cosmetics increase skin evenness: Evidence from perceptual and physical measures.

BACKGROUND: Cosmetics are commonly attributed with increasing skin evenness, yet little published data characterizes the effect, either perceptually or physically. We therefore investigated whether makeup increases skin evenness using a perceptual measurement and two physical measurements of color and luminance homogeneity. MATERIALS AND METHODS: Twenty-two French women (aged 29-45 years) were photographed without cosmetics, with self-applied cosmetics, and with professionally-applied cosmetics. In Study 1, 143 participants rated skin evenness. In Study 2, each face was delineated to create regions of interest (ROI) in the cheek and forehead areas. Both ROIs were then analyzed for luminance homogeneity using an established measure (Haralick homogeneity) and a new measure that incorporates chromaticity (H76 ). RESULTS: In Study 1, the faces were rated as having more even-looking skin with either self-applied cosmetics or professionally-applied cosmetics than without cosmetics. In Study 2, the luminance homogeneity measure found that the cheek ROI, but not the forehead ROI, was more homogeneous after both self-applied cosmetics and professionally-applied cosmetics when compared to without cosmetics. The new measure incorporating chromaticity found greater homogeneity in both ROIs in the two cosmetics conditions. The new measure incorporating chromaticity also better predicted the perceived skin evenness ratings from Study 1. CONCLUSION: These results provide systematic empirical evidence that makeup increases perceived skin evenness, and that these increases are partly predicted by physical measurements of skin luminance and color. The data also indicate that H76 -the new measure of skin evenness that incorporates chromaticity-better predicts perceived skin evenness.

A genome wide association study identifies new genes potentially associated with eyelid sagging.

Sagging eyelid is considered as an outward of skin ageing and may cause medical issues. However, little is known about the factors involved in sagging eyelid. The study, which aims at determining genetic risk factors for eyelid sagging, was conducted in a cohort of 502 unrelated Caucasian women living in the Paris region. All included participants were aged between 44 and 70 years old (mean age, 57.6 years old). The severity of sagging eyelid was graded in 6 categories by a dermatologist using standardized photographs of the face. A genome wide association study adjusted on potential risk factors (including age and smoking habits) was conducted to identify genetic associations. Two single nucleotide polymorphisms in total linkage disequilibrium on chromosome 10, rs16927253 (P = 7.07 × 10-10 ) and rs4746957 (P = 1.06 × 10-8 ), were significantly associated with eyelid sagging severity. The rs16927253-T and rs4746957-A alleles showed a dominant protective effect towards eyelid sagging. These polymorphisms are located in intronic parts of the H2AFY2 gene which encodes a member of the H2A histone family and very close to the AIFM2 gene that induces apoptosis. Additionally, single nucleotide polymorphisms with a false discovery rate below 0.25 were located nearby the type XIII collagen COL13A1 gene on chromosome 10 and in the ADAMTS18 gene on chromosome 16. Several relevant genes were identified by the genome wide association study for their potential role in the sagging eyelid severity.

Differential effects of makeup on perceived age.

Makeup accentuates three youth-related visual features - skin homogeneity, facial contrast, and facial feature size. By manipulating these visual features, makeup should make faces appear younger. We tested this hypothesis in an experiment in which participants estimated the age of carefully controlled photographs of faces with and without makeup. We found that 40- and especially 50-year-old women did appear significantly younger when wearing makeup. Contrary to our hypothesis, 30-year-old women looked no different in age with or without makeup, while 20-year-old women looked older with makeup. Two further studies replicated these results, finding that makeup made middle-aged women look younger, but made young women look older. Seeking to better understand why makeup makes young women look older, we ran a final study and found evidence that people associate makeup use with adulthood. By activating associations with adulthood, makeup may provide an upward bias on age estimations of women who are not clearly adult. We propose that makeup affects social perceptions through bottom-up routes, by modifying visual cues such as facial contrast, facial feature size, and skin homogeneity, and also through top-down routes, by activating social representations and norms associated with makeup use.

Evidence That the Hormonal Contraceptive Pill Is Associated With Cosmetic Habits.

Hormonal contraception is known to cause subtle but widespread behavioral changes. Here, we investigated whether changes in cosmetic habits are associated with use of the hormonal contraceptive pill. We photographed a sample of women (N = 36) who self-reported whether or not they use the contraceptive pill, as well as their cosmetic habits. A separate sample of participants (N = 143) rated how much makeup these target women appeared to be wearing. We found that women not using the contraceptive pill (i.e., naturally cycling women) reported spending more time applying cosmetics for an outing than did women who use the contraceptive pill. We also found that the faces of these naturally cycling women were rated as wearing more cosmetics than the faces of the women using the contraceptive pill. Thus, we found clear associations between contraceptive pill use and makeup use. This provides evidence consistent with the possibility that cosmetic habits, and grooming behaviors more generally, are affected by hormonal contraception.

Small extracellular vesicles and their miRNA cargo are anti-apoptotic members of the senescence-associated secretory phenotype.

Loss of functionality during aging of cells and organisms is caused and accompanied by altered cell-to-cell communication and signalling. One factor thereby is the chronic accumulation of senescent cells and the concomitant senescence-associated secretory phenotype (SASP) that contributes to microenvironment remodelling and a pro-inflammatory status. While protein based SASP factors have been well characterized, little is known about small extracellular vesicles (sEVs) and their miRNA cargo. Therefore, we analysed secretion of sEVs from senescent human dermal fibroblasts and catalogued the therein contained miRNAs. We observed a four-fold increase of sEVs, with a concomitant increase of >80% of all cargo miRNAs. The most abundantly secreted miRNAs were predicted to collectively target mRNAs of pro-apoptotic proteins, and indeed, senescent cell derived sEVs exerted anti-apoptotic activity. In addition, we identified senescence-specific differences in miRNA composition of sEVs, with an increase of miR-23a-5p and miR-137 and a decrease of miR-625-3p, miR-766-3p, miR-199b-5p, miR-381-3p, miR-17-3p. By correlating intracellular and sEV-miRNAs, we identified miRNAs selectively retained in senescent cells (miR-21-3p and miR-17-3p) or packaged specifically into senescent cell derived sEVs (miR-15b-5p and miR-30a-3p). Therefore, we suggest sEVs and their miRNA cargo to be novel, members of the SASP that are selectively secreted or retained in cellular senescence.

Blocking negative effects of senescence in human skin fibroblasts with a plant extract.

There is increasing evidence that senescent cells are a driving force behind many age-related pathologies and that their selective elimination increases the life- and healthspan of mice. Senescent cells negatively affect their surrounding tissue by losing their cell specific functionality and by secreting a pro-tumorigenic and pro-inflammatory mixture of growth hormones, chemokines, cytokines and proteases, termed the senescence-associated secretory phenotype (SASP). Here we identified an extract from the plant Solidago virgaurea subsp. alpestris, which exhibited weak senolytic activity, delayed the acquisition of a senescent phenotype and induced a papillary phenotype with improved functionality in human dermal fibroblasts. When administered to stress-induced premature senescent fibroblasts, this extract changed their global mRNA expression profile and particularly reduced the expression of various SASP components, thereby ameliorating the negative influence on nearby cells. Thus, the investigated plant extract represents a promising possibility to block age-related loss of tissue functionality.

Validation of digital photographic reference scales for evaluating facial aging signs.

BACKGROUND: Validated tools are essential to evaluate facial skin aging for both dermatological and cosmetic investigations. While many visual aging scales have been developed, few have been validated and none in terms of degree of distinguishability (DD). We developed and validated a series of visual scales using a novel digital interface for scoring facial skin aging in Caucasian women. MATERIALS AND METHODS: Three dermatologists independently established scales for 12 distinct aging signs from high-definition facial photographs of 400 adult women (Fitzpatrick phototypes I-IV) taken under standardized conditions. They then selected a consensus scale for each individual sign with a representative photo per grade. Scales were integrated into a digital interface allowing simultaneous viewing of all grades of each scale alongside the photograph of a test subject. Next, scales were validated by a different dermatologist, a general practitioner and a non-medical expert skin evaluator using photos of 350 women which had not been used for establishing the scales. RESULTS: Kappa estimates showed almost perfect agreement for wrinkle and skin aging scales (≥0.85) and moderate to substantial agreement for scales relating to color irregularities (telangiectasia, solar lentigines, freckles) for both inter- and intra-observer reproducibility. Intra-observer DD estimates were mostly high. Non-dermatologists performed well on reproducibility for both Kappa (from 0.6 to 0.9) and DD estimates. CONCLUSION: Our work demonstrates that the digital interface scales for 12 distinct aging features are highly suitable for use in clinical and epidemiological studies on skin aging by both dermatologists and non-dermatologists.

Facial Contrast Is a Cross-Cultural Cue for Perceiving Age.

Age is a fundamental social dimension and a youthful appearance is of importance for many individuals, perhaps because it is a relevant predictor of aspects of health, facial attractiveness and general well-being. We recently showed that facial contrast-the color and luminance difference between facial features and the surrounding skin-is age-related and a cue to age perception of Caucasian women. Specifically, aspects of facial contrast decrease with age in Caucasian women, and Caucasian female faces with higher contrast look younger (Porcheron et al., 2013). Here we investigated faces of other ethnic groups and raters of other cultures to see whether facial contrast is a cross-cultural youth-related attribute. Using large sets of full face color photographs of Chinese, Latin American and black South African women aged 20-80, we measured the luminance and color contrast between the facial features (the eyes, the lips, and the brows) and the surrounding skin. Most aspects of facial contrast that were previously found to decrease with age in Caucasian women were also found to decrease with age in the other ethnic groups. Though the overall pattern of changes with age was common to all women, there were also some differences between the groups. In a separate study, individual faces of the 4 ethnic groups were perceived younger by French and Chinese participants when the aspects of facial contrast that vary with age in the majority of faces were artificially increased, but older when they were artificially decreased. Altogether these findings indicate that facial contrast is a cross-cultural cue to youthfulness. Because cosmetics were shown to enhance facial contrast, this work provides some support for the notion that a universal function of cosmetics is to make female faces look younger.

A genome-wide association study in Caucasian women suggests the involvement of HLA genes in the severity of facial solar lentigines.

Solar lentigines are a common feature of sun-induced skin ageing. Little is known, however, about the genetic factors contributing to their development. In this genome-wide association study, we aimed to identify genetic loci associated with solar lentigines on the face in 502 middle-aged French women. Nine SNPs, gathered in two independent blocks on chromosome 6, exhibited a false discovery rate below 25% when looking for associations with the facial lentigine score. The first block, in the 6p22 region, corresponded to intergenic SNPs and also exhibited a significant association with forehead lentigines (P = 1.37 × 10(-8) ). The second block, within the 6p21 HLA region, was associated with decreased HLA-C expression according to several eQTL databases. Interestingly, these SNPs were also in high linkage disequilibrium with the HLA-C*0701 allele (r(2)  = 0.95). We replicated an association recently found by GWAS in the IRF4 gene. Finally, a complementary study on 44 selected candidate SNPs revealed novel associations in the MITF gene. Overall, our results point to several mechanisms involved in the severity of facial lentigines, including HLA/immunity and the melanogenesis pathway.

Facial contrast is a cue for perceiving health from the face.

How healthy someone appears has important social consequences. Yet the visual cues that determine perceived health remain poorly understood. Here we report evidence that facial contrast-the luminance and color contrast between internal facial features and the surrounding skin-is a cue for the perception of health from the face. Facial contrast was measured from a large sample of Caucasian female faces, and was found to predict ratings of perceived health. Most aspects of facial contrast were positively related to perceived health, meaning that faces with higher facial contrast appeared healthier. In 2 subsequent experiments, we manipulated facial contrast and found that participants perceived faces with increased facial contrast as appearing healthier than faces with decreased facial contrast. These results support the idea that facial contrast is a cue for perceived health. This finding adds to the growing knowledge about perceived health from the face, and helps to ground our understanding of perceived health in terms of lower-level perceptual features such as contrast. (PsycINFO Database Record

Coloration in different areas of facial skin is a cue to health: The role of cheek redness and periorbital luminance in health perception.

Looking healthy is a desirable trait, and facial skin color is a predictor of perceived health. However, skin conditions that cause dissatisfaction with appearance are specific to particular facial areas. We investigated whether color variation in facial skin is related to perceived health. Study 1 defined three areas based on color differences between faces perceived as healthy or unhealthy: the forehead, periorbital areas, and the cheeks. Periorbital luminance and cheek redness predicted perceived health, as did global skin yellowness. In Study 2, increased luminance and redness caused faces to be perceived as healthier, but only when the increase was in the periorbital and cheek areas, respectively. Manipulating each area separately in Study 3 revealed cheek redness and periorbital luminance equally increased perceived health, with low periorbital luminance more negatively affecting perceptions. These findings show that color variation in facial skin is a cue for health perception in female faces.

A genome-wide association study in Caucasian women points out a putative role of the STXBP5L gene in facial photoaging.

A genome-wide association study (GWAS) was conducted on 502 French middle-aged Caucasian women to identify genetic factors that may affect skin aging severity. A high-throughput Illumina Human Omni1-Quad beadchip was used. After single-nucleotide polymorphism (SNP) quality controls, 795,063 SNPs remained for analysis purposes. Possible stratification was first examined using the Eigenstrat method, and then the relationships between genotypes and four skin aging indicators (global photoaging, lentigines, wrinkles, and sagging) were investigated separately by linear regressions adjusted on age, smoking habits, lifetime sun exposure, hormonal status, and the two main Eigen vectors. One signal passed the Bonferroni threshold (P=1.53 × 10(-8)) and was significantly associated with global photoaging. It was also correlated with the wrinkling score and the sagging score. According to HapMap, this SNP, rs322458, was in linkage disequilibrium (LD) with intronic SNPs of the STXBP5L gene, which is expressed in the skin. In addition, it was also in LD with another SNP that increases the expression of the FBXO40 gene in the skin. These two genes, which were not previously described in the context of aging, may constitute good candidates for the investigation of molecular mechanisms of skin photoaging.

Skin surface hydration decreases rapidly during long distance flights.

INTRODUCTION: Dehydration of the stratum corneum leads to sensations and symptoms of 'dry skin' such as skin tightness and itchiness. As these complaints are frequently experienced by airline travellers, the aim of this study was to investigate the changes in skin surface hydration during long distance flights. METHODS: The study was performed on four healthy Caucasian, and on four Japanese women aged 29-39 years, travelling on long distance flights. They had stopped using skin care products at least 12 h before, and did not apply them during the flights. The air temperature and relative humidity inside the cabin, as well as skin capacitance of the face and forearm of participants, were registered at several time points before and during the flights. RESULTS: Relative humidity of the aircraft cabin dropped to levels below 10% within 2 h after take-off and stayed at this value throughout the flight. Skin capacitance decreased rapidly on both the face and forearms with most pronounced changes on the cheeks where it decreased by up to 37%. CONCLUSION: Our results demonstrate that during long distance flights, the aircraft cabin environment leads to a rapid decrease in stratum corneum hydration, an alteration, which probably accounts for the discomfort experienced by long distance aircraft travellers.

Influence of skin colour on the detection of cutaneous erythema and tanning phenomena using reflectance spectrophotometry.

BACKGROUND/PURPOSE: This research aims at assessing the influence of baseline skin colour on the ability of reflectance spectrophotometry to detect cutaneous erythema induced by a low concentration of methyl nicotinate (2.5 mM) (first objective), and to detect tanning induced by ultraviolet rays (UVA+UVB) at infra-erythemal doses (second objective). METHODS: Two independent studies were conducted to reach their respective objectives, on 27 women for the first study and on 12 women for the second study. Skin colour measurements were expressed in two different ways: percentages of reflected light at increasing wavelengths lambda (400 nm or =40 degrees. The assumption is that in the darkest skins, the emitted light is mainly absorbed by the melanin in the epidermis. Otherwise, after UV irradiation, the tanning was detectable only for individuals with fair to dark skin defined by ITA <50 degrees. This can be explained by the fact that UV stimulation of the fairest skin subjects, known to be melano-compromised individuals, can only produce a weak tanning that our study did not succeed in detecting.

Effect of hormonal replacement therapy on skin biophysical properties of menopausal women.

BACKGROUND/PURPOSE: Hormonal replacement therapy (HRT) has been shown in the past to influence well being as well as several somatic features in menopausal women. The aim of this analysis was to study the effect of HRT on various biophysical properties of the skin of menopausal women. Two sub-samples were built to test the effect (1) of 'short-term' HRT in recently menopausal women (n=15), and (2) the effect of 'medium- and long-term' HRT in menopausal women (n=78). METHODS: The analysis was performed on data from a study of 106 menopausal Caucasian women with apparent healthy skin. Self-report data on the menopausal status, the duration of the menopause, whether or not HRT has been taken and the duration of the HRT were collected. A series of biophysical measurements on the cheek, the forehead and the inner forearm were performed under controlled environmental conditions. Twenty women were menopausal for less than 5 years (eight with HRT for at least 1 year and seven who had never taken HRT) and 86 women for at least 5 years (35 with HRT for at least 5 years and 43 who had never taken HRT). Analyses of covariance with adjustment for age were performed. RESULTS: With regard to skin colour, the measurements in women treated for at least 1 year were significantly higher for red intensity (a*) and lower for brightness (L*) on the forearm as compared with the non-treated women. Furthermore, yellow intensity (b*) mean values were higher on the forehead and the forearm in women treated for at least 5 years compared with the non-treated women. Concerning sebum casual level, the mean values were significantly higher on the forehead and the cheek in women treated for at least 5 years. Regarding skin surface parameters reflecting hydration, the mean values for capacitance on the forehead and the cheek were significantly higher in women treated for at least 5 years. Finally, as regards with skin relief, parameters mean values for amplitude and roughness on the forehead were significantly higher in women treated for at least 5 years. CONCLUSION: The skin colour parameters showed a higher red intensity value in menopausal women who had been treated for at least 1 year. In menopausal women who had been treated for at least 5 years, the biophysical measurements were significantly higher for the parameters reflecting hydration and sebum secretion, which generally decrease after the menopause. These features were associated with higher values for the yellow intensity parameter and the skin relief parameters on the forehead. Our results support the subjective impression and the clinical evaluation concerning the impact of HRT on the development and the evolution of some skin properties after menopause.

Relative contribution of intrinsic vs extrinsic factors to skin aging as determined by a validated skin age score.

OBJECTIVE: To assess the relative contribution of intrinsic aging vs lifestyle factors to facial skin age. DESIGN: Prospective analysis of a cohort. SETTING: Skin research institute. STUDY SUBJECTS: A cohort of 361 white women (age range, 18-80 years) with apparently healthy skin. MEASUREMENTS: Visual and tactile assessment of facial skin features. RESULTS: Twenty-four skin characteristics were used to build a skin age score (SAS). The relationship between the SAS and chronological age followed a linear model with 2 plateaus--1 before age 30 years and 1 after age 71 years. An analysis was performed to determine whether certain lifestyle habits known to have effects on skin aging were related to the discrepancies between chronological age and the SAS. Significant effects were identified for phototype, body mass index, menopausal status, degree of lifetime sun exposure, and number of years of cigarette smoking. However, these factors accounted for only 10% of the discrepancies. Moreover, most skin characteristics used reflected changes understood to represent intrinsic aging rather than photodamage or other extrinsic factors. CONCLUSIONS: An SAS can be generated from multiple discrete signs evaluated on facial skin and is an informative tool for quantifying skin aging. The SAS is influenced by factors already recognized to affect the aging phenotypes; however, factors related to the rate of intrinsic aging, presumably genetic in character, seem to play a larger role than previously suspected.

Transepidermal water loss, temperature and sebum levels on women's facial skin follow characteristic patterns.

BACKGROUND/AIMS: The aim of this study was to compare the biophysical properties of different facial zones. METHODS: We investigated transepidermal water loss (TEWL), skin temperature and sebum casual level (CL) on 90 adjacent test sites distributed on the forehead, cheeks and chin of five women. RESULTS: All three parameters showed a symmetrical distribution around the facial median line. Only minor variations of individual values were found within the forehead and the chin areas. In contrast, the cheeks exhibited a distinct gradient with highest values in the paranasal zones and lowest on the cheek bones for all of the three parameters. The mean values on both cheeks of a given individual were nearly identical, and the patterns within the two cheeks were superimposable. Both CL and skin temperature distributions pointed out a "T-zone" with highest values on the forehead, on the chin and on the median part of the cheek. CONCLUSIONS: Our data demonstrate that biophysical skin properties differ considerably between different facial areas but that they follow a characteristic distribution.