An analysis of shapes and location of anterior hairline in Asian men.

An individual's first impression can be influenced by a number of esthetic factors, one of which is a natural hairline. The anterior hairline can be affected by various factors, such as race, age, forehead shape, and hair loss progression, similar to androgenetic alopecia. Information on the shape, hairline location on the forehead, and race are required to classify the anterior hairline. According to previous studies, the shape of the anterior hairline can be classified as round, M-shaped, rectangular, bell-shaped, or triangular. This study aimed to analyze the type of anterior hairline in Asian males without androgenetic alopecia. The subjects consisted of 461 males in China, Japan, Kazakhstan, Mongolia, the Philippines, South Korea, and Taiwan. We classified the shape of the anterior hairline into four types: M-shaped (M), rectangular (R), round (O), and wave-shaped (W). We classified the location of the anterior hairline into three types: high (H), middle (M) and low (L), according to the subjects' forehead ratio. Finally, we combined the shape and location types to devise a novel classification system for the anterior hairline type. The most common hairline types in our newly developed classification were MM (23.94%), RM (18.00%) and ML (11.04%). The least common types were OL (0.58%), OH (2.34%) and WH (2.74%). Our results will set standards for hairline shapes and locations, which will be helpful for evaluating hair loss treatment, establishing criteria and designs for reconstructive surgery, and developing camouflage makeups for the forehead hairline.

Dynamic Evaluation of Microwound Healing Induced by a Fractional CO2 Laser Using Reflectance Confocal Microscopy.

Background: A model for evaluating the in-vivo skin wound healing process over time is needed. Wound healing can be evaluated using reflectance confocal microscopy (RCM), which permits the dynamic characterization of the skin in a noninvasive manner. Objective: The aim of this study was to analyze the healing process of fractionally induced microwounds using RCM. Methods: Eight healthy volunteers had a fractional carbon dioxide (CO2) laser applied to the healthy skin of their inner arm in a single session. The wound healing of the skin at the stratum spinosum and stratum basale layers was examined using RCM. Two dermatologists evaluated the changes in the ablative zone using a grading system (ranging from completely recovered to slightly enlarged ablative zone) at four temporal time points: 3, 7, 14, and 28 days after laser application. Results: The ablative zone in the epidermis was 75 percent of the baseline after seven days and 25 percent of the area after 14 days compared to the baseline. The expanded ablative zones in the epidermis were observed in some subjects resulting from contraction between Day 3 and Day 7. The ablative zone completely healed 28 days after laser application in both the spinous and basal layers of the epidermis. Conclusion: The healing process in the ablative zone of the fractional CO2 laser-induced microwounds was observed over 2 to 4 weeks, revealing a regenerated epidermis of replaced keratinocytes from the basal layer through RCM.

Early detection of microcomedones induced by cocoa butter using reflectance confocal microscopy.

BACKGROUND: Many ingredients used in cosmetics evoke a comedogenic response. The concept of ''acne cosmetica'' was developed to link the use of certain ingredients to comedone formation. Various clinical research methods have been suggested for the effective screening of an ingredient that can worsen acne or acneiform eruption and confirm its clinical relevance as to whether it is used as a non-comedogenic claim. Although comedogenicity assessment has not yet been established, attempts have been made to evaluate the comedogenicity of cosmetic ingredients and find the most appropriate method to evaluate comedogenicity in human skin. MATERIALS AND METHODS: Total 6 participants were included in the study. Each participant received patches on the upper back containing cocoa butter. We used reflectance confocal microscopy (RCM) to count the number of microcomedones and follicles induced by cocoa butter. RESULTS: The mean value change of microcomedone/follicle by the comedogenic substance was significantly higher than that of the non-applied site for 2 weeks (p = 0.0419). The mean value changes of the microcomedone diameter caused by the comedogenic substance were significantly larger than those found at the non-applied site at 2 and 4 weeks (p = 0.0026 and p = 0.0038, respectively). CONCLUSIONS: We recommend RCM as a non-invasive real-time method that is useful for evaluating comedogenicity and early detection of a microcomedone.

4-n-butylresorcinol dissolving microneedle patch for skin depigmentation: a randomized, double-blind, placebo-controlled trial.

BACKGROUND: For effective skin depigmentation, the skin depigmentation agent must be delivered to melanocytes, where melanin is synthesized. Although dissolving microneedle (DMN) is one of the best transdermal drug delivery systems to deliver the active compound, no clinical trial has been conducted in terms of safety and efficacy. OBJECTIVES: To assess the clinical efficacy and safety of a DMN patch that contained 4-n-butylresorcinol, a skin depigmentation agent. METHODS: In the safety assessment, 31 subjects were selected for primary skin irritation test using Frosch & Kligman's method and 50 women for the cumulative irritation test and sensitization potential test using a modification of the Shelanski-Shelanski method. In the efficacy assessment, the 4-n-butylresorcinol DMN patch was compared with a control (DMN without 4-n-butylresorcinol) in our double-blind, placebo-controlled study with 45 subjects by measuring two parameters, the melanin index and individual typology angle value, during 8 weeks of administration. RESULTS: The 4-n-butylresorcinol DMN patch was shown to be safe based on the results of the safety assessment and was more than two times effective than the control patch. CONCLUSION: The 4-n-butylresorcinol DMN patch was effective and safe for skin depigmentation through targeting melanocytes and could be a useful functional cosmetic product.

Clinical Evidence of Effects of Lactobacillus plantarum HY7714 on Skin Aging: A Randomized, Double Blind, Placebo-Controlled Study.

The beneficial effects of probiotics are now widely reported, although there are only a few studies on their anti-aging effects. We have found that Lactobacillus plantarum HY7714 (HY7714) improves skin hydration and has anti-photoaging effects, and in the present study, we have further evaluated the anti-aging effect of HY7714 via a randomized, double blind, placebo-controlled clinical trial. The trial included 110 volunteers aged 41 and 59 years who have dry skin and wrinkles. Participants took 1 × 10(10) CFU/day of HY7714 (probiotic group) or a placebo (placebo group) for 12 weeks. Skin hydration, wrinkles, skin gloss, and skin elasticity were measured every 4 weeks during the study period. There were significant increases in the skin water content in the face (p < 0.01) and hands (p < 0.05) at week 12 in the probiotic group. Transepidermal water loss decreased significantly in both groups at weeks 4, 8, and 12 (p < 0.001 compared with baseline), and was suppressed to a greater extent in the face and forearm in the probiotic group at week 12. Volunteers in the probiotic group had a significant reduction in wrinkle depth at week 12, and skin gloss was also significantly improved by week 12. Finally, skin elasticity in the probiotic group improved by 13.17% (p < 0.05 vs. controls) after 4 weeks and by 21.73% (p < 0.01 vs. controls) after 12 weeks. These findings are preliminary confirmation of the anti-aging benefit to the skin of L. plantarum HY7714 as a nutricosmetic agent.

Identification of a possible susceptibility locus for UVB-induced skin tanning phenotype in Korean females using genomewide association study.

A two-stage genomewide association (GWA) analysis was conducted to investigate the genetic factors influencing ultraviolet (UV)-induced skin pigmentation in Korean females after UV exposure. Previously, a GWA study evaluating ~500 000 single nucleotide polymorphisms (SNPs) in 99 Korean females identified eight SNPs that were highly associated with tanning ability. To confirm these associations, we genotyped the SNPs in an independent replication study (112 Korean females). We found that a novel SNP in the intron of the WW domain-containing oxidoreductase (WWOX) gene yielded significant replicated associations with skin tanning ability (P-value = 1.16 × 10(-4) ). To understand the functional consequences of this locus located in the non-coding region, we investigated the role of WWOX in human melanocytes using a recombinant adenovirus expressing a microRNA specific for WWOX. Inhibition of WWOX expression significantly increased the expression and activity of tyrosinase in human melanocytes. Taken together, our results suggest that genetic variants in the intronic region of WWOX could be determinants in the UV-induced tanning ability of Korean females. WWOX represents a new candidate gene to evaluate the molecular basis of the UV-induced tanning ability in individuals.

A study of the human skin-whitening effects of resveratryl triacetate.

Resveratrol has a variety of bioactivities that include its anti-melanogenic effects, but its use in cosmetics has been challenging partly because of its chemical instability. Resveratryl triacetate (RTA) is a prodrug that can enhance stability. The purpose of this study was to examine the skin safety and whitening effects of RTA in human subjects. The primary skin irritation potentials of RTA and resveratrol were tested at 0.1 and 0.5 % on human subjects. Resveratrol at a concentration of 0.5 % induced weak skin irritation, whereas RTA did not induce any skin responses. The skin-whitening efficacy of a cosmetic formulation containing 0.4 % RTA was evaluated in two different test models. In the artificial tanning model, the test product and the control product were applied twice daily to the skin of the forearms of 22 human subjects after pigmentation induction by ultraviolet irradiation. Applying the test and the control products to the artificial tanning model for 8 weeks increased the individual topology angles (ITA°) by 17.06 and 13.81 %, respectively, a difference that was statistically significant (p < 0.05). In the hyperpigmentation model, the test product and the control product were applied twice daily to the faces of 21 human subjects. The averaged intensity of the hyperpigmented spots decreased by 2.67 % in the test group and 1.46 % in the control group, a difference that was statistically significant (p < 0.05). Therefore, RTA incorporated into cosmetic formulations can whiten human skin without inducing skin irritation.

The Atopic Dermatitis Antecubital Severity score: validity, reliability, and sensitivity to change in patients with atopic dermatitis.

BACKGROUND: The Atopic Dermatitis Antecubital Severity (ADAS) score is a new objective scale for the assessment of the severity of atopic dermatitis (AD). It is calculated by multiplying the intensity of inflammatory signs by the size of an antecubital eczema lesion. AIM: To test the validity, reliability, and sensitivity to changes of the ADAS score compared with those of the Eczema Area and Severity Index (EASI) score. METHODS: Forty patients with AD were enrolled and treated with a moisturizer. At baseline, and in weeks 1 and 2, two independent evaluators measured the ADAS score, the EASI score, and the investigator's global assessment score rated on a six-point scale. RESULTS: The ADAS score showed a higher validity than the EASI score. The superiority of the ADAS to the EASI was prominent in mild AD. Inter-evaluator reliability was excellent in both the ADAS score and the EASI. The sensitivity to changes was higher in the ADAS score than in the EASI score. CONCLUSIONS: The ADAS score may be used as a simple scoring system with good validity, reliability, and sensitivity to changes, especially in patients with mild-to-moderate AD.

A validation study to find highly correlated parameters with visual assessment for clinical evaluation of cosmetic anti-cellulite products.

BACKGROUND/PURPOSE: There has been growing interest in cellulite on parts of the body; however, no objective assessment has been specifically established. This study aims to demonstrate an optimized method by comparing the existing assessments of cellulite. METHODS: In Test 1, for subjects of 20 healthy females who have cellulite, we measured volume and roughness of cellulite using fringe projection method, roughness using replica method, dermo-subcutaneous interface length and subcutaneous thickness using ultrasonography and skin temperature using infrared ray, elasticity and blood flow. In Test 2, we applied an anti-cellulite cosmetic to 28 subjects for 6 weeks and observed if they have any changes. RESULTS: In Test 1, the effective parameter that is the most correlated with visual assessment was volume of skin measured using fringe projection method (r = 0.780). Dermo-subcutaneous interface length (r = 0.355) and subcutaneous thickness (r = 0.502) measured using ultrasonography followed in order. In Test 2, after applying a tested product, the correlation coefficient of volume of skin, of dermo-subcutaneous interface length and of subcutaneous thickness are 0.409 (P = 0.000), 0.275 (P = 0.016) and 0.311 (P = 0.012) respectively. CONCLUSION: We conclude that visual assessment, volume of skin (cavities), dermo-subcutaneous interface length and subcutaneous thickness are optimized methods for assessing an effect of cosmetics on cellulite.

Protective effects of skin permeable epidermal and fibroblast growth factor against ultraviolet-induced skin damage and human skin wrinkles.

BACKGROUND: Epidermal and fibroblast growth factor (EGF and FGF1) proteins play an important role in the regeneration and proliferation of skin cells. EGF and FGF1 have considerable potential as possible therapeutic or cosmetic agents for the treatment of skin damage including wrinkles. OBJECTIVES: Using protein transduction domains (PTD), we investigated whether PTD-EGF and FGF1 transduced into skin cells and tissue. Transduced proteins showed protective effects in a UV-induced skin damage model as well as against skin wrinkles. METHODS: Transduced PTD-EGF and FGF1 proteins were detected by immunofluorescence and immunohistochemistry. The effects of PTD-EGF and FGF1 were examined by WST assay, Western blotting, immunohistochemistry, and skin wrinkle parameters. RESULTS: The PTD-EGF and FGF1 increased cell proliferation and collagen type 1 alpha 1 protein accumulation in skin tissue. Also, PTD-EGF and FGF1 inhibited UV-induced skin damage. Furthermore, topical application of PTD-EGF and FGF1 contained ampoules which were considered to improve the wrinkle parameters of human skin. CONCLUSION: These results show that PTD-EGF and FGF1 can be a potential therapeutic or cosmetic agent for skin damaged and injury including wrinkles and aging.

Characterization of microthermal zones induced by fractional radiofrequency using reflectance confocal microscopy: a preliminary study.

BACKGROUND: The microthermal zone (MTZ) produced by fractional device have been mostly evaluated through histopathologic analysis. Study of ablative type MTZ created by fractional device and skin thermal interaction using in vivo reflectance confocal microscopy (RCM) has been rarely reported. OBJECTIVES: We evaluated the in vivo human RCM skin images of ablated fractional radiofrequency (RF) treatment to demonstrate the thermal interaction between RF and skin tissue. SUBJECTS AND METHOD: Three healthy male subjects underwent a single irradiation of fractional RF on the upper back area. The diameter, area, and depth of the MTZ was measured using RCM. Three female domestic swine were used for comparative histopathological study. RESULTS: After RF treatment, crusted holes surrounding pale edematous rings were visible in RCM images. The empty spaces correlated to the location of ablative channels, and the presence of thermal modification zones corresponded to areas with a whitish ring showing high reflectance. Ablation depth increased in proportion to the energy of fractional RF. However, the diameter of the thermal modified zone was relatively constant regardless of RF energy. CONCLUSIONS: RCM successfully identified in vivo acute thermal changes after fractional RF. The results obtained in this study provide a clearer picture of tissue-thermal interactions in the skin.

Analysis of the temporal change in biophysical parameters after fractional laser treatments using reflectance confocal microscopy.

BACKGROUND: Fractional photothermolysis is a popular treatment option for photorejuvenation. Previous literature studies have demonstrated the clinical effectiveness of fractional photothermolysis on cutaneous photoaging; however, the associated changes in biophysical properties of the skin following fractional photothermolysis have not been fully elucidated. This study was conducted to investigate the temporal changes in biophysical parameters after fractional laser treatment on Asian skin. MATERIALS AND METHODS: Eleven female subjects underwent a single treatment with an erbium glass fractional laser. Skin roughness, elasticity, transepidermal water loss (TEWL), dermal thickness were evaluated before and immediately after treatment and 3 days, 1 week, 2 weeks, and 4 weeks after treatment. The changes in the dermal papilla were analyzed using a reflectance confocal microscopy (RCM). RESULTS: Skin roughness showed the greatest improvement at the first week and net elasticity was most improved at the second week. TEWL and the percentage of melanized and active dermal papillae (DP) were mostly increased for 3 days. At 4 weeks after treatment, the number of total dermal papillae showed a significant increase compared with pretreatment. CONCLUSION: This is the first study of the characterization and quantification of dermal papilla reflecting the dermal repair process after fractional photothermolysis through an RCM.

Reduction of facial wrinkles depth by sleeping on copper oxide-containing pillowcases: a double blind, placebo controlled, parallel, randomized clinical study.

BACKGROUND: Copper up-regulates the secretion of extracellular skin proteins and stabilizes the extracellular matrix once formed. As copper can be absorbed through intact skin, we reasoned that sleeping on pillowcases containing copper-impregnated fibers would reduce skin wrinkles. OBJECTIVE: Demonstrate that sleeping on pillowcases containing copper-impregnated fibers reduce facial skin wrinkles. PATIENTS/METHODS: An 8-week, double blind, parallel, randomized study was carried out, in which healthy volunteers, aged 30-60, used either copper oxide-containing pillowcases (1% weight/weight) (test group, n = 30) or control pillowcases without copper (control group, n = 31). Skin conditions of the subjects were evaluated by visual grading by two expert graders and by 3D Image Analysis GFM PRIMOS(®) at baseline (before treatment) and following 4 and 8 weeks of sleeping on the pillowcases. RESULTS: The use of the copper oxide-containing pillowcase resulted in significant decrease of crow's feet after 4 (P = 0.01) and 8 (P = 0.002) weeks, but none was observed in the control group, as determined by the expert graders. On the basis of the 3D measurements, three roughness (R) parameters were improved after 4 and 8 weeks (P < 0.02) and the Rmax parameter at 8 weeks (P = 0.016) in the test group, but there were no changes in the R-parameters during the course of the study in the control group. The average reduction per month in the R-parameters was approximately 9%. No adverse reactions were observed or reported during the 8 weeks study. CONCLUSIONS: Sleeping on copper oxide-containing pillowcases results in reduction of wrinkles depth and overall improvement of skin appearance.

Cosmetic efficacy evaluation of an anti-acne cream using the 3D image analysis system.

BACKGROUND/PURPOSE: Acne is a skin disease which accompanies pathological and morphological changes. Although acne severity is scored by clinicians based on pathological status, aesthetic aspect of acne symptom is also concerned by patients. This study was conducted to examine the usefulness of a 3D image analysis method for the cosmetic efficacy evaluation of an anti-acne cream. METHODS: Twenty-one healthy volunteers with acne lesions were recruited for the study and treated with an 'anti-acne cream' for 4 weeks. Acne symptoms on the facial skin were graded by the visual evaluation of photographs taken before and after the treatment. Skin color of acne lesions was and measured by a spectrophotometer. In addition, a 3D image analysis system was used to quantify skin surface roughness and acne volumes. RESULTS: Both visual and spectrophotometric assessments of acne lesions provided similar results indicating that the cream treatment improved acne symptoms significantly. The 3D image analysis of acne lesions confirmed that the cream treatment decreased skin surface roughness and acne volumes. CONCLUSION: The current study demonstrated that the 3D image-based analysis of the skin may be useful for the quantification of acne symptoms of cosmetic relevance.

Use of non-melanocytic HEK293 cells stably expressing human tyrosinase for the screening of anti-melanogenic agents.

Tyrosinase (TYR) from mushrooms has been inappropriately used in the screening assay for hypopigmenting agents even though its biochemical properties are different from those of human TYR. Cell-free extracts of human epidermal melanocyes (HEMs) could be another choice for the assay, but HEMs grow too slowly to get a sufficient amount of cell-free extracts. In the present study, human embryonic kidney (HEK) 293 cells were transfected with a human TYR construct to establish a cell line that grows rapidly and expresses human TYR constitutively. Cell-free extracts of the established cell line, HEK293-TYR, were tentatively used in the screening assays for 11 phenylpropanoids that have chemical structures similar to that of L-tyrosine, the substrate of TYR. Of the 11 compounds, the strongest inhibition of TYR activity was shown by p-coumaric acid (IC50, 3 µM), followed by 3-(4-hydroxyphenyl)propionic acid (50 µM) and 3-(4-hydroxyphenyl)lactic acid (70 µM). The results indicate that p-coumaric acid has an optimal chemical structure for the inhibition of TYR. The effects of these phenylpropanoids on melanin synthesis in HEMs correlated well with their effects on TYR activity in vitro. This study demonstrated that HEK293-TYR cells can be a good source of the human TYR enzymes needed in the screening assay of anti-melanogenic agents.

Comparison of the antimelanogenic effects of p-coumaric acid and its methyl ester and their skin permeabilities.

BACKGROUND: p-Coumaric acid (PCA) inhibits human tyrosinase (TYR) activity and melanin synthesis in human epidermal melanocytes. OBJECTIVE: The purpose of the current study was to examine the potential of PCA and its hydrophobic derivative, methyl p-coumarate (MPC), as hypopigmenting agents for topical use. METHODS: PCA and MPC were comparatively tested against in vitro human TYR enzyme activity and cellular melanin synthesis in human epidermal melanocytes. Permeation studies were undertaken using an artificial lipophilic membrane and an excised porcine skin. In vivo hypopigmenting efficacy was assessed on the skin of melanin-possessing hairless mice exposed to UVB. RESULTS: Although PCA was a stronger inhibitor than MPC against TYR activity in vitro, the former inhibited cellular melanin synthesis less effectively than the latter. A non-cell based permeability assay indicated that PCA was practically impermeable through the lipophilic barrier while MPC was highly permeable. In contrast, an ex vivo skin permeation study demonstrated that topically applied PCA in the form of a cream can diffuse into the aqueous medium underneath the skin. No MPC was released from a MPC cream but PCA was released instead as a bio-converted product. Topical application of PCA cream attenuated the UVB-induced erythema formation and pigmentation in mice models, more effectively compared with MPC cream. CONCLUSION: PCA may be useful as an active ingredient for topical applications for a hypopigmenting effect. MPC has potential as a hypopigmenting agent but requires rather invasive methods for its delivery to the target cells.

p-coumaric acid not only inhibits human tyrosinase activity in vitro but also melanogenesis in cells exposed to UVB.

Tyrosinase (TYR) catalyzes rate-limiting steps of melanogenesis and thus its inhibitors are potentially useful as hypopigmenting agents. Recently, p-coumaric acid (p-CA) has been suggested to interfere with the pro-melanogenic actions of tyrosine due to its structural similarity with tyrosine (An SM et al., Br J Dermatol 2008. 159: 292). In this study, we compared the inhibitory effects of p-CA and two other well known TYR inhibitors used in cosmetics--arbutin and kojic acid--on the catalytic activities of mushroom, murine and human TYRs in vitro, using tyrosine and 3,4-dihydroxyphenylalanine (DOPA) as substrates. The results showed that p-CA is a weaker inhibitor of mushroom TYR but much stronger inhibitor of human or murine TYR in comparison with kojic acid and arbutin. In addition, p-CA inhibited human TYR at much lower concentrations than those required for the inhibition of murine or mushroom TYRs. Enzyme kinetics analysis indicated that p-CA is a mixed type (for tyrosine) or competitive inhibitor (for DOPA) of human TYR. Potent antimelanogenic effects of p-CA were observed in human epidermal melanocytes exposed to UVB. The present study demonstrated that p-CA is a potent and selective inhibitor of human TYR and is potentially useful as a hypopigmenting agent.

Inhibition of melanogenesis by tyrosinase siRNA in human melanocytes.

Tyrosinase (TYR) plays a critical role in cellular melanogenesis and, thus, has been the major target of pharmacological approaches for the control of skin pigmentation. This study examined an alternative molecular approach using TYR-small interfering RNA (siRNA) to control melanogenesis in the human melanocytes. Both the mRNA and protein levels of TYR were significantly lowered by TYR-siRNA treatment, whereas TYR-related protein 1 and TYR-related protein 2 displayed no such changes. TYR-siRNA treatment inhibited the cellular melanin synthesis from the externally supplied TYR substrate L-tyrosine. TYR-siRNA also suppressed melanin synthesis and decreased the viability of cells exposed to ultraviolet radiation, supporting a critical role of melanin in protection against ultraviolet radiation. These results suggest that molecular approaches using siRNA targeted to the enzymes of melanogenic pathway may provide a novel strategy for the control of cell pigmentation.

Cigarette smoking associated with premature facial wrinkling: image analysis of facial skin replicas.

BACKGROUND: Despite the obvious relation between smoking and facial wrinkling, grossly undetectable wrinkling and the consequences of smoking on the face have been poorly studied. OBJECTIVE: To assess the risk factor of cigarette smoking on the development of premature facial wrinkling. METHODS: One hundred and twenty-three nonsmokers, 160 current smokers, and 67 past smokers, aged 20-69 years, were studied. Cigarette smoking status, weight changes, average sun exposure time (recreational and occupational) in 1 month, and past medical and facial cosmetic surgery were quantified by self-questionnaire. Computerized image analysis of silicone skin replicas was used in addition to clinical visual measurement, and a severity score based on predetermined criteria was assigned to each patient. RESULTS: Current smokers have a higher degree of facial wrinkling than nonsmokers and past smokers. Past smokers who smoked heavily at a younger age show less facial wrinkling than current smokers. In the analysis, which was adjusted for age group, the relative risk of moderate to severe wrinkling for current smokers compared with nonsmokers was 2.72 (confidence interval, CI: 1.32-3.21, P < 0.05). In current smokers, the relative risks associated with more than 19 pack-years and 11-19 pack-years of smoking compared with nonsmokers were 2.93 (CI: 1.14-4.1, P < 0.05) and 1.75 (CI: 1.54-3.67, P < 0.05), respectively. On image analysis of facial skin replicas, the mean values of Ra (arithmetic average roughness), Rz (average roughness), and Rt (distance between the highest and lowest values) of current smokers were higher than those of nonsmokers and past smokers in all age groups. This indicates a strong correlation between cigarette smoking and skin wrinkling. In addition, microscopic superficial wrinkling (Ra and Rt) was noted in current smokers in the younger age group (20-39 years). CONCLUSIONS: This study suggests that attention should be paid to smoking-associated facial wrinkling (not evident from a visual assessment) in young people and added to the list of disorders seemingly caused by smoking.