New finding that might explain why the skin wrinkles more on various parts of the face.

The mechanism of formation of facial wrinkles has not been fully clarified due to the existence of many distinct influential factors. To clarify the relationship between facial wrinkles and structures in the skin, especially sebaceous glands, image analysis was performed on the forehead and lateral canthus regions of cadaveric skin specimens; 58 male and female donated cadavers (age range at death 20s - 90 s) were included in the study. Specimens were obtained from forehead and lateral canthus region after measuring wrinkle depth. Then tissue slices were prepared to observe the sebaceous gland and its density was measured and analyzed in relation to wrinkle depth, retinacula cutis density, dermal thickness, and solar elastosis degree. A correlation was found between sebaceous gland density and wrinkle depth in forehead specimens with a lower retinacula cutis density. Wrinkles were shallower in specimens with a higher sebaceous gland density. However, no such correlation was found in lateral canthus wrinkles, presumably due to the lack of sebaceous glands in that region. In addition, specimens with a higher sebaceous gland density tended to have a thicker dermis and/or less solar elastosis. Sebaceous gland density seems to be one of the multiple factors that prevent wrinkle deepening, and that is why wrinkles are deeper in the lateral canthus area than in the forehead. Functional studies will elucidate the mechanism of wrinkle formation in the future.

Improvement of crow's feet lines by topical application of 1-carbamimidoyl-L-proline (CLP).

Wrinkles, one of the characteristics of chronic sun-damaged and/or aged skin, are associated with psychological distress. Apart from the deterioration of collagen and elastic fibers in the dermis, which induces the loss of skin elasticity, it has been recently proposed that decreased flexibility or elasticity of the stratum corneum (SC) is also correlated with wrinkle formation. The elasticity of the SC has been shown to be regulated, at least in part, by the amounts and types of amino acids. To evaluate the ability of our newly developed amino acid-derivative (1-carbamimidoyl-L-proline; CLP), which recovers the elastic properties of the SC ex vivo, to improve wrinkles, a clinical test was performed with 126 Japanese female subjects aged 32-50 years who had crow's feet lines on their faces. Three eligible dermatologists evaluated the study according to authorized grades by the Japanese Cosmetic Science Society and scored the subjects who were much improved or improved as 29.7% and 57.8% of all CLP-treated subjects at 4 and 8 weeks, respectively. In contrast, only 1.5% and 8.1% of subjects improved with the placebo lotion at 4 and 8 weeks, respectively. These results suggest a significant efficacy of CLP to improve wrinkles. In parallel with the dermatologists' assessments, skin surface roughness in the CLP-treated group was significantly reduced after treatment with CLP for 4 and 8 weeks compared to the placebo-treated group. The sum of these data suggests that CLP is a promising and useful ingredient for the improvement of wrinkles through its ability to enhance the elasticity of the SC.

Seasonal and annual variation in the intensity of facial wrinkles.

BACKGROUND/AIMS: We studied annual, seasonal and gender-dependent changes in the intensity of facial wrinkles. METHODS: The study included 32 Japanese subjects (16 men, 16 women, aged 35-47). Wrinkles on the forehead, corner of the eye, lower eyelid and nasolabial groove of each subject were captured in photos and in replicas at 3-month intervals from March, 2009 to March, 2010. At each occasion, skin conductance in the stratum corneum and skin color were measured on the cheek. The photos obtained were used for visual scoring of wrinkle intensity, and the quantitative 3-D data of replicas were obtained. RESULTS: An apparent annual variation in skin conductance, which is supposed to be caused by skin dryness, and in the level of skin brightness, which is supposed to be caused by sun tan, was found for both genders. In contrast, no seasonal changes were determined in wrinkle intensity. Three-D analysis revealed no annual variation in women but revealed annual variation in the depths of eye wrinkles in men. CONCLUSION: There was a gender-dependent difference in the annual variation of wrinkle depth in subjects around 40 years of age. Seasonal environmental changes may affect on the eye wrinkles depth in men within a year.

Gender-dependent differences in degree of facial wrinkles.

BACKGROUND/PURPOSE: This study aimed to reveal gender-dependent differences in the degree of facial wrinkles. METHODS: Subjects comprised 173 Japanese men and women, divided into four groups according to age. Photographs were taken from nine facial regions and used to classify the intensity of wrinkles into five grades. In addition, replicas were taken from five facial sites and used to measure surface roughness. Data were compared between men and women within each age group. RESULTS: In all age groups, men showed increased forehead wrinkles compared with women. In contrast, no gender-dependent differences were found in upper eyelid wrinkles. Other facial wrinkles were greater in men than in women in all except the oldest group (age, 65-75 years), in which wrinkles in women were greater than or equal to those in men. CONCLUSION: Our results showed that gender-dependent differences exist in the degree of facial wrinkles. In general, men tend to have more severe wrinkles than women. This tendency disappeared or was reversed in some regions of the face and in individuals more than 60 years old.

Morphological study of the relationship between solar elastosis and the development of wrinkles on the forehead and lateral canthus.

OBJECTIVE: To identify whether there is a relationship between solar elastosis and the development of wrinkles in human skin. DESIGN: Wrinkle depth was measured on the forehead and lateral canthus of human cadavers using image analysis. The thickness of the dermis was measured in skin sections obtained around wrinkles and stained with Elastica-van Gieson. SETTING: Gross Anatomy Section, Kagoshima University Graduate School of Medical and Dental Sciences, Kagoshima, Japan. SUBJECTS: Fifty-eight male and female cadavers (age range at death, 29-93 years). MAIN OUTCOME MEASURES: The ratio of solar elastosis dermal thickness to full dermal thickness (elastosis ratio) was calculated and compared between the deepest point of a wrinkle (wrinkle point) and a point within 1 mm where no wrinkle existed (nonwrinkle point). The relationship between elastosis ratios and wrinkle depths was investigated. RESULTS: Advanced solar elastosis was present at nonwrinkle points but was present a little bit at wrinkle points. On the forehead, a positive correlation between elastosis ratios and wrinkle depths was observed at nonwrinkle points but not at wrinkle points. On the lateral canthus, a positive correlation between elastosis ratios and wrinkle depths was observed at nonwrinkle points, as well as at wrinkle points until the wrinkle became deeper than one-half of the original dermal thickness (0.6 mm). Solar elastosis on the lateral canthus ceased developing at this point, but the wrinkle developed further. CONCLUSIONS: Solar elastosis tends to commence with the development of a wrinkle until the wrinkle becomes deeper than 0.6 mm. This tendency is less evident at wrinkle points than at nonwrinkle points.

Relationship between the depth of facial wrinkles and the density of the retinacula cutis.

OBJECTIVE: To identify whether there is a relationship between the depth of facial wrinkles and the density of the retinacula cutis in the subcutaneous tissue of the skin. DESIGN: Wrinkle depth was assessed with image analysis on the forehead and the lateral canthus of human cadavers. The density of the retinacula cutis was measured in Azan-Mallory-stained skin sections obtained around the wrinkles. SETTING: Gross Anatomy Section, Kagoshima University Graduate School of Medical and Dental Sciences. PARTICIPANTS: Fifty-five male and female cadavers (35-93 years old). MAIN OUTCOME MEASURES: The maximum depth of each wrinkle was used to represent the wrinkle's degree. In the skin sections, the density of the retinacula cutis was measured around the deepest point of each wrinkle in a 1-mm-wide area (the wrinkle-specific area) and a 10-mm-wide area that included the wrinkle (the wrinkle-inclusive area). RESULTS: In both the wrinkle-specific and wrinkle-inclusive areas, the retinacula cutis densities became lower in the forehead and in the lateral canthus areas. When a wrinkle was shallow, the density was lower in the wrinkle-specific area than in the wrinkle-inclusive area. With wrinkle progression, the density difference between the wrinkle-specific and the wrinkle-inclusive areas gradually decreased until there was no apparent difference. CONCLUSIONS: Facial wrinkles seem to develop above sites of reduced lower retinacula cutis density. As a wrinkle develops, the density decreases in both the wrinkle-specific and the wrinkle-inclusive areas, whereas the density difference between those areas vanishes.

The relationship between wrinkle depth and dermal thickness in the forehead and lateral canthal region.

OBJECTIVE: To identify a relationship between dermal thinning and wrinkle formation. DESIGN: We assessed the wrinkle depth of the forehead and lateral canthus of 58 male and female human cadavers (range of age at death, 29-93 years) using image analysis and measured the dermal thicknesses in Azan-Mallory-stained skin sections obtained around the wrinkles. SETTING: Gross Anatomy Section, Kagoshima University Graduate School of Medical and Dental Sciences, Kagoshima, Japan. MAIN OUTCOME MEASURES: The maximum depth of the wrinkle was obtained from the forehead and lateral canthus. The dermal thickness was measured at the deepest point of the wrinkle (wrinkle point) and at a location where no wrinkle existed within 1 mm of its surface (nonwrinkle point). The ratio of the dermal thickness at the wrinkle point to the dermal thickness at the nonwrinkle point was calculated. RESULTS: The dermal thickness underneath a wrinkle decreased as the depth of the wrinkle increased (P < .001). When the dermis became thinner than one-half of its original thickness, the dermis stopped thinning. Microscopic observations revealed that the junction between the dermis and subcutaneous layers under advanced wrinkles curved downward with invaginations of the dermis into the subcutaneous layer. CONCLUSIONS: The dermis under a wrinkle becomes thinner in association with the progression of wrinkles until the dermis becomes thinner than one-half of its original thickness. When the dermis stops thinning, wrinkles develop further by dermal invagination into the subcutaneous layer.

Relationship between the echogenicity of subcutaneous tissue and the depth of eye corner wrinkles.

BACKGROUND/PURPOSE: A close relationship was found between the internal structure of subcutaneous tissue and wrinkle depth in our previous study of human forehead. The present study examined the relationship in the eye corner. METHODS: One hundred and seventy-three Japanese subjects were divided into four age groups (21-28 years, 35-41 years, 47-59 years and 65-75 years). The maximum wrinkle depth (Rmax) of an eye corner wrinkle was obtained. On an ultrasound image, which was captured by scanning perpendicular to the wrinkle, we set a 12.5-mm-wide region of interest (ROI) by aligning the deepest point of the wrinkle at its center and then obtained the ratio of the area exceeding an echogenicity threshold to the total area as the echogenicity. RESULTS: No correlation was found between the Rmax and echogenicity. The relationship was further analyzed by dividing the ROI into two: one above and the other below the deepest point of the wrinkle. A negative correlation was detected in the lower ROI of the oldest group. The difference in echogenicity between the upper and the lower ROIs of the oldest group showed a positive correlation with the Rmax. CONCLUSION: These results suggest that the subcutaneous tissue structure is related to wrinkle depth.

Relationship between the echogenicity of subcutaneous tissue and the depth of forehead wrinkles.

BACKGROUND/PURPOSE: Subcutaneous tissue is rarely studied in research on wrinkles. We used diagnostic ultrasonography to produce images of subcutaneous tissue of the forehead, one of the areas where age-related wrinkles form. We quantified the resulting echogenicity and investigated its relationship with wrinkle depth. METHODS: One hundred and seventy-three Japanese subjects were divided into four age groups (21-28 years, 35-41 years, 47-59 years, 65-75 years). Maximum wrinkle depth (Rmax) was measured in the forehead. On an ultrasound image, which was captured by scanning perpendicular to the deepest wrinkle, we set a 12.5-mm-wide region of interest by aligning the deepest point of the wrinkle at its center and then obtained the ratio of the area exceeding this threshold to the total area as the echogenicity. RESULTS: R max increased and the echogenicity decreased with age. A negative correlation was found in the oldest group but was not found in other groups, presumably due to individual differences in the echogenicity among younger subjects. CONCLUSION: This study suggests that the internal structure of subcutaneous tissue, as observed by ultrasonography, is related to wrinkle depth. The relationship between wrinkle depth and echogenicity of subcutaneous tissue is particularly pronounced in the elderly.

Findings of unique small muscle fibers at the superficial portion of the orbicularis oculi in the lateral canthal region of Japanese adult cadavers.

This study reports the existence of previously unknown muscle fascicles in Japanese adult cadavers. A bundle of these muscle fascicles diverged from the pretarsal portion of the orbicularis oculi muscle and coursed in a lateral direction superficial to this muscle. When observed with the naked eye, the bundle seemed to originate at the medial canthus and run along or near the edge of the upper eyelid. However, its boundary with the orbicularis oculi muscle was indistinguishable until it crossed superficial to this muscle. Throughout our observations, the thin muscle bundle was identified with high frequency (94%, 49 of 52 individual cadavers), and is thus unlikely to be an artifact. Light microscopy revealed that, in sagittal sections, the thin muscle bundle was located on the superficial side of the lateral portion of the orbicularis oculi muscle, while in horizontal sections, it ran in a superficial plane to the orbicularis oculi muscle in a medial to lateral direction. Despite having some similarity to a muscular raphe, the lateral canthal band, and to one of the previously known inferior muscles of the orbicularis oculi muscle, the results of our anatomical study suggest that the bundle is none of these. Rather, it is a previously unreported muscle that likely contributes to the surface morphology at the lateral canthus.

The effect of eye opening and closing on the result of facial wrinkle assessment.

BACKGROUND/PURPOSE: Since there is no standard protocol that specifies the opening or closing of eyelids in wrinkle assessment, the eyelid position of subjects at the time of assessment varies from study to study. This study aimed to reveal the effect of eye opening and closing on the result of wrinkle assessment. The study also attempted to analyze the age-, site-, and ethnicity-dependent variations in the effect. METHODS: Open- and closed-eyed photographs were obtained from 87 Japanese women of ages ranging from 21 to 73 years, and also from 80 American women comprising of Asians, Caucasians, Africans and Hispanics in their 30s. Their wrinkle intensities were scored separately at nine facial sites using a five-point photo scale. The obtained scores were compared between open- and closed-eyed photographs. The differences were then compared across age- and ethnic-groups. RESULTS: In Japanese subjects, a significant difference was found at the forehead wrinkle in all age groups, and also at the glabella, nasal root and eye corner in the middle and old age groups. In American subjects, significant differences were found at the forehead, nasal root, eye-corner, and upper and lower eyelids. The differences were more prominent in the Hispanic and Caucasian Americans than in the Asian and African Americans. CONCLUSION: These results suggest that the eyelid position should be considered as a variable in wrinkle assessment.

The characteristics of aromatase deficient hairless mice indicate important roles of extragonadal estrogen in the skin.

The roles of extragonadal estrogen in the skin are poorly understood, due to the lack of proper animal models. We examined the skin phenotypes of aromatase-knockout hairless (ArKO) mice and wild-type hairless (WT) mice, both of which were obtained through crossbreeding of Ar+/- mice and hairless mice. Differences in the skins of ArKO and WT mice were compared with those of ovariectomized (OVX) and control (Sham) mice. A difference was observed in the skin tone of ArKO mice, which is pale white and differs from the pinkish tone of all other mice. However, both ArKO and OVX mice similarly exhibited deteriorations of skin properties as compared to their respective controls. Furthermore, all the deteriorations were similarly amplified by chronic UVB irradiation in both ArKO and OVX mice as compared to their respective controls. The unique skin phenotype of ArKO mice was observed in sunburn reactions. Specifically, skins of ArKO mice showed no reaction after an acute UVB irradiation at dose intensities caused sunburn in others. However, follow-up observation found delayed reactions associated with brownish skin color and swelling only in ArKO mice, thereby suggesting that the role of extragonadal estrogen may be connected with the protective reactions of skin.

Comparison of age-related changes in facial wrinkles and sagging in the skin of Japanese, Chinese and Thai women.

BACKGROUND: Differences in skin aging features between Asians and Caucasians are commonly known, whereas little is known about such differences in various Asian populations. OBJECTIVE: A survey was carried out in Tokyo, Shanghai and Bangkok to identify specific features of skin aging in each population and to evaluate whether our conventional photo scale is an appropriate tool for this type of comparative study. METHODS: Eighty-seven women residing in Tokyo, 100 women residing in Shanghai, and 90 women residing in Bangkok were examined by a specialist. Facial wrinkles (forehead, glabella, upper eyelid, crow's feet, lower eyelid, cheek, nasolabial groove and mouth corner) and cheek sagging were evaluated using photo scales previously obtained from Japanese subjects. Comparisons were made according to 10-year age groups. RESULTS: Women in Bangkok showed the most severe level of wrinkles, followed by those in Shanghai in the three groups. Significant differences were observed between Thai and Japanese women in the intensity of wrinkles at many facial sites. Chinese women had significantly more severe wrinkles in the area around the eyes compared to Japanese women, while Thai women had significantly more severe wrinkles in the lower halves of their faces compared to Chinese women. In cheek sagging scores, significant differences were observed between Japanese and Thai women in their 30s and 50s, but not between Japanese and Chinese women or between Chinese and Thai women in all age groups. CONCLUSION: These results indicate variations in skin aging features among women from three Asian cities thereby suggesting the diversity of Asian skin. Our scaling method proved to be appropriate for facial wrinkles, but required modification to compare cheek sagging among Asian populations.

Vestiges of vibrissal capsular muscles exist in the human upper lip.

In nonhuman mammals, sinus hairs grow around the nostrils and serve for tactile sensation. The hair follicles of sinus hairs are rooted in vibrissal capsular muscles connected to the underlying, deeper orbicularis oris, thereby enabling the voluntary movement of sinus hairs. These vibrissal capsular muscles as well as the sinus hairs are believed to have been lost during human evolution, and no previous reports can be found on the existence of vestiges of sinus hairs in humans. Our study, however, has now verified the existence of vestigial muscles of vibrissae in the human upper lip. Using conventional histological techniques, microscopic observations were made on specimens obtained from the upper lips of human adult cadavers. In 35% of these individuals, several striated muscle fascicles diverging from the underlying orbicularis oris and reaching the hair follicles were observed. Histological findings of the vibrissal capsular muscle complex suggest that these fascicles are a vestigial remnant of the sinus hair muscle.

Effect of room humidity on the formation of fine wrinkles in the facial skin of Japanese.

BACKGROUND/PURPOSE: Changes in humidity are commonly known to influence the condition of the skin. Previous studies of the skin dealt with variations in relative humidity (RH) either through statistical analysis or by maintaining room humidity at a constant level; however, the range of humidity and the length of acclimation varied in each study. This study aimed to determine whether the generally used ranges of RH are truly acceptable for studies of human skin. METHODS: Skin conductance, elasticity and fine wrinkles were assessed on the eyelids of 20 volunteers, first after acclimation for 30 min in a high-humidity room (70% RH) and again after acclimation for 30 min in a low-humidity room (40% RH). RESULTS: The study found significant decreases in skin conductance and elasticity and significant increases in fine wrinkles after acclimation to low humidity compared with high humidity. CONCLUSION: These results indicate that even a 30% difference in RH can affect skin properties in 30 min. The importance of humidity stabilization and the necessity of acclimation to the humidity, particularly when the study concerns wrinkles, were thus confirmed.

A horse chestnut extract, which induces contraction forces in fibroblasts, is a potent anti-aging ingredient.

Contraction forces generated by non-muscle cells, such as fibroblasts, play important roles in determining cell morphology, vasoconstriction, and/or wound healing. We have searched among various plant extracts for ingredients that generate cell contraction forces using fibroblast-populated collagen gels. Using that model, we found that an extract of horse chestnuts (Aesculus hippocastanum) is able to generate such contraction forces in fibroblasts. The involvement of stress fiber formation in that response is suggested by the inhibition of such force generation by cytochalasin D and rhodamine phalloidin stain. Clinical testing of the extract was carried out using 40 healthy female volunteers. A gel formulation that included 3% of the extract was applied topically to the skin around the eye three times daily for nine weeks. The efficacy of the extract to diminish wrinkles was evaluated by visual scoring based on photo scales. After six weeks, significant decreases in the wrinkle scores at the corners of the eye or in the lower eyelid skin were observed compared with controls. After nine weeks, similar results were obtained. Taken together, our results suggest that an extract of horse chestnuts can generate contraction forces in fibroblasts and is a potent anti-aging ingredient.

Inhibition of ultraviolet-B-induced wrinkle formation by an elastase-inhibiting herbal extract: implication for the mechanism underlying elastase-associated wrinkles.

BACKGROUND: Previously, we have demonstrated that fibroblast-derived elastase plays an essential role in the increased three-dimensional tortuosity of elastic fibers, contributing to the loss of skin elasticity in UV-B-exposed skin. This decrease in skin elasticity is closely associated with the formation of wrinkles induced by UV exposure. OBJECTIVE: To further clarify the role of elastase in the formation of wrinkles induced by UV exposure, we assessed the effects of an extract of Zingiber officinale (L.) Rose (which inhibits fibroblast-derived elastase) on the wrinkle formation induced by chronic UV-B irradiation. RESULTS: Topical application of an extract of Zingiber officinale (L.) Rose to rat or hairless mouse skin significantly inhibited the wrinkle formation induced by chronic UV-B irradiation at a suberythemal dose, which was accompanied by a significant prevention of the decrease in skin elasticity in both types of animal skin. In the rat hind limb skin, consistent with the inhibition of reduced skin elasticity, wrinkle prevention occurred concomitantly with a significant decrease in the curling and three-dimensional tortuosity of dermal elastic fibers. CONCLUSION: Our results indicate that herbal extracts with an ability to inhibit fibroblast-derived elastase may prove to be effective as anti-wrinkling agents, confirming the important role of elastase in UV-B-induced wrinkle formation.

The effect of sunscreen on skin elastase activity induced by ultraviolet-A irradiation.

It has been reported that application of sunscreens prevents the photoaging of skin in animal models and in humans. We irradiated the dorsal skin of hairless mice with ultraviolet-A (UVA), and investigated the effects of sunscreens on skin elastase activity and on skin properties. Six-week-old female HR/ICR hairless mice were used in these experiments. After being treated with either a UVA sunscreen (also containing ultraviolet-B (UVB) sunscreen to eliminate any slight UVB in the UVA lamps; Protection Factor of UVA (PFA)=6, Sun Protection Factor (SPF)=20) or a vehicle, the dorsal skins of mice were irradiated with the UVA lamps at 22.3 J/cm(2)/d, 5 times a week. At the end of 15 weeks skin properties were evaluated and elastase activities were measured. In the vehicle control group, UVA irradiation increased the brightness and yellowing of the skin, decreased the water content of the stratum corneum, increased skin thickness, decreased skin elasticity, increased skin elastase activity, and decreased the ability of the skin to recover in a pinch test, as compared to an unirradiated group. All these differences were statistically significant. In the UVA sunscreen group, both the UVA induced skin damage and the increase in skin elastase activity were significantly inhibited, as compared to the vehicle group. However, as compared to the unirradiated group, skin elastase activity was significantly increased and immediate extensibility of skin (Ue) was significantly decreased, thereby indicating that the UVA sunscreen did not prevent photoaging to the same level as the unirradiated group. These results suggest the partial efficacy of the topical photoprotection from UVA by the sunscreen in inhibiting elastase activation, and also suggest the possibility of reducing photoaging.

Comparison of age-related changes in wrinkling and sagging of the skin in Caucasian females and in Japanese females.

We compared age-related changes in wrinkles in eight areas of facial skin (forehead, glabella, upper eyelid, corner of the eye, lower eyelid, nasolabial groove, cheek, and corner of the mouth) and sagging in the subzygomatic area of Caucasian females and of Japanese females. The subjects studied included 85 healthy Caucasian females (ages 20-69 years) living in Cincinnati in the U.S. and 70 Japanese females (ages 20-69 years) living in Tokyo. Photos of the face in frontal and in oblique 45 degrees views were analyzed. Wrinkles in the face and sagging in the subzygomatic area were graded on Japanese photoscales, respectively, by the same experienced observer. The wrinkle score increased with age in all eight areas of the face examined in Caucasian females as well as in Japanese females. In the group aged 20-29 years, the wrinkle score in each area was significantly higher in Caucasian females than in Japanese females. The wrinkle scores in the forehead, glabella, upper eyelid, and corner of the eye were similar at advanced ages between the two groups, while the wrinkle scores in lower areas of the face (lower eyelid, nasolabial groove, cheek, and corner of the mouth) were markedly higher in Caucasian females than in Japanese females in each age group, and reached an upper limit at advanced ages in Caucasian females. The sagging score also increased with age in Caucasian females as well as in Japanese females. The sagging score was significantly higher in Caucasian females than in Japanese females in the groups aged 40 years or more. These results suggest more marked wrinkle formation in all areas of the face in younger age groups of Caucasian females living in North America than in Japanese females living in Tokyo. In particular, Caucasian females showed marked age-related wrinkle formation in the lower areas of the face, probably due to sagging in the subzygomatic area, which suggests a higher susceptibility to sagging in the subzygomatic area of Caucasian females.

A study of diurnal variation in wrinkles on the human face.

We recently evaluated diurnal variations in facial skin elasticity and thickness and reported a decrease in skin thickness and an increase in skin elasticity in the afternoon compared with the morning. This phenomenon may be associated with the effect of gravity on dermal fluids. There have been no reports on the diurnal variation in wrinkles on the face, and we now describe such a study. The subjects of this study included 38 healthy Japanese males and females (mean age 34 years). Wrinkles on their foreheads, in the corners of their eyes and in their nasolabial grooves were evaluated. Replicas were obtained and skin thickness and skin elasticity measured in the morning (0830-1000 hours) and in the afternoon (1530-1700 hours) of the same day. Image analysis of the replicas showed significant aggravation of wrinkles in all areas of the face in the afternoon compared with the morning. The results of skin thickness and skin elasticity were in agreement with those of our last study. Swelling tended to occur in the morning due to the effects of gravity during sleep. Therefore, wrinkles may be swollen in the morning. Furthermore, repeated movements of the face due to changes of facial expression may gradually increase wrinkle formation and depth from the morning to the afternoon.