Ceramide synthase 4 is highly expressed in involved skin of patients with atopic dermatitis.

BACKGROUND: Ceramide is a crucial lipid in the stratum corneum (SC) which maintains the barrier function and hydration of the skin. In atopic dermatitis (AD) patients who have defective skin barrier function, ceramide levels are altered. We previously reported that although the amount of total ceramide was lower in involved skin compared with uninvolved skin of AD patients and with healthy control skin, the amounts of smaller ceramide species of Cer[NS] (<40 total carbons, which are total carbons of both sphingoid base and amide-linked fatty acid), especially Cer[NS] with 34 total carbons (C34-Cer[NS]), were higher. However, the enzyme(s) that produces the higher levels of smaller ceramide species in involved skin of AD patients was unclear. OBJECTIVE: To identify the enzyme(s) that produces higher levels of smaller ceramide species of Cer[NS] in the involved skin of AD patients. METHODS: Eight female Caucasian subjects who were diagnosed with AD on their arms (age range: 21-45 years) were enroled in this study. We compared ceramide levels in the SC and the expression levels of enzymes involved in ceramide metabolism using real-time PCR and immunohistochemistry between involved and uninvolved skin of AD patients. RESULTS: Level of mRNA encoding ceramide synthase 4 (CERS4), which is one of the enzymes that synthesize ceramide from a sphingoid base and an amide-linked fatty acid, was significantly higher in involved skin than in uninvolved skin (P < 0.01). Additionally, the protein expression level of CERS4 in the epidermis was also higher in involved skin compared with uninvolved skin. The expression level of CERS4 correlated with the amount of C34-Cer[NS] (P < 0.01) and the skin hydration value (P < 0.05). CONCLUSIONS: The elevated expression level of CERS4 contributes to the increase of C34-Cer[NS] and the impaired SC barrier function in involved skin of AD patients.

The relationship between skin aging and steady state ultraweak photon emission as an indicator of skin oxidative stress in vivo.

BACKGROUND/PURPOSE: Ultraweak photon emission (UPE) is one potential method to evaluate the oxidative status of the skin in vivo. However, little is known about how the daily oxidative stress of the skin is related to skin aging-related alterations in vivo. We characterized the steady state UPE and performed a skin survey. METHODS: We evaluated the skin oxidative status by UPE, skin elasticity, epidermal thickness and skin color on the inner upper arm, the outer forearm, and the buttock of 70 Japanese volunteers. RESULTS: The steady state UPE at the three skin sites increased with age. Correlation analysis revealed that the steady state UPE only from the buttock was related to skin elasticity, which showed age-dependent changes. Moreover, analysis by age group indicated that b* values of the inner upper arm of subjects in their 20s were inversely correlated with UPE as occurred in buttock skin. In contrast, photoaged skin did not show a clear relationship with steady state UPE because the accumulation of sun-exposure might influence the sensitivity to oxidative stress. CONCLUSION: These results suggest that steady state UPE reflects not only intrinsic skin aging and cutaneous color but also the current oxidative status independent of skin aging.

Eucalyptus increases ceramide levels in keratinocytes and improves stratum corneum function.

The objectives of this study were to identify a plant extract that would improve stratum corneum functions and to elucidate the mechanism(s) involved. Based on the information that stratum corneum functions depend on the level of ceramide in the stratum corneum, we identified a Eucalyptus extract that was able to increase the level of ceramide in human keratinocytes in culture and in human stratum corneum and that improves the stratum corneum water holding and barrier functions. Addition of the Eucalyptus extract to human keratinocytes in culture increased the level of ceramide in a dose-dependent manner and also increased the biosynthesis of ceramide, glucosylceramide and sphingomyelin. Topical application of the Eucalyptus extract on the dry skin of human subjects induced by acetone and diethylether treatment resulted in a significant increase in ceramide level in the stratum corneum, a significant improvement in its water-holding function and an improvement in its barrier function. The addition of macrocarpal A, one of the main components of the Eucalyptus extract, to human keratinocytes in culture increased the level of ceramide and the mRNA expression of serine palmitoyltransferase, acid sphingomyelinase, neutral sphingomyelinase, glucosylceramide synthase and glucocerebrosidase in a dose-dependent manner. Our results indicate that the increased content of ceramides in the stratum corneum may underlie the therapeutic effect of the Eucalyptus extract. Our results also indicate the possibility that macrocarpal A is the key component that stimulates the synthesis of ceramide in the stratum corneum.

Real-life use of underwear treated with fabric softeners improves skin dryness by decreasing the friction of fabrics against the skin.

Scientific evaluation of fabric softener use on the friction of fabrics and on dry skin in real life is very limited. The coefficient of friction of fabrics was measured with a surface tester using model skin and cotton fabrics. The effects and safety of fabric softener-treated T-shirts on subjects with dry skin were evaluated in real-life conditions in the cold and arid environment. Twenty male volunteers with dry skin used T-shirts without softener for 1 week before day 0 and then wore softener-treated T-shirts for 14 days. Effects were evaluated by clinical grading, subjective symptoms, stratum corneum water content (SCWC) and transepidermal water loss (TEWL). A significant decrease in the coefficient of friction of fabrics was detected. A significant improvement in SCWC was observed on the torso, waist and shoulder, but no improvement was found on the inner forearm. There was a slight change in TEWL. The visual grading of dryness improved significantly at all sites, but no significant change was found in subjective symptoms. Our results suggest that fabric softener provides benefits to individuals with dry skin because of the decreased friction of the garments against the skin. It is possible that the softener treatment of fabrics contributes to the state of moisturization of the skin and may work in collaboration with the application of moisturizers.

Gene transfer in human skin with different pseudotyped HIV-based vectors.

Pseudotyping lentiviral vector with other viral surface proteins could be applied for treating genetic anomalies in human skin. In this study, the modification of HIV vector tropism by pseudotyping with the envelope glycoprotein from vesicular stomatitis virus (VSV), the Zaire Ebola (EboZ) virus, murine leukemia virus (MuLV), lymphocytic choriomeningitis virus (LCMV), Rabies or the rabies-related Mokola virus encoding LacZ as a reporter gene was evaluated qualitatively and quantitatively in human skin xenografts. High transgene expression was detected in dermal fibroblasts transduced with VSV-G-, EboZ- or MuLV-pseudotyped HIV vector with tissue irregularities in the dermal compartments following repeated injections of EboZ- or LCMV-pseudotyped vectors. Four weeks after transduction, double-labeling immunofluorescence of beta-galactosidase and involucrin or integrin beta1 demonstrated that VSV-G-, EboZ- or MuLV-pseudotyped HIV vector effectively targeted quiescent epidermal stem cells which underwent terminal differentiation resulting in transgene expression in their progenies. Among the six different pseudotyped HIV-based vectors evaluated, VSV-G-pseudotyped vector was found to be the most efficient viral glycoprotein for cutaneous transduction as demonstrated by the highest level of beta-galactosidase expression and genome copy number evaluated by TaqMan PCR.

Ovariectomy is sufficient to accelerate spontaneous skin ageing and to stimulate ultraviolet irradiation-induced photoageing of murine skin.

BACKGROUND: Wrinkling and sagging of the skin during photoageing is physiologically associated with diminished elasticity, which can be attributed to increased fibroblast-derived elastase activity. This degrades the dermal elastic fibres needed to maintain the three-dimensional structure of the skin. We previously reported that ovariectomy accelerates ultraviolet (UV)B-induced wrinkle formation in rat hind limb skin by altering the three-dimensional structure of elastic fibres. OBJECTIVES: In this study, we used hairless mice to assess the effects of ovariectomy with or without chronic UVA or UVB radiation on sagging and wrinkling of skin, on the elasticity of skin, as well as on matrix metalloproteinase activities in the skin. METHODS: Ovariectomies or sham operations were performed on 6-week-old female ICR/HR hairless mice. RESULTS: Even in the ovariectomy group without UV irradiation, the skin elasticity was significantly decreased during the 3-13 weeks after ovariectomy, which was accompanied by a significant increase in elastase activity in the skin. After UVA or UVB irradiation, skin elasticity was significantly decreased to a greater extent in the ovariectomy group than in the sham operation group, and this was accompanied by a reciprocal increase in elastase activity but not in the activities of collagenases I or IV in the skin. Consistent with the decreased skin elasticity, UVA irradiation for 12 weeks elicited more marked sagging in the ovariectomy group than in the sham operation group. UVB irradiation for 12 weeks also induced more marked wrinkle formation in the ovariectomy group than in the sham operation group. CONCLUSIONS: These results suggest that ovariectomy alone is sufficient to accelerate skin ageing and to increase UV sensitivity, which results in the further deterioration of the skin and photoageing, and may account for the accelerated skin ageing seen in postmenopausal women.

Age-related changes in skin wrinkles assessed by a novel three-dimensional morphometric analysis.

BACKGROUND: A system has been developed whereby the morphology of the skin surface can be evaluated directly in three dimensions. This system employs a non-invasive device that utilizes white light of halogen origin, and which allows the computation of wrinkle depth and width, and other parameters of skin surface morphology. Using innovative engineering, an optical system has been devised so that light is transmitted via a slit and can be used to measure not only replicas of the skin but also the skin surface directly. The measurement area is 6.4 x 6.4 mm, and the theoretical resolution with a x 50 magnification lens is within 12.5 micro m. OBJECTIVES: To use this system to study age-related changes in the morphology of wrinkles at the eye corner areas of women of varying ages. METHODS: One hundred and one healthy women (age range 20-80 years) residing in the Tokyo area were the subjects used in this study. RESULTS: Wrinkles demonstrated a rapid increase in depth in women aged 40 years or older, and plateaued at the age of 60 years. Surface morphology parameters yielded results similar to those of age-related changes in wrinkles. CONCLUSIONS: This new analytical system provides a rapid and convenient non-invasive method to evaluate skin surface morphology in three dimensions, especially with respect to wrinkle formation. The results obtained using this system provide a deeper insight into the mechanistic relationship between wrinkles and skin elasticity.

Quantitative two-dimensional analysis of facial wrinkles of Japanese women at various ages.

To evaluate individual differences in the recognition of facial wrinkles, we asked 40 Japanese female observers to identify wrinkles using transparent sheets over frontal facial photos of four females aged 20, 39, 55 or 75 years. We then measured the number and length of those wrinkles by image analysis. Wrinkles identified by those 40 observers showed aged-related increases in the standard deviation (SD) values for number and length but age-related decreases in the coefficient of variation (CV)%. Therefore, to clarify factors affecting the degree of wrinkle detection, wrinkles were identified by two groups of age-matched male and female observers, by two groups that differed by age, and by two other groups, one of which who felt that there was an improvement in their wrinkles after application of an antiwrinkle agent and another group who did not feel that there was any improvement after the same treatment. Improvement was observed by replica image analysis in all groups. The degree of wrinkles identified was not affected by the age or by the sex of the observer group. However, the group who felt that there was an improvement in their wrinkles after treatment with the antiwrinkle agent identified a significantly higher number of wrinkles than did the group who did not feel that there was an improvement. These results suggest marked individual differences in the recognition of wrinkles. Fine wrinkles in relatively young subjects are difficult to detect, but moderate to marked wrinkles in middle-aged and in aged subjects can easily be detected. Concerning the cause of individual differences in the extent of wrinkle detection, observers who identified a large number of wrinkles tended to recognize not only pronounced wrinkles but also recognized fine wrinkles as 'wrinkles'. This seems to have also affected their feelings about the success of treatment with the antiwrinkle agent.

Dermal fluid translocation is an important determinant of the diurnal variation in human skin thickness.

BACKGROUND: Ultrasonography has been used as a non-invasive approach to measure skin thickness. To date there have been no studies on diurnal variations in skin thickness. OBJECTIVES: To evaluate diurnal variations in skin thickness and to compare these with corresponding echogenicity and skin elasticity. METHODS: Measurements by ultrasonography B-mode and by Cutometer SEM 575 were carried out in the morning and in the afternoon on 20 men and 20 women (mean age 30 years) on three areas of the face (forehead, corner of the eye and cheek), the forearm and the upper arm, and the flank, thigh and calf. RESULTS: From the morning to the afternoon, the skin thickness in both sexes significantly decreased on three areas of the face, the forearm and the upper arm, but significantly increased on the thigh and calf. In parallel, the echogenicity significantly increased from the morning to the afternoon on the three areas of the face, the forearm and the upper arm, but decreased significantly on the thigh and calf. Measurements of mechanical properties at four sites demonstrated that from the morning to the afternoon, the major parameters of skin elasticity Ue* and Uf* increased significantly in both sexes on two areas of the face and slightly on the forearm, but decreased significantly on the calf. CONCLUSIONS: The diurnal profiles of skin thickness and skin elasticity in the upper half of the body are the reverse of those in the lower half of the body. These findings suggest that shifts of dermal fluid from the face to the leg by gravity during the day cause the diurnal variation in skin thickness.

The role of elastases secreted by fibroblasts in wrinkle formation: implication through selective inhibition of elastase activity.

We have previously demonstrated that decreases in skin elasticity, accompanied by increases in the tortuosity of elastic fibers, are important early events in wrinkle formation. In order to study the role of elastases in the degeneration of elastic fibers during wrinkle formation we examined the effects of an inhibitor of skin fibroblast elastase, N-phenethylphosphonyl-L-leucyl-L-tryptophane (NPLT), on wrinkle formation in hairless mice skin following UV irradiation. Dorsal skins of hairless mice were exposed daily to UV light for 18 weeks at doses of 65-95 mJ/cm2 and treated topically with 100 microL of 1 mM NPLT immediately after each UV irradiation. Wrinkles on dorsal skins were evaluated from week 6 through week 18. The daily exposure of mouse skin to UV light with less than 1 minimal erythemal dose significantly enhanced the activity of elastase in the exposed skin by week 4, and the elevated levels of elastase activity were significantly reduced by the in vitro incubation with NPLT in a dose-dependent manner to a level similar to that in unexposed mice skin, indicating that NPLT can efficiently inhibit the UV-inducible elastase activity. Topical application of NPLT significantly suppressed wrinkle formation when compared with vehicle controls by week 15 of treatment (P < 0.05). Histochemistry of elastic fibers with Orcein staining demonstrated that there were no obvious decreases of the fine elastic fibers in UV-exposed NPLT-treated skin in contrast to their marked decreases in the UV-exposed vehicle-treated skin. These findings suggest that skin fibroblast elastase plays a decisive role in wrinkle formation through the degeneration of elastic fiber.

Selective inhibition of skin fibroblast elastase elicits a concentration-dependent prevention of ultraviolet B-induced wrinkle formation.

We previously reported that wrinkle formation in the skin following long-term ultraviolet B irradiation is accompanied by decreases in skin elasticity and the curling of elastic fibers in the dermis. We further showed that wrinkles could be repaired by treatment with retinoic acid and that this was concomitant with the recovery of skin elasticity ascribed to the repair of damaged elastic fibers. Those studies suggested that decreasing the tortuosity of dermal elastic fibers is an important factor involved in inhibiting or repairing wrinkle formation. Therefore, it is of particular interest to determine whether the inhibition of elastase activity in vivo would prevent the damage of dermal elastic fibers and might abolish wrinkle formation associated with the loss of skin elasticity. Because the major elastase in the skin under noninflammatory conditions is skin fibroblast elastase, we used a specific inhibitor of that enzyme to assess its biologic role in wrinkle formation. The hind limb skins of Sprague-Dawley rats were irradiated with ultraviolet B at a suberythemal dose three times a week for 6 wk. During that period, 0.1-10.0 mM N-phenetylphosphonyl-leucyl-tryptophane, an inhibitor of skin fibroblast elastase, was applied topically five times a week. N-phenetylphosphonyl-leucyl-tryptophane application at concentrations of 0.1-1.0 mM abolished wrinkle formation in a concentration-dependent manner, with a peak for inhibition at 1.0 mM. This inhibition was accompanied by a continued low tortuosity of dermal elastic fibers and a maintenance of skin elasticity. Measurement of elastase activity after 6 wk of ultraviolet B irradiation demonstrated that whereas phosphoramidon-sensitive elastase activity was significantly enhanced in the ultraviolet B-exposed skin, there was no significant increase in that activity in the ultraviolet B-exposed, N-phenetylphosphonyl-leucyl-tryptophane-treated skin. These findings suggest that skin fibroblast elastase plays an essential part in the degeneration and/or tortuosity of elastic fibers induced by cumulative ultraviolet B irradiation.

Inhibitory effect of an extract of Sanguisorba officinalis L. on ultraviolet-B-induced photodamage of rat skin.

We previously reported that chronic Ultraviolet-B (UVB) irradiation causes wrinkle formation, decreases skin elasticity, and damages/curls dermal elastic fibers. Those UVB-induced wrinkles can be improved by treatment with retinoic acid or with a CO2 laser which results in a recovery of skin elasticity and a repair of elastic fiber linearity. We showed further that topical application of N-phenetyl-leucyl-tryptophane, an agent that specifically inhibits fibroblast-derived elastase, immediately after UVB irradiation inhibited UVB-induced wrinkle formation, maintained skin elasticity, and inhibited changes in the three-dimensional structure of dermal elastic fibers in a dose-dependent manner. In this study, the effects of an extract of Sanguisorba officinalis L., which also inhibits fibroblast-derived elastase, was evaluated for possible inhibition of UVB induced wrinkle formation, maintenance of skin elasticity, and prevention of damage to the 3-dimensional structure of dermal elastic fibers. Hind limb skins of 3-week-old Sprague-Dawley rats were irradiated with UVB at a suberythemal dose 3 times a week for 6 weeks. Simultaneously, an extract of Sanguisorba officinalis L. (at 0.2% (v/v) or 1% (v/v)) was topically applied 5 times per week immediately following each UVB irradiation and 1 d later. The extract of Sanguisorba officinalis L. inhibited wrinkle formation, maintained skin elasticity, and inhibited the decrease of dermal elastic fiber linearity in the rat hind limb skin in a dose-dependent manner. We have confirmed that the inhibition of elastase activity in fibroblasts immediately after UVB irradiation using an extract of Sanguisorba officinalis L. prevents chronic photodamage following UVB irradiation.

The inhibitory effect of an extract of Sanguisorba officinalis L. on ultraviolet B-induced pigmentation via the suppression of endothelin-converting enzyme-1alpha.

Endothelin-1 (ET-1) has been reported to be expressed in human epidermis at both the gene and protein levels. ET-1 plays a pivotal role in ultraviolet B (UVB)-induced pigmentation due to its accentuated secretion after UVB irradiation and its function as a mitogen and as a melanogen for human melanocytes. We have recently found that endothelin-converting enzyme (ECE)-1alpha plays a constitutive role in the secretion of ET-1 by human keratinocytes and that an extract of Sanguisorba officinalis L. inhibits ECE activity in human endothelial cells, which predominantly express ECE-1alpha. In this report, to clarify the potential use of this botanical extract as a whitening agent, we examined whether this extract inhibits UVB-induced pigmentation in vivo. When this extract was applied to human keratinocytes after UVB irradiation, secretion of ET-1 by those cells was reduced, and this was accompanied by a concomitant increase in the secretion of inactive precursor Big endothelin-1. When hairless mice were exposed to UVB light and were treated with the extract, it suppressed the induction of ET-1 in the UVB-irradiated epidermis. In the course of UVB-induced pigmentation of brownish guinea pig skin, this extract significantly diminished pigmentation in UVB-exposed areas. These findings indicate that ECE-1alpha in keratinocytes plays a pivotal role in the induction of pigmentation following UVB irradiation and that an extract of S. officinalis, which inhibits ET-1 production in human keratinocytes, is a good ingredient for a whitening agent.

Analytical method to examine the effects of carbon dioxide lasers on skin: a study using wrinkles induced in hairless mice.

BACKGROUND AND OBJECTIVE: Investigation of the wrinkle smoothing process elicited by CO(2) laser treatment is important for understanding the mechanism involved in their repair. STUDY DESIGN/MATERIALS AND METHODS: Hairless mice with wrinkles induced in their dorsal skin by long-term exposure to ultraviolet radiation in the wavelength range of 290-320 nm were treated with a CO(2) laser. By using this model, we investigated the external appearance, histologic changes, and the mechanical properties of the skin during the wrinkle repair. RESULTS: Laser treatment with an appropriate intensity caused wrinkles to smooth completely. In the healing process, reepithelialization and collagen tissue regeneration in the upper dermis was observed. However, marked changes in the skin were noted, such as increases in the collagen layer and in the skin thickness, and changes in the mechanical properties of the skin, despite the favorable external appearance. CONCLUSIONS: An abnormal state characterized by excessive collagen regeneration and other changes in the dermis occur concomitantly with wrinkle smoothing.

Ovariectomy accelerates photoaging of rat skin.

We have previously reported the formation of wrinkles, a decrease in skin elasticity and a loss in the linearity of dermal elastic fibers in rat hind limb skin irradiated with ultraviolet radiation in wavelength ranging 290-320 nm (UVB) at a suberythemal dose for 6 weeks. Estrogens are considered effective in preventing photoaging in postmenopausal females, but the role of estrogen in the skin remains unclear. In this study we have evaluated the influence of short-term chronic UVB irradiation at a suberythemal dose on the skin of ovariectomized rats. An ovariectomy or a sham operation was performed on each 3 week-old female Sprague-Dawley rat. Starting 1 week after the operation the hind limb skin of each rat was irradiated with UVB at a suberythemal dose (130 mJ/cm2) three times a week for 3 or 6 weeks. Decreases in elasticity and wrinkle formation in the skins of ovariectomized animals were induced more quickly than in the skins of sham-operated animals following UVB irradiation. The linearity of elastic fibers in the ovariectomy group decreased significantly compared with the sham-operation group, but erythema in the ovariectomy group was induced more readily than in the sham-operation group following UVB irradiation. These findings suggest that decreases in the estrogen levels after ovariectomy accelerate photoaging in terms of the morphology and physical properties of the skin surface and the three-dimensional structure of elastic fibers.

The paracrine role of stem cell factor/c-kit signaling in the activation of human melanocytes in ultraviolet-B-induced pigmentation.

The interaction of stem cell factor with its receptor, c-kit, is well known to be critical to the survival of melanocytes. Little is known about the role(s) of the stem cell factor/c-kit interaction in epidermal pigmentation, however. To clarify whether the stem cell factor/c-kit signaling has a paracrine role in ultraviolet-B-induced pigmentation, we determined whether the exposure of human keratinocytes, melanocytes, and the epidermis to ultraviolet B light stimulates the expression of stem cell factor or c-kit at the gene and/or protein levels. We further examined whether interrupting the binding of stem cell factor to c-kit by subepidermal injection of a monoclonal antibody to c-kit affects ultraviolet-B-induced pigmentation in brownish guinea pig skin. When human keratinocytes and melanocytes in culture were exposed to ultraviolet B light, transcripts of stem cell factor and c-kit (as assessed by reverse transcription polymerase chain reaction) and expression of those proteins (by enzyme-linked immunosorbent assay and western blotting) increased significantly and peaked at a dose of 20-40 mJ per cm2. In ultraviolet-B-exposed human epidermis, stem cell factor transcripts and protein expression were also markedly enhanced compared with the nonexposed epidermis. Immunohistochemistry with antibodies to stem cell factor revealed an increased staining in the ultraviolet-B-exposed epidermis, which was accompanied by a slight epidermal hyperplasia. In the course of ultraviolet-B-induced pigmentation of brownish guinea pig skin, the subepidermal injection of c-kit inhibitory antibodies completely abolished the induction of pigmentation in the ultraviolet-B-exposed area, and there was no increase in the number of dihydroxyphenylalanine-positive melanocytes. These findings indicate that the stem cell factor/c-kit signaling is critically involved in the biologic mechanism of ultraviolet-B-induced pigmentation.

Carbon dioxide laser treatment promotes repair of the three-dimensional network of elastic fibres in rat skin.

BACKGROUND: We have previously reported that ultraviolet (UV) B irradiation induces a loss of linearity in the three-dimensional structure of dermal elastic fibres, which results in the reduction of elastic properties of the skin and leads to wrinkle formation. We further reported that repair of wrinkles by all-trans retinoic acid is accompanied by recovery of the linearity of elastic fibres. Carbon dioxide (CO2) lasers are widely used for treating wrinkles in cosmetic surgery. OBJECTIVES: To perform CO2 laser treatment of wrinkles induced in rat skin by UVB irradiation and to evaluate changes in the three-dimensional structure of dermal elastic fibres during wrinkle repair. METHODS: Wrinkles were induced in the hind limb skin of Sprague-Dawley rats by UVB irradiation (130 mJ cm-2 three times weekly for 6 weeks), followed by CO2 laser treatment (11.3 J cm-2). The surface appearance of the skin was evaluated by replica observation 6 and 10 weeks after CO2 laser treatment followed by measurement of mechanical properties using a Cutometer. Subsequently, perfusion fixation and digestion with formic acid were performed and elastic fibres were observed by scanning electron microscopy (SEM). Image analysis of SEM micrographs was carried out to evaluate the linearity in the three-dimensional structure of elastic fibres. RESULTS: Six weeks after CO2 laser treatment, all parameters of skin mechanical properties in the UVB-irradiated group recovered to levels of the control non-irradiated group, accompanied by repair of wrinkles and a significant increase in linearity of the three-dimensional structure of elastic fibres. CONCLUSIONS: These findings indicate that CO2 laser treatment has a therapeutic potential to repair wrinkles to non-irradiated levels through recovery of the three-dimensional structure of elastic fibres.

Diurnal variation affects age-related profile in skin thickness.

We have previously demonstrated that over the course of each day there are changes in skin thickness that can be measured by B-mode ultrasonography. This suggests that there is a shift in dermal fluid from the face toward the legs by gravity, resulting in a diurnal variation in skin thickness. Therefore, age-dependent profiles in skin thickness were evaluated by B-mode ultrasonography in the morning or in the afternoon for 130 normal Japanese females aged 18-83 years. Three areas of the face (the forehead, the corners of the eye, and the cheeks) were measured as distinctively sun-exposed areas while the flexion side of the forearm was measured as a weakly sun-exposed area. A weak correlation between skin thickness and age was found in all areas measured (positive for the forehead, the corners of the eye, and the cheeks; negative for forearms) in the morning but not in the afternoon, when only a weak positive correlation was observed in the cheek. These results indicate that when measuring skin thickness, an appropriate time for taking measurements should be selected with consideration of the movements of dermal fluid over the course of each day.

Epidermal change can alter mechanical properties of hairless mouse skin topically treated with 1alpha, 25-dihydroxyvitamin D(3).

Wrinkle formation caused by photoaging clearly involves changes of extracellular matrix components and mechanical properties of the skin. Recently, it was reported that the topical application of 1alpha, 25-dihydroxyvitamin D(3) to hairless mouse skin induced wrinkling. Here we have evaluated the effect of topical application of 1alpha, 25-dihydroxyvitamin D(3), which causes skin wrinkling, on the mechanical properties of the hairless mouse (HR/ICR) skin, using a commercially available non-invasive in vivo instrument. The elasticity element of the skin was unchanged, but the viscosity element significantly increased. Histologically, the epidermis became remarkably thick, but no conspicuous changes were observed in the dermis. Changes in the mechanical properties of the skin after 1alpha, 25-dihydroxyvitamin D(3) treatment take place through epidermal physical variation, especially changes of viscosity elements. It is suggested that the visco-elastic properties of the epidermis are also attributable to the morphology as well as the mechanical properties of the skin.