A simple experimental method to study depigmenting agents.

The first objective of the study was to verify that a controlled UV exposure of four areas of the forearms together with randomized product application enabled to compare treatment efficacy and then to compare the depigmenting efficacy of different products with a simple experimental method. Sixteen volunteers received 0.7 minimal erythermal dose for four consecutive days. Products tested were ellagic acid (0.5%), vitamin C (5%) and C8-LHA (2%). Product application started 72 h post last exposure, was repeated for 42 days, the control zone being exposed, non-treated. Colour measurements included Chromameter, Chromasphere, Spectro-colorimeter and visual assessment. Comparison of colour values at day 1 and at day 7 showed that all zones were comparably tanned, allowing a rigorous comparison of the treatments. We report a new simple experimental model, which enables the rapid comparison of different depigmenting products. The efficacy and good tolerance of C8-LHA make it an excellent candidate for the treatment of hyperpigmentory disorders.

Mexoryl SX: a broad absorption UVA filter protects human skin from the effects of repeated suberythemal doses of UVA.

There is now considerable evidence that chronic UVA exposure induces damage in animal and human skin; however, little is known about UVA protection of human skin. The aim of this study is to evaluate the efficacy of Mexoryl SX, a broad UVA absorber (lamada max = 345 nm) against UVA-induced changes in human skin. The regimen of UVA exposure (13 weeks with increasing suberythemal doses) induces intense pigmentation with no erythema. Skin hydration and elasticity decrease, whereas total skin thickness, assessed by echography, remains unchanged. Irradiated epidermis reveals a significant thickening of the stratum corneum, an absence of hyperplasia and an increase in the expression of the protective iron-storage protein ferritin. No significant alterations are seen using antisera against type IV collagen or laminin, suggesting that the dermal-epidermal junction (DEJ) is mainly preserved. In dermis, enhanced expression of tenascin is seen just below the DEJ but type I procollagen, which is localized at the same site, is unaltered. Although we are unable to visualize any changes in elastic network organization using Luna staining or specific antiserum directed against human elastin, we notice an increased deposition of lysozyme or alpha-1 antitrypsin on elastin fibres. Mexoryl SX (5%) efficiently prevents these alterations. Thus, these results suggest that UVA photoprotection can prevent early putative alterations leading to photoageing.

Echographic assessment of corticosteroid-induced skin-thinning, including echogenicity measurement.

BACKGROUND/AIMS: Skin atrophy is one of the main side effects of long-term topical corticosteroid therapy. It has already been studied through ultrasound skin-thickness measurement. In this study, the quantification of dermal echogenicity was introduced to provide new information on this phenomenon. METHODS: Skin thinning induced by topical application (without occlusion) of the superpotent corticosteroid clobetasol propionate (0.05% lotion), was assessed by means of ultrasonography in terms of thickness and echogenicity. 15 healthy volunteers were treated for 6 weeks, 1 daily, 5 days a week on the forearms. RESULTS: The thinning showed a biphasic pattern, with a 1st period of rapid change (about 15% of thinning in a week) followed by a period of slower but significant change. Skin thickness returned to baseline values 3 weeks after the end of treatment. Dermal echogenicity, which represents the mean intensity of the ultrasound signals reflected by the dermis, was found to follow the same variations, increasing strongly during the 1st week, then more slowly. The 2 parameters are correlated and probably reflect the same physiological modifications responsible for skin thinning, i.e., a reduction in glycosaminoglycan synthesis (leading to a drastic fall in dermal water content) and vasoconstriction. CONCLUSION: This sensitive and non invasive method enables us to identify the effects of clobetasol propionate on the healthy dermis in the absence of any clinical signs of thinning.

The effects of aging on the cutaneous microvasculature.

BACKGROUND: Few studies have attempted to quantitatively assess in vivo changes in the microvasculature with age. OBJECTIVE: The objective was to assess in vivo structural and functional changes in the cutaneous microvasculature with aging and to analyze the contribution of the microvasculature to skin color. METHODS: Video capillaroscopy, in conjunction with fluorescein angiography, and laser-Doppler flowmetry were used to compare elderly and young normal volunteers. Skin color differences were assessed with a handheld color reflectance meter. A photoexposed site, the forehead, and the relatively photoprotected ventral forearm were studied to differentiate photoaging from chronologic aging. RESULTS: Dermal papillary loops were significantly reduced in old skin compared with young skin (forehead by 40%; forearm by 37%). Horizontal vessels showed increased volume fraction in elderly forehead and forearm skin. Laser-Doppler studies demonstrated no significant differences between young and old skin; indeed, hyperemic responsiveness appeared more rapid in the elderly. Color measurements showed elderly skin, particularly in men, to be significantly darker and redder. CONCLUSION: A marked loss in dermal nutritional vessel density and surface area for exchange is a feature of both chronologic aging and photoaging.

Noninvasive measurement of the effect of lifetime exposure to the sun on the aged skin.

The cumulative and comparative effects of chronological and actinic changes to the skin were studied in an elderly population. Two adjacent sites with different degrees of exposure to the sun were examined to distinguish the contribution of each factor, by means of noninvasive measurements. The following parameters were measured on the skin of the low neck (an exposed site), as well as on the adjacent clothes-protected skin of 30 elderly women: electrical conductance, color, microrelief, biomechanical properties and the thickness of both the full skin and the subepidermal nonechogenic band (SENEB), using ultrasound measurements. Persistent exposure to the sun accentuated most of the age-induced modifications. The following changes were significantly more marked in exposed skin: SENEB thickening, full-skin thinning, loss of extensibility and elasticity, and color heterogeneity. The cumulative effects of sun exposure and chronological aging lead to atrophy in elderly people. These results suggest that, if there is an autoprotective skin reaction, such as thickening in young adults, it is eliminated in old people.

How does sodium lauryl sulfate alter the skin barrier function in man? A multiparametric approach.

The effects of topically applied sodium lauryl sulfate were studied in vivo, in man, through noninvasive methods and compared with those obtained in vitro, using human stratum corneum as a model. The results show that the very marked increase of transepidermal water loss obtained in vivo cannot be related to any removal of epidermal lipids. The most likely explanation of our findings lies in the hyperhydration of stratum corneum consecutive to the inflammation process and a possible disorganization of the lipid bilayers.

Influence of age and sun exposure on the biophysical properties of the human skin: an in vivo study.

The physical properties of the skin were measured by using noninvasive methods on 72 people displaying various levels of solar elastosis on the neck. The physical parameters measured were the skin extensibility, the elastic recovery, the skin colour, the skin thickness and the electrical conductance. The correlation between the above parameters, the clinical grades of elastosis and the chronological age of each subject were studied using two different statistical approaches. They both showed that elastotic skin is less elastic, dryer, darker, more erythematous and less yellowish than the nonexposed skin. The similarities and differences between the properties of elastotic skin and purely chronologically aged skin are discussed.

In vivo biophysical characterization of skin physiological differences in races.

The role of race in modulating skin responses has been investigated. Several parameters (skin thickness, transepidermal water loss, water content of the stratum corneum and skin biomechanics) have been measured using noninvasive tools in whites, Hispanics and blacks to assess whether the melanin content could induce changes in skin biophysical properties. Marked differences between races appear in stratum corneum water content and in skin extensibility, recovery and elastic modulus. Measurements done in different sun-exposed sites highlight the effects of solar irradiation on the skin and the role of melanin in preventing skin damage. The study shows that racial differences in skin physiology exist and are mainly related to the protective role of melanin present in races with darker skin. Moreover, differences in skin hydration are not fully explained according to the site and presence of hair.

Effect of topical corticosteroids on human sebum production assessed by two different methods.

Topical corticosteroids are widely used in cutaneous diseases. Although their mode of action on different skin compartments has been documented, little is known about their effects on the human sebaceous gland. We investigated the effects of two corticosteroids of differing potency on the excretion of sebum by means of two validated techniques: the Sebutape and the Lipometre. This study was conducted on the forehead skin of normal healthy subjects. The results obtained with both techniques correlated well. The application of both corticosteroids during a 4-week period led to a significant decrease in sebum excretion. This decrease was more pronounced with the more potent corticosteroid (Dermovate). In the light of these findings, it is likely that topical corticosteroids exert an anti-proliferative action upon the sebaceous cells in a similar manner to their effect in other skin compartments.

In vivo study of scleroderma by non-invasive techniques.

Sixteen patients with localized scleroderma (LS) and 11 with progressive systemic sclerosis (PSS) were studied using non-invasive techniques to determine skin thickness, skin extensibility, transcutaneous PO2 and cutaneous blood flow and were compared with normal controls. LS was characterized by a decreased skin extensibility with thickening of the skin in progressive lesions. There was an increase in the cutaneous microcirculation with a decrease in the PO2. In PSS, there was a decrease in the skin extensibility caused by thickening of the skin, but no change in cutaneous microcirculation or PO2 values. These results indicate that skin thickness and extensibility are the most useful parameters in the study of the progression of LS and PSS.

Assessment of aging of the human skin by in vivo ultrasonic imaging.

The ultrasonic imaging technique that we have developed provides cross-sectional images of human skin in vivo with a resolution of about 80 microns axially (i.e., deep into the skin) and 250 microns lateral (parallel to the surface). In order to study aging skin, we obtained ultrasonic images from the mid-forearm (volar and dorsal sides) of 142 women. Ultrasonically, on the images, the dermis appears composed of two bands: a dark superficial one where the ultrasonic waves are propagated in a relatively homogeneous or non-echogenic medium, and a deeper one, which is lighter in color, suggesting a heterogeneous medium. Our results show that skin is thicker on the dorsal than on the volar forearm. In contrast to previously published results, skin thickness remains constant until the seventh decade of life, diminishing thereafter. The relative thickness of the two bands show marked variations with age: a progressive thickening of the dark band, from zero in infants to approximately 75% of total skin thickness in aged subjects, while the light band shows the inverse trend. Comparing the amplitude of the bands on the volar and dorsal forearm, the relative thickness of the dark band is larger on the dorsal (exposed) side and increases with age. These findings and the analysis of variously stained biopsies taken in some of our patients lead us to assign this dark band to a zone in the upper dermis where the collagen network is delicate, dense, and well organized. This is supported by some data in the literature. The thickness of this subepidermal non-echo-genic band appears to be a far more sensitive marker of skin aging at the dermal level than is the measurement of skin thickness.

Age-related mechanical properties of human skin: an in vivo study.

We have investigated in vivo how various viscoelastic parameters that describe the mechanical properties of the human skin may vary with age. Accordingly, we have used a mechanical device that records the torsional extensibility of the skin. When submitted to a low torque, the time-response curve of the skin affords the determination of the immediate extensibility (UE), the immediate recovery (UR), the viscoelastic part of the deformation (UV), the elastic recovery (UR/UE), and the creep relaxation time (tau). Because the skin thickness varies with age and primarily governs the mechanical properties, it was measured through an ultrasound technique at the same sites (forearm) where the torque was applied. The results show that the skin maintains its thickness and extensibility up to the seventh decade as opposed to its elasticity or recovery capacities, which decrease from an early age. The viscous part of the deformation is constant through life, whereas the creep relaxation time decreases linearily with age. Except for skin thickness, no differences in these parameters between men and women were detected. The significance of these results are discussed in terms of structure alterations. The determination of the elastic recovery (UR/UE) appears to be a parameter of choice for illustrating skin aging.

In vivo cross-sectional ultrasonic imaging of human skin.

This paper describes a new experimental set-up allowing routine cross-sectional ultrasonic images of human skin to be obtained in vivo and presents the first high-resolution images it gives. This system is characterized by the utilization of damped, high-frequency ultrasonic transducers. This type of transducer is longitudinally driven along the skin by a stepping motor under the control of a computer, which at the same time stores the digitized reflected signals. After the acquisition sequence, the image of the skin is displayed on a video monitor after data processing of the ultrasonic signals. On the images thus obtained, fascia, hypodermis, reticular dermis, pilosebaceous units, and adventitial dermis can be visualized. Due to a lack of resolution (about 80 microns) epidermis can only be viewed on certain zones like the hand.

Are corneocytes elastic?

By using a microhandling system, we have recorded the force necessary to elongate isolated corneocytes. According to certain hypothesis, it appears that the elastic modulus of corneocyte is far higher than that of stratum corneum itself. Aggregates of corneocytes are disassembled by forces with a wide range of intensity. In the light of these experiments, stratum corneum appears as a very supple material where the deformations occur more by a plastification of the intercellular medium and an unfolding of the microrelief lines than an elongation of the corneocytes themselves. Moreover, desmosomes remnants could play a certain role in the corneocytes cohesiveness.

New experimental approach to measure the skin-reflected light. Application to cutaneous erythema and blanching.

By modifying the blood supply in the upper part of the dermis, the colour of the skin is changed by any material in contact with it. This is a great difficulty when we want to measure the colour intensity in erythema or blanching. To avoid this problem, we designed and built a device which measures the skin reflectance value without any contact with the skin. The use of this device on man to study the skin blanching after application of dermocorticoids allows us to determine the more potent drug among a series (clobetasol propionate) and, for a given drug (betamethasone), the more efficient preparation (oil-in-water emulsion). The study of the effect of increasing irradiation doses of ultraviolet light show that the sensitivity of the device is equivalent to that of the clinical assessment and that the relationship between the irradiation dose and the skin reflectance is sigmoidal.

In vivo studies of the evolution of physical properties of the human skin with age.

In vivo measurements taken by valid methods, help to interpret the phenomena associated with the aging process. The authors developed some routine techniques to measure physical properties of the skin both deep in the dermis-epidermis and at the surface (stratum corneum). Measurements on the forearm of 150 people yielded the following results regarding the dermis-epidermis level: (1) skin thickness begins to decrease at 45 years of age for men and women, when women's skin becomes thinner than men's skin; (2) torsion extensibility, normalized for a given skin thickness, sharply decreases after 35 years of age; (3) skin optical properties are modified, ie the photoplethysmographic signal, measured on the forehead, greatly increase after 60 years of age; (4) at the cutaneous surface level, the main modifications are increased shedding of stratum corneum after 60 years of age in parallel with a modification of the stratum corneum cohesivity, altered skin microrelief during the aging process, and slightly decreased transepidermal water loss while the corneocyte projected size increases. These findings allow consideration of the aging process at the skin level as a phased process. The alterations of the physical properties and their kinetics suggest a various process according to the different cutaneous sections.

[Study of the correlation between transepidermal water loss and percutaneous penetration of steroid (author's transl)]. / Etude comparée de la perte insensible d'eau et de la pénétration cutanée des corticoïdes.

Modifications of transepidermal water loss due to repeated topical applications of water/oil emulsion, salicylic acid and urea, with and without occlusion, were studied on the forearms of healthy volunteers. Percutaneous absorption of three different corticosteroids was evaluated by means of the bleaching test on each treated area. It was shown that only salicylic acid significantly increased transepidermal water loss. This increase was positively correlated with an increase in the percutaneous absorption of corticosteroids. Thus it seems that a comparative evaluation of the percutaneous absorption of topical treatments in normal and lesional skin may be obtained, in vivo in man, by nonaggressive comparative evaluation of transepidermal water loss.

Mechanical properties and Young's modulus of human skin in vivo.

The mechanical properties of the in vivo dermis were measured by means of a torque applied to the skin. The resulting deformation of 2-6 degrees, including the immediate and delayed visco-elastic components, as well as the relaxation were measured, and the raw values corrected for a constant skin thickness. The experiment performed on 138 individuals from 3 to 89 years old revealed a diminished elasticity and stretchability after the age of 30, associated with an increase in the visco-elastic component. The Young's modulus doubles with age. The results are discussed in terms of the various models proposed to explain dermal structure.

Influence of ageing on the in vivo extensibility of human skin at a low stress.

The response of the forearm skin to a 9 . 10(-3) N . m torque was measured on 141 subjects aged 3-89 years. The results for each age class were adjusted to a skin thickness of 1 mm by use of a skinfold caliper. Results indicate a maximum skin thickness around the age of 40 years. Standardised skin extensibility decreases with age, the skin of females being less extensible than that of males. These results, which should be considered in relation to the total water content of the skin, are compared to those of other papers dealing with this subject.