Unexpected distribution of surface hydration level of the lip.

BACKGROUND: The functional properties of human lips depend on their hydration level. Limited data are, however, available. OBJECTIVE: To investigate water distribution through the lower lip surface. METHODS: Images from the surface of the lip were recorded using a new capacitance imaging technique. Capacitance and conductance measurements were also carried out in various points of lower lip and on adjacent skin. RESULTS: Data clearly show that the closest part of the mucosa of the lip is less hydrated than the external part. These unexpected findings could be related to the presence of a premucosa area whose structure is clearly different from that of vermilion zone.

Oriented structure in human stratum corneum revealed by X-ray diffraction.

Various types of human stratum corneum (sheets or callus) were exposed, in parallel and perpendicular geometry, to the high flux of X rays produced by a synchrotron radiation source. Under these conditions, very clear and rich diffraction patterns, corresponding to the supramolecular organization of stratum corneum proteins and lipids, were obtained. The comparative study of normal or delipidized stratum corneum sheets and membrane couplets allows one to attribute certain diffraction features to lipids. Our results in the 3-7-nm range show two different distances for lipid bilayers. Concerning the protein nature of normal stratum corneum, the results show that keratin would occur in the beta form, whereas for callus it is in the alpha form. Indeed, normal stratum corneum sheets never display the 0.514-nm characteristic of alpha keratin. This result means that the supramolecular organization of keratin could depend on the keratinization process. Finally, our studies also confirm the presence of a still-unknown protein component existing in the beta form that would be located either inside the corneocytes or in some dilatated zones of the intercellular spaces.

In vivo proton relaxation times analysis of the skin layers by magnetic resonance imaging.

If in vivo magnetic resonance imaging is nowadays a powerful non-invasive method in medical diagnosis, its application in order to study the skin in vivo is not yet in common use because skin imaging requires a high resolution, at least in the direction perpendicular to the skin surface. We have therefore designed a specific imaging module, which, connected to a standard whole-body imager at 1.5 Tesla, allows us to obtain in vivo magnetic resonance images of skin on most parts of the body. With a depth resolution of about 70 microns, we are able to differentiate the skin layers: epidermis, dermis, subcutaneous fat, and even a thickened stratum corneum on palm as well as on heel. This paper reports the T1 and T2 water proton relaxation times of the different skin layers, in vivo, which are magnetic resonance parameters extracted from the images. Results show that skin layers are characterized by shorter T2 relaxation times than other biologic soft tissues. On the contrary, the measured T1 values are in the same range as in other tissues. These short T2 values may be assigned to the fibrous protein content of the skin and particularly of the dermis. This study on normal skin is the precursor of further works such as the influence of aging. As regards skin pathologies, it will be a powerful tool to follow the evolution of skin diseases under treatment.

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.

Measurement of mechanical forces generated by skin fibroblasts embedded in a three-dimensional collagen gel.

Mechanical activities developed by cells play a significant role in the embryogenesis, development, and physiopathology of pluricellular organisms. A technique is described to measure in vitro the traction force developed by cells seeded into a three-dimensional polymeric collagen lattice. It is based on the use of strain gauges generating an electrical signal upon tension that is amplified and recorded. The intensity of the signal depends on the number and type of cells, cytoskeleton integrity, concentration of collagen in the lattice support, and fetal calf serum in the culture medium. Skin fibroblasts from humans and animals produce traction forces ranging from 100 to 1000 mg per million cells. In the gel under tension, the cells are in mechanical dynamic equilibrium with their support. It is suggested that the mechanical activity of fibroblasts and the control of the tension that they operate on the lattice support participate in the structural organization of the dermis and in its physiologic tension.

Changes in tactile spatial discrimination and cutaneous coding properties by skin hydration in the elderly.

Neurosensory tactile functions were investigated in human subjects by two different and complementary experimental approaches. First, a conventional psychophysical method (two-point gap discrimination) was used to determine the tactile discrimination threshold by analyzing the subjects' ability to detect a gap of variable width between two contact points when a series of stimuli was applied to the skin. Using this method we confirmed the marked degradation of tactile spatial acuity with age and showed that skin discriminative function was partially restored after hydration of the skin with a moisturizer. The second approach consisted of a microneurographic recording of tactile afferent fibers in response to two types of mechanical stimuli applied reproducibly to the corresponding receptive fields. With this method, we found that the afferent messages were depressed following hydration of the skin surface. Thus, partial restoration of tactile spatial acuity after hydration appears to be due to both a softening of the stratum corneum permitting better localization of the stimulus and a weaker transfer of the stimulus toward the sensory receptors.

Reversal effects of topical retinoic acid on the skin of kidney transplant recipients under systemic corticotherapy.

The systemic long-term corticosteroid treatment administered to kidney graft recipients (KGR) within the framework of the required immunosuppressive therapy induces an atrophy of the skin, from the sixth month onwards. We studied the effect of topical all-trans retinoic acid (0.05%; Galderma Labs.) applied to the forearms of 27 KGR (14 men, 13 women) over a 6-month period. Twenty-four subjects completed the trial. The following results were obtained in the treated forearm versus the untreated forearm (excipient alone): clinically, an increase in skin thickness; by noninvasive techniques, an increase in skin thickness, skin elasticity, skin conductance, and TEWL, and a reduction in the size of the corneocytes. No change in stratum corneum lipid content was observed. A sex-related difference was noted in the response to treatment under our experimental conditions, the female patients responding better. A punch biopsy (4 mm) was performed on both forearms of four patients after the 6-month period. Histologic and ultrastructural examination revealed epidermal and dermal changes evoking increased cellular metabolism in the retinoic acid-treated forearms.

Characterization of the skin in vivo by high resolution magnetic resonance imaging: water behavior and age-related effects.

The age-related modifications of magnetic resonance imaging parameters in the skin have been studied in vivo. Modification of these parameters should provide important information about alterations in water structure and content in aged skin. Relaxation times, T1 and T2, and relative proton density, which corresponds to the mobile water proton fraction of tissues, have been measured on people under age 40 and over 70 on a sun-protected area. Results have confirmed in vivo skin layer differentiation through relaxation times performed in a previous study. Moreover, relative proton density quantification has shown that epidermal mobile water is at least twice as abundant as dermal mobile water. No significant age-related T1 and T2 modification could be established, basically because of a large dispersion of values. The main result concerns the upper part of dermis (about 200 microns in thickness) which contains significantly more mobile water protons in chronologic aged skin than in young adult skin. This increase has been related both to an increase of total water content in dermis with age and to an apparent decrease of collagen and proteoglycan content. Associated with alterations of their structure, this decrease reduces macromolecular-water interaction sites. This finding has to be compared with ultrasound evaluation of aged skin, which is characterized by modifications of the echogenicity, related to collagen bundles size and density, in the outer part of dermis, too. Both of the imaging techniques tend to consider the outer part of dermis as one of the privileged sites of skin aging.

Morphometry of human epidermis in vivo by real-time confocal microscopy.

Real-time confocal microscopy has brought substantial improvements to the imaging of the human skin in vivo. On early images, the stratum corneum could be distinguished from the living epidermis and the circulatory network of the superficial dermis. We have adapted the Tandem Scanning Microscope to obtain images of the living skin, showing thinner structures such as the stratum lucidum and the dermo-epidermal junction, both of which are essential markers for micron-order measurements of the thickness of the stratum corneum and living epidermis. The measurements were corrected for the differences in the refractive index of the various cutaneous layers, and the undulation of the dermo-epidermal junction. Furthermore, nucleus size and number could be assessed from horizontal optical sections. To illustrate the sensitivity of the thickness measurements, changes in the thickness of the epidermis were recorded during and after stripping of the horny layers. This non-invasive methodology is a very promising tool for morphometric studies of the living human skin at the cellular level.

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.

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.

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.

Assessment of topical non-steroidal anti-inflammatory drugs.

A new non-invasive technique for assessing the efficacy of topical non-steroidal anti-inflammatory drugs (NSAID) in man is proposed. The NSAID are initially applied to the skin under occlusion before inflammation is induced by a methyl nicotinate solution. The inflammatory response is quantified in terms of cutaneous blood flow by a laser Doppler velocimeter (LDV). The efficacy of NSAID preparations is calculated by comparing the responses of the LDV to the methyl nicotinate challenge on the pretreated and the non-treated skin sites. This protocol has been used to investigate the effect of three different NSAID preparations (indomethacin, niflumic acid, palmitoyl collagenic acid) and the influence of the vehicle on the efficacy of indomethacin. The three preparations tested gave positive results but with different amplitudes in response. The efficacy of indomethacin varied with the vehicle used.

Quantitative assessment of skin aging.

Noninvasive methods have allowed physicians to give an objective description of aged skin in terms of functional and esthetic properties. The relative influence of environment (mainly sun) on the true aging process can be assessed through the obtained data. It is also possible to measure the efficacy of topical preparations (cosmetics or drugs) designed for treating the various cutaneous aging marks.

In vivo confocal microscopy of human skin: a new design for cosmetology and dermatology.

In-depth exploration of cellular structures in living human skin in situ is possible with the tandem scanning microscope (TSM). However, the rigid design of the microscope limited observations to the arms, hands, and fingers. A mobile version allowing the investigation of any parts of the body has been designed. The head containing the Nipkow disk and the optical path were the only part saved from the original TSM. This prototype can be used to observe, in real time, the different skin structures down to a depth of 200 microns and to measure the thickness of the different layers with micron precision level. The hydration of the stratum corneum (SC) could be assessed. For example, lengthy immersion of the hand in water led to an increase in SC thickness without affecting that of the living epidermis. Occlusive patch tests also showed that water and, even more so. propylene glycol, led to transient swelling of the SC. In dermatology, the example of psoriasis illustrated the value of the TSM for describing, measuring, and assessing pathologic skin changes. The availability of this noninvasive method for observing changes with time in a given skin site should prove useful for monitoring treatment efficacy. This tool opens up new insight for the investigation of cutaneous pathophysiology.

Influence of hairdressing on the psychological mood of women.

A questionnaire, specially designed to measure the mood change was filled in by 359 women at their arrival in different hair-salons and after having received various hair care services. The results show that hair treatments increase all the positive dimensions of mood and decrease all the negative ones. Only the improvement in sociability appears to be linked to age.

Do cosmetics enhance female Caucasian facial attractiveness?

This study sought to investigate whether cosmetics do improve female facial attractiveness, and to determine whether the contribution of different cosmetic products are separable, or whether they function synergistically to enhance female beauty. Ten volunteers were made up by a beautician under five cosmetics conditions: (i) no make-up; (ii) foundation only; (iii) eye make-up only; (iv) lip make-up only; and (v) full facial make-up. Male and female participants were asked to view the 10 sets of five photographs, and rank each set from most attractive to least attractive. As predicted, faces with full make-up were judged more attractive than the same faces with no make-up. Sex differences within the results were also apparent. Women judged eye make-up as contributing most to the attractiveness. Men rated eye make-up and foundation as having a significant impact on the attractiveness of a full facial makeover. Surprisingly, lipstick did not appear to contribute to attractiveness independently.

Frictional properties of skin: proposal of a new approach.

Phenomena occurring at the interface between skin and a glass pad, alternatively moved at its surface, are described by direct observation and measured by recording the force on the pad induced by the skin displacement. Static and dynamic friction coefficients are determined as a function of the load applied on the pad. Influence on the recordings of the friction coefficient of some cosmetic treatments applied on the skin are described. Results show that discontinuous phenomena, due to compression and distortion of the skin microrelief, occur during the beginning of the sliding process. Friction coefficients are depending on the load applied on the pad. Our studies confirm the increase of friction coefficient after hydration of the skin surface and suggest that the fluctuations in the frictional force amplitude could supply an objective, although indirect, measurement of skin smoothness.