Ultrastructural morphometry of normal human dermal-epidermal junction. The influence of age, sex, and body region on laminar and nonlaminar components

To obtain baseline data for future studies on such processes as wound healing, carcinogenesis, and blistering, a morphometric analysis of the dermal-epidermal junction was undertaken on normal skin from 3 or 4 standard sites on the arm and leg of 12 subjects aged 20-60 years. Lamina densa was thinner in females than in males (p less than 0.01) but no sex difference was apparent for lamina lucida. Both laminar elements were thinner beneath melanocytes than beneath keratinocytes. Sex, age, and body region had no apparent influence on numbers of hemidesmosomes or basal cell plasmalemmal vesicles, nor was there a significant variation of these structures among individuals. Numbers of dermal microfibril bundles diminished with age (p less than 0.01). Anchoring fibril counts varied widely both among individuals (p less than 0.025) and within the same subject; there were fewer in the upper arm compared with different parts of the leg (p less than 0.005). These results emphasize the importance of appropriate controls in studies of physiologic and pathologic conditions involving the dermal-epidermal junction.     

Effects of corticosteroids on the proliferation of normal and abnormal human connective tissue cells

Four corticosteroids were tested in vitro for effect on the proliferation of four strains of fibroblasts from scleroderma skin, four strains from normal adult skin and four strains of rheumatoid synovial cells. Significant effects on fibroblasts occurred only at the highest steroid concentration tested (10 microgram/ml) where the inhibitory ranking of the steriods was clobetasol propionate greater than clobetasone butyrate greater than betamethasone valerate greater than hydrocortisone. Hydrocortisone and betamethasone valerate stimulated proliferation of two normal strains, had no certain effect on the scleroderma group, and inhibited growth of synovial cells. Clobetasone butyrate and clobetasol propionate inhibited growth of all cells. All four steroids substantially reduced acid mucopolysaccharide secretion by scleroderma fibroblasts. These results suggest that fibroblasts from normal and abnormal skin show only small differences in their responses to corticosteroids in vitro, but contrast sharply with the mouse L-929 fibroblasts previously used in some assays of topical corticosteroid potency.     

Chronological ageing and photoageing of the fibroblasts and the dermal connective tissue

In recent years, the exposure of human skin to environmental and artificial UV irradiation has increased dramatically. This is due not only to increased solar UV irradiation as a consequence of stratospheric ozone depletion, but also to inappropriate social behaviour with the use of tanning salons still being very popular in the public view. Besides this, leisure activities and a lifestyle that often includes travel to equatorial regions add to the individual annual UV load. In addition to the common long-term detrimental effects such as immunosuppression and skin cancer, the photo-oxidative damage due to energy absorption of UV photons in an oxygenized environment leads to quantitative and qualitative alterations of cells and structural macromolecules of the dermal connective tissue responsible for tensile strength, resilience and stability of the skin. The clinical manifestations of UV/reactive oxygen species (ROS)-induced disturbances result in photoaged skin with wrinkle formation, laxity, leathery appearance as well as fragility, impaired wound healing capacities and higher vulnerability. Strategies to prevent or at least minimize ROS-induced photo-ageing and intrinsic ageing of the skin necessarily include protection against UV irradiation and antioxidant homeostasis.     

Elastic properties of human skin: relation to age,sex, and anatomical region

Summary Using a recently developed noninvasive, in vivo suction device for measuring skin elasticity, we evaluated age, sex, and regional differences in the viscoelastic properties of skin. A total of 33 volunteers participated in the study consisting of (a) 8 young females, (b) 9 old females, (c) 8 young males and (d) 8 old males. Measurements were performed on 11 anatomical regions; three different loads were applied: 100,200, and 500 mbar. The parameters used were: immediate distension (Ue); delayed distension (Uv); immediate retraction (Ur); and, final deformation (Uf). To compare between subjects and anatomical regions, relative parameters independent of skin thickness were calculated: Uv/Ue, the ratio between the viscoelastic properties of skin and immediate distension, and Ur/Uf, which measures the ability of the skin to regain its initial position after deformation. Generally, Uv/Ue increased while Ur/Uf decreased with aging. Responses were variable with respect to load applied. Variability within anatomical regions was also noted. However, differences between the sexes were not statistically significant for most regions. These findings are in congruence with earlier studies suggesting the differences are mainly attributable to alterations in the elastic fiber network. This procedure provides a simple, quantitative assessment of elastic properties of the skin. Its application may help in future investigations of other connective tissue disorders.     

Effect of dermal thickness, tissue composition, and body site on skin biomechanical properties

BACKGROUND/PURPOSE: Quantitative measurement of skin biomechanical properties has been used effectively in the investigation of physiological changes in tissue structure and function and to determine treatment efficacy. As the methods are applied to new questions, tissue characteristics that may influence the resultant biomechanical properties are important considerations in the research design. For certain applications, variables such as dermal thickness and subdermal tissue composition, as well as age and/or solar exposure, may influence the skin biomechanics. METHODS: We determined the influence of dermal thickness, tissue composition, and age on the skin biomechanical properties at the shoulder, thigh, and calf among 30 healthy females. We compared two devices, the Biomechanical Tissue Characterization System and the Cutometer SEM 575 Skin Elasticity Meter , to determine the effect of tissue sampling size. Dermal thickness was measured with 20 MHz ultrasound (Dermascan C) and tissue composition was inferred from anthropomorphic data. RESULTS: Skin thickness was significantly correlated with stiffness, energy absorption, and U(r)/U(f) for the shoulder. Body mass index (BMI) was significantly correlated with stiffness (negative correlation), energy absorption (positive), and skin thickness (negative) for the shoulder. Significant differences across body sites were observed. The calf was significantly different from the thigh and shoulders for all parameters (P<0.05, one-way anova). The calf had significantly lower laxity, laxity%, elastic deformation, energy absorption, elasticity, elasticity %, U(r), U(f), and U(r)/U(f) and significantly higher stiffness compared with the thighs and shoulders. sites. The thigh and shoulder sites were significantly different for all parameters except U(r)/U(f), elasticity %, laxity%, and stiffness. The dominant and non-dominant sides were significantly different. The dominant side (right for 90% of the subjects) had increased stiffness and decreased energy absorption (tissue softness, compliance) compared with the left side. A significant (P< or =0.02) negative relationship with age was seen for all biomechanical measures except stiffness at the shoulder. For the thigh and calf sites, significant negative correlations with age were found for elasticity %, U(r), and U(r)/U(f). Age and skin thickness were not correlated in this population. Skin thickness and age influenced the energy absorption at the shoulder site. The biological elasticity at the calf site could be predicted by age and BMI. The biological activity at the thigh site could be predicted by skin thickness and BMI. CONCLUSIONS: Significant regional variations in biomechanical properties and dominant side effects were observed. The biomechanical properties were significantly influenced by age. Certain properties varied with dermal thickness and tissue composition. The parameters were well correlated between the two instruments. The Cutometer, with its smaller aperture, was found to be more sensitive to age relationships.     

Ultrastructural observations on the effect of azelaic acid on normal human melanocytes and a human melanoma cell line in tissue culture

Azelaic acid has been shown clinically to have a cytotoxic effect on the abnormally active and malignant human melanocyte, but it has no apparent effect upon normal melanocytes. This difference in reactivity between normal and abnormal cells in vivo is further examined here in vitro. The disodium salt of azelaic acid (C₉2Na) was added to pure and mixed cultures of normal human melanocytes and to cultured human melanoma cells, at 10^?3M, 10^?2M, 5×10^?2M, and 10^?1 M for 1 and 6 h. Control cultures and cultures exposed to the same concentrations of the disodium salt of adipic acid (C₆2Na) were also examined. No damage to cells of any line was observed with diacids at 10^?3 M or 10^?2 M up to 6 h. At 5×10^?2M some mitochondria of melanoma cells appeared swollen. With C₆2Na at 10^?1 M for 1 and 6 h, minimal swelling of mitochondria was observed in some cells of all lines. Pure normal melanocytes and melanocytes of mixed cultures exhibited greater swelling of mitochondria with 10^?1M C₉2Na at 1 and 6 h, but the mitochondria of the malignant melanocytes were massively swollen with destruction of cristae. Plasma and nuclear membranes and membranes of rough endoplasmic reticulum were intact, but Golgi membranes exhibited vesiculation. These results provide further evidence that azelaic acid damages the human malignant melanocyte and that one of its targets is the mitochondrion. Damage to normal melanocytes, found here, may be due to the fact that, in culture, they are more active than in intact epidermis.     

The influence of age and sex on skin thickness, skin collagen and density

Forearm skin collagen, dermal thickness and collagen density were measured in a large number of normal subjects as a standard reference for future studies. Skin collagen decreased with age and was less in the females at all ages. There is a direct relationship between skin collagen and dermal thickness but variations in collagen density in disease limit the use of dermal thickness as a guide to changes in its collagen content.     

经皮水分丢失与健康人性别、年龄、解剖部位的关系

通过经皮水分丢失（TEWL）来评价皮肤的屏障功能，并研究经皮水分丢失与性别，年龄，解剖部位的关系，参加本次实验的为健康志愿者共104名，其中女性53名，男性51名，分为5个不同年龄组，用与计算机相连的蒸发测定仪Darmalab TEWL探头测量。结果发现不同年龄组男女性别之间TEWL值均无显著性差异（P＞0．05），新生儿组TEWL值明显高于其他各年龄组（P＜0．05），老年组TEWL值低于其他年龄组。在8个不同部位测量中，经统计得到TEWL值顺序为：手掌＞额部＞颊部＝手背＞小腿＝背部＞前臂＝胸部。说明TEWL与性别无明显相关性，而与年龄相关，以新生儿最高，老年人最低，在身体各部位的测量中，四肢末端和暴露部位经皮水分丢失较高。     

Patch testing in children,adults, and the elderly: influence of age and sex on sensitization patterns

Patch testing was done on 2776 consecutive patients (76.5% female) with a locally revised standard series of 34 contact allergens and the results analyzed for age- and gender-specific differences. At least one positive epicutaneous test reaction occurred in 48.9% of patients. Nickel (20.9%), ethylmercuric chloride (13.2%), thimerosal (11.8%), fragrance mix (9.3%), metallic mercury (8.9%), palladium (5.8%), balsam of Peru (3.8%), copper (3.7%), cobalt (3.3%), and chromium (2.3%) were the 10 most important sensitizers. The following tested allergens with sensitization rates of more than 1% were not part of the usual standard series: ethylmercuric chloride, metallic mercury, copper, propolis (1.3%), propylene glycol (1.0%). Reactions to nickel, cobalt, and palladium, but not to chromium, were significantly more abundant in females (p < 0.002, chi-squared test). The overall sensitization rate was highest in children less than 10 years old (62%) and decreased steadily, to be lowest among patients more than 70 years old (34.9%). The rate of positive reactions to nickel and thimerosal decreased with age, while fragrance mix and metallic mercury stayed at the same level through all age groups.     

Frictional properties of human skin: relation to age, sex and anatomical region, stratum corneum hydration and transepidermal water loss

Differences in the dynamic skin friction coefficients (mu) were investigated with respect to age, sex, and anatomical region. A total of 29 volunteers consisting of seven young females, seven old females, seven young males, and eight old males participated in the study. Measurements were obtained from II anatomical regions, namely, the forehead, upper arm, volar and dorsal forearm, postauricular, palm, abdomen, upper and lower back, thigh, and ankle. The friction data were compared with stratum corneum hydration and transepidermal water loss (TEWL). The dynamic friction coefficient did not vary significantly between age and sex groups but varied considerably among the anatomical regions of the body. The forehead and postauricular had the highest mu (0.34 +/- 0.02) while the abdomen had the lowest (0.12 +/- 0.01); the remaining regions had an average mu value of 0.21 +/- 0.01. Similarly, no sex differences were observed for TEWL and stratum corneum hydration. Capacitance was only significantly lower on the palms of the elderly. Regional differences showed a higher state of hydration on the forehead and postauricular as well as the upper arm, upper and lower back when compared with the volar forearm. TEWL was generally lower in the elderly on all anatomical regions except the postauricular and palm. A significant correlation was established between mu and capacitance for most regions. Between mu and TEWL significant correlation was observed only on the palm and thigh. These findings suggest that frictional properties of skin are dependent on more than water content or non-apparent sweating and the role of sebum secretion is suggested as one possible factor.     

Prostaglandin E1 in normal human skin: Methodological evaluation,topographical distribution and data related to sex and age

Summary A methodological evaluation of a radioimmunoassay technique for PGE₁ measurement was applied to normal human skin. The detection limit of the assay was 15 pg and recovery (mean±SD) was 90±6%. The mean value of PGE₁ (±SEM) in nine punch biopsy specimens (4 mm in diameter) from a piece of skin surgically removed from one person was 117±14 pg/mg dry weight. The individual variation of PGE₁ activity in biopsy specimens from the same topographical area in ten persons belonging to the same sex and age group was 33±4 pg/mg dry weight. The influence has been elucidated of temperature and tissue processing, local anaesthesia, sex, age and topographical distribution on endogenous PGE₁ activity.Supported by grant 512–8125, 512–15539 and 12–1690 from the Danish Medical Research Council     

Ultrastructural abnormalities in the dermal papillae of both lesional and clinically normal follicles from alopecia areata scalps

Summary Alopecia areala is a form of balding whose aetiology is uncertain. Although the dermal papilla in the hair bulb regulates the iollicie and may play a part in the pathogenesis of alopecia areata. Its ultrastructure has not been well described. As clinically normal, i.e. non-balding, follicles from alopecia areata scalps show abnormalities at the light microscope level, it would be expected that they should exhibit the earliest pathological changes involved in the dysfunction of the follicle. This study was designed to investigate the ultrastructure of normal human scalp follicular dermal pap-illae and to see if changes occurred in the ultrastructure of dermal papillae from either lesional or non-balding regions of alopecia areata. Normal dermal papillae contained well formed fibroblast-like cells with large, oval nuclei and well-developed endoplasmic reticulum; the cells were separated from each other by extracellular matrix containing small pieces of collagen and basal lamina-like material. Dermal papille from both clinically normal and lesional alopecia areata follicles were less well organized and the dermal papilla cells exhibited signs of cell injury and contained abnormal amounts of pigment; an increased concentration of fibrous material in the extracellular matrix and thickening of the dermal papilla-epithelial junction were also seen. Follicles from lesional areas showed more pronounced changes than clinically normal ones. Ultrastructural abnormalities in the dermal papillae of clinically normal scalp follicles support the study of these follicles as a prime research target. The changes detected suggest that dermal papilla cells in alopecia areata would be less able to synthesize regulatory factors and that these may have more difficulty crossing into the epithelial compartment. They are consistent with an early pathological role for the dermal papilla in alopecia areata, but do not distinguish whether this is a primary aetiological role or a secondary response to an insult elsewhere in the follicle.