Increase In Epidermal Thickness Research Articles

Modulation of ultraviolet light-induced epidermal damage: beneficial effects of tocopherol.

Oxygen free radicals have been shown to result from and mediate deleterious effects of ultraviolet radiation on the skin. The purpose of this study was to determine if topical DL-alpha-tocopherol (vitamin E) could reduce ultraviolet-induced damage to the epidermis. Twenty mice were treated with either ethanol or a 1:1 mixture of tocopherol and ethanol. Treatments consisted of once-daily 0.1-ml topical applications for 1 week, followed by irradiation with 0.30 mW/cm2 of ultraviolet B irradiation. A statistically significant decrease in Schiff base formation was noted between tocopherol-treated animals and their controls. Histologic study revealed a statistically significant increase in epidermal thickness in tocopherol-treated skin versus controls or vehicle alone. The thicker epidermis was accompanied by the presence of parakeratosis, implicating increased proliferation as the cause of the increasing thickness. The number of sunburn cells was decreased by tocopherol treatment. Tocopherol protection from ultraviolet irradiation may have been due to both direct protection from free radicals and indirect protection by means of increased epidermal thickness. The demonstration of beneficial effects of tocopherol administration suggests that further studies in clinically relevant models to define optimal dosage, frequency of administration, vehicle, and quantitation of the possible protective effects afforded to Langerhans cells may be useful.

Topical vitamin D3 derivatives impair the epidermal permeability barrier in normal mouse skin.

Calcipotriol, 1,25(OH)2D3 and 1,24(OH)2D3 are potent drugs for the treatment of psoriasis. It has recently been published that these compounds induce epidermal hyperproliferation in hairless mouse skin. The aim of our study was to examine the effect of vitamin D3 derivatives on epidermal growth, keratinization and permeability barrier function in vivo. Calcipotriol, 1,25(OH)2D3 and 1,24(OH)2D3 in isopropanol or in an ointment formula were applied to normal hairless mouse skin. Transepidermal water loss (TEWL), a marker of cutaneous barrier function, and epidermal proliferation were determined at different time points 0-264 h after treatment. In addition, light and electron microscopy studies were performed. A single treatment in solution led to a transient (2- to 3-fold) increase in TEWL after application of calcipotriol or 1,25(OH)2D3 and to a 3- to 6-fold increase in epidermal proliferation after application of each of the compounds. Repeated applications also resulted in an up to 3-fold increase in TEWL which persisted for 3 days after the end of the treatment. By light microscopy an increase in epidermal thickness was observed. There was no sign of inflammation. Electron microscopy studies showed the formation of a transitional cell zone as a sign of a premature keratinization. These results demonstrate that in normal mouse skin vitamin D3 and its analogues disrupt the epidermal permeability barrier by induction of epidermal proliferation and premature keratinization but without morphological signs of inflammation.

Topical Retinaldehyde Increases Skin Content of Retinoic Acid and Exerts Biologic Activity in Mouse Skin

Retinaldehyde, a natural metabolite of beta-carotene and retinol, has been proposed recently for topical use in humans. Because retinaldehyde does not bind to retinoid nuclear receptors, its biologic activity should result from enzymatic transformation by epidermal keratinocytes into ligands for these receptors, such as all-trans retinoic acid and 9-cis-retinoic acid. In this study, we analyzed by high performance liquid chromatography the type and amounts of tissue retinoids as well as several biologic activities resulting from topical application of either retinaldehyde or all-trans retinoic acid on mouse tail skin. Biologic activities of all-trans retinoic acid and retinaldehyde were qualitatively identical in metaplastic parameters (induction of orthokeratosis, reduction of keratin 65-kDa mRNA, increase in filaggrin and loricrin mRNAs) and hyperplastic parameters (increase in epidermal thickness, increase in bromodeoxyuridine (BrdU)-positive cells, increase in keratin 50-kDa mRNA, and reduction in keratin 70-kDa mRNA). Some quantitative differences, not all in favor of all-trans retinoic acid, were found in several indices. Cellular retinoic acid-binding protein II and cellular retinol-binding protein I mRNAs were increased by both topical retinaldehyde and all-trans retinoic acid. Whereas all-trans retinoic acid, 9-cis-retinoic acid, and 13-cis-retinoic acid were not detectable (limit 5 ng/g) in vehicle-treated skin, 0.05% retinaldehyde-treated skin contained 13 +/- 6.9 ng/g wet tissue of all-trans retinoic acid (mean +/- SD), 12.6 +/- 5.9 ng/g 13-cis-retinoic acid, and no 9-cis-retinoic acid. In contrast, 9-cis-retinoic acid was detectable in 0.05% of all-trans retinoic acid-treated skin, which also contained 25-fold more all-trans retinoic acid and 5-fold more 13-cis-retinoic acid than retinaldehyde-treated skin. Our results show that topical retinaldehyde is transformed in vivo into all-trans retinoic acid by mouse epidermis. The small amounts of ligand for retinoic acid nuclear receptors thus produced are sufficient to induce biologic effects similar to those resulting from the topical application of the ligand itself in much higher concentration.

Changes in leaf expansion and epidermal screening effectiveness in Liquidambar styraciflua and Pinus taeda in response to UV‐B radiation

Absorption or screening of ultraviolet‐B (UV‐B) radiation by the epidermis may be an important protective method by which plants avoid damage upon exposure to potentially harmful UV‐B radiation. In the present study we examined the relationships among epidermal screening effectiveness, concentration of UV‐absorbing compounds, epidermal anatomy and growth responses in seedlings of loblolly pine (Pinus taeda L.) and sweetgum (Liquidambar styraciflua L.). Seedlings of each species were grown in a greenhouse at the University of Maryland under either no UV‐B radiation or daily supplemental UV‐B radiation levels of 4, 8 or 11 kJ m−2 of biologically effective UV‐B (UV‐BBE) radiation. Loblolly pine seedlings were subsequently grown in the field under either ambient or supplemental levels of UV‐B radiation. At the conclusion of the growing season, measurements of epidermal UV‐B screening effectiveness were made with a fiber‐optic microprobe. In loblolly pine, less than 0.5% of incident UV‐B radiation was transmitted through the epidermis of fascicle needles and about 1% was transmitted in primary needles. In contrast, epidermal transmittance in sweetgum ranged from about 20% in leaves not preconditioned to UV‐B exposure, to about 10% in leaves grown under UV‐B radiation. The concentration of UV‐absorbing compounds was unaffected by UV‐B exposure, but generally increased with leaf age. Increases in epidermal thickness were observed in response to UV‐B treatment in loblolly pine, and this accounted for over half of the variability in UV‐B screening effectiveness. In spite of the low levels of UV‐B penetration into the mesophyll, delays in leaf development (both species) and final needle size (loblolly pine) were observed. Seedling biomass was reduced by supplemental UV‐B radiation in loblolly pine. We hypothesize that the UV‐induced growth reductions were manifested by changes in either epidermal anatomy or epidermal secondary chemistry that might negatively impact cell elongation.

Bacterial lipopolysaccharide (LPS) and interleukin 1 (IL-1) exert multiple physiological effects in the tilapia Oreochromis mossambicus (Teleostei).

To gain insight in immuno-endocrine communication in teleosts the physiological effects of interleukin 1 and bacterial lipopolysaccharide in teleosts were investigated. Tilapia (Oreochromis mossambicus) were treated with murine interleukin 1 and E. coli lipopolysaccharide in vivo, and lipopolysaccharide was administered to pituitary lobes and head kidneys in vitro. The integument of the fish appeared to be a sensitive target for the preparations tested, since proliferation of chloride cells and of epidermal mucous cells was observed as well as an increase in epidermal thickness. These effects may relate to an acute phase-like reaction caused by the treatments. Lipopolysaccharide administration furthermore resulted in an increase in plasma free fatty acids levels. Lipopolysaccharide, but not interleukin 1, stimulated the interrenal axis of the fish, as judged by the increase in cortisol production measured in superfusion of head kidneys. In addition to these in vivo effects, lipopolysaccharide also displayed several effects in vitro. Pituitary adrenocorticotropic hormone, as well as alpha-melanocyte stimulating hormone, release was inhibited, and the head kidney responsiveness to adrenocorticotropic hormone was inhibited after pretreatment of the tissue with the E. coli product. This latter effect coincided with the release of an unidentified alpha-melanocyte stimulating hormone immunoreactive fraction by the head kidneys which could be stimulated by lipopolysaccharide. The data strongly support the notion that the immune system is involved in adaptive regulations in teleosts, and that immunoendocrine interactions are phylogenetically old mechanisms.

Skin ultrastructure in relation to prolactin and MSH function in rainbow trout (Oncorhynchus mykiss) exposed to environmental acidification

The present study describes the effects of 14 days exposure to acidified (pH 4.0) soft water in the absence of aluminium, on the ultrastructure of the skin in rainbow trout (Oncorhynchus mykiss). Compared to control fish, there was a moderate increase in the incidence of necrosis in the filament cells of the fish exposed to pH 4.0, but since the integrity of the tissue appeared to be maintained, most of the ultrastructural changes observed may be considered to be adaptive. There was an increase in epidermal thickness, a higher frequency of electrondense vesicles in filament cells, an increase in the undulation of the basal lamina, and the penetration of the epidermis by cytoplasmatic processes of melanocytes in acid-exposed specimens. An infiltration of leucocytes into the epidermis, and the appearance of serous mucous cells, was also evident. Whether these events were under the control of prolactin and/or α-MSH, was also investigated, but no indication for activation or inhibition of either prolactin or α-MSH producing cells was obtained. Since a previous study (Balm and Pottinger 1993) had demonstrated that plasma cortisol levels were also identical in control and low pH treated trout throughout a 14 day experimental period, it is concluded that under conditions of environmental acidification, the integument autonomically maintains the adjustments necessary for successful acclimation, presumably via paracrine regulatory circuits.

Variations in epidermal thickness in expanded human breast skin.

Tissue expansion generates metabolic and morphological changes in the expanded tissues. The alteration of epidermal and dermal thickness has been investigated in many animal studies, but few human studies have been published. We have studied 11 women whose breasts were reconstructed with tissue expansion after mastectomy. The expanded breast skin was biopsied on three occasions: before expansion, at the time of maximal expansion, and six months after the completed reconstruction. Specimens were prepared for light microscopy and epidermal thickness was assessed. There was a significant increase in epidermal thickness (p < 0.05) at the time of maximal expansion. The values before and at six months after reconstruction were not significantly different from each other. We conclude that the epidermis increases in thickness as a result of expansion and returns to normal thickness six months after cessation of expansion.

Skin ultrastructure in relation to prolactin and MSH function in rainbow trout ( Oncorhynchus mykiss ) exposed to environmental acidification

The present study describes the effects of 14 days exposure to acidified (pH 4.0) soft water in the absence of aluminium, on the ultrastructure of the skin in rainbow trout (Oncorhynchus mykiss). Compared to control fish, there was a moderate increase in the incidence of necrosis in the filament cells of the fish exposed to pH 4.0, but since the integrity of the tissue appeared to be maintained, most of the ultrastructural changes observed may be considered to be adaptive. There was an increase in epidermal thickness, a higher frequency of electrondense vesicles in filament cells, an increase in the undulation of the basal lamina, and the penetration of the epidermis by cytoplasmatic processes of melanocytes in acid-exposed specimens. An infiltration of leucocytes into the epidermis, and the appearance of serous mucous cells, was also evident. Whether these events were under the control of prolactin and/or α-MSH, was also investigated, but no indication for activation or inhibition of either prolactin or α-MSH producing cells was obtained. Since a previous study (Balm and Pottinger 1993) had demonstrated that plasma cortisol levels were also identical in control and low pH treated trout throughout a 14 day experimental period, it is concluded that under conditions of environmental acidification, the integument autonomically maintains the adjustments necessary for successful acclimation, presumably via paracrine regulatory circuits.

Topical Retinaldehyde on Human Skin: Biologic Effects and Tolerance

The present study was designed to explore if *etinaldehyde, a natural metabolite of vitamin A, has any biologic activity and is tolerated by human skin. Biologic activity was shown by the induction of cellular retinoic acid-binding protein type 2 (CRABP 2) mRNA and protein; the rank order for CRABP-2 increase was retinoic acid > retinaldehyde > 9 cis retinoic acid > retinol > beta carotene. In volunteers treated 1-3 months with 0.5, 0.1, and 0.05% retinaldehyde, there was a dose-dependent and significant increase in epidermal thickness, keratin 14 immunoreactivity, and Ki67-positive cells. The area of distribution of involucrin, transglutaminase, and filaggrin immunoreactivity was also increased in a dose-dependent manner, and keratin 4 immunoreactivity was induced in the upper epidermis. In pilot clinical tolerance studies, 229 patients received topical retinaldehyde at different concentrations; the 1% preparation was tolerated by up to 70% of the treated subjects; tolerance of the 0.5% preparation was slightly better, whereas both 0.1 and 0.05% preparations applied on facial skin were well tolerated and allowed prolonged use (up to 3 years) in patients with inflammatory dermatoses. These findings indicate that topical retinaldehyde has biologic activity and is well tolerated on human skin.

UV-induced alterations in skin and lymphocytes during a one-week holiday in the Canary Islands in May.

The effect of solar exposure during a one-week sunbathing vacation in May at 29 degrees N latitude was investigated in 22 volunteers. The following end-points were measured before and after the holiday: skin reflectance at 507 nm, transmission of radiation from 289 nm to 448 nm through the epidermis, epidermal thickness, minimal erythema dose (MED), total white cell, neutrophil, lymphocyte, and lymphocyte subpopulation counts in blood, spontaneous DNA synthesis, DNA strand breaks and sister-chromatid exchange in lymphocytes, and the UVC tolerance of lymphocytes. There was a statistically significant decrease in skin reflectance (p < 0.001) and epidermal transmission (p < 0.01) after the holiday, reflecting increased pigmentation and epidermal thickening. There was a statistically significant increase in epidermal thickness (p < 0.001), MED (p < 0.05), spontaneous DNA synthesis (p < 0.01) and DNA strand breaks in lymphocytes (p < 0.02) after the holiday. The other end-points were not significantly changed. We report that changes in skin pigmentation and epidermal thickness occur after one week of UV exposure. It was also observed that a one-week UV exposure increased both spontaneous DNA repair and the amount of DNA strand breaks in the lymphocytes of the volunteers, while no changes in T-cell subsets were detected.

Targeting expression of keratinocyte growth factor to keratinocytes elicits striking changes in epithelial differentiation in transgenic mice.

Keratinocyte growth factor (KGF) is a member of the fibroblast growth factor (FGF) family. Synthesized by cells of the dermal component of skin, KGF's potent mitogenic activity is on the epidermal component, which harbors the receptors for this factor. To explore the possible role of KGF in mesenchymal-epithelial interactions in skin, we used a human keratin 14 promoter to target expression of human KGF cDNA to the stratified squamous epithelia of transgenic mice. Mice expressing KGF in their epidermis typically appeared frail and weak, and often had grossly wrinkled skin. These mice exhibited a gross increase in epidermal thickness accompanied by alterations in epidermal growth and differentiation. Most remarkably, animals displayed several striking and unexpected changes, including a marked suppression of hair follicle morphogenesis and suppression of adipogenesis. With age, some animals developed gross transformations in the tongue epithelium and in epidermis. In addition, they exhibited elevated salivation and their salivary glands showed signs of altered differentiation. Collectively, our findings provide new and important insights into the roles of KGF, implicating this potent growth factor in eliciting global effects not only on growth, but also on development and differentiation, of skin and other tissues. In particular, KGF seems to interfere with signalling of some mesenchymal-epithelial interactions.

Quantitative Determination of EGF-R During Epidermal Wound Healing

Little is known about the intrinsic regulation of growth factors of cytokines during the normal epidermal wound-healing processes in skin. A simplified model of wounding (tape stripping to remove the stratum corneum) was used to study the role of epidermal growth factor receptors (EGF-R) in this process. Although the dynamics of EGF-R in epidermal wound healing have not been determined, the immunoreactive EGF-R that are present presumably play an active role. Prior studies show that 1) EGF-R are present in increased numbers in proliferative skin diseases; 2) a hypertrophic epidermis, closely resembling normal wound healing, is induced in mouse skin by EGF injections; and 3) exogenous topical EGF potentiates wound healing. The number of immunoreactive receptors as measured by an enzyme-linked immunosorbent assay (ELISA) and histologic methods increased prior to an increase in epidermal thickness, total protein, and DNA content. This early increase in the levels of EGF-R was followed by a sharp decline in EGF-R and subsequent decline in epidermal thickness (hypertrophy), total protein, and DNA levels. Alterations in the temporal sequence in these parameters indicate that the EGF-R-mediated signaling systems play an active role in epidermal wound repair.

Effects of the Phthalazinone Azelastine on Epidermal Metabolism after Mechanical Skin Irritation

In female NMRI mice, the phthalazinone azelastine was administered orally once daily over 7 days. The drug influenced the epidermal thymidine triphosphate and amino acid incorporation rates at doses between 1 and 5 mg/kg. In control mice, an epidermal hyperproliferation induced by abrasion of superficial epidermal layers was characterized by enhanced prostaglandin and leukotriene concentrations in epidermal homogenate, an increase in thymidine triphosphate and amino acid incorporation and an increase in epidermal thickness. In mice treated with 1 mg/kg azelastine HCl, this epidermal reaction was changed. Compared to controls, the increase in leukotriene concentration was diminished, and that of prostaglandins was enhanced. The incorporation of thymidine triphosphate and of amino acids as well as the epidermal thickness and the ratio cell count/epidermal thickness were increased in irritated skin of azelastine-treated mice. In conclusion, azelastine influences the epidermal metabolism in irritated and unirritated skin. Therefore, a beneficial role of this phthalazinone in the treatment of psoriasis and related skin disorders seems to be possible.

Aged versus young skin before and after transplantation onto nude mice

The behaviour of aged skin transplanted onto nude mice was investigated to determine whether the skin maintains its histological features. Split-thickness skin grafts obtained from the unexposed skin on the thighs of healthy aged and young volunteers were grafted onto nude mice. A significant difference between the mean thickness of young versus aged epidermis was noted before transplantation (P less than 0.001). The epidermis of aged and young skin showed an increase in thickness following engraftment with a mean increase in epidermal thickness of 18.8% in the young (P less than 0.01) and 142.5% in aged skin (P less than 0.001). The number of blood vessels in the aged skin was significantly lower than in the young skin, but a remarkable increase was found post-transplantation. These findings indicate that part of the typical histological changes of unexposed aged skin are reversible.

Quantification of Epidermal Histological Changes Induced by Topical Retinoids and CD271 in the Rhino Mouse Model Using a Standardized Image Analysis Technique

The rhino mouse has been used as an experimental model to screen topically active comedolytic agents. Adult rhino mice were treated on the back once daily for 5 consecutive days per week during 3 weeks. Skin histological preparations were analyzed by image analysis techniques to quantify the number of epidermal comedones, comedo profile and epidermal thickness. Using both a negative (treated with acetone) and a positive (treated with Aberel gel 0.025%) control group of animals in all experiments conducted over a period of about 3 years, we defined the upper and lower limit of acceptability of the results. Topical treatment with an acetone solution of all-trans retinoic acid (0.01, 0.03, 0.1%) and 13-cis-retinoic acid (0.1%) induced comedolysis and a marked increase in epidermal thickness. Commercial preparations of all-trans retinoic acid (Aberel lotion, gel and cream, Retin A cream, Retacnyl cream) presented a similar comedolytic activity. However, the epidermal thickening was higher with Retin A and weaker with Retacnyl. CD271, a new modulator of cell differentiation, applied either in acetone solution (0.01, 0.1%) or in lotion, gel or cream formulations (0.1%) also demonstrated a marked activity (i.e. comedolysis and epidermal thickening). These data confirm that the rhino mouse model can be used to assay drugs applied either in solvent or in topical formulations. Activity in this model compares favorably with published clinical observations in the treatment of acne.

Morphometric and statistical analyses describing the in utero growth of human epidermis.

Epidermal development of human embryonic and fetal skin from the lower limb was studied using morphometric and statistical methods. Epidermal growth, as defined by an increase in epidermal thickness and the number of cell layers, occurred in three distinct stages during the first and second trimesters. The first growth spurt occurred between 5 and 13 weeks estimated gestational age (EGA) and was followed by a plateau phase with little change in epidermal thickness from 14 to 21 weeks, after which the epidermis began to increase in height again. The periderm reached its maximal height by approximately 13 weeks EGA, and by 25 weeks was shed into the amniotic fluid. Thus, within a five-month period (5 to 25 weeks EGA) the epidermis changed from a single cell layer less than 10 micron thick to a 10 to 12-cell layer, keratinized epithelium greater than 60 micron thick. In contrast, epidermis from adult lower limb consisted of about 25 cell layers and was almost 75 micron in thickness. The age-related differences in epidermal thickness probably reflect changes in cell size and shape more than changes in the directional movement (apically vs. laterally) of proliferating keratinocytes, because the addition of cell layers throughout development was relatively constant. During the plateau phase, when there is a rapid increase in fetal growth rate, the suprabasal keratinocytes become more flattened, thereby allowing for the addition of new cell layers while maintaining a relatively constant epidermal thickness.(ABSTRACT TRUNCATED AT 250 WORDS)

Sn-1,2-didecanoylglycerol effectively induces epidermal ornithine decarboxylase but only weak hyperplasia in mouse skin.

The abilities of sn-1,2-didecanoylglycerol (sn-1,2-DDG) to induce epidermal ornithine decarboxylase (ODC) activity and epidermal hyperplasia were tested using SENCAR, DBA/2 and C57BL/6 mice. Following a single application of 5000 nmol of sn-1,2-DDG, ODC activity reached a maximum at 4 h after treatment with a peak activity of 6.03 nmol CO2/mg protein/60 min in C57BL/6, 1.50 in SENCAR and 0.73 in DBA/2, respectively. The time course and magnitude for induction of ODC activity after multiple treatments was very similar to that after a single application in these three mouse lines. Interestingly, the induced ODC activity in C57BL/6 was always higher than that in SENCAR and DBA/2 mouse epidermis regardless of the treatment protocol. Induction of hyperplasia and dark basal keratinocytes (DCs) and changes in the labeling index (LI) of basal keratinocytes in DBA/2 and C57BL/6 mice following treatment with sn-1,2-DDG were investigated. Multiple treatments (twice weekly for 2 weeks) of 5000 nmol sn-1,2-DDG did not induce substantial increases in epidermal thickness or DCs 24 or 48 h after the last treatment. In contrast, TPA induced a marked increase in epidermal thickness in DBA/2 rather than C57BL/6 and a considerably higher induction of DCs in DBA/2 (37.3 +/- 2.2%) than in C57BL/6 (9.6 +/- 2.5%) 48 h after the last treatment. The LIs after topical application of sn-1,2-DDG were elevated at 24 h, but returned to basal levels by 48 h in both strains, whereas TPA treatment significantly elevated the LI in both strains at 48 h after the last application. In addition, the effects of various doses and frequencies of application of sn-1,2-DDG were investigated using SENCAR mice. High doses (20,000 nmol) or more frequent applications (5000 nmol once daily for 7 days) of sn-1,2-DDG still produced only weak hyperplasia. These results suggest that the induction of epidermal ODC activity can be dissociated from the induction of epidermal hyperplasia and may provide an explanation for the lack of complete promoting activity presently observed with membrane permeable diacylglycerol derivatives.

Role of platelet-derived growth factor in wound healing: synergistic effects with other growth factors.

Platelet-derived growth factor (PDGF) in vitro stimulates DNA synthesis and chemotaxis of fibroblasts and smooth muscle cells and stimulates collagen, glycosaminoglycan, and collagenase production by fibroblasts. These in vitro properties suggest that PDGF, delivered by platelets to the site of injury in vivo, may play an important role in the initiation of the wound repair process. Studies presented here show that the addition of pure PDGF to a wound site involving the epidermis and dermis has little effect on the morphology or biochemistry of the healing wound. In contrast, the addition of partially purified PDGF resulted in significant dose-dependent increases in the width of the newly synthesized connective tissue and epidermal layers. Autoradiography using [3H]thymidine revealed increased numbers of labeled cells in the new connective tissue and epithelial layers. Furthermore, addition of partially purified PDGF resulted in significant increases in the rate of protein and DNA synthesis and the total content of these components in biopsies taken from the wound site. Similar effects were obtained when insulin-like growth factor I was added in combination with pure PDGF. This combination of factors caused a 2.4-fold increase in the width of the newly formed connective tissue layer and a 95% increase in epidermal thickness compared with controls. Insulin-like growth factor I alone caused no significant morphologic changes. Epidermal growth factor alone or in combination with PDGF resulted in a thickening only of the epidermis. These results indicate that the synergistic actions of other factors with PDGF are important in the modulation of the wound healing process.

Induction of ornithine decarboxylase activity in hairless rat epidermis as a pharmacological model: validation of the animal model.

We use a mutant hairless Sprague Dawley rat to evaluate the capacity of retinoids to inhibit the epidermal ornithine decarboxylase activity induced by sellotape stripping. In order to minimize the variability introduced by the animals in our model we decided to validate the hairless rats used. A number of animal parameters were examined using a single lot of 50 males and 50 females aged from 4 to 11 weeks and acclimatized to laboratory conditions. The body weight growth curves were established. Nude animals present two periods of hair growth, the first at 6-7 weeks and the second at about 10-11 weeks. Hair development is more pronounced in males. No histological change was observed in the stratum corneum but an increase in epidermal thickness was noted in males aged 9 weeks. Removal of the stratum corneum by sellotape stripping was more effective and reproducible in the females, as determined histologically. Sellotape-stripping induction of ornithine decarboxylase in the epidermis was higher in rats aged 5-6 weeks and reached a plateau in animals aged 6-12 weeks. Individual variations obtained were lower in females (about 5%-10% in females and 10%-20% in males). The present research suggests that female rats aged about 8 weeks provide maximum reproducibility of response and ease of use.

Nuclear changes and morphology of the epidermis in the hibernating frog

Cytochemical changes of chromatin and DNA in frog epidermal cells were correlated with some morphological features to investigate the skin physiology during hibernation in comparison with the active period. The epidermal cells of hibernating frogs showed less condensed chromatin in all the layers; a greater loss of DNA was found during the transition from the middle to the superficial layer. In the germinative layer, a lesser frequency of hyperdiploid cells and a remarkably low amount of mitoses were detected; this is accompanied by the increase of epidermal thickness and the presence of two layers of cornified cells. The slowing of tissue differentiation and cell renewal kinetics during hibernation can be related to lowered activity of the frog skin. Further, the smaller intercellular spaces as well as the scarcity of puffed ER and vacuoles may be indicative of a lower ion transport in epidermal cells during hibernation.