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Biology of human skin transplanted to the nude mouse: I. Response to agents which modify epidermal proliferation.

To accept human skin transplanted to the congenitally athymic (nude) mouse as a system to study human skin and its physiologic and pathologic states, it must be demonstrated that skin so maintained retains its function as a biologic unit. We have found that responses of grafted human skin and nude mouse skin to various agents differ. This difference in response has been utilized to assess barrier function and proliferative capacity of human skin grafts. Human skin grafts undergo a proliferative response when 10 ng of the tumor promoter, 12-O-tetradecanoyl phorbol 13-acetate (TPA) is applied. Nudes do not respond to this dose. Increasing the dose to 100 ng of TPA evokes a response in both. However, only in the human skin grafts can this response be blocked with betamethasone valerate (BV). In that human skin grafts do not take on their hosts' responsiveness, and the response of domestic pig skin to these agents before and after grafting is identical, the conclusion is reached that human skin appears to retain its inherent biologic unit function. The data also demonstrate some of the potential of this system to study kinetics of the epidermis of human skin.

Animals↗

Topical corticosteroid allergy in an urban Australian centre.

The reported prevalence of allergic contact dermatitis from topical corticosteroids in clinical populations, in the period 1993-2002, varied from 0.55 to 5.98%. This study is a retrospective analysis of 1153 individuals undergoing routine patch testing in an Occupational Dermatology Clinic in Melbourne, Australia. We report a rate of 0.52% for positive patch test reactions to 5 corticosteroids. Corticosteroids tested were betamethasone-17-valerate, budesonide, Diprosone cream (betamethasone diproprionate 0.05%) (Essex-Pharma, a division of Schering-Plough Pty Ltd, Sydney, Australia), tixocortol-21-pivalate and triamcinolone acetonide. Population characteristics were described using the MOAHL (M = percentage of males tested; O = occupational; A = atopics; H = patients with hand eczema; L = patients with leg ulcers or stasis eczema) index. Prescribing patterns, rate of referral and rate of relevant positive patch test reactions were characterized for the region. These results were compared to the rates of corticosteroid allergy and patch testing methodologies from published international studies. It was noted that many high-sensitization potential corticosteroids were not available in our region. Although a low percentage of leg ulcers and stasis dermatitis may be associated with a lower rate of corticosteroid allergy, this association may be confounded by regional factors such as prescribing habits and the local availability of corticosteroids. We conclude that the low rate of topical corticosteroid contact allergy reported by our clinic is associated with regional availability and prescribing practices and the scarcity of stasis dermatitis and leg ulcers in our clinic population.

Administration, Topical↗

A study of systemic and topical effects of topical steroid application through the comparison of two application schedules.

We compared two application schedules of a topical steroid, BV (Betamethasone valerate). The schedules were once daily and twice a week on juvenile Wistar rats treated for four weeks. Steroid actions were assessed topically (skin thickness and epidermal Langerhans cell density) and systemically (body weight and internal organ wet weights). The results revealed some rather severe systemic effects caused by frequent steroid application. And we also found a uniquely weak BV effect with Langerhans cells compared to obvious systemic effects on rats. We came to the conclusion that frequent steroid application to the young should be avoided.

Administration, Topical↗

An improved method of assessing topical corticosteroid activity.

Topical application of ointment bases causes varying degrees of epidermal thickening in guinea-pigs. This is reproducible and can be accurately measured. Suppression of these changes was produced by addition of beta-methasone 17-valerate, fluocinolone acetonide, fluocinonide and hydrocortisone acetate. The inhibitory effect of these steroids was related to the type of corticosteriod, its concentration and the ointment base. The effect was still evident at extremely high dilutions of the steroids and could be measured at concentrations as low as 5 X 10(-5) % of fluocinonide in FAPG. This technique has the merits of being reproducible, sensitive and accurate. It should find a place among existing methods in assaying the efficacy of topical corticosteroids and in aiding in the selection of bases most suitable for formulation of these preparations.

Administration, Topical↗

The treatment of vitiligo with topical corticosteroids. Light and electronmicroscopic studies.

Twenty patients with lesions of vitiligo were treated either with 0.I% betamethasone valerate (BV) or with 0.05% clobetasol propionate (CP) creams and similar control areas with placebo preparations. After 3 months of treatment three of the ten patients treated with BV showed a significant amount of repigmentation only in the steroid-treated areas. Only one of the ten patients treated with CP showed partial repigmentation. Using both light and electron microscopy, histological studies of biopsies taken from control- and steroid-treated areas showed a marked repopulation with functional melanocytes in the repigmenting vitiliginous skin. The melanocytes in these steroid-treated repigmented areas were more dendritic and dopa-positive and, unlike melanocytes in the pigmented margins of untreated areas of vitiligo, contained many melanosomes of normal size, shape and melanization. It appears that both topical corticosteroids are of use for the treatment of selected patients with vitiligo and can induce repigmentation of the skin.

Administration, Topical↗

The effect of percutaneously absorbed steroids on hypothalamic--pituitary--adrenal function after intensive use in in-patients.

Hypothalamic-pituitary-adrenal (HPA) axis function has been monitored in adults and children who required intensive treatment of their skin disease with topical corticosteroid preparations while in hospital. Evidence of mild suppression of the HPA axis was seen in adults when the more potent topical steroids were used, but recovery of function was rapid when the intensive treatment ceased. In children suppression was still present in twelve of sixteen cases on the 2nd day after treatment with 0-1% betamethasone 17-valerate ointment had stopped, yet in nine cases treated in a comparable manner with 1% hydrocortisone acetate ointment, there was no evidence of impaired HPA axis function.

17-Hydroxycorticosteroids↗

Essential fatty acid deficient hairless mouse: the effects of topical agents on the epidermis.

Essential fatty acid deficient (EFA deficient) hairless mice were used to test the ability of topical agents to reduce the high epidermal DNA synthesis and acanthosis present in EFA deficiency. An established therapeutic agent known to be effective in psoriasis (betamethasone 17-valerate) significantly reduced the epidermal DNA synthesis, mitotic count and epidermal cell thickness. Linoleic acid reduced DNA synthesis and returned epidermal histology towards that of normal skin. Prostaglandin E2 analogue (15 (S)-15-methyl PGE2 methyl ester) reduced epidermal DNA synthesis both in treated and distant skin sites. Prostaglandin E2 was less effective than the analogue but slightly reduced epidermal DNA synthesis and thickness. Prostaglandin F2alpha had no obvious effect on the epidermis. Chlorambucil reduced epidermal DNA synthesis and thickness. Vitamin A acid increased DNA synthesis and epidermal cell thickness but reduced the hyperkeratosis of EFA deficiency. A rapid epidermal cell transit was found in EFA deficient mice, approximately three times faster than in normal mouse epidermis.

Administration, Topical↗

Utilization of epidermal phospholipase A2 inhibition to monitor topical steroid action.

The effect of several steroid creams on epidermal phospholipase A2 (PLA2) activity in symptomless psoriatic and normal epidermis was studied. The magnitude of PLA2 inhibition produced by the steroids was directly proportional to the initial level of the enzyme activity. This differential inhibition resulted in PLA2 activity approaching or attaining the normal range regardless of its initial level. Clobetasol propionate 0.05% (Dermovate) produced more enzyme inhibition than betamethasone valerate 0.1% (Betnovate) but there was no difference in inhibition between this latter steroid and clobetasone butyrate 0.05% (Eumovate). All were more inhibitory than hydrocortisone I% (Efcortelan).

Administration, Topical↗

Mechanism of anthralin inflammation. 2. Effect of pretreatment with glucocorticoids, anthralin and removal of stratum corneum.

The inflammatory dose-response to anthralin was measured in human skin 24 h after pretreatment with topical corticosteroids and anthralin, and 48 h after removal of the stratum corneum with adhesive tape. Anthralin inflammation was increased after 1% hydrocortisone application and decreased by 0.1% betamethasone valerate and 0.05% clobetasol propionate; although the difference between these effects was not significant, the difference between the effect of hydrocortisone and clobetasol propionate was. Anthralin inflammation was not significantly affected by pretreatment with anthralin and was reduced, although not significantly by removal of the stratum corneum. The finding that anthralin inflammation is not altered in skin in which aryl hydrocarbon hydroxylase (AHH) activity is increased and that anthralin inflammation may be altered in situations in which AHH activity is unchanged, excludes a direct relationship between anthralin inflammation and AHH activity.

Administration, Topical↗

Effects of topical corticosteroid therapy on Langerhans cell antigen presenting function in human skin.

We have investigated the mechanisms by which topical corticosteroids modulate cutaneous immune reactions in man. Volunteers applied clobetasone butyrate 0.05% (Eumovate; EV), betamethasone valerate 0.1% (Betnovate; BV), clobetasol propionate 0.05% (Dermovate; DV), and control vehicles twice daily to forearm skin for 7 days. Steroid therapy significantly decreased the number of HLA-DR/T6 (CD1a) positive Langerhans cells (LCs) per mm2 in suction blister-derived epidermal sheets, expressed as a mean percentage of controls, as follows: EV 69.2%; BV 67.3%; DV 37.8%. LC antigen presenting capacity was determined in the allogeneic and autologous epidermal cell-lymphocyte reactions. The LC-dependent allostimulatory capacity of epidermal cells, expressed as a mean percentage of controls, was also significantly reduced by steroid therapy: EV 45.1%; BV 41.9%; DV 23.4%. Following therapy with clobetasol propionate 0.05%, the capacity of epidermal cells to present tetanus toxoid to, and to augment concanavalin A mediated lymphocyte stimulation of, autologous lymphocytes was reduced to 33.6% and 19.7% respectively of controls. Depression of epidermal cell allostimulatory capacity was not the result of a steroid-induced decrease in the production of epidermal cell-derived thymocyte activating factor (ETAF)/interleukin 1 by keratinocytes, since it could not be reversed by addition of exogenous interleukin 1. Indomethacin, added to block any potential prostaglandin synthesis during the culture period, did not restore the allostimulatory capacity of epidermal cells from steroid-treated sites. Addition of epidermal cells from DV-treated sites depressed the capacity of control epidermal cells to stimulate lymphocytes in the allogeneic epidermal-lymphocyte reaction. Our results demonstrate that the anti-inflammatory action of topical corticosteroids in man is associated not only with a reduction in the number of HLA-DR/T6 positive LCs, but also with a marked decrease in Langerhans cell-dependent T lymphocyte activation. The effects of the different steroids on both of these parameters correlated with their potency as determined in the standard occlusive vasoconstrictor assay. Topical corticosteroids are widely used for the treatment of inflammatory skin disorders, and inhibit not only the elicitation phase, but also the induction phase, of allergic contact dermatitis reactions.(ABSTRACT TRUNCATED AT 400 WORDS)

Antigen-Presenting Cells↗

Sensitivity of different areas of the flexor aspect of the human forearm to corticosteroid-induced skin blanching.

The intensity of corticosteroid-induced blanching has been found to vary at different areas of the flexor aspect of the human forearm. A retrospective analysis of 38,880 observations of skin blanching in 56 volunteers was conducted to assess the sensitivity of forearm skin to betamethasone 17-valerate. The mid-forearm appears to be more sensitive to the blanching response than do the areas close to the wrist or elbow. These results indicate that each preparation under evaluation should be applied to several sites along the forearm when using the human skin blanching assay in order to obtain an accurate comparative assessment of corticosteroid release from topical delivery vehicles.

Administration, Topical↗

The cutaneous corticosteroid vasoconstriction assay: a reflectance spectroscopic and laser-Doppler flowmetric study.

Cutaneous vasoconstriction induced by topical corticosteroids was investigated using non-invasive bioengineering techniques. Corticosteroids of different potency in alcoholic solution were applied topically, under occlusion, and cutaneous blanching was investigated using visual scoring, reflectance spectroscopy (RS) and laser-Doppler flowmetry (LDF). The RS technique allowed separation of cutaneous haemoglobin content into arterial oxygenated (OH) and venous deoxygenated haemoglobin (DOH) components. Application of alcohol decreased total haemoglobin by 10%, with a corresponding 8% increase in blood flow (BF). Clobetasol propionate was the most potent vasoconstrictor, inducing significant visible blanching and decreasing DOH (30%), OH (33%) and BF (18%) (P < 0.01). Fluocinolone acetonide, betamethasone-17-valerate and dexamethasone also caused visible blanching (P < 0.01). There was no significant decrease in BF, but reflectance spectroscopy showed a decrease in DOH (P < 0.01). Tixocortol, CMJ and hydrocortisone acetate did not produce significant blanching, although DOH was decreased compared with the alcohol control. Measured by reflectance spectroscopy, corticosteroid-induced blanching was predominantly venoconstriction and only the most potent corticosteroid caused a significant decrease in OH and blood flow. This may explain why previous attempts to improve cutaneous vasoconstriction assays using laser-Doppler flowmetry have been unsuccessful.

Adrenal Cortex Hormones↗

A time correlation study between reflectance spectroscopic cutaneous vasoconstriction and plasma corticosteroid concentration.

Although cutaneous vasoconstriction assays are used as a primary screen for ranking the in vivo efficacy of new corticosteroids and in vivo human drug delivery studies, little is known about the relationship between the blanching reaction and corticosteroid tissue or plasma concentrations. We measured cutaneous vascular reactions in five volunteers, using an improved reflectance spectroscopic method, and a sensitive radioimmunoassay technique was employed to measure plasma betamethasone concentrations. Using a specially developed betamethasone-17-valerate patch prepared in BIO-PSA, constant corticosteroid release was ensured, and correlations between cutaneous blanching and plasma corticosteroid concentrations were calculated. Maximal skin blanching was documented 12 h post-application, whereas plasma corticosteroid concentrations peaked later, at 32 h post-application, when a paradoxical telangiectatic vasodilatation occurred. At 72 h post-application, when the plasma corticosteroid concentration was still above the 12 h level, this paradoxical vasodilatation was maximal. The corticosteroid-induced vascular reactions were mainly due to arterial haemoglobin (Oxy Haem), and both vasoconstriction and vasodilatation were related to changes in Oxy Haem. Our results suggest a dual, probably both time and concentration related, interaction between corticosteroids and dermal vessels in which lower concentrations at 6-12 h exposure caused vasoconstriction, but as the exposure time increased (> or = 24 h) paradoxical vasodilatation was induced, although plasma corticosteroid concentrations were still rising.

Adult↗

The effect of topical steroid application on natural killer cell activity.

Peripheral blood natural killer (NK) cell activity of a group of 10 healthy non-atopic volunteers was reduced following the topical application of 15 g of 0.1% betamethasone valerate ointment to the skin nightly for 1 week. In contrast, no such effect was observed when the inactive base of the steroid ointment was used. NK cell activity dropped significantly by day 7 (P less than 0.05) and then recovered, although NK cell activity at day 22 was still lower than that observed at the start of the experiment. These findings suggest that topically applied steroid is absorbed in sufficient amounts to cause a systemic effect on NK cell function. This may have implications in a number of dermatological disorders, including atopic dermatitis, where topical steroids form the mainstay of treatment.

Administration, Topical↗

Ranking of the antipsoriatic effect of various topical corticosteroids applied under a hydrocolloid dressing--skin-thickness, blood-flow and colour measurements compared to clinical assessments.

In 10 patients with chronic plaque type psoriasis one or two plaques affected equally with psoriasis were chosen for study. Five punched out rings of a hydrocolloid dressing were applied to the psoriasis plaque(s). In each circular test area 20 mg of one of the following creams was applied: base, 1% hydrocortisone (DAK), 0.1% triamcinolone acetonide (Kenalogue), 0.1% betamethasone-17-valerate cream (Betnovate), and 0.05% clobetasol proprionate cream (Dermovate). The areas were occluded with a thin film of transparent hydrocolloid dressing (Comfeel Transparent Dressing), for 1 week. Non-invasive measurements (ultrasound skin thickness, laser-Doppler flowmetry, colorimetry) were performed before and after treatment. Therapeutic response was evaluated blindly by clinical score. The measurements showed a decline in blood flow, a decrease in skin thickness, and normalization of colour approaching that of normal skin, the more potent the corticosteroid used. The clinical scores showed the same: the more potent a corticosteroid used, the closer to the score of normal skin. Data on variability and applications of the methods are presented. The study concludes that potent corticosteroids occluded with a hydrocolloid dressing can clear psoriasis in 1 week. Short-course corticosteroid therapy appears harmless and relevant for clinical dermatology.

Administration, Topical↗

The stability and blanching efficiency of some Betnelan-V cream dilutions.

In this study Betnelan-V cream was diluted with Beeler's basis and Cold cream. Cold Cream reduced the skin-blanching activity of the original cream more than Beeler's base. The stability of betamethasone-17-valerate was better in a Beeler's basis dilution than in a Cold cream dilution.

Administration, Topical↗

Multicenter trial of fluocinonide in an emollient cream base.

Working independently, eight investigators evaluated the clinical effectiveness of fluocinonide 0.05% in a three-week double-blind paired comparison trial of 240 patients. Half the investigators compared fluocinonide emollient cream with betamethasone valerate 0.1% cream in the treatment of atopic dermatitis, while the remaining four compared the study drug with triamcinolone acetonide 0.1% cream in the treatment of psoriasis. Statistical analysis showed clinical responses favored fluocinonide p = 0.021 (Wilcoxon Signed Rank) for the atopic dermatitis group and p = 0.001 (Wilcoxon Signed Rank) for the psoriasis group.

Adolescent↗