Effects of site and menstrual cycle on barrier function and stratum corneum water-holding capacity

Background/aims: Areas of the skin with similar anatomical structure may have different functional behaviour. In vivo barrier function and stratum corneum water-holding capacity during the menstrual cycle on two sites of the volar forearm (upper and lower) and on the anterior aspect of the thigh were evaluated using the plastic occlusion stress test (POST).
Methods: 13 healthy women (age 31 ± 4) with regular menses entered the study. POST was performed by applying a plastic chamber (1.8 cm diameter) on the skin for 24 h. After removal the skin surface water loss (SSWL) was measured using an evaporimeter (EP1-Servomed, Sweden) every 5 min for 30 min. Measurements were taken on the 10th and 25th day of the menstrual cycle. Statistical analysis was performed using one-factor A nova for repeated measures. Free and bound water compartments of evaporation were also analysed.
Results: Higher hydration and SSWL were detectable on the 25th day of the cycle. However, no significant influence of menstrual cycle was found. Significant differences between the upper and lower volar foream were detected ( P < 0.001).
Conclusion: The differences observed were mainly related to the evaporation of bound water, confirming a different barrier function at the sites investigated; therefore, it must be taken into account that adjacent skin sites with equal structure may have different functional behaviour.     

Electrical impedance and other physical parameters as related to lipid content of human stratum corneum

Background/aims: In previous studies we have shown that variations in the properties of the stratum corneum are reflected by alterations in electrical impedance. The aim of this study was to explore the ability of the electrical impedance technique to detect changes in the lipid content of the stratum corneum, and to compare It with the other non-invasive methods, measurement of transepidermal water loss and of skin moisture. Methods: Twenty-two healthy test subjects were recruited. Transepidermal water loss was measured at standard sites on the forearms and wrists, followed by skin moisture estimation by electrical capacitance, and finally by the recording of electrical impedance spectra in the frequency range 1 kHz to 1 MHz. Readings by all three methods were taken before the start of each series of test procedures, as well as after cyclohexane swabbing, a skin stripping procedure and lipid extraction, and also during the recovery process. A mixture of hexane:isopro-panol was used for lipid extraction of the skin, and the extracts were evaluated using HPLC/LSD and GC/MS/FID analysis. Biopsy samples for light and electron microscopy were obtained after lipid extraction. Results: Electrical impedance results showed greater changes after lipid extraction than either transepidermal water loss or skin moisture content. Baseline values varied from the cubital fossa to the wrist, both for the non-invasive methods and for lipid composition. Conclusions: The electrial impedance is dependent on the lipid content of the stratum corneum, as studied by lipid extraction experiments.     

Intra-individual variation of irritant threshold and relationship to transepidermal water loss measurement of skin irritation

Irritant susceptibility studies have used either visual assessment or transepidermal water loss (TEWL) to determine subject response. We have developed a visual assessment method which determines subject irritant threshold. We examined the relationship between sodium lauryl sulfate (SLS) irritant threshold and TEWL measurements from normal skin and SLS patch tests. 19 subjects were recruited. The irritant threshold of each subject was measured and TEWL measurements made from the applied SLS patch tests. Individuals with a lower irritant threshold (easily irritated skin) had elevated TEWL levels compared to those with higher thresholds. The irritant threshold test had a low intraindividual variation. This study showed that the 2 methods grouped patients in a similar manner. The variation seen may reflect the different outcomes measured: irritant threshold visually assesses the skin inflammatory response while TEWL measures skin barrier modification.     

Modulation of stratum corneum lipid composition and organization of human skin equivalents by specific medium supplements

Our in‐house human skin equivalents contain all stratum corneum (SC) barrier lipid classes, but have a reduced level of free fatty acids (FAs), of which a part is mono‐unsaturated. These differences lead to an altered SC lipid organization and thereby a reduced barrier function compared to human skin. In this study, we aimed to improve the SC FA composition and, consequently, the SC lipid organization of the Leiden epidermal model (LEM) by specific medium supplements. The standard FA mixture (consisting of palmitic, linoleic and arachidonic acids) supplemented to the medium was modified, by replacing protonated palmitic acid with deuterated palmitic acid or by the addition of deuterated arachidic acid to the mixture, to determine whether FAs are taken up from the medium and are incorporated into SC of LEM. Furthermore, supplementation of the total FA mixture or that of palmitic acid alone was increased four times to examine whether this improves the SC FA composition and lipid organization of LEM. The results demonstrate that the deuterated FAs are taken up into LEMs and are subsequently elongated and incorporated in their SC. However, a fourfold increase in palmitic acid supplementation does not change the SC FA composition or lipid organization of LEM. Increasing the concentration of the total FA mixture in the medium resulted in a decreased level of very long chain FAs and an increased level of mono‐unsaturated FAs, which lead to deteriorated SC lipid properties. These results indicate that SC lipid properties can be modulated by specific medium supplements.     

Repair of acetone- and sodium lauryl sulphate-damaged human skin barrier function using topically applied emulsions containing barrier lipids

BACKGROUND: It is generally acknowledged that well-formulated moisturizing skin care products can restore disturbed barrier function that can be assessed by transepidermal water loss (TEWL) measurements. When ceramides and/or other barrier lipids are incorporated, it is, however, not always clearly demonstrated which ingredients of the formulation exert the beneficial effects. OBJECTIVES: In this study the effects of topically applied ceramide-containing mixtures on the barrier repair of sodium lauryl sulphate (SLS)- and acetone-induced skin damage have been studied in human volunteers. TEWL and stratum corneum hydration measurements were carried out. The emulsions applied contained either a mixture of two types of ceramides, CerIII and CerIIIB (emulsion 1) or a complete mixture of ceramides III, IIIB and VI together with phytosphingosine, cholesterol and the free fatty acid linoleic acid (emulsion 2). RESULTS: After SLS damage, it was observed that barrier recovery was significantly accelerated by topical application (14 days, 2 x/d) of emulsion 2 compared with the results obtained with emulsion 1. Corneometrical results were not relevant due to the occurrence of scaly fissured skin, failing to provide a good skin/probe contact. Although no effect on TEWL could be observed, the improvement of skin hydration after acetone treatment and a single application of the emulsions, was significantly more positive for emulsion 2 than for emulsion 1. CONCLUSIONS: The investigative methods used in this study show that ceramides combined with other skin lipids can improve barrier repair after damage.     

Effect of Aqueous Cream BP on human stratum corneum in vivo

Background Aqueous Cream BP is widely prescribed to patients with eczema to relieve skin dryness. The formulation contains sodium lauryl sulphate (SLS), a chemical that is a known skin irritant and a commonly used excipient in personal care and household products. The chronic effects of Aqueous Cream BP application on skin barrier function have not been determined. Objectives To characterize and assess skin barrier function of healthy skin after application of Aqueous Cream BP and to study the physical effects of the formulation on the stratum corneum (SC). Methods The left and right volar forearms of six human volunteers were each separated into treated and control sides. The treated sides of each forearm were subjected to twice daily applications of Aqueous Cream BP for 4 weeks at the end of which concomitant tape stripping and transepidermal water loss (TEWL) measurements were made. The untreated sides of the forearms were not exposed to any products containing SLS during the study period. Results Changes in SC thickness, baseline TEWL and rate of increase in TEWL during tape stripping were observed in skin treated with Aqueous Cream BP. The mean decrease in SC thickness was 1·1 μm (12%) (P = 0·0015) and the mean increase in baseline TEWL was 2·5 g m^?2 h^?1 (20%) (P < 0·0001). Reduced SC thickness and an increase in baseline TEWL, as well as a faster rate of increase in TEWL during tape stripping, were observed in 16 out of 27 treated skin sites. Conclusions The application of Aqueous Cream BP, containing ～1% SLS, reduced the SC thickness of healthy skin and increased its permeability to water loss. These observations call into question the continued use of this emollient on the already compromised barrier of eczematous skin.     

Molecular interactions of plant oil components with stratum corneum lipids correlate with clinical measures of skin barrier function

Plant‐derived oils consisting of triglycerides and small amounts of free fatty acids (FFAs) are commonly used in skincare regimens. FFAs are known to disrupt skin barrier function. The objective of this study was to mechanistically study the effects of FFAs, triglycerides and their mixtures on skin barrier function. The effects of oleic acid (OA), glyceryl trioleate (GT) and OA/GT mixtures on skin barrier were assessed in vivo through measurement of transepidermal water loss (TEWL) and fluorescein dye penetration before and after a single application. OA's effects on stratum corneum (SC) lipid order in vivo were measured with infrared spectroscopy through application of perdeuterated OA (OA‐d₃₄). Studies of the interaction of OA and GT with skin lipids included imaging the distribution of OA‐d₃₄ and GT ex vivo with IR microspectroscopy and thermodynamic analysis of mixtures in aqueous monolayers. The oil mixtures increased both TEWL and fluorescein penetration 24 h after a single application in an OA dose‐dependent manner, with the highest increase from treatment with pure OA. OA‐d₃₄ penetrated into skin and disordered SC lipids. Furthermore, the ex vivo IR imaging studies showed that OA‐d₃₄ permeated to the dermal/epidermal junction while GT remained in the SC. The monolayer experiments showed preferential interspecies interactions between OA and SC lipids, while the mixing between GT and SC lipids was not thermodynamically preferred. The FFA component of plant oils may disrupt skin barrier function. The affinity between plant oil components and SC lipids likely determines the extent of their penetration and clinically measurable effects on skin barrier functions.     

Functional properties of the stratum corneum in patients with diabetes mellitus: similarities to senile xerosis

BACKGROUND: Although diabetes mellitus is known to induce many pathophysiological changes in the skin, none of the earlier studies has focused on the state of the stratum corneum (SC) in patients with diabetes. In our previous report on a diabetic mouse model, we described a decreased SC hydration state and abnormalities in the function of the sebaceous gland together with a normally retained barrier function of the SC. OBJECTIVES: To examine the functional changes of the SC in patients with diabetes. METHODS: The following values were measured in 49 patients with diabetes: fasting plasma glucose (FPG) and haemoglobin A1c (HbA1c); skin surface lipid level on the forehead; and high-frequency conductance (HFC) and transepidermal water loss (TEWL) of the SC of the volar forearm and extensor surface of the lower leg. These functions of the SC were compared in two pairs of patient groups, i.e. between groups with high and low FPG, and between groups with high and low HbA1c. RESULTS: The HFC levels on the extensor leg and volar forearm were significantly lower in the group with high FPG (FPG > 110 mg dL(-1)) than in the group with low FPG (FPG < 110 mg dL(-1)), although TEWL values were almost the same in these two groups. The skin surface lipids on the forehead were significantly lower in the former than in the latter. The HFC did not differ significantly between the group with high HbA1c (HbA1c > 5.8%) and the group with low HbA1c (HbA1c < 5.8%), whereas the TEWL of the volar forearm was slightly lower in the former than in the latter. CONCLUSIONS: These new results as well as our earlier results with diabetic mice suggest that patients with diabetes mellitus tend to show a reduced hydration state of the SC together with decreased sebaceous gland activity, without any impairment of the SC barrier function.     

Biophysical changes after mechanical injury of the stratum corneum in normal skin

Scrubbing off the stratum corneum with a rough towel after soaking in warm water is a bathing custom unique to Korea. However, Korean dermatologists have advised against this practice due to the potential harm that it may cause, though there is little data to support this advice. This study aimed to evaluate the changes in biophysical characteristics after such mechanical injury of stratum corneum, as reflected by water-holding capacity, stratum corneum barrier function, and stratum corneum turnover time (SCTT). 10 Korean female volunteers (aged 19-34 years) were enrolled in this study to investigate the acute effects and 32 volunteers (aged 19-49 years) the chronic effects. After soaking in warm water (36 degrees C) for 5 min, the flexor surfaces of the forearms were scrubbed with a rough towel. The volunteers' forearms were divided into 4 compartments, and each compartment was scrubbed 0, 5, 10, and 15x, respectively. After a single injury of the stratum corneum, there was an initial decrease in water-holding capacity until 6 h. Thereafter, it increased until day (D) 3, when it began to descend to the normal range. The transepidermal water loss (TEWL) values also peaked at D2 and declined thereafter. However, there were no changes in either water-holding capacity or skin barrier function, and only the SCTT was significantly shortened, after chronic injury of the stratum corneum.     

Increased stratum corneum serine protease activity in acute eczematous atopic skin

Background Atopic dermatitis (AD) is a chronic inflammatory disease associated with changes in stratum corneum (SC) structure and function. The breakdown of epidermal barrier function in AD is associated with changes in corneocyte size and maturation, desquamation, lipid profiles, and some protease activities. Objectives The purpose of this study was: (i) to examine physiological changes in lesional (L) skin of acute eczematous AD, compared with nonlesional (NL) AD skin and healthy (H) skin, using sequential tewametry and SC protein analysis to estimate SC thickness; and (ii) to assess which serine proteases might be involved in pathogenesis. Methods Six subjects with H skin, six AD patients with NL skin and six AD patients with mild to moderate eczema (L skin) were enrolled. Skin was assessed using several noninvasive techniques but SC thickness was estimated using tewametry and SC protein content of D‐Squame strippings. SC integrity was determined by sequential tape stripping (D‐Squame) and infrared densitometry. Kallikreins, plasmin, urokinase and leucocyte elastase protease activities together with a novel SC tryptase‐like enzyme activity were quantified. Results Transepidermal water loss (TEWL) levels after D‐Squame stripping were elevated in L compared with NL and H skin at all sampling points (P < 0·05). Conversely, the amount of SC removed by sequential tape stripping was decreased in L skin, indicating increased intracorneocyte cohesion (P < 0·05). By correlating 1/TEWL values and SC removed as an estimate of SC thickness, a significantly thinner SC was observed in L compared with NL and H skin (P < 0·05). Elevated extractable serine protease activity was measured in AD skin in the order: SC tryptase‐like enzyme (45×), plasmin (30×), urokinase (7·1×), trypsin‐like kallikreins (5·8×) and chymotrypsin‐like kallikreins (3·9×). Leucocyte elastase activity was not detected in H and NL skin but was observed in AD SC samples (L skin). All enzymes were elevated in the deeper layers of L SC compared with NL and H SC samples. All consistently elevated SC protease activities were significantly correlated with the bioinstrumental data. Conclusions We report increased serine protease activities in acute eczematous AD, especially in deeper layers of the SC, including SC tryptase‐like enzyme, plasmin, urokinase and leucocyte elastase activities. These elevations in protease activities were associated with impaired barrier function, irritation, and reduced skin capacitance. Increased SC cohesion was apparent despite elevated TEWL during tape stripping, which would indicate reduced SC thickness in acute eczematous lesions of AD. Indeed, this was observed using an estimate of SC thickness.     

Stratum corneum hydration and flexibility are useful parameters to indicate clinical severity of congenital ichthyosis

To determine any correlation between the stratum corneum barrier function and the phenotypic severity of congenital ichthyosis, we studied stratum corneum hydration, flexibility, thickness and transepidermal water loss (TEWL) in patients with congenital ichthyosis. Seven patients with congenital ichthyosis aged 2-46 years and age-matched controls were included in the present study. We divided seven patients into two groups; patients with non-bullous type (non-bullous congenital ichthyosiform erythroderma patients) and patients with the bullous type of congenital ichthyosis (bullous congenital ichthyosiform erythroderma and ichthyosis bullosa of Siemens). Stratum corneum hydration, thickness and flexibility were measured using a Corneometer ASA-M2. The stratum corneum thickness was also examined using a skin biopsy technique. TEWL was measured using Evaporimeter AS-TW1. The clinical severity of ichthyosis phenotype was evaluated using a visual analogue scale (VAS). Stratum corneum hydration and flexibility were significantly reduced in both congenital ichthyosis patient groups. Stratum corneum thickness was significantly increased in both groups. In the patient group with non-bullous congenital ichthyosis, significant negative correlations were confirmed between the VAS score and stratum corneum hydration and between the VAS score and flexibility. A significant, positive correlation was also observed between the VAS score and stratum corneum thickness. There was a positive correlation between the VAS score and TEWL on both the extensor and flexor sides of the forearm and back. We conclude that stratum corneum hydration, flexibility and thickness measured by the corneometer, and TEWL on the arm may be a useful indicator of the severity of ichthyosis phenotype.     

Physical and physiological effects of stratum corneum tape stripping

Background/aims: Tape stripping of human stratum corneum has been performed to measure stratum corneum mass, barrier function, drug reservoir and percutaneous penetration. However, the technique itself requires further development to facilitate interpretation.
Methods: In this study we quantified stratum corneum (SC) tape stripping and water kinetic parameters utilizing three types of adhesive tapes, in an in vivo randomized clinical trial. Stratum corneum was tape stripped, and the mass of SC removed by each tape was quantified utilizing a protein assay. Transepidermal water loss (TEWL) was measured and barrier disruption and SC water kinetics calculated. Three commonly utilized acrylate adhesive tapes were utilized and a comparison made between them.
Results: Each type of tape successfully stripped the stratum corneum, but the rayon tape did not induce SC barrier disruption. Neither the type of tape nor the site stripped significantly influenced the mass of SC removed. Water kinetic parameters did not differ significantly for the tapes that did induce barrier disruption. Individual variation in barrier disruption to water following tape stripping was demonstrated.
Conclusion: The tapes utilized removed a similar amount of SC. The tapes have a different propensity to cause barrier disruption. Some individuals do not demonstrate increased TEWL despite an equivalent mass of SC being removed compared to those who do show a response.     

Knock‐down of filaggrin does not affect lipid organization and composition in stratum corneum of reconstructed human skin equivalents

Human skin mainly functions as an effective barrier against unwanted environmental influences. The barrier function strongly relies on the outermost layer of the skin, the stratum corneum (SC), which is composed of corneocytes embedded in an extracellular lipid matrix. The importance of a proper barrier function is shown in various skin disorders such as atopic dermatitis (AD), a complex human skin disorder strongly associated with filaggrin (FLG) null mutations, but their role in barrier function is yet unclear. To study the role of FLG in SC barrier properties in terms of SC lipid organization and lipid composition, we generated an N/TERT‐based 3D‐skin equivalent (NSE) after knock‐down of FLG with shRNA. In these NSEs, we examined epidermal morphogenesis by evaluating the expression of differentiation markers keratin 10, FLG, loricrin and the proliferation marker ki67. Furthermore, the SC was extensively analysed for lipid organization, lipid composition and SC permeability. Our results demonstrate that FLG knock‐down (FLG‐KD) did not affect epidermal morphogenesis, SC lipid organization, lipid composition and SC permeability for a lipophilic compound in NSEs. Therefore, our findings indicate that FLG‐KD alone does not necessarily affect the functionality of a proper barrier function.     

International guidelines for the in vivo assessment of skin properties in non‐clinical settings: Part 2. transepidermal water loss and skin hydration

Background

There is an emerging perspective that it is not sufficient to just assess skin exposure to physical and chemical stressors in workplaces, but that it is also important to assess the condition, i.e. skin barrier function of the exposed skin at the time of exposure. The workplace environment, representing a non‐clinical environment, can be highly variable and difficult to control, thereby presenting unique measurement challenges not typically encountered in clinical settings.

Methods

An expert working group convened a workshop as part of the 5th International Conference on Occupational and Environmental Exposure of Skin to Chemicals (OEESC) to develop basic guidelines and best practices (based on existing clinical guidelines, published data, and own experiences) for the in vivo measurement of transepidermal water loss (TEWL) and skin hydration in non‐clinical settings with specific reference to the workplace as a worst‐case scenario.

Results

Key elements of these guidelines are: (i) to minimize or recognize, to the extent feasible, the influences of relevant endogenous‐, exogenous‐, environmental‐ and measurement/instrumentation‐related factors; (ii) to measure TEWL with a closed‐chamber type instrument; (iii) report results as a difference or percent change (rather than absolute values); and (iv) accurately report any notable deviations from this guidelines.

Conclusion

It is anticipated that these guidelines will promote consistent data reporting, which will facilitate inter‐comparison of study results.     

The effects of protective occlusive gloves on stratum corneum barrier properties *

Data suggests that protection From gloves is paradoxically reduced became of effects of occlusion on the skin. The aim of this study was to characterize these effects on physical and functional properties of stratum corneum. Volunteer trials were conducted using patches of polyvinyl chloride (PVC) glove material on the skin. Impairment of barrier function -was assessed by measuring transepidermal water loss (TEWL). Hydration and water sorption-desorption was assessed using skin conductance. The major finding was a short-term impairment of barrier function, measured as an increase in TEWL of 1.7 gn ^?2 h ^?1 (95% C.I. 0.4 to 2.6 gn ^?2, h ^?1, p < 0.01) Temporary increases in hydration and water sorption were also observed. On this basis, the effects of wearing PVC gloves over 2 days on stratum corneum barrier properties on the dorsum of the hand were studied. TEWL measurements remained elevated by 1.5 gn ^?2 h ^?1 the day after occlude glove removal (95% C.I 0.2 to 2.8 gn ^?2 h ^?1, p < 0.01), indicating a possible cumulative effect. In conclusion, our studies demonstrate a potential hazard resulting from the repeated use of protective gloves.     

Efficacy of skin barrier creams (IV). The repetitive irritation test (RIT) with a set of 4 standard irritants

An improved human model for the quantification of skin barrier creams (BCs) is described. In contrast to the previously published procedure, the back, instead of the forearm, and a total of 4 irritants are used. Due to the larger area, 3 BC formulations can be simultaneously compared to the control field, which receives the irritant only, without BC-pretreatment. On 10 human volunteers, the irritants 10% sodium lauryl sulfate (SLS), 1% sodium hydroxide (NaOH), 30% lactic acid (LA) and undiluted toluene (TOL) were applied via large Finn Chambers for 30 min, 5 × during the 1st week and 4 × during the 2nd week. Taktosan Salbe (water-in-oil emulsion) and RAWI Speerschutzcreme (oil-in-water emulsion) were applied 30 min before contact with the irritants. In order to asses reproducibility and interindividual variation, the BC RAWI was tested in duplicate. Irritant cutaneous reactions were quantified by 4 parameters: erythema score, transepidermal water loss, blood flow volume and stratum corneum hydration by measuring capacitance. The results showed marked differences in efficacy. Taktosan significantly suppressed irritation by SLS, NaOH and LA, which was apparent in nearly all parameters. RAWI caused significant inhibition of SLS irritation, and a positive trend against NaOH and LA was observed. Both BCs failed against TOL. The results of duplicate testing with RAWI showed good reproducibility. The dogma that oil-in-water emulsions are primarily effective against lipophilic irritants, and water-in-oil emulsions against hydrophilic irritants, needs to be re-evaluated on the basis of our findings. This model seems to have potential for further studies on BCs and might elucidate the complex interaction of BCs with irritants.