Development of a read-across workflow for skin irritation and corrosion predictions.

We developed a read-across workflow using the OECD QSAR Toolbox for the prediction of skin irritation and corrosion. In the workflow, we gathered analogues using an improved profiler for skin irritation and corrosion to define valid categories. In addition, we refined categories by removing chemicals based on melting points and structural features. Finally, prediction results were obtained using our self-determined rule for read-across. In this rule, we decided the number of analogues from which the read-across is performed, analogue selection criteria (i.e. high similarity vs. near log Pow) and prediction rule (i.e. majority vs. unanimity). We created a program for the optimization of read-across workflows. We applied this program to 313 chemicals in the training set and sought the optimized workflows among >1000 possible choices of profilers and ways of subcategorization and data gap filling. Use of the optimized workflows provided highly accurate, unbiased, user-independent and reproducible read-across predictions. The prediction results obtained from read-across workflows can be used for the selection of in vitro test methods or as part of the weight-of-evidence approaches in the Integrated Approach on Testing and Assessment for skin irritation and corrosion. Moreover, these results can be used for screening purposes and/or preliminary hazard assessment.

A whole-cell biosensor as in vitro alternative to skin irritation tests.

This study presents the time-resolved detection of chemically induced stress upon intracellular signaling cascades by using genetically modified sensor cells based on the human keratinocyte cell line HaCaT. The cells were stably transfected with a HSP72-GFP reporter gene construct to create an optical sensor cell line expressing a stress-inducible reporter protein. The time- and dose-dependent performance of the sensor cells is demonstrated and discussed in comparison to a label-free impedimetric monitoring approach (electric cell-substrate impedance sensing, ECIS). Moreover, a microfluidic platform was established based on µSlidesI(0,4)Luer to allow for a convenient, sterile and incubator-independent time-lapse microscopic observation of the sensor cells. Cell growth was successfully achieved in this microfluidic setup and the cellular response to a cytotoxic substance could be followed in real-time and in a non-invasive, sensitive manner. This study paves the way for the development of micro-total analysis systems that combine optical and impedimetric readouts to enable an overall quantitative characterization of changes in cell metabolism and morphology as a response to toxin exposure. By recording multiple parameters, a detailed discrimination between competing stress- or growth-related mechanisms is possible, thereby presenting an entirely new in vitro alternative to skin irritation tests.

Review of methods used for quantifying excess water in over-hydrated skin using evaporimetry.

BACKGROUND: Advances in diapers and skin barrier products are often aimed at reducing water penetration of the skin to prevent diaper dermatitis and evaporimetry has commonly been measured to quantify excess water in the skin. The aim of this study was to critically review the methods used to measure water vapour flux density (WVFD) using evaporimetry in order to identify a standardised methodology. METHODS: We used MEDLINE (1980-2008) and hand searching to identify published papers that used evaporimetry to measure WVFD when the skin has been exposed to water/saline/urine. We compared the papers with respect to subjects, sites, methods of hydrating the skin, the conditions of logging, timing and analysing the evaporimetry data. RESULTS: We identified 10 papers. Methods and techniques for measuring WVFD and analysing data varied considerably between studies and it was not possible to identify a standardised method. The main sources of error and variation are discussed. CONCLUSION: Little work has been carried out to establish the optimum methods and techniques needed to minimise variation in measurements of WVFD using evaporimetery. There is a need to develop more robust, standardised methods and to demonstrate their reliability for further work.

Is it possible to characterize objectively sensitive skin?

BACKGROUND/PURPOSE: Sensitive skin is a subject of intense research work. However, its contours have not been defined and properly investigated so far. The aim of this work was to characterize objectively the consumers of cosmetic products, which referred skin susceptibility to several agents or conditions. METHODS: Twenty-four healthy female volunteers, mean age 38.9+/-13 years were recruited. The volunteers were fully informed about the study having previously expressed their consent, and were grouped in to I: individuals without any skin sensitivity complaint and II: individuals with self-reported sensitive skin to regular contact with household cleaning products. Sodium lauryl sulphate was used as a 'provocative' agent and it was applied on the back of the volunteers' hands, as an occlusive patch for 24 h. After the patch removal (30 min, 7 and 14 days), the variables erythema, transepidermal water loss, stratum corneum hydration and blood perfusion were measured. RESULTS: No evidence of a statistical difference was found between the biomechanical behaviour of the skin of the two groups. CONCLUSION: The results of our study did not enable us to establish a clear discrimination between sensitive and non-sensitive skin, which once again underlines the subjective nature of this condition.

Electrical impedance model for evaluation of skin irritation in rabbits and humans.

BACKGROUND/AIM: The electrical impedance method has been used as a quantitative technique for evaluating changes in the skin during irritation within the invisible range. The purpose of this study was to apply an electrical model of skin to the interpretation of impedance data after the application of an irritant (SLS) in the skin of humans and rabbits. METHODS: Investigations were performed on 12 humans and 15 albino rabbits. Responses were evaluated by measuring electrical impedance before irritant exposure and 24 h after its removal and also by visual inspection. Using the raw impedance parameter, a novel index was developed and its value was correlated with visual scoring. RESULTS: The derived impedance index showed a significant correlation with visual scores and its value decreased (P<0.05) after irritant removal even without macroscopic signs of irritation. CONCLUSION: The proposed electrical model of skin seems to be suitable for the detection and interpretation of changes in the impedance characteristics of skin induced by SLS in rabbits and humans.

Use of the cytosensor microphysiometer to predict results of a 21-day cumulative irritation patch test in humans.

The cytosensor microphysiometer (mu phi) was investigated as a rapid, relatively inexpensive test to predict performance of skin cleansing wipes on the human 21-day cumulative irritation patch test (21CIPT). It indirectly measures metabolic rate changes in L929 cells as a function of test article dose, by measuring the acidification rate in a low-buffer medium. The dose producing a 50% reduction in metabolic rate (MRD50), relative to the baseline rate, is used as a measure of toxicity. The acute toxicity of the mu phi assay can be compared to the chronic toxicity of the 21CIPT, which is based largely on the exposure of test agents to the epidermal cells, resulting in damage and penetration of the stratum corneum leading to cell toxicity. Two series of surfactant-based cleansing wipe products were tested via the mu phi assay and 21CIPT. The first series, consisting of 20 products, was used to determine a prediction model. The second series of 38 products consisted of routine product development formulas or marketed products. Comparing the results from both tests, samples with an MRD50 greater than 50 mg/ml provided a 21CIPT score consistent with a product that performs satisfactorily in the market. When the MRD50 was greater than 78 mg/ml, the 21CIPT score was usually zero. The mu phi may be more sensitive than the 21CIPT for ranking minimally irritating materials. The mu phi assay is useful as a screen for predicting the performance of a wet wipes formula on the 21CIPT, and concurrently reduces the use of animals for safety testing in a product development program for cleansing wipes.

The Behind-the-Knee test: an efficient model for evaluating mechanical and chemical irritation.

BACKGROUND/PURPOSE: The 'Behind-the-Knee' method (BTK test), using the popliteal fossa as a test site, evaluates both the inherent chemical irritation, and the potential for mechanical irritation of substrates and products. This approach eliminates some of the difficulties of in-use clinical test systems while still providing reliable results. In this publication, examples of the results of BTK tests on several materials are presented with direct comparisons, where possible, with results of in-use clinical testing conducted on the same materials. METHODS: In in-use clinical tests, volunteer panelists were provided with catamenial products to use in place of their normal product. In the BTK test, samples were applied daily to the popliteal fossa using an elastic athletic band. In both studies, irritation reactions were scored visually. RESULTS: Levels of irritation in the BTK test are consistently higher than those of standard patch tests, illustrating the contribution of mechanical irritation to the overall irritant potential of materials and products. Repeated tests on identical test materials demonstrated that the BTK test results are reproducible. Side-by-side comparisons of the BTK test and in-use clinical tests demonstrated that the BTK test produces results of similar quality to the in-use clinical. By using several concurrent panels with a common test material, it is possible to compare the irritant properties of several materials at once. CONCLUSIONS: We have tested over 25 different materials in over 35 BTK studies. The test method has proven reliable and versatile in testing a wide variety of materials, including menstrual pads, topsheets, interlabial pads, pantiliners, tampons and lotion coatings on products. Unlike in-use clinicals, the BTK test allows the direct comparison of two products at one time on the same individual, and is easily adapted to investigative programs. It is subject to fewer confounding factors, is much easier to implement, has a shorter turnaround time, and is less expensive than in-use clinical testing. Importantly, unlike standard patch tests, the BTK test evaluates both the inherent chemical irritation associated with materials and the mechanical irritation owing to friction. Although the BTK test was developed using catamenial products, the test system provides a valuable alternative for evaluating any material where mechanical irritation may play a role, including textiles, facial tissues, baby and adult diapers, and laundry products that may leave residues on fabrics.

Additive impairment of the barrier function and irritation by biogenic amines and sodium lauryl sulphate: a controlled in vivo tandem irritation study.

Biogenic amines are potential irritants e.g. in fish-, meat-, milk- and egg-processing professions like cooks, butchers and bakers. The aim of this study was to test the irritative and barrier-disrupting properties of the biogenic amines ammonium hydroxide (AM), dimethylamine (DMA) and trimethylamine (TMA). A repeated sequential irritation of 30 min twice per day was performed over a total of 4 days (tandem repeated irritation test) on the back of 20 healthy volunteers of both sexes with AM, DMA, TMA and sodium lauryl sulphate (SLS). The epidermal barrier function was assessed with a Tewameter TM 210, stratum corneum surface pH was measured with a Skin-pH-Meter 900, inflammation was assessed with a Chromameter CR-300 on the a* axis for redness and a visual score was recorded. All tested biogenic amines (AM, DMA and TMA) induced a barrier disruption and a pH increase paralleled with a 1-day-delayed onset of inflammatory signs. These effects were further enhanced and accelerated by a sequential application of SLS together with the biogenic amines, and inflammation occurred earlier than with the single compounds. Acetic acid (AA) in contrast did only show mild barrier disruption and no significant inflammatory signs. Our system allowed a ranking of the different compounds in their irritative potential in the tandem irritation with SLS: SLS > NaOH > TMA > AA > AM > DMA. The results are suggestive that in the food-processing industry the simultaneous contact with biogenic amines and harmful detergents like SLS should be minimized.