In vivo monitoring of epidermal absorption of hazardous substances by confocal Raman micro-spectroscopy.

BACKGROUND: Presently, percutaneous absorption of potentially hazardous chemicals in humans can only be assessed in animal experiments, in vitro, or predicted mathematically. Our aim was to demonstrate the proof-of-principle of a novel quantitative in vivo assay for percutaneous absorption: confocal Raman micro-spectroscopy (CRS). The advantages and limitations of CRS for health risk assessments are discussed. PATIENTS AND METHODS: 2-butoxyethanol, toluene, and pyrene were applied in pure form, diluted in water, or in ethanol on the skin of three healthy volunteers. CRS measurements were done following application for 15 min and 3 hours. The concentrations of the three substances as a function of distance to the skin surface were calculated and further analyzed with regard to mass transport into the stratum corneum (µg/cm(2)) and the flux through the stratum corneum (µg/cm(2)h). The results were compared with the available data from literature. RESULTS: Considering the preliminary nature of these data, good accordance with data from the literature was observed. In addition, we observed that 2-butoxyethanol penetrates markedly faster when dissolved in water as compared to ethanol. This observation is also in agreement with previous results. CONCLUSIONS: CRS has the potential to provide fast, accurate and reliable results for advanced studies of in vivo percutaneous absorption kinetics of hazardous substances in human skin. This will require further research with other substances and under differing conditions.

Depth profiling of Stratum corneum hydration in vivo: a comparison between conductance and confocal Raman spectroscopic measurements.

The high-frequency electrical conductance of tape-stripped human skin in vivo can be used to evaluate the hydration profile of Stratum corneum (SC). Tape-stripping provides access to the underlying SC layers, and the conductance of these layers (as measured by the Skicon instrument) correlates well with their water content, as demonstrated by independent confocal Raman spectroscopic measurements. The correlation shows high inter-individual variance and is not linear over the full measurement range of the instrument, but is helpful to discriminate between dry, normal and highly hydrated SC. The depth profile of hydration in tape-stripped SC corresponds to the one in intact SC only if the barrier function of the skin is not impaired. Thus, conductometry of tape-stripped skin must be used in conjunction with a method that allows to estimate the barrier damage inflicted to SC during the tape-stripping procedure, for example, measurement of the trans-epidermal water loss. The methodology described here is simple, rapid and minimally invasive, and it employs commercially available instrumentation that is cheap, portable and easy to use. This approach is applicable to in vivo estimation of the SC hydration in studies in the areas of dermatology, skin care and transdermal drug delivery.

Lipid uptake and skin occlusion following topical application of oils on adult and infant skin.

BACKGROUND: Topical application of oils and oil-based formulations is common practice in skin care for both adults and infants. Only limited knowledge however is available regarding skin penetration and occlusive potential of oils and common methods for measuring skin moisturization fall short when it comes to the moisturizing effect of oils. OBJECTIVE: In this study we used in vivo confocal Raman microspectroscopy to test the efficacy of paraffin oil (mineral oil) and two vegetable oils in terms of skin penetration and occlusion. Petrolatum was used as a positive control. METHODS: The products were applied topically on the forearms of nine volunteers and seven infants and Raman spectra were acquired before and at 30 and 90 min following application. Depth concentration profiles for lipid and water were calculated from the Raman spectra. Skin occlusion was assessed from the amount of stratum corneum (SC) swelling measured from the water concentration profiles. RESULTS: The paraffin oil and the vegetable oils penetrate the top layers of the SC with similar concentration profiles, a result that was confirmed both for adult and infant skin. The three oils tested demonstrated modest SC swelling (10-20%) compared to moderate swelling (40-60%) for petrolatum. CONCLUSION: These data indicate that there is no statistical difference between the paraffin oil and vegetable oils in terms of skin penetration and skin occlusion. The results for petrolatum show that in vivo confocal Raman microspectroscopy is sensitive and specific enough to measure both lipid uptake and skin occlusion events following topical application.

Potential of short-wave infrared spectroscopy for quantitative depth profiling of stratum corneum lipids and water in dermatology.

We demonstrate the feasibility of short wave infrared (SWIR) spectroscopy combined with tape stripping for depth profiling of lipids and water in the stratum corneum of human skin. The proposed spectroscopic technique relies on differential detection at three wavelengths of 1720, 1750, and 1770 nm, with varying ratio of the lipid-to-water absorption coefficient and an 'isosbestic point'. Comparison of the data acquired using SWIR spectroscopy with that obtained by a gold standard for non-invasive quantitative molecular-specific skin measurements, namely confocal Raman spectroscopy (CRS), revealed specificity of the proposed modality for water and lipid quantification. At the same time, we provide evidence showing aberrant sensitivity of Corneometer hydration read-outs to the presence of skin surface lipids, and a lack of sensitivity of the Sebumeter when attempting to measure the lipids of the cornified lipid envelope and intracellular lipid layers. We conclude that a spectroscopic SWIR-based spectroscopic method combined with tape stripping has the potential for depth profiling of the stratum corneum water and lipids, due to superior measurement sensitivity and specificity compared to the Corneometer and Sebumeter.

An evaluation of barrier repair foam on the molecular concentration profiles of intrinsic skin constituents utilizing confocal Raman spectroscopy.

For decades, transepidermal water loss and corneometry have been accepted as measures of skin barrier function. However, these tests are not capable of informing clinicians of the biochemical constituents and biophysical status of the stratum corneum. Knowledge of how the stratum corneum reacts to topical agents is important, as it reveals significant detail regarding the composition and function of this vital skin layer. Furthermore, transepidermal water loss and corneometry serve only as surrogate markers of barrier function. A more precise method of assessing stratum corneum hydration and lipid levels is emerging; in vivo confocal Raman spectroscopy is able to detect and quantify specific biochemical constituents in skin. This information then allows for assessment of the actual physiological status of this vital layer of the skin. This pilot study sought to elucidate a biophysical rationale for the clinical improvement achieved by hyaluronic acid/ceramide barrier repair foam in prior studies as measured by in vivo confocal Raman spectroscopy. Study results include increased lipid and hydration levels in the stratum corneum to depths of 25µm and 40µm, respectively, at the 2-hour, 48-hour, and 7-day time points.

Assessment of the "skin reservoir" of urea by confocal Raman microspectroscopy and reverse iontophoresis in vivo.

PURPOSE: To demonstrate the "skin reservoir" of urea by confocal Raman microspectroscopy in vivo and to evaluate its impact on the non-invasive monitoring of the analyte by reverse iontophoresis. METHODS: Urea was extracted iontophoretically over a 2-h period across the skin of adult volunteers and patients with chronic kidney disease. Confocal Raman microspectroscopic profiles of skin were recorded before and after 30 min of current application. RESULTS: Urea extraction was higher at the beginning of current passage, but then decreased to achieve stable values after 2 h of iontophoresis. After 30 min of iontophoresis, the Raman spectra highlighted a clear depletion of urea at the surface of the skin. Lactate distribution was also modified both at the surface and deeper into the skin. CONCLUSIONS: A source of urea in the skin, unrelated to the concentration circulating in the blood, was strongly suggested by extracted urea flux observed over time and by the Raman spectroscopy. This "urea reservoir" must be removed before systemic urea levels can be non-invasively monitored by reverse iontophoresis.