H.-J. Weigmann

The tape stripping procedure ― evaluation of some critical parameters

Tape stripping is a simple and efficient method for the assessment of quality and efficacy of cosmetical and dermatological formulations. After topical application and penetration of formulations, the cell layers of the stratum corneum are successively removed from the same skin area using adhesive films. The tape strips contain the amount of corneocytes and the corresponding amount of the penetrated formulation, which can be determined by classical analytical chemical methods. Different formulations can strongly influence the amount of stratum corneum removed with every tape strip. Therefore, it is essential for the comparison of the penetration of different formulations that the amount of formulation detected on the single tape strip is not related to the tape strip number as a relative measure of the penetration depths, but to their standardized real position in the stratum corneum. Therefore, different methods are reported for the determination of the amount of stratum corneum removed with every tape strip. The tape stripping method in its standardized form is well-suited to determine the dermatopharmacokinetics of topically applied substances. Additionally, the method can be used to obtain information about the homogeneity and the distribution of formulations on the skin and in the stratum corneum. This is used, e.g., for the determination of the homogeneity of the distribution and the ex vivo determination of a universal sun protection factor (USPF) characterizing the efficacy of sunscreens.

Estimation of the relative stratum corneum amount removed by tape stripping.

Background/purpose: The tape stripping procedure is a suitable minimal invasive tool to study, e.g. the penetration and dermatopharmacokinetics of topically applied substances. In the present study, this procedure was used to remove the stratum corneum (SC) completely and to study the penetration of the UVA filter substance butyl methoxydibenzoylmethane after application in two different vehicles.

Comparison of Transepidermal Water Loss and Spectroscopic Absorbance to Quantify Changes of the Stratum corneum after Tape Stripping

The objective and quantitative application of tape stripping in pharmaceutics and dermatopharmacokinetics requires the determination of the exact position of each removed tape strip inside the stratum corneum (SC) and/or the determination of the relative SC thickness. In this study, transepidermal water loss (TEWL) and the optical spectroscopic data of the corneocytes were measured simultaneously during the complete removal of the SC by tape stripping. The spectroscopic data quantitatively reflect the amount of corneocytes removed by the individual tape strips, whereas TEWL and 1/TEWL are not sensitive enough to measure the relatively small changes in the SC thickness realized by the removal of the individual strips. The relative SC thickness can be determined directly by the spectroscopic data, while the 1/TEWL values require a second independent method. The results demonstrate the importance of tape stripping characterizing the behaviour of topically applied substances.

In vivo methods for the analysis of the penetration of topically applied substances in and through the skin barrier.

The efficacy of a drug is characterized by its action mechanism and its ability to pass the skin barrier. In this article, different methods are discussed, which permit this penetration process to be analysed non‐invasively. Providing qualitative and quantitative information, tape stripping is one of the oldest procedures for penetration studies. Although single cell layers of corneocytes are removed from the skin surface, this procedure is considered as non‐invasive and is applicable exclusively to the stratum corneum. Recently, optical and spectroscopic methods have been used to investigate the penetration process. Fluorescence‐labelled drugs can be easily detected in the skin by laser scanning microscopy. This method has the disadvantage that the dye labelling changes the molecular structures of the drug and consequently might influence the penetration properties. The penetration process of non‐fluorescent substances can be analysed by Raman spectroscopy, electron paramagnetic resonance, CARS and multiphoton microscopic measurements. Using these methods, the concentration of the topically applied formulations in different depths of the stratum corneum can be detected by moving the laser focus from the skin surface deeper into the stratum corneum. The advantages and disadvantages of these methods will be discussed in this article.

Penetration studies of topically applied substances: Optical determination of the amount of stratum corneum removed by tape stripping.

Tape stripping is a standard measuring method for the investigation of the dermatopharmacokinetics of topically applied substances using adhesive films. These tape strips are successively applied and removed from the skin after application and penetration of topically applied substances. Thus, layers of corneocytes and some amount of topical applied substances are removed. The amount of substances and the amount of stratum corneum removed with a single tape strip has to be determined for the calculation of the penetration profile. The topically applied substances removed from the skin can be determined by classical analytical methods like high-pressure liquid chromatography, mass spectroscopy, and spectroscopic measurements. The amount of corneocytes on the tape strips can be easily detected by their pseudoabsorption. In the present paper, an easy and cheap corneocyte density analyzer is presented that is based on a slide projector. Comparing the results of the measurements obtained by the corneocyte density analyzer and by uv-visible spectrometry, identical results were obtained.

Reservoir Function of the Stratum corneum: Development of an in vivo Method to Quantitatively Determine the Stratum corneum Reservoir for Topically Applied Substances

Investigations on the stratum corneum (SC) reservoir for topically applied substances are of importance in dermatologic science in order to assess the pharmacokinetics of these substances. In the present study, an in vivo method was developed to determine the SC reservoir quantitatively and to investigate the temporal behavior of this reservoir. Therefore, increasing amounts of an oil-in-water emulsion (o/w emulsion) containing 4% of a chemical UV filter were topically applied onto the flexor forearms of 5 healthy volunteers. The saturation of the SC reservoir was determined utilizing the tape stripping technique 1 and 6 h after application. The capacity of the SC reservoir for the o/w emulsion was found to be approximately 2.7 mg/cm2. Furthermore, a correlation of the capacity of the SC with transepidermal water loss was observed. Extending the time between the topical application and SC removal did not affect the distribution or the recovery rate of the UV filter in the SC. The results indicate that the reservoir of the SC is limited. This is reflected by the saturation level, which depends on the individual volunteer and, presumably, the topically applied substances and formulations used. The results show that the method developed is suited to quantitatively determine in vivo the SC reservoir for topically applied substances.

Comparison of human and porcine skin for characterization of sunscreens

The universal sun protection factor (USPF) characterizing sunscreen efficacy based on spectroscopically determined data, which were obtained using the tape stripping procedure. The USPF takes into account the complete ultraviolet (UV) spectral range in contrast to the classical sun protection factor (SPF). Until now, the USPF determination has been evaluated only in human skin. However, investigating new filters not yet licensed excludes in vivo investigation on human skin but requires the utilization of a suitable skin model. The penetration behavior and the protection efficacy of 10 commercial sunscreens characterized by USPF were investigated, comparing human and porcine skin. The penetration behavior found for typical UV filter substances is nearly identical for both skin types. The comparison of the USPF obtained for human and porcine skin results in a linear relation between both USPF values with a correlation factor R(2)=0.98. The results demonstrate the possibility for the use of porcine skin to determine the protection efficacy of sunscreens.

Methods for the Evaluation of the Protective Efficacy of Sunscreen Products

The objective of the present investigation was to examine the utilization of optical and spectroscopic methods for the noninvasive characterization of Anthelios XL Fluide Extreme (SPF 50+), an exemplary sunscreen, concerning its homogeneity of distribution on the skin, its spectroscopic properties and its overall protective efficacy. The homogeneity of the distribution of the sunscreen on the skin was investigated with a multiphoton tomography microscope. Additionally, the sum transmission spectrum was determined using tape stripping and spectroscopic measurements. The results revealed a very homogeneous distribution of the sunscreen on the skin surface and also in the deep furrows. The sum transmission spectrum reflects a high protective efficacy of the sunscreen in both the UVA and UVB ranges. The sunscreen Anthelios XL Fluide Extreme (SPF 50+) generates a comfortable feeling on the skin and can be easily distributed. The presented optical methods have been shown to be suitable to investigate the overall protective efficacy of sunscreen products objectively, noninvasively and quickly.

Pathways of lateral spreading.

In the case of topically applied substances, usually both lateral spreading and competitive penetration into the skin occur in parallel. In the present study, the pathways of lateral spreading were studied quantitatively and visually. The local distribution and lateral spreading of the UV filter substance butyl methoxydibenzoylmethane applied in an o/w emulsion was studied on the forearm and the back. The tape stripping procedure was used to determine the recovery rates inside and outside the area of application. The skin characteristics of transepidermal water loss, pH value, hydration of the stratum corneum and sebum rate were determined at both anatomic sites. Photography and laser scanning microscopy were used to visually investigate the lateral spreading of topically applied dyes. On the back, a preferred direction of lateral spreading parallel to the body axis was observed. This result was caused by differences in the network of furrows. The furrows functioned as a pathway for lateral spreading, whereas the follicles formed a reservoir for the topically applied substance.

Gain or Loss? Sunscreen Efficiency after Cosmetic Pretreatment of the Skin

Sunscreens are a key pillar of the multimodal protection strategy against short- and long-term impacts of intermittent and continuous UV exposure. Hitherto, an unanswered part of current scientific discourse is the question whether a cosmetic pretreatment has an impact on distribution and adhesiveness of sunscreens on the skin and therefore affects UV protection. In order to evaluate the homogeneity of sunscreen filter distribution, water resistance as a parameter of adhesiveness and effective UV protection of sunscreens after a pretreatment with cream or lotion was investigated in 18 volunteers who were examined before and after swimming, using the established combination of the tape stripping procedure and UV/VIS spectroscopy. It was shown that a cosmetic skin pretreatment affects neither filter homogeneity nor effective UV protection prior to water contact. However, compared to nonpretreated skin, a considerable loss of water resistance is caused. Therefore, using a cream or lotion before application of sunscreens is not to be recommended.