Modified Corneosurfametry as a new accelerated high-throughput ex vivo methodology for predicting cleanser effects towards human skin.

BACKGROUND: Corneosurfametry (CSM) was originally developed as a tool to predict irritation potential of cleansers. In this method, surface skin stripped using cyanoacrylate is contacted with surfactants/products, rinsed and stained with toluidine blue and basic fuschin dyes. The intensity of staining increases with increases in irritation potential of surfactant. Our objective was to modify the CSM technique to achieve better control of the tape stripping process. Another objective was to correlate the modified CSM (MCSM) with a traditional in-vivo forearm controlled application test (FCAT) for mildness and to explore its utility to assess the state of corneum after a clinical test. METHODS: Surface skin cells were tape stripped from forearms of volunteers with D-Squame Adhesive Discs. Discs were treated with a 10% solution of the product in a 96-plate well for 10 min, rinsed, dried and treated with basic fuschin-toluidine blue dye solution, rinsed and dried again. Forearm Controlled Application Test (FCAT) was based on a published protocol. Tape strips obtained after product treatment were also analyzed by the MCSM procedure without additional product treatment. RESULTS: Mildness/barrier damage assessed from in-vivo FCAT showed a similar ranking to the MCSM results. MCSM, TEWL and Erythema analysis of between-treatment differences showed a good correlation indicating that barrier damage seen in in-vivo studies can be predicted from ex-vivo MCSM studies. MCSM analysis of tape strips after the FCAT study showed that the damage decreased with increase in tape strip number. A moisturizing body wash (MBW) with mild surfactants showed the least damage in all layers. In contrast, harsh dish washing liquid showed significantly higher damage down to several layers. Another MBW with petrolatum in a harsher base showed damage almost similar to that of the harsh dish washing liquid in the surface layers. Thus, the MCSM was able to show underlying damage which would have been normally masked by the deposited petrolatum. CONCLUSION: The MCSM assay was shown to be a valuable tool for accelerated high throughput evaluation of mildness of surfactants and fully formulated products. MCSM can also be used to assess the state of the corneum after a product treatment.

Characteristic differences in barrier and hygroscopic properties between normal and cosmetic dry skin. II. Depth profile of natural moisturizing factor and cohesivity.

OBJECTIVE: In a previous paper (Lu et al., International Journal of Cosmetic Science, in press), we reported that in comparison with normal skin, cosmetic dry skin characteristically has a thicker stratum corneum (SC) with weaker barrier quality. Our goal here was to obtain a more complete picture of the difference between normal and cosmetic dry skin, by further analysing the data collected from the same clinical study to address two additional aspects of skin: the amount and depth profile of natural moisturizing factor (NMF) and the cohesivity of the SC. METHOD: The SC samples were collected by sequential tape stripping in a clinical study consisting of 64 healthy Caucasian female subjects with either normal or cosmetic dry skin. Protein and free amino acids (FAA) on tape strips were extracted and analysed using high-throughput methods. The level of pyrrolidone carboxylic acid (PCA), the prominent component of NMF, was detected using HPLC analysis. RESULTS: It was found that dry skin had significantly lower ratios of FAA/protein and PCA/protein across the stripped SC depth, corresponding to a lower hydration level compared with the normal skin. Our results indicated that filaggrin to FAA hydrolysis occurred deeper in dry skin and at a lower level compared with normal skin. In addition, the mass of proteins stripped from dry skin subjects was significantly higher than that from the normal skin counterpart, especially for the first 10-12 tape strips. CONCLUSION: Our results showed that, compared with normal skin, cosmetic dry skin had a lower NMF level across the SC depth and a lower cohesivity. Such results suggested that NMF levels are critical for cosmetic skin conditions, and lower NMF production may be associated with dry skin. In addition, the NMF level and its depth profile and the SC cohesivity are useful clinical end points for assessing the efficacy of dry skin treatments such as skin moisturizers.

Characteristic differences in barrier and hygroscopic properties between normal and cosmetic dry skin. I. Enhanced barrier analysis with sequential tape-stripping.

OBJECTIVE: Cosmetic dry skin often has a lower hydration level but a similar apparent barrier function, as measured by transepidermal water loss (TEWL), than that of the normal skin. To investigate the intrinsic difference in barrier property and moisture-holding ability between the cosmetic dry and normal skin, we developed a new clinical and data analysis procedure based on sequential tape-stripping with TEWL measurement, coupled with chemical analysis for protein and natural moisturizing factors (NMF) in the stratum corneum. METHODS: A clinical study consisting of 64 healthy Caucasian female subjects with normal and cosmetic dry skin was conducted according to our clinical and data collection protocols. After the baseline visual dryness assessment, 20 tape-strips were placed and removed on each test site using D-Squame tapes. TEWL was measured at baseline and after the 5th, 10th, 15th and 20th tape-strips. All tapes were analysed for protein mass via chemical extraction and the Pierce BCA protein assay, as well as using an infrared densitometry device SquameScan 850A. The stratum corneum thickness and barrier quality (water transport resistance per thickness of the stratum corneum) were decoupled from the apparent barrier function using the TEWL and protein data. RESULTS: A linear relationship between 1/TEWL and cumulative protein removal was observed for both normal and cosmetic dry skin. However, the slope of the linear relation was significantly steeper for normal skin, and significantly more protein was removed from cosmetic dry skin. The results showed that on average, the barrier quality of the stratum corneum of the normal skin is about 40% higher than that of the dry skin, whereas the stratum corneum of the dry skin is about 30% thicker than that of the normal skin. In addition, the amount of SC removal in sequential tape-stripping is generally non-uniform. CONCLUSION: Our results demonstrated that there are characteristic differences in the barrier property between normal and cosmetic dry skin. In comparison to the normal skin, the stratum corneum of the cosmetic dry skin is considerably thicker, however, with a lower barrier quality. The results also showed that the amount of the SC removal in sequential tape-stripping is generally non-uniform. Therefore, the number of tape strips is not a good indicator for the tape-stripping depth.