Interaction of mineral salts with the skin: a literature survey.

There is growing scientific evidence that the health, well-being and the attractiveness of the skin are strongly influenced by nutrition. Consumers recognize this and have supported the creation of a global cosmeceuticals market estimated in 2010 at $27.2 billion. Early in 2011, the US Department of Health and Human Services and Department of Agriculture issued the Dietary Guidelines for Americans, 2010. Twelve vitamins and nine minerals were recognized as essential. The minerals include calcium, copper, iron, magnesium, phosphorus, selenium, zinc, potassium and sodium. Although the topical benefits of several minerals such as zinc, magnesium and iron are recognized and, in some cases, approved by the FDA, the topical benefits of the others to the skin are largely unexplored and unexploited. This review attempts to summarize what has been published in the literature on the interactions of the eight of the nine essential elements with the skin.

Grey hair: clinical investigation into changes in hair fibres with loss of pigmentation in a photoprotected population.

Loss of pigmentation in hair fibres is one of the most obvious phenotypic changes with ageing and has been a topic of increasing interest in the study of follicle biology. The onset of greying brings cosmetic complaints that grey fibres are wild or difficult to manage. Of course, these perceptions may be the consequence of visual obviousness rather than underlying physical or chemical differences. Although several studies have compared pigmented and unpigmented fibres, few have tried to control genetic and ethnic difference as well as extrinsic factors such as photoexposure and chemical treatment. We have recruited subjects with salt-and-pepper hair from a population of Old Order Mennonites who, for cultural reasons, are not only prohibited from chemically treating their hair but also limit their exposure to sunlight. Hair samples were examined for elemental composition, surface energy, Young's modulus, break stress, bending modulus, shear modulus and water sorption/desorption isotherm. The parameters were evaluated statistically for global differences, individual differences and typical individual differences. Consistent with previous published literature, few global differences were found between pigmented and unpigmented hair across the population. We do find that many individual subjects had differences between pigmented and unpigmented fibres. These differences tend to be more pronounced in bulk than in surface properties. The small differences in mechanical properties and moisture uptake and loss lend support to the perception by consumers that grey hair is wilder, drier and less manageable.

RAPA: a novel in vitro method to evaluate anti-bacterial skin cleansing products.

Development of efficacious anti-bacterial skin cleansing products has been limited by the availability of a pre-clinical (in vitro) method to predict clinical efficacy adequately. We report a simple and rapid method, designated as rapid agar plate assay (RAPA), that uses the bacteriological agar surface as a surrogate substrate for skin and combines elements of two widely used in vivo (clinical) methods (Agar Patch and Cup Scrub). To simulate the washing of the human hand or forearm skin with the test product, trypticase soy agar plates were directly washed with the test product and rinsed under running tap water. After air-drying the washed plates, test bacteria (Staphylococcus aureus or Escherichia coli) were applied and the plates were incubated at 37 degrees C for 18-24 h. Using S. aureus as the test organism, anti-bacterial bar soap containing triclocarbanilide showed a strong linear relationship (R(2) = 0.97) between bacterial dose and their per cent reduction. A similar dose-response relationship (R(2) = 0.96) was observed for anti-bacterial liquid hand soap against E. coli. RAPA was able to distinguish between anti-bacterial products based on the nature and level of actives in them. In limited comparative tests, results obtained by RAPA were comparable with the results obtained by clinical agar patch and clinical cup scrub methods. In conclusion, RAPA provides a simple, rugged and reproducible in vitro method for testing the relative efficacy of anti-bacterial skin cleansing products with a likelihood of comparable clinical efficacy. Further testing is warranted to improve the clinical predictability of this method.

Fluorescence and coloration of grey hair.

Grey hair samples were collected from 11 individuals and separated into un-pigmented and pigmented fibres (International Hair Importers). Fluorescence measurements were obtained by using a double-grating fluorescence spectrophotometer and a bifurcated fibre optics accessory to measure the spectra directly from the surface of hair at various distances from the fibre root. Colour measurements were carried out by using a Hunter colorimeter. The fluorescence spectra of un-pigmented hair obtained by the excitation at 290 nm show a peak at 356 nm [tryptophan (Trp)], and multi-peak emissions in the range from 395 to 500 nm. A significant variation in the Trp emission intensity at 356 nm vs. the intensity of emission in the 395-500 nm range was observed for hair collected from various individuals with yellow coloured hair producing stronger relative emission in 395-500 nm range. Quantitative measurements of coloration and the calculation of the Yellowness Index (YI) showed linear correlation between YI and the ratio of fluorescence intensities I(440)/I(356) The spectra obtained by excitation at 320 nm showed the emission peaks at 395 nm (unidentified), 420 nm (N-formylkynurenine), 460 nm (kynurenine), and 495 nm (3-hydroxykynurenine), which are the products of oxidative or metabolic conversion of tryptophan. Un-pigmented, yellow hair showed a build-up of the fluorescence band corresponding to 3-hydroxykynurenine at 495 nm. The data also showed the fluorescence quenching effect of melanin resulting in the lowering of the fluorescence intensity of pigmented hair. The spectra obtained at various positions along the fibres demonstrated gradual photo-decomposition of hair chromophores during their lifetimes. This was indicated by a decrease of Trp fluorescence intensity, which was relatively fast (8.10(-4)-1.5.10(-3) [day(-1)] as calculated for hair obtained from various individuals) for un-pigmented hair and slower for pigmented hair. A decrease in Trp emission was accompanied by an increase in the yellow coloration toward the ends of un-pigmented fibres.

Confocal Raman microspectroscopy of stratum corneum: a pre-clinical validation study.

Skin moisturization is not only important for maintaining skin functional properties but also has great impact on the skin's aesthetic properties. The top layer of the skin, the stratum corneum (SC), plays a key role in protecting and preventing against external aggressions as well as in regulating water flux in and out. Confocal Raman microspectroscopy is the first commercially available technique that provides a non-invasive, in vivo method to determine depth profiles of water concentration in the skin, however, in this case it was applied in an in vitro setting. As the first phase of validating the usefulness of confocal Raman microspectroscopy, we used porcine skin as a surrogate for human skin. Water concentration profiles were obtained using confocal Raman microspectroscopy from isolated pigskin SC and compared with that using the Karl Fischer titration method. The two methods correlated very well with a regression coefficient of 1.07 as well as a correlation coefficient, R(2) = 0.989, which demonstrated the consistency and accuracy of confocal Raman microspectroscopy for water concentration determination. To evaluate the instrument's response to different skin care/cleansing products, a wide range of products were tested to compare their skin moisturization ability. Among those tested were a lotion, commercial soap bar, syndet bar, traditional non-emollient shower gel (water, Sodium Laureth Ether Sulfate (SLES), cocamidopropyl betaine system) and emollient containing shower gel (water, sunflower oil, SLES, cocamidopropyl betaine, glycerin, petrolatum). The results were consistent with what was expected. The water content on skin treated with (A) lotion was significantly higher than the non-treated control; (B) syndet bar-treated skin had a significantly higher water content than soap-based bar-treated sites; (C) non-emollient shower gel washed sites were more moisturized than soap-based bar-treated samples; and (D) emollient shower gel-treated skin was significantly more hydrated than non-emollient shower gel washed skin. The unique and direct quantitative water content information provided by confocal Raman microspectroscopy offers a whole new perspective for fundamental skin moisturization studies and will play an important role in evaluating moisturizing profiles and the hydration potential of products designed for personal care in the cosmetic industry.

Effects of environmentally realistic levels of ozone on stratum corneum function.

The impact of troposphere ozone (O(3)), the major oxidant in photochemical smog, on the overall wellbeing of skin is of considerable interest. To date, limited information is available on the impact of O(3) on human skin. Using a specially designed O(3) exposure chamber, we provide the first evidence that exposure of human skin to O(3) (0.8 ppm, 2-h time-weighted average) significantly reduced vitamin E by 70% and concomitantly increased lipid hydroperoxides by 2.3 fold in the superficial stratum corneum (SC). Although the dose of O(3) used here reduced the resident microflora population by 50% and created a state of oxidative stress within the SC, it did not affect several key enzymes involved in SC homeostasis including the redox-sensitive transglutaminase and the SC tryptic (KLK5) and chymotryptic (KLK7) proteases. Importantly, no signs of skin dryness or erythema were observed. We hypothesize that the limited effects of low doses of O(3) on SC function is attributable to several factors including: (i) protection provided by the anti-oxidant defence system; (ii) inability of O(3) to penetrate the SC; and (iii) limited water available to catalyse the Criegee reaction. Although chronic exposure to O(3) may produce a different outcome than that reported here, our data suggest that exposure to environmentally relevant doses of O(3), at best, induces a moderate state of oxidative stress, without producing a visible clinical response. In our opinion, exposure of skin to UV radiation is a much more significant threat than exposure to ground-level O(3).

Lectin-receptor interactions in liposomes. II. Interaction of wheat germ agglutinin with phosphatidylcholine liposomes containing incorporated monosialoganglioside.

Bovine brain gangliosides incorporated into phospholipid liposomes provide receptors for wheat germ agglutinin. Purified monosialogangliosides were mixed with egg phosphatidylcholine, and unilamellar liposomes were generated. Addition of wheat germ agglutinin induced the liposomes to fuse, and gel filtration analysis revealed that the lectin was incorporated into the fused liposomes. The fusion process was studied by following the changes in the 90 degrees light scattering. Increasing the proportion of the monosialoganglioside in the liposomes was found to increase both the extent of the lectin-induced liposome fusion and the rate of the reaction; below a threshold of approx. 5 mol%, the process was extremely slow. The increase in light scattering could be prevented by the addition of the hapten inhibitor, N-acetyl-D-glucosamine (1 mM). Addition of the inhibitor, subsequent to the lectin, caused a partial decrease in light scattering due to the dissociation of unfused vesicle aggregates. Electron microscopic examination revealed that the ganglioside-containing liposomes were vesicles, 244 +/- 25 A (S.D.) in diameter. Upon addition of wheat germ agglutinin, the vesicles appeared to fuse to form larger vesicles, corresponding to dimers and trimers of the initial vesicles. Inhibition studies with a variety of monosaccharides indicated that the sialic acid moieties present in the ganglioside acted as the lectin-receptor sites. This was confirmed by the observation that wheat germ agglutinin did not interact with phosphatidylcholine vesicles containing desialyated ganglioside.

Permeability of Novikoff hepatoma cells to water and monohydric alcohols.

The permeability coefficients of Novikoff hepatoma ascites cell membranes for tritiated water (3HHO) and for a homologous series of monohydric alcohols (methanol through hexanol) were deduced from linear diffusion coefficients by means of a series-parallel pathway model (Redwood et al. (1974) J. Gen. Physiol. 64, 706-729). Membrane permeability coefficients for 3HHO at 20, 30 and 37 degrees C were (all x 10(-5)) 97, 125, and 163 cm . s-1, respectively, and were significantly smaller than the corresponding values for the alcohols tested. In the alcohols series, ethanol had the lowest permeability coefficient 198 x 10(-5) cm . s-1 at 20 degrees C. The apparent activation energy for water permeation was 6.7 +/- 1.9 S.E. kcal . mol-1. The apparent membrane diffusion coefficients for the alcohols were a complex function of molecular properties with less diffusional membrane resistance to the alcohols in the middle of the homologous series than would have been expected on the basis of oil-water partitioning or molar volume considerations. The conventional parallel aqueous lipophilic pathway model is not consistent with the present data which can be interpreted by consideration of parallel lipophilic pathways through the Novikoff hepatoma cell membrane.

Osmotic permeability of Novikoff hepatoma cells.

The osmotic permeability coefficient (Pf) for water movement across Novikoff hepatoma cells was found to be 82 +/- 3 (S.E.) x 10(-5) cm . s-1 at 20 degrees C. The corresponding diffusional permeability coefficient for 3HHO (Pd) was 97 +/- 10 (S.E.) . 10(-5) cm . s-1, therefore the ratio Pf/Pd is close to unity. The apparent activation energy for water filtration was 10.4 +/- 0.4 (S.E.) kcal . mol-1. This value is significantly greater than the activation energy for the self diffusion of water. The product of the hydraulic permeability coefficient and the viscosity coefficient for water was temperature-dependent. However, the product of the hydraulic permeability coefficient and the viscosity coefficient for membrane lipid did not vary with temperature. These data are interpreted as evidence for water movement across a lipid membrane barrier rather than through aqueous channels.

Ozone-exposure depletes vitamin E and induces lipid peroxidation in murine stratum corneum.

The presence of ozone (O(3)) in photochemical smog is an important health concern. We hypothesized that the stratum corneum (SC), as the outermost skin layer and the permeability barrier of the skin, represents a sensitive target for O(3)-induced oxidative stress. To test this hypothesis, SKH-1 hairless mice were anesthetized and exposed for 2 h to O(3) by using two strategies: (i) single exposures to 0 (n = 12), 1 (n = 4), 5 (n = 4), and 10 (n = 4) ppm; and (ii) repeated daily exposures to 0 ppm (controls; n = 4) and 1 ppm (n = 4) for six consecutive days. New techniques based on the removal of SC by tape stripping were used to analyze the biologic effects of O(3) with respect to vitamin E depletion and lipid peroxidation. SC tissue was extracted from the tape and immediately analyzed by HPLC for vitamin E and malondialdehyde (MDA) concentrations. After in vivo exposure to increasing O(3) doses, vitamin E was depleted and MDA formation was increased, both in a dose-dependent manner. Remarkably, repeated low-level O(3) exposures resulted in cumulative oxidative effects in the SC: As compared with O(3) exposures of 0 ppm (alpha-tocopherol, 8.95 +/- 1.3 pmol per mg; gamma-tocopherol, 3.00 +/- 0.3 pmol per mg; MDA, 3.69 +/- 0.3 pmol per mg), vitamin E was depleted (alpha-tocopherol, 2.90 +/- 0.6 pmol per mg, p < 0.001; gamma-tocopherol, 0.5 +/- 0.1 pmol per mg, p < 0.001) and MDA levels were increased (4.5 +/- 0.2; p < 0.01). This report demonstrates the unique susceptibility of the SC to oxidative damage upon exposure to O(3).

Assessment of the effects of dentifrices on plaque acidogenesis via intra-oral measurement of plaque acids.

Dionex ion chromatography was used for identification and quantification of the plaque acids (formate, lactate, acetate, and propionate) formed in vivo following a sucrose rinse. Consistent with other literature reports, lactate and acetate were the predominant acid species generated by plaque. This micro-analytical technique was used for investigation of the effects of dentifrice formulations containing NaF, xylitol, or NaF/xylitol on plaque acid metabolism. Neither a single nor multiple exposures (three times daily for one week) to the experimental dentifrices significantly altered the profiles of the individual acids or total plaque acidity relative to the placebo dentifrice. The lack of an effect of fluoride and xylitol on plaque acid metabolism is attributed to the variability of dental plaque and the low concentrations of active ingredients reaching the plaque. However, we believe that, with further improvement in sampling technique and study design, this approach may prove useful for the study of the effects of active ingredients (i.e., anticaries agents) on plaque acidogenesis.

Vitamin E delivery to human skin by a rinse-off product: penetration of alpha-tocopherol versus wash-out effects of skin surface lipids.

alpha-Tocopherol, the major biologically active form of vitamin E, represents a frequently added lipophilic compound of skin care products. Despite its emerging use in rinse-off formulations, little is known on its efficacy with respect to its deposition or its antioxidant potential in human skin. The objective of this study was to investigate whether the single use of an alpha-tocopherol-enriched rinse-off product provides effective deposition of alpha-tocopherol on human stratum corneum. To test this, forearm skin of 13 volunteers was washed either with an alpha-tocopherol-enriched rinse-off product (test product, TP) or with an alpha-tocopherol-free vehicle control (control product, CP) (contralateral arm) using a standardized wash protocol. Thereafter, skin surface lipids were extracted with pure ethanol after the wash procedure as well as after 24 h. Additionally, one group of volunteers was subjected to irradiation of their forearms with low-dose UVA (8 J/cm(2)) prior to lipid extraction. Skin lipid extracts were analyzed by high performance liquid chromatography using electrochemical detection for vitamin E and UV detection for squalene (SQ) and squalene monohydroperoxide. The results of this in vivo study demonstrated that (1) while CP treatment lowers, TP treatment strongly increases alpha-tocopherol levels of skin barrier lipids; (2) increased vitamin E deposition levels were maintained for a period of at least 24 h, and (3) TP treatment significantly inhibited photooxidation of SQ. In conclusion, the use of alpha-tocopherol-enriched rinse-off products may help to maintain the integrity of the skin barrier by providing protection against photooxidative stress at the level of skin surface lipids.

Water permeability of alveolar macrophages.

The hydraulic conductivity coefficient (Lp) of alveolar macrophages, recovered by lavage from dog lungs, was determined by following volume changes induced by changes of nonpermeating solute concentrations of suspending fluid as a function of time at 20 degrees C. The volume changes were monitored as changes in absorbance of the suspended cells at 600 nm. Cell surface area was calculated from cell volume and diameter. Linear relationships between cell volume and solution osmolality changes were found over the range of 320-520 mosmol/kg; beyond these ranges the macrophages did not respond with swelling or shrinking. Lp and the filtration coefficient (Pf) were calculated from the total volume change over time. At 20 degrees C these were, respectively, 15.7 X 10(-10) cm X cmH2O-1 X s-1 and 217 X 10(-5) cm/s. Comparison of Pf and the diffusional permeability coefficient (Pd) for water of 70 X 10(-5) cm/s, yields a Pf-to-Pd ratio of 3.1. The hypothesis of water passage through aqueous membrane pores is compatible with these data. However, diffusion of water in the glycocalyx of the pericellular domain could be restricted. Pd would then be underestimated, and a falsely high ratio would be calculated. We have no evidence to support this possibility.

The effects of prolonged use of surfactants on the skin of normal and photo-exposed hairless mice.

Laboratory tests to assess the irritant potential of materials, such as skin cleansers, which are normally used over a long period by humans, fail to mimic actual use. Most washing tests last a few days or at most a few weeks. Skin sites and techniques are often not standardized. The more standardized patch test involves occlusion and results in exaggerated reactions, since even water and blank patches produce visible and pathophysiologic changes. All of these tests rely on visual assessment despite strong evidence that similarly appearing skin can be very different histologically. The primary objective of this study was to use a well-defined animal model to evaluate the cumulative effects of repeated skin exposure to low levels of surfactants of varying skin irritation potential. A secondary aim was to examine whether or not surfactant-induced skin changes were exacerbated by suberythemal UV radiation. Test materials were applied topically, 2x daily to the dorsal areas of normal and low-dose solar simulator exposed mice for 15 weeks. Our results show that, with conditions mimicking typical normal use, these surfactants and skin cleansers produce little or very mild histological changes in the skin. UV irradiation alone produced the greatest change in all histological parameters examined, with no synergistic or additive effects with the topical treatments.

Solute-excluded volumes near the Novikoff cell surface.

A differential centrifugation technique, in which all extracellular water except that intimately associated with the cell (pericellular domain) is removed, has been applied to isolated Novikoff hepatoma cells. The pericellular volumes accessible to albumin, inulin, raffinose, and sucrose were inversely related to the molecular weights of the test solutes. This phenomenon was not detectable in erythrocytes or in fat cells. Selective removal of cell surface components by enzymatic treatment produced proportional changes in the relative volumes of distribution accessible to the solutes. This discrimination in the volume accessible to each of the solutes is analogous to that obtained in gel chromatographic separation and represents, in effect, excluded volumes which are inversely related to solute size. This exclusion is associated with components of the Novikoff cell surface, including the surface coat and the microvilli that cover the Novikoff cell. These structures provide an additional level of discrimination for the Novikoff cell not seen in certain other cell types.