Two fluorescent wavelengths, 440(ex)/520(em) nm and 370(ex)/440(em) nm, reflect advanced glycation and oxidation end products in human skin without diabetes.

BACKGROUND: Advanced glycation end products (AGEs) and oxidation products (OPs) play an important role in diabetes complications, aging, and damage from sun exposure. Measurement of skin autofluorescence (SAF) has been promoted as a noninvasive technique to measure skin AGEs, but the actual products quantified are uncertain. We have compared specific SAF measurements with analytically determined AGEs and oxidative biomarkers in skin collagen and determined if these measurements can be correlated with chronological aging and actinic exposure. METHODS: SAF at four excitation (ex)/emission (em) intensities was measured on the upper inner arm ("sun protected") and dorsal forearm ("sun exposed") in 40 subjects without diabetes 20-60 years old. Skin collagen from the same sites was analyzed by liquid chromatography-tandem mass spectrometry for three AGEs-pentosidine, carboxymethyllysine (CML), and carboxyethyllysine (CEL)-and the OP methionine sulfoxide (MetSO). RESULTS: There was poor correlation of AGE-associated fluorescence spectra with AGEs and OP in collagen, with only pentosidine correlating with fluorescence at 370(ex)/440(em) nm. A little-studied SAF (440(ex)/520(em) nm), possibly reflecting elastin cross-links, correlated with all AGEs and OPs. Levels of CML, pentosidine, and MetSO, but not SAF, were significantly higher in sun-exposed skin. These AGEs and OPs, as well as SAF at 370(ex)/440(em) nm and 440(ex)/520(em) nm, increased with chronological aging. CONCLUSIONS: SAF measurements at 370(ex)/440(em) nm and 335(ex)/385(em) nm, except for pentosidine, which correlated with fluorescence at 370(ex)/440(em), correlate poorly with glycated and oxidatively modified protein in human skin and do not reflect actinic modification. A new fluorescence measurement (440(ex)/520(em) nm) appears to reflect AGEs and OPs in skin.

Structural and functional differences in barrier properties of African American, Caucasian and East Asian skin.

BACKGROUND: Differences in structural and functional skin characteristics have been linked with ethnical background. But racial differences in skin have not been thoroughly investigated by objective methods and the data are often contradictory. OBJECTIVES: This study was undertaken to compare skin barrier-related parameters of the stratum corneum on African American, Caucasian and East Asian skin by objective measurements. METHODS: Baseline values of trans epidermal water loss were collected on the face. Consecutive stratum corneum D-squame tape strippings were collected on the panelist's ventral forearm and face to evaluate skin barrier strength and cohesion. Stratum corneum ceramides, maturation, measured as the transglutaminase-mediated cross-linking of stratum corneum proteins, and stratum corneum trypsin like enzyme activity were measured on the D-squame tape strippings. RESULTS: East Asian and to some extent Caucasian skin was characterized by low maturation and relatively weak skin barrier. African American skin was characterized by low ceramide levels and high protein cohesion in the uppermost layers of the stratum corneum. These data can be interpreted in terms of the high prevalence of xerosis in black skin and increased skin sensitivity in East Asian skin. CONCLUSION: These results demonstrate that skin properties at the level of the stratum corneum vary considerably among these ethnic groups. This contributes to an improved understanding of physiological differences between these study populations.

Modification of skin discoloration by a topical treatment containing an extract of Dianella ensifolia: a potent antioxidant.

Skin hyperpigmentation, and the reactions that precipitate it, have been linked to free radicals by the fact that free radical scavengers or antioxidants can slow that hyperpigmentation. We have screened several hundred plant extracts for antioxidants and discovered one that is both a strong antioxidant and can reduce skin hyperpigmentation. Extracts of Dianella ensifolia contain 1-(2,4-dihydrophenyl)-3-(2,4-dimethoxy-3-methylphenyl) propane (DP), which was found to inhibit the free radical 1-1-diphenyl-2-picryl-hydrazyl (DPPH) with an EC(50) value of 78 mum. DP was also found to inhibit Ultraviolet (UV)C-induced lipid oxidation with an EC(50) of about 30 mum. We next investigated the effects of this antioxidant on skin hyperpigmentation. The reduction of discoloration by different topical treatments has been assessed in human volunteers using an in vivo assay for the rate of fading of UVB-induced tan. Two pharmaceutical formulas containing 4% hydroquinone (HQ) were used as positive controls, and we tested the ability of DP, a plant-derived amphoteric antioxidant, to increase performance of non-HQ cosmetic formulations. We found that the cosmetic formula containing DP produced an increase in the rate of fading compared to the two pharmaceutical treatments containing HQ.

Calculation of apparent age by linear combination of facial skin parameters: a predictive tool to evaluate the efficacy of cosmetic treatments and to assess the predisposition to accelerated aging.

The estimated apparent age (EAA) was estimated by a panel of trained experts, for the individuals in a cohort. Twelve independent clinical, biophysical and biochemical parameters measured on facial skin, have been identified by multiple regression analysis, which influence the EAA of a person of chronological age (CA) (under eye lines, clinically assessed crow's feet, age spots, clinically evaluated firmness, forehead lines, pores, lip lines, instrumentally evaluated firmness, instrumentally evaluated crow feet, skin texture, in vivo fluorescence related to proliferation and glycation). An algorithm has been devised to obtain the calculated age score (CAS) in a cohort of 452 female volunteers, as CAS(n) = ∑RCiPi(n) (i = 1-13, n = 1-452 and P13 = 1) where the coefficients Ci are obtained by minimizing the difference EAA - CAS, and Pi(n) are the experimental values of the i-th parameter for the n-th volunteer. The determination of CAS before and after a specific cosmetic or pharmacological anti-aging treatment can be used to objectively assess the efficacy of the treatment. The comparison of EAA(n) and of CAS(n) with CA(n) allows one to predict the susceptibility of an individual's face to undergo aging. It has been observed that the biophysical and biochemical parameters play a relevant role in the assessment of the predisposition of skin to undergo accelerated aging.

Antioxidant enzyme activity in human stratum corneum shows seasonal variation with an age-dependent recovery.

The stratum corneum, as the body's principal barrier to the environment, is continuously exposed to environmental sources of reactive oxygen species like ultraviolet light, ozone, and pollution. Reactive oxygen species are believed to be involved in cancer, aging, and inflammatory skin disorders. We have developed a method to measure catalase and superoxide dismutase activity on tape strippings from the human stratum corneum and demonstrated a gradient of antioxidant enzyme activity across the stratum corneum with decreasing levels towards the skin surface. Sun exposure resulted in a seasonal variation of the catalase activity in stratum corneum, with low activities in summer and higher activities in winter for the same person, whereas superoxide dismutase activity in stratum corneum did not seem to vary in those conditions. Exposure of human skin to broadband ultraviolet-A resulted in a dose-dependent deactivation of the catalase activity in stratum corneum within 24 h, whereas exposure to ultraviolet-B had no effect. Superoxide dismutase activity in stratum corneum was not affected by ultraviolet-A or ultraviolet-B irradiation within 24 h. After exposure to a dose of 15 J per cm2 broadband ultraviolet-A, full recovery of the catalase activity occurred in 3-4 wk at an age-dependent rate. We conclude that sun exposure results in a disturbed catalase to superoxide dismutase ratio in the stratum corneum. This may lead to an increased vulnerability to oxidative damage in stratum corneum barrier components. These results therefore stress the importance of providing efficient protection for this internal defense mechanism in sun-exposed areas of the skin.