The present study examines the cumulative effects of sub-erythema application of squalene-monohydroperoxide (Sq-OOH), the initial products of UV-peroxidated squalene, to the skin of hairless mice. Sq-OOH was isolated by the methanol extraction and preparative HPLC method. Repeated topical application of 10 mM Sq-OOH to hairless mice for 3 weeks induced definite skin roughness and crinkle formation. 3-D surface parameter analysis revealed changes in all roughness parameters (number of furrows and crests, distance between a furrow and next crest, and irregularity) of the group treated with more than 3 mM Sq-OOH compared to the control group. These skin surface changes were not induced by squalene, squalene-monohydroxide (Sq-OH) or organic hydroperoxides such as tert-butyl hydroperoxide and cumene-hydroperoxide at 10 mM. Similarly, such changes were not induced by primary irritants, such as sodium lauryl sulfate and n-tetradecane under the same experimental conditions. Skin conductance decreased, following application of 10 mM Sq-OOH. Histological observation revealed that application of 10 mM Sq-OOH induced slight hyperkeratosis, moderate epidermal thickening and slight hyperplasia of sebaceous glands.

We have studied the effect of squalene monohydroperoxides (Sq-OOH), initial products of UV-peroxidated squalene, on the skin of hairless mice. Repeated topical application of 10 mM Sq-OOH to hairless mice for 15 weeks induced definite skin wrinkling. When image analysis was used to compare wrinkle formation induced by ultraviolet B (UVB) irradiation and Sq-OOH treatment, the degree of wrinkling in exposed skin was seen to be similar. However, the characteristics of wrinkles induced by either method differed markedly with regard to direction and distribution. Biochemical analysis revealed a significant decrease in collagen content per unit area and mass in Sq-OOH-treated skin, whereas no changes per unit area and decrease in collagen per unit mass were observed in UVB-irradiated skin. As for glycosaminoglycan (GAG) content per unit area, significant increases were observed in both Sq-OOH-treated skin and UVB-irradiated skin. These changes were not induced by organic hydroperoxides such as tert-butylhydroperoxide or cumene hydroperoxide treatment. Histological observation revealed epidermal hyperplasia and dermal alterations such as collagen degradation and GAG increases in Sq-OOH-treated skin. Histological changes induced by Sq-OOH were not as pronounced as those induced by UVB irradiation. These results clearly suggest that the wrinkling and changes in dermal collagen content induced by Sq-OOH are qualitatively different to those induced by UVB exposure. This may provide a useful model for the study of skin aging, particularly with regard to collagen content.

A double-blind, placebo-controlled, randomized trial was conducted to investigate the beneficial effect of probiotic and prebiotic fermented milk on the skin of healthy adult women. Forty healthy Japanese adult female volunteers with healthy skin randomly received either a bottle of probiotic and prebiotic fermented milk containing Bifidobacterium breve strain Yakult and galacto-oligosaccharides (GOS) (active group) or a non-fermented placebo milk containing neither probiotics nor GOS (placebo group) daily for 4 weeks. Before and after intake, hydration levels and cathepsin L-like activity in the stratum corneum and phenol levels in the serum and urine were determined. After intake, the hydration level of the stratum corneum decreased significantly in only the placebo group and was significantly lower than in the active group (p=0.031). Cathepsin L-like activity, an indicator of keratinocyte differentiation, was significantly increased in the active group (p=0.027). Serum and urine phenol levels decreased significantly in the active group (p=0.014, p=0.002, respectively), and serum phenol levels were significantly lower in the active group compared with the placebo group (p=0.006). The consecutive intake of probiotic and prebiotic fermented milk can benefit skin condition without dryness and decrease the levels of phenol production by gut bacteria in healthy adult women.

Recent studies have shown that some probiotics affect not only the gut but also the skin. However, the effects of probiotics on ultraviolet (UV)-induced skin damage are poorly understood. In this study, we aim to examine whether oral administration of live Bifidobacterium breve strain Yakult (BBY), a typical probiotic, can attenuate skin barrier perturbation caused by UV and reactive oxygen species (ROS) in hairless mice. The mice were orally supplemented with a vehicle only or BBY once a day for nine successive days. Mouse dorsal skin was irradiated with UV from days 6 to 9. The day after the final irradiation, the transepidermal water loss (TEWL), stratum corneum hydration, and oxidation-related factors of the skin were evaluated. We elucidated that BBY prevented the UV-induced increase in TEWL and decrease in stratum corneum hydration. In addition, BBY significantly suppressed the UV-induced increase in hydrogen peroxide levels, oxidation of proteins and lipids, and xanthine oxidase activity in the skin. Conversely, antioxidant capacity did not change regardless of whether BBY was administered or not. In parameters we evaluated, there was a positive correlation between the increase in TEWL and the oxidation levels of proteins and lipids. Our results suggest that oral administration of BBY attenuates UV-induced barrier perturbation and oxidative stress of the skin, and this antioxidative effect is not attributed to enhancement of antioxidant capacity but to the prevention of ROS generation.

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