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Antioxidant property of fullerene is effective in skin whitening
DERMATOPHARMACOLOGY/COSMECEUTICALS P1600 Antioxidant property of fullerene is effective in skin whitening Takahiro Fujimoto, MD, PhD, MBA, Clinic F, Chiyoda-ku, Tokyo, Japan; Hisae Aoshima, Vitamin C60 BioResearch, Chuo-Ku, Tokyo, Japan; Ken Kokubo, PhD, Osaka University, Suita, Osaka, Japan; Masayuki Ito, Vitamin C60 BioResearch, Chuo-Ku, Tokyo, Japan; Nobuhiko Miwa, PhD, Prefectural University of Hiroshima, Shobara, Hiroshima, Japan Ultraviolet (UV) rays generate reactive oxygen species, which can cause a series of biologic effects in human skin cells, resulting in cosmetic skin damage such as pigmentation. Antioxidants have been reported to inhibit the progression of UVinduced skin damage. Fullerene, a carbon allotrope, is characterized as an antioxidant and is reported to react with various reactive chemical species, such as free radicals. Because of certain properties of fullerene, such as poor water solubility and stability, the cosmetic formulation of fullerene presents some technical and formulating challenges. Therefore, we used polyvinylpyrrolidone (PVP), a water soluble polymer commonly used in cosmetic products, as the physical trapping material. PVP-wrapped fullerene was found to be a water soluble and very stable cosmetic ingredient. In this investigation, we extensively evaluated the antioxidant ability and whitening effects of PVPefullerene in vitro by testing its formulated sample in our study subjects. We aimed to verify the performance of PVPefullerene and to compare it with that of other major antioxidants; hence, we used a simple and conventional method that employs beta-carotene as an indicator to assess antioxidant ability of fullerene. The results indicated that PVPefullerene can inhibit radicals, such as liporadicals, hydroxyl radicals, and superoxide radicals. The ability of PVPefullerene to inhibit ultraviolet A (UVA)-induced melanogenesis was tested in human melanocytes. Cells were treated with or without PVPefullerene, arbutin, and ascorbic acid for 3hours and irradiated by weak UVA rays for 24hours. PVPefullerene significantly reduced UVA-induced melanogenesis in a dose dependent manner, and melanogenesis was reduced to a larger extent in PVPefullerenetreated cells than in ascorbic acide and arbutin-treated cells. To prove the practical effectiveness of PVPefullerene, clinical tests were performed on 32 women volunteers. A gel containing PVPefullerene (C60, 2ppm) was prepared and applied on the face twice daily. Whitening efficiency was evaluated by L* value measurement. After 6 weeks of application, 94% subjects became fairer than before treatment without any inflammation or irritation. These data suggest that PVPefullerene has a whitening effect because of its ability to scavenge UV-induced free radicals.
P1602 A new body wash provides all-day protection against UV light Sergio Nacht, PhD, Riley-Nacht, Las Vegas, NV, United States; Clay J. Cockerell, MD, Cockerell and Associates Dermatopathology, Dallas, TX, United States Introduction: The incidence of skin cancer continues to increase and is reaching epidemic proportions. In spite of knowing that UV exposure can lead to skin cancer, most individuals either do not routinely apply sunscreens before sun exposure or they do it deficiently. Skin cancers also pose a significant public health problem that amounts to billions of dollars per year in the United States alone. An SPF 5 sunscreen reduces UV exposure by 80%. The daily application of such a sunscreen starting at age 10 could reduce the overall life UV exposure by 65% and, presumably, the incidence of skin cancer by at least the same percentage. Objective: To develop a body cleansing product to be incorporated in a daily routine containing sunscreens in a formulation substantive to the skin to provide an SPF of at least 8 following rinsing. The product contains zinc oxide, titanium dioxide, octocrylene, and red petrolatum in a cleansing vehicle with natural melanin to provide antioxidant and free radical quenching properties to further protect skin cell structures. Two product forms were developed: a body lotion and an aerosol ‘‘mousse.’’ Methods: The SPF of all formulations was determined according to the guidelines in the FDA OTC Tentative Final Monograph for sunscreen products. This document does not provide a procedure for cleansing products; therefore, a suitable in vivo test protocol was developed. Two 50cm2 areas were defined on the lower back of each subject. The SPF as a sunscreen was determined on one site using a dosage of 2mg/cm2. The other site was wetted with plain water and the same amount of product was applied. Product was rubbed in for 1minute followed by rinsing with water at about 328C. The area was patted dry and SPF was determined. ‘‘Real life’’ studies were conducted by asking volunteers to use the product while they showered daily. Results: The lotion has an SPF 21 as a sunscreen and an SPF 19 as a wash while the ‘‘mousse’’ gave SPFs of 20 and 19, respectively. When used in the shower, the SPF immediately after usage were 20 and 19 for the lotion and mousse and when retested 8hours later, the SPFs were 19 and 18 respectively. Finally, when the products were used in the shower for 5 days, the SPF at day 5 was similar to those determined without washing. Conclusions: We developed a product that could be used as a daily shower cleanser and provide significant long-lasting UV protection. Commercial support: Riley-Nacht.
Commercial support: None identified.
P1603 P1601 Antiitching properties of patented avocado peptides Stephanie Bredif, Laboratoires Expanscience, Epernon, Eure Et Loir, France; Caroline Baudouin, PhD, Laboratoires Expanscience, Epernon, Eure Et Loir, France; Philippe Msika, PhD, Laboratoires Expanscience, Epernon, Eure Et Loir, France Objectives: Itch is defined as an unpleasant sensation evoking the desire to scratch. Cross-talk between C neuron terminals and the spatially closely related dermal mast cells seems to play an important part in the pruritus pathogenesis. Indeed, mast cell mediators, such as histamine and tryptase (via proteinase-activated receptor 2 [PAR2]) are able to stimulate C neuron terminals, therefore inducing the sensation of itch. Moreover, PAR2 is also expressed by keratinocytes, where its activation initiates inflammatory response. Although the itch pathogenesis in dry skin is not fully understood, the clinical association between itch and dry skin is well established. In this study, we investigated the antiitching and moisturizing properties of a patented natural extract, avocado peptides (AP). Methods: Mast cells were treated with calcium ionophore or substance P in order to induce the release of tryptase and histamine, respectively, measured according to spectrophotometric and spectrofluorometric methods. Gene expression of PAR2 was analyzed in a reconstructed human epidermis model using DNA microarrays. Epidermal lipid neosynthesis was followed in reconstructed human epidermis thanks to lipids radiolabeling and thin layer chromatography analysis. Hyaluronic acid release by normal human epidermal keratinocytes (NHEK) was measured by ELISA. Glycosaminoglycans (GAGs) synthesis by NHEK was evaluated by measuring incorporation of 35S-sulfate. Results: Avocado peptides (AP) were able to significantly inhibit the release of histamine (up to -53%; P \.01) and tryptase (-30%; P \.01) by mast cells. Epidermal gene expression of PAR2 was reduced by 52% after treatment with AP. AP positively affected epidermal ceramides and free fatty acid synthesis. Finally, AP significantly enhanced NHEK production of hyaluronic acid and GAGs by 31% (P \.05) and 34% (P \.05), respectively.
Effects of galactomyces ferment filtrate on epidermal barrier marker caspase-14 in human skin cells Kenji Hattori, PhD, MS, Keio University Faculty of Pharmacy, Tokyo, Japan; Akiko Nakajima, MS, Procter & Gamble Japan, Kobe, Hougo, Japan; Akira Date, PhD, MS, MPH, Procter & Gamble Japan, Kobe, Hyougo, Japan; Hiroomi Tamura, PhD, MS, Keio University Faculty of Parmacy, Tokyo, Japan; Kyoko Ishida, MS, Procter & Gamble Japan, Kobe, Hougo, Japan Background: The stratum corneum of human epidermis is composed of terminally differentiated keratinocytes serving as an essential barrier to environmental stresses, such as UV-induced photodamage and water loss. The regulatory and proteolytic events that coordinate barrier formation are tightly controlled. Caspase-14 belongs to a conserved family of aspartate-specific proteases. Its epidermal expression is restricted almost exclusively to the suprabasal layers, implicating this protease in keratinocyte terminal differentiation and cornification. Therefore, caspase-14 is a useful biomarker to monitor formation and homeostasis of the stratum corneum barrier. Objective: Determine the in vitro effects on barrier formation and hydration of topically applied galactomyces ferment filtrate (GFF) via expression of epidermal late differentiation biomarkers (caspase-14 and related genes). Methods: In vitro human skin models, including skin keratinocytes and skin equivalents (SE) with partially (EPI-201) or completely formed (EPI-200) stratified, cornified epidermis were treated topically with GFF or dexamethasone (Dex). Caspase-14 and related barrier biomarkers were assayed via enzyme activity (caspase-14), mRNA expression by RT-PCR and protein levels by Western blot analysis. Histologic evaluation was conducted also in the human SE models. Results: GFF increased expression of caspase-14 and peptidylarginine deiminases (PAD1, PAD3) in human SE cultures (EPI-200 and -201) and also increased the expression of transglutaminase (TGM1) and involucrin in a human SE model of earlier stage corneocyte differentiation (EPI-201). Dex increased only the expression of caspase-14 in both human SE models. RU-486, a glucocorticoid receptor antagonist, inhibited the effects of Dex but not GFF, suggesting independent pathways such as MAPK may be involved.
Conclusion: The antiitching activities of these patented avocado peptides have been shown on mast cells, which are at the heart of pruritus reaction. Moreover, its moisturizing and relipidizing properties were suggested by in vitro assays. AP could therefore be of particular interest in the management of cutaneous disorders associated with itchy and dry skin.
Conclusions: These results indicate that GFF increases caspase-14 expression by epidermal cells, specifically during late stage differentiation. The positive effect of GFF on late differentiation biomarkers supports previous observations that GFFcontaining moisturizers help protect the skin against damage, such as dryness, from environmental stress. These results provide a compelling reason to further understand the nature of GFF and its skin benefits.
Commercial support: None identified.
Commercial support: None identified.
AB54
J AM ACAD DERMATOL
MARCH 2010
P1602 A new body wash provides all-day protection against UV light Sergio Nacht, PhD, Riley-Nacht, Las Vegas, NV, United States; Clay J. Cockerell, MD, Cockerell and Associates Dermatopathology, Dallas, TX, United States Introduction: The incidence of skin cancer continues to increase and is reaching epidemic proportions. In spite of knowing that UV exposure can lead to skin cancer, most individuals either do not routinely apply sunscreens before sun exposure or they do it deficiently. Skin cancers also pose a significant public health problem that amounts to billions of dollars per year in the United States alone. An SPF 5 sunscreen reduces UV exposure by 80%. The daily application of such a sunscreen starting at age 10 could reduce the overall life UV exposure by 65% and, presumably, the incidence of skin cancer by at least the same percentage. Objective: To develop a body cleansing product to be incorporated in a daily routine containing sunscreens in a formulation substantive to the skin to provide an SPF of at least 8 following rinsing. The product contains zinc oxide, titanium dioxide, octocrylene, and red petrolatum in a cleansing vehicle with natural melanin to provide antioxidant and free radical quenching properties to further protect skin cell structures. Two product forms were developed: a body lotion and an aerosol ‘‘mousse.’’ Methods: The SPF of all formulations was determined according to the guidelines in the FDA OTC Tentative Final Monograph for sunscreen products. This document does not provide a procedure for cleansing products; therefore, a suitable in vivo test protocol was developed. Two 50cm2 areas were defined on the lower back of each subject. The SPF as a sunscreen was determined on one site using a dosage of 2mg/cm2. The other site was wetted with plain water and the same amount of product was applied. Product was rubbed in for 1minute followed by rinsing with water at about 328C. The area was patted dry and SPF was determined. ‘‘Real life’’ studies were conducted by asking volunteers to use the product while they showered daily. Results: The lotion has an SPF 21 as a sunscreen and an SPF 19 as a wash while the ‘‘mousse’’ gave SPFs of 20 and 19, respectively. When used in the shower, the SPF immediately after usage were 20 and 19 for the lotion and mousse and when retested 8hours later, the SPFs were 19 and 18 respectively. Finally, when the products were used in the shower for 5 days, the SPF at day 5 was similar to those determined without washing. Conclusions: We developed a product that could be used as a daily shower cleanser and provide significant long-lasting UV protection. Commercial support: Riley-Nacht.
Commercial support: None identified.
P1603 P1601 Antiitching properties of patented avocado peptides Stephanie Bredif, Laboratoires Expanscience, Epernon, Eure Et Loir, France; Caroline Baudouin, PhD, Laboratoires Expanscience, Epernon, Eure Et Loir, France; Philippe Msika, PhD, Laboratoires Expanscience, Epernon, Eure Et Loir, France Objectives: Itch is defined as an unpleasant sensation evoking the desire to scratch. Cross-talk between C neuron terminals and the spatially closely related dermal mast cells seems to play an important part in the pruritus pathogenesis. Indeed, mast cell mediators, such as histamine and tryptase (via proteinase-activated receptor 2 [PAR2]) are able to stimulate C neuron terminals, therefore inducing the sensation of itch. Moreover, PAR2 is also expressed by keratinocytes, where its activation initiates inflammatory response. Although the itch pathogenesis in dry skin is not fully understood, the clinical association between itch and dry skin is well established. In this study, we investigated the antiitching and moisturizing properties of a patented natural extract, avocado peptides (AP). Methods: Mast cells were treated with calcium ionophore or substance P in order to induce the release of tryptase and histamine, respectively, measured according to spectrophotometric and spectrofluorometric methods. Gene expression of PAR2 was analyzed in a reconstructed human epidermis model using DNA microarrays. Epidermal lipid neosynthesis was followed in reconstructed human epidermis thanks to lipids radiolabeling and thin layer chromatography analysis. Hyaluronic acid release by normal human epidermal keratinocytes (NHEK) was measured by ELISA. Glycosaminoglycans (GAGs) synthesis by NHEK was evaluated by measuring incorporation of 35S-sulfate. Results: Avocado peptides (AP) were able to significantly inhibit the release of histamine (up to -53%; P \.01) and tryptase (-30%; P \.01) by mast cells. Epidermal gene expression of PAR2 was reduced by 52% after treatment with AP. AP positively affected epidermal ceramides and free fatty acid synthesis. Finally, AP significantly enhanced NHEK production of hyaluronic acid and GAGs by 31% (P \.05) and 34% (P \.05), respectively.
Effects of galactomyces ferment filtrate on epidermal barrier marker caspase-14 in human skin cells Kenji Hattori, PhD, MS, Keio University Faculty of Pharmacy, Tokyo, Japan; Akiko Nakajima, MS, Procter & Gamble Japan, Kobe, Hougo, Japan; Akira Date, PhD, MS, MPH, Procter & Gamble Japan, Kobe, Hyougo, Japan; Hiroomi Tamura, PhD, MS, Keio University Faculty of Parmacy, Tokyo, Japan; Kyoko Ishida, MS, Procter & Gamble Japan, Kobe, Hougo, Japan Background: The stratum corneum of human epidermis is composed of terminally differentiated keratinocytes serving as an essential barrier to environmental stresses, such as UV-induced photodamage and water loss. The regulatory and proteolytic events that coordinate barrier formation are tightly controlled. Caspase-14 belongs to a conserved family of aspartate-specific proteases. Its epidermal expression is restricted almost exclusively to the suprabasal layers, implicating this protease in keratinocyte terminal differentiation and cornification. Therefore, caspase-14 is a useful biomarker to monitor formation and homeostasis of the stratum corneum barrier. Objective: Determine the in vitro effects on barrier formation and hydration of topically applied galactomyces ferment filtrate (GFF) via expression of epidermal late differentiation biomarkers (caspase-14 and related genes). Methods: In vitro human skin models, including skin keratinocytes and skin equivalents (SE) with partially (EPI-201) or completely formed (EPI-200) stratified, cornified epidermis were treated topically with GFF or dexamethasone (Dex). Caspase-14 and related barrier biomarkers were assayed via enzyme activity (caspase-14), mRNA expression by RT-PCR and protein levels by Western blot analysis. Histologic evaluation was conducted also in the human SE models. Results: GFF increased expression of caspase-14 and peptidylarginine deiminases (PAD1, PAD3) in human SE cultures (EPI-200 and -201) and also increased the expression of transglutaminase (TGM1) and involucrin in a human SE model of earlier stage corneocyte differentiation (EPI-201). Dex increased only the expression of caspase-14 in both human SE models. RU-486, a glucocorticoid receptor antagonist, inhibited the effects of Dex but not GFF, suggesting independent pathways such as MAPK may be involved.
Conclusion: The antiitching activities of these patented avocado peptides have been shown on mast cells, which are at the heart of pruritus reaction. Moreover, its moisturizing and relipidizing properties were suggested by in vitro assays. AP could therefore be of particular interest in the management of cutaneous disorders associated with itchy and dry skin.
Conclusions: These results indicate that GFF increases caspase-14 expression by epidermal cells, specifically during late stage differentiation. The positive effect of GFF on late differentiation biomarkers supports previous observations that GFFcontaining moisturizers help protect the skin against damage, such as dryness, from environmental stress. These results provide a compelling reason to further understand the nature of GFF and its skin benefits.
Commercial support: None identified.
Commercial support: None identified.
AB54
J AM ACAD DERMATOL
MARCH 2010