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Photoprotective Effects of Isoflavone precursor (DOB-3-A) against UV-induced Damages in Skin fibroblasts
JSID Abstracts / Journal of Dermatological Science 69 (2013) e47–e93
2-fold, respectively. A similar enhancement was shown for flux profiles. Surprisingly, a lower UVA dose revealed greater enhancement compared to the higher dose. The skin deposition and flux of tetracycline both decreased with UVB exposure. UVB also significantly reduced quercetin flux. The skin absorption behavior of chronologically aged skin approximated that of the UVA group, with photoaged skin showing higher enhancement. UV generally exhibited a negligible effect on modulating oxybenzone permeation. http://dx.doi.org/10.1016/j.jdermsci.2012.11.566 P12-14 Ultraviolet irradiation generates low molecular weight hyaluronan and decreases the ability of hyaluronan metabolism in aged skin Miho Narita ∗ , Midori Tanaka, Satoshi Morita Research and Development Department, Naris Cosmetics Co., LTD., Osaka, Japan Hyaluronan (HA), a well-known moisturizer, has been reported to have some biological activities in the epidermis and dermis, but not much is available on HA’s contribution to the stratum corneum (SC). In this study, we investigated the relationship of the SC conditions to the quantitative and qualitative variations of HA with age or the site in the human SC, as well as the influence of HA on human keratinocytes. Consequently, in the human SC, transepidermal water loss (TEWL) in the sun-exposed site was remarkably greater than that in the sun-protected site, whereas variations in TEWL with age were not detected in both sites. Also, the water content of the SC of the elderly subjects was significantly low in the sun-exposed site. The quantity of HA was larger in the sun-exposed site than in the sunprotected site, and the quantity of HA decreased with age in both sites. The molecular weight of HA was smaller in the sun-exposed site, whereas it showed no changes with age. In human keratinocytes obtained from an elderly subject, ultraviolet (UV) irradiation suppressed mRNA expressions of hyaluronan synthase (HAS)-1, HAS-3, hyaluronoglucosaminidase (HYAL)-1, HYAL-2, and CD44 antigen that contribute to the biosynthesis and the degradation of HA. Additionally, smallermolecular-weight HA inhibited the differentiation of keratinocytes regardless of the donor’s age. These results suggest that smaller-molecular-weight HA, which is generated by UV-irradiation, suppresses keratinocytes differentiation, thus inhibiting barrier formation in the sun-exposed site. Furthermore, a decline in the HA metabolizing capacity in the sunexposed aged skin induces the decrease in HA due to aging, which might be causing the lower water content of elderly subjects’ SC. http://dx.doi.org/10.1016/j.jdermsci.2012.11.567
e87
P12-15 A New Objective Histological Scale for Studying Human Photoaged skin Keigo Kawabata 1,∗ , Masaki Kobayashi 1 , Ayumi KusakaKikushima 1 , Emiko Akasaka 2 , Tomotaka Mabuchi 2 , Tsuyoshi Fukui 3 , Yoshinori Sugiyama 1 , Susumu Takekoshi 4 , Muneo Miyasaka 3 , Akira Ozawa 2 , Shingo Sakai 1 1
Innovative Beauty Science Laboratory, Kanebo Cosmetics Inc. Department of Dermatology, Tokai University, School of medicine 3 Department of Plastic Surgery, Tokai University, School of medicine 4 Department of Cell Biology, Tokai University, School of medicine 2
Chronic sun exposure induces photoaging of the skin, including wrinkle formation. A quantitative understanding of the histological alteration of photoaged skin is important for assessing the severity of photoaging. In this study, we evaluated the histological alteration of elastic and collagenic fibers in pre/post-auricular skin sections from 36 Asian subjects using light microscopy. Initial alteration of elastic fibers was observed in the deep dermis. In immunohistochemical studies, signals of Decorin, a modulator for collagen fibrillogenesis, were not detected in very severely altered skin. Based on the histological changes observed in the elastic fibers, decorin, and collagen fibers, we categorized altered skin tissues into 6 stages of severity (stage I to VI). The objectivity of the scale was evaluated based on the degree of agreement in stage determination by 11 inter-observers with blind procedure. A weighted kappa statistic analysis showed almost perfect agreement between observers. We detected a significant positive correlation between stage and wrinkle score as well as between stage and surface roughness parameter (Sm) in 26 Caucasian subjects. Altogether, we propose an objective histological scale that is useful for assessing the severity of photoaging. http://dx.doi.org/10.1016/j.jdermsci.2012.11.568 P12-16 Photoprotective Effects of Isoflavone precursor (DOB-3-A) against UV-induced Damages in Skin fibroblasts Nan-Lin Wu 1,∗ , Jia-Ying Lin 2 , Chi-Feng Hung 2 1 Department of Dermatology, Mackay Memorial Hospital, Hsinchu, Taiwan 2 School of Medicine, Fu Jen Catholic University, New Taipei City, Taiwan
Deoxybenzoins (DOBs) are the precursors of synthetic isoflavone compounds and share similar structures with isoflavones. Recent investigations have suggested that DOBs can be potential pharmacologic agents for the treatment of breast cancer and hormone replacement therapy. DOBs also have various biologic activities such as anti-oxidation and anti-inflammation, and even have stronger ability to scavenge free radicals than vitamin E and vitamin C. However, the studies about the roles of Deoxybenzoin structural compound, DOB-3-A, in photobiology of skin are still limited. Herein, we used the fibroblast as a model to explore the photoprotective effects of DOB-3-A in UVA-induced damages in skin. Our results showed that pretreatment of DOB3-A could reduce the UV-induced cell death. Further exploration revealed DOB-3-A could attenuate UV-triggered phosphorylation of mitogen activation protein kinases (MAPKs) including ERK, p38 and JNK. Additionally, DOB-3-A could also inhibit UV-induced COX-2 expression in fibroblasts. Furthermore, we also demonstrated DOB-3-A could dose-dependently diminish ROS generation after UVA exposure, which implies the photoprotective effects of
e88
JSID Abstracts / Journal of Dermatological Science 69 (2013) e47–e93
DOB-3-A can be related to its ROS scavenger activity. Collectively, we conclude that DOB-3-A can protect the fibroblasts against UV-induced toxic effects and can be applied in our daily life to prevent UV-induced damages in skin in the future.
P13-02[C06-01] Melanocyte stem cell activities and pigment pattern formation in regenerating feathers Sung-Jan Lin 1,2,3,4,∗ , John Foley 5 , Cheng-Ming Chuong 3,4
http://dx.doi.org/10.1016/j.jdermsci.2012.11.569 P13-01[C02-02] CD10-armored melanoma cells acquire highly-potent tumorigenic activity -A plausible explanation of its significance in poor prognosisJunna Oba 1,∗ , Takeshi Nakahara 1 , Chie Kamata 1 , Min Liu 2 , Takehiko Yokomizo 2 , Masutaka Furue 1 , Yoichi Moroi 1 1
Department of Dermatology, Kyushu University, Fukuoka, Japan Department of Biochemistry, Juntendo University School of Medicine, Tokyo, Japan 2
Introduction: In the previous study, we demonstrated that expression of CD10 in malignant melanoma accelerated tumor progression and was associated with poor patient survival. However, the mechanism how CD10 plays a role in melanoma progression remains unclear. The purpose of this study was to elucidate the genetic and pathophysiological function of CD10 in malignant melanoma. Methods: Human CD10 cDNA was amplified and sub-cloned into a vector pCXN2.1. A375 melanoma cell line, which originally lacks CD10 expression at protein level, was transfected with CD10-pCXN2.1. A375 cells transfected with empty vector served as control. By using DNA microarrays, the three cell types: wild type-, CD10- and mock-A375 were profiled. Cell proliferation assay, mouse xenograft model, apoptosis assay against chemotherapeutic drugs, scratch assay, and RT-PCR of array-targeted mRNA were performed using these cell lines. Results: CD10-A375 showed significantly greater cell proliferation in vitro and tumor growth in vivo; CD10 augmented melanoma cell resistance to apoptosis mediated by etoposide, gemcitabine and doxorubicin. CD10 inhibitors, thiorphan and phosphoramidon, significantly blocked the tumor growth of CD10-A375 in mice. In scratch assay, however, CD10-A375 displayed lower migratory capacity than mock-A375. DNA microarray analysis revealed that up-regulated genes in CD10-A375 were mostly involved in angiogenesis, cytokine signaling, and blood coagulation; downregulated genes mostly belonged to cell adhesion and migration. The microarray results coincided with those of the cell proliferation and migratory scratch assay. Conclusion: CD10 may promote tumor progression by increasing cell proliferation, resistance to anti-cancer drugs and angiogenesis. http://dx.doi.org/10.1016/j.jdermsci.2012.11.570
1
Department of Dermatology, National Taiwan University Hospital and College of Medicine, Taipei, Taiwan 2 Institute of Biomedical Engineering, National Taiwan University, Taipei, Taiwan 3 Research Center for Developmental Biology and Regenerative Medicine, National Taiwan University, Taipei, Taiwan 4 Department of Pathology, University of Southern California, Los Angeles, CA, USA 5 Medical Sciences, Indiana University School of Medicine, Bloomington, IN, USA The diversity of feather pigment patterns has amazed many, but the identity of feather melanocyte stem cell (McSC) and the mechanisms regulating pigment patterns have been unveiled little. Here we show that McSCs are arranged as a ring around the proximal collar bulge, continuously sending out progeny distally to paint the keratinocytes in growing stage. In resting stage, this circular niche descends to the lowest tips of papilla ectoderm and McSCs become quiescent. The unique cylindrical plane formed by McSCs and their progeny yields new dimensions of regulatory possibilities that are constrained by the highly localized McSC niche topology in mammalian hair follicles. For pigment switch, all whites observed in various pigment patterns are not equal. They are created by several basic cellular mechanisms including McSC removal, suppressed melanocyte emigration or inhibited differentiation. Variation in temporal and spatial employment of these cellular mechanisms helps to create pigment patterns. We also found an unexpected role of feather mesenchymal pulp cells in regulating pigment patterns by inhibiting melanocyte differentiation through patterned expression of inhibitors. Feather pulp cells are also able to respond to physiological changes, such as sexual maturity, to change pigment patterns in regenerating feathers through varying temporal and spatial inhibitor expression patterns. Hence, the complex feather pigment patterning on the cylindrical epithelial canvas is conferred by the unique niche topology through multiple-dimensional cooption of regulatory mechnisms during evolution. http://dx.doi.org/10.1016/j.jdermsci.2012.11.571 P13-03[C06-02] Liver X receptor activation inhibits melanogenesis through the acceleration of ERK-mediated MiTF degradation Kyung-Min Lim 1,2,∗ , Chang Seok Lee 1 , Miyoung Park 1 , Jiwon Han 1 , Ji-hae Lee 1 , Il-Hong Bae 1 , Hyunjung Choi 1 , Eui Dong Son 1 , Young-Ho Park 1 1 2
AmorePacific R&D center, Yongin-si, Republic of Korea College of Pharmacy, Gachon University, Incheon, Republic of Korea
Liver X receptors(LXRs) are nuclear receptors that act as ligandactivated transcription factors regulating lipid metabolism and inflammation. In the skin, activation of LXRs stimulates differentiation of keratinocytes and augments lipid synthesis in sebocytes. However, function of LXRs in melanocytes has not been elucidated. Here we investigated if LXR activation might affect melanogenesis, the representative function of melanocytes. In human primary melanocytes and B16 melanoma cells, TO901317, a synthetic LXR ligand inhibited melanin synthesis. Enzymatic activities of tyrosinase were unaffected but the expression of melanogenesis-related proteins such as tyrosinase, tyrosinase related protein(TRP)-1 and
2-fold, respectively. A similar enhancement was shown for flux profiles. Surprisingly, a lower UVA dose revealed greater enhancement compared to the higher dose. The skin deposition and flux of tetracycline both decreased with UVB exposure. UVB also significantly reduced quercetin flux. The skin absorption behavior of chronologically aged skin approximated that of the UVA group, with photoaged skin showing higher enhancement. UV generally exhibited a negligible effect on modulating oxybenzone permeation. http://dx.doi.org/10.1016/j.jdermsci.2012.11.566 P12-14 Ultraviolet irradiation generates low molecular weight hyaluronan and decreases the ability of hyaluronan metabolism in aged skin Miho Narita ∗ , Midori Tanaka, Satoshi Morita Research and Development Department, Naris Cosmetics Co., LTD., Osaka, Japan Hyaluronan (HA), a well-known moisturizer, has been reported to have some biological activities in the epidermis and dermis, but not much is available on HA’s contribution to the stratum corneum (SC). In this study, we investigated the relationship of the SC conditions to the quantitative and qualitative variations of HA with age or the site in the human SC, as well as the influence of HA on human keratinocytes. Consequently, in the human SC, transepidermal water loss (TEWL) in the sun-exposed site was remarkably greater than that in the sun-protected site, whereas variations in TEWL with age were not detected in both sites. Also, the water content of the SC of the elderly subjects was significantly low in the sun-exposed site. The quantity of HA was larger in the sun-exposed site than in the sunprotected site, and the quantity of HA decreased with age in both sites. The molecular weight of HA was smaller in the sun-exposed site, whereas it showed no changes with age. In human keratinocytes obtained from an elderly subject, ultraviolet (UV) irradiation suppressed mRNA expressions of hyaluronan synthase (HAS)-1, HAS-3, hyaluronoglucosaminidase (HYAL)-1, HYAL-2, and CD44 antigen that contribute to the biosynthesis and the degradation of HA. Additionally, smallermolecular-weight HA inhibited the differentiation of keratinocytes regardless of the donor’s age. These results suggest that smaller-molecular-weight HA, which is generated by UV-irradiation, suppresses keratinocytes differentiation, thus inhibiting barrier formation in the sun-exposed site. Furthermore, a decline in the HA metabolizing capacity in the sunexposed aged skin induces the decrease in HA due to aging, which might be causing the lower water content of elderly subjects’ SC. http://dx.doi.org/10.1016/j.jdermsci.2012.11.567
e87
P12-15 A New Objective Histological Scale for Studying Human Photoaged skin Keigo Kawabata 1,∗ , Masaki Kobayashi 1 , Ayumi KusakaKikushima 1 , Emiko Akasaka 2 , Tomotaka Mabuchi 2 , Tsuyoshi Fukui 3 , Yoshinori Sugiyama 1 , Susumu Takekoshi 4 , Muneo Miyasaka 3 , Akira Ozawa 2 , Shingo Sakai 1 1
Innovative Beauty Science Laboratory, Kanebo Cosmetics Inc. Department of Dermatology, Tokai University, School of medicine 3 Department of Plastic Surgery, Tokai University, School of medicine 4 Department of Cell Biology, Tokai University, School of medicine 2
Chronic sun exposure induces photoaging of the skin, including wrinkle formation. A quantitative understanding of the histological alteration of photoaged skin is important for assessing the severity of photoaging. In this study, we evaluated the histological alteration of elastic and collagenic fibers in pre/post-auricular skin sections from 36 Asian subjects using light microscopy. Initial alteration of elastic fibers was observed in the deep dermis. In immunohistochemical studies, signals of Decorin, a modulator for collagen fibrillogenesis, were not detected in very severely altered skin. Based on the histological changes observed in the elastic fibers, decorin, and collagen fibers, we categorized altered skin tissues into 6 stages of severity (stage I to VI). The objectivity of the scale was evaluated based on the degree of agreement in stage determination by 11 inter-observers with blind procedure. A weighted kappa statistic analysis showed almost perfect agreement between observers. We detected a significant positive correlation between stage and wrinkle score as well as between stage and surface roughness parameter (Sm) in 26 Caucasian subjects. Altogether, we propose an objective histological scale that is useful for assessing the severity of photoaging. http://dx.doi.org/10.1016/j.jdermsci.2012.11.568 P12-16 Photoprotective Effects of Isoflavone precursor (DOB-3-A) against UV-induced Damages in Skin fibroblasts Nan-Lin Wu 1,∗ , Jia-Ying Lin 2 , Chi-Feng Hung 2 1 Department of Dermatology, Mackay Memorial Hospital, Hsinchu, Taiwan 2 School of Medicine, Fu Jen Catholic University, New Taipei City, Taiwan
Deoxybenzoins (DOBs) are the precursors of synthetic isoflavone compounds and share similar structures with isoflavones. Recent investigations have suggested that DOBs can be potential pharmacologic agents for the treatment of breast cancer and hormone replacement therapy. DOBs also have various biologic activities such as anti-oxidation and anti-inflammation, and even have stronger ability to scavenge free radicals than vitamin E and vitamin C. However, the studies about the roles of Deoxybenzoin structural compound, DOB-3-A, in photobiology of skin are still limited. Herein, we used the fibroblast as a model to explore the photoprotective effects of DOB-3-A in UVA-induced damages in skin. Our results showed that pretreatment of DOB3-A could reduce the UV-induced cell death. Further exploration revealed DOB-3-A could attenuate UV-triggered phosphorylation of mitogen activation protein kinases (MAPKs) including ERK, p38 and JNK. Additionally, DOB-3-A could also inhibit UV-induced COX-2 expression in fibroblasts. Furthermore, we also demonstrated DOB-3-A could dose-dependently diminish ROS generation after UVA exposure, which implies the photoprotective effects of
e88
JSID Abstracts / Journal of Dermatological Science 69 (2013) e47–e93
DOB-3-A can be related to its ROS scavenger activity. Collectively, we conclude that DOB-3-A can protect the fibroblasts against UV-induced toxic effects and can be applied in our daily life to prevent UV-induced damages in skin in the future.
P13-02[C06-01] Melanocyte stem cell activities and pigment pattern formation in regenerating feathers Sung-Jan Lin 1,2,3,4,∗ , John Foley 5 , Cheng-Ming Chuong 3,4
http://dx.doi.org/10.1016/j.jdermsci.2012.11.569 P13-01[C02-02] CD10-armored melanoma cells acquire highly-potent tumorigenic activity -A plausible explanation of its significance in poor prognosisJunna Oba 1,∗ , Takeshi Nakahara 1 , Chie Kamata 1 , Min Liu 2 , Takehiko Yokomizo 2 , Masutaka Furue 1 , Yoichi Moroi 1 1
Department of Dermatology, Kyushu University, Fukuoka, Japan Department of Biochemistry, Juntendo University School of Medicine, Tokyo, Japan 2
Introduction: In the previous study, we demonstrated that expression of CD10 in malignant melanoma accelerated tumor progression and was associated with poor patient survival. However, the mechanism how CD10 plays a role in melanoma progression remains unclear. The purpose of this study was to elucidate the genetic and pathophysiological function of CD10 in malignant melanoma. Methods: Human CD10 cDNA was amplified and sub-cloned into a vector pCXN2.1. A375 melanoma cell line, which originally lacks CD10 expression at protein level, was transfected with CD10-pCXN2.1. A375 cells transfected with empty vector served as control. By using DNA microarrays, the three cell types: wild type-, CD10- and mock-A375 were profiled. Cell proliferation assay, mouse xenograft model, apoptosis assay against chemotherapeutic drugs, scratch assay, and RT-PCR of array-targeted mRNA were performed using these cell lines. Results: CD10-A375 showed significantly greater cell proliferation in vitro and tumor growth in vivo; CD10 augmented melanoma cell resistance to apoptosis mediated by etoposide, gemcitabine and doxorubicin. CD10 inhibitors, thiorphan and phosphoramidon, significantly blocked the tumor growth of CD10-A375 in mice. In scratch assay, however, CD10-A375 displayed lower migratory capacity than mock-A375. DNA microarray analysis revealed that up-regulated genes in CD10-A375 were mostly involved in angiogenesis, cytokine signaling, and blood coagulation; downregulated genes mostly belonged to cell adhesion and migration. The microarray results coincided with those of the cell proliferation and migratory scratch assay. Conclusion: CD10 may promote tumor progression by increasing cell proliferation, resistance to anti-cancer drugs and angiogenesis. http://dx.doi.org/10.1016/j.jdermsci.2012.11.570
1
Department of Dermatology, National Taiwan University Hospital and College of Medicine, Taipei, Taiwan 2 Institute of Biomedical Engineering, National Taiwan University, Taipei, Taiwan 3 Research Center for Developmental Biology and Regenerative Medicine, National Taiwan University, Taipei, Taiwan 4 Department of Pathology, University of Southern California, Los Angeles, CA, USA 5 Medical Sciences, Indiana University School of Medicine, Bloomington, IN, USA The diversity of feather pigment patterns has amazed many, but the identity of feather melanocyte stem cell (McSC) and the mechanisms regulating pigment patterns have been unveiled little. Here we show that McSCs are arranged as a ring around the proximal collar bulge, continuously sending out progeny distally to paint the keratinocytes in growing stage. In resting stage, this circular niche descends to the lowest tips of papilla ectoderm and McSCs become quiescent. The unique cylindrical plane formed by McSCs and their progeny yields new dimensions of regulatory possibilities that are constrained by the highly localized McSC niche topology in mammalian hair follicles. For pigment switch, all whites observed in various pigment patterns are not equal. They are created by several basic cellular mechanisms including McSC removal, suppressed melanocyte emigration or inhibited differentiation. Variation in temporal and spatial employment of these cellular mechanisms helps to create pigment patterns. We also found an unexpected role of feather mesenchymal pulp cells in regulating pigment patterns by inhibiting melanocyte differentiation through patterned expression of inhibitors. Feather pulp cells are also able to respond to physiological changes, such as sexual maturity, to change pigment patterns in regenerating feathers through varying temporal and spatial inhibitor expression patterns. Hence, the complex feather pigment patterning on the cylindrical epithelial canvas is conferred by the unique niche topology through multiple-dimensional cooption of regulatory mechnisms during evolution. http://dx.doi.org/10.1016/j.jdermsci.2012.11.571 P13-03[C06-02] Liver X receptor activation inhibits melanogenesis through the acceleration of ERK-mediated MiTF degradation Kyung-Min Lim 1,2,∗ , Chang Seok Lee 1 , Miyoung Park 1 , Jiwon Han 1 , Ji-hae Lee 1 , Il-Hong Bae 1 , Hyunjung Choi 1 , Eui Dong Son 1 , Young-Ho Park 1 1 2
AmorePacific R&D center, Yongin-si, Republic of Korea College of Pharmacy, Gachon University, Incheon, Republic of Korea
Liver X receptors(LXRs) are nuclear receptors that act as ligandactivated transcription factors regulating lipid metabolism and inflammation. In the skin, activation of LXRs stimulates differentiation of keratinocytes and augments lipid synthesis in sebocytes. However, function of LXRs in melanocytes has not been elucidated. Here we investigated if LXR activation might affect melanogenesis, the representative function of melanocytes. In human primary melanocytes and B16 melanoma cells, TO901317, a synthetic LXR ligand inhibited melanin synthesis. Enzymatic activities of tyrosinase were unaffected but the expression of melanogenesis-related proteins such as tyrosinase, tyrosinase related protein(TRP)-1 and