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P6417 Down-regulation of type I collagen expression in silibinin-treated human skin fibroblasts through blocking Smad2/3-dependent signaling pathway: Its potential therapeutic use in the chemoprevention of keloid Kyu-Suk Lee, MD, PhD, Department of Dermatology, Keimyung University School of Medicine, Daegu, South Korea; Jae-We Cho, MD, PhD, Department of Dermatology , School of Medicine, Keimyung University, Daegu, South Korea; Jun-Il Kwon, MD, Department of Dermatology, School of Medicine, Keimyung University, Daegu, South Korea Inhibition of the Smad2/3 pathway plays a key role on down-regulation of type I collagen synthesis, eventually prevention of keloid formation in tissue. In this study, we investigated the effect of silibinin on the proliferation of human skin fibroblasts (HSFs), expressions of type I collagen, matrix metalloproteinase-1, and Smad2/3. Our data show that proliferation rates of fibroblasts were not markedly decreased in a dose- and time-dependent manner of silibinin. Even though silibinin did not exert cytotoxic effect in HSF, the expression of type I collagen was markedly decreased by dose- and time-dependent manner in silibinin-treated HSF. Consistent with, de- creased promoter activity of type I collagen was observed in HSF by silibinin treatment. MMP-1 and MMP-2 expressions were increased in silibinin-treated HSF. Moreover, silibinin-induced down-regulation of type I collagen were associated with inhibition of Smad2/3 activation in TGF-b1etreated HSF. We further demonstrated that silibinin attenuated the translocation of Smad2/3 into nucleus in TGF- b1etreated HSF. Taken together, our data indicated that silibinin has the potential to protect fibrotic change of skin through inducing down-regulation of type I collagen expression, which is partly mediated by blocking Smad2/3-dependent signaling pathway in HSF. Commercial support: None identified. P6184 Evidence from gene-expression profiling of the inhibition by niacinamide of both innate and adaptive immune response processes and pathways associated with skin photoaging Michael K. Robinson, PhD, The Procter and Gamble Company, Cincinnati, OH, United States; Jay P. Tiesman, PhD, The Procter and Gamble Company, Cincinnati, OH, United States; Kenton D. Juhlin, PhD, The Procter and Gamble Company, Cincinnati, OH, United States; Kevin J. Mills, PhD, The Procter and Gamble Company, Cincinnati, OH, United States; Robert L. Binder, PhD, The Procter and Gamble Company, Cincinnati, OH, United States; Rosemarie Osborne, PhD, The Procter and Gamble Company, Cincinnati, OH, United States; Scott M. Hartman, The Procter and Gamble Company, Cincinnati, OH, United States Background: Gene expression profiling of photoaged skin revealed strong up- regulation of biologic themes related to immune and inflammatory processes and their connection to processes of extracellular matrix, oxido-reductase activity, and protease activity. This suggests that immune and inflammatory processes contribute to the skin photoaging phenotype. A separate gene expression profiling of skin treated with the known skin benefit agent, niacinamide, showed evidence of a subtle reversal of these or related themes. The purpose of this study was to apply additional informatics analyses to these datasets to determine the potential relevance of innate and adaptive immune response processes and pathways in skin photoaging and the effect on these of niacinamide treatment. Objective: To determine the potential role of innate and adaptive immune responses in skin photoaging, and the effects of repeat topical application of niacinamide, via gene expression profiling analysis. Method: Gene expression datasets were analyzed from 2 studies: (1) a photoaging phenotype study of UV exposed (outer forearm) and UV protected (buttock) skin from young (age 18-20) and older (age 60-67) female subjects, and (2) a study of forearm skin (females age 35-55) treated with 3.5% niacinamide-containing or vehicle lotion formulations twice a day for up to 15 days. In both studies, full- thickness skin biopsies were obtained and total RNA was extracted and labeled for Affymetrix GeneChip analysis. The analysis was based on hybridization to Affymetrix microarrays, containing up to 54,613 probe sets. The data were subjected to rigorous statistical quality control and analysis, gene expression profiling, and gene set and subnetwork enrichment analysis. The latter was specifically focused on pathways and processes related to innate and adaptive immunity. Results: Significant up-regulation of various immune pathways and processes were noted in older photoaged versus younger UV protected skin. These included general processes of innate and adaptive immunity and more specific processes such as cellular immunity, T cell function, NK cell function, leukocyte chemotaxis, toll-like receptor processes, etc. Many of these processes were down-regulated in niacin- amide-treated versus vehicle-treated (or untreated) skin, providing additional mechanistic insights into the known antiinflammatory effects of this skin benefit agent. 100% funded by The Procter and Gamble Company. P6248 In vivo laser speckle measurement of skin roughness versus profilometric measurement of silicone replicas Gurbir Dhadwal, MD, Photomedicine Institute, Department of Dermatology and Skin Science, University of British Columbia, Vancouver, Canada; Bernadette Ortiz- Policarpio, MD, Photomedicine Institute, Department of Dermatology and Skin Science, University of British Columbia, Vancouver, Canada; David McLean, MD, Photomedicine Institute, Department of Dermatology and Skin Science, University of British Columbia, Vancouver, Canada; Diana Diao, MD, Photomedicine Institute, Department of Dermatology and Skin Science, University of British Columbia, Vancouver, Canada; Harvey Lui, MD, Photomedicine Institute, Department of Dermatology, UBC; Cancer Control Research and Integrative Oncology, BC Cancer Agency, Vancouver, Canada; Liuodmila Tchvialeva, PhD, Photomedicine Institute, Department of Dermatology and Skin Science, University of British Columbia, Vancouver, Canada; Tim Lee, PhD, Photomedicine Institute, Dermatology, University of British Columbia; Cancer Control Research and Integrative Oncology, BC Cancer; Computing Science, Simon Fraser University, Vancouver, Canada Background: Roughness is a property used in helping make the diagnosis of many skin conditions, such as verrucae, actinic keratoses, and skin cancers, and is also used to assess the response to treatment in cosmetic dermatology. We developed a novel device and are conducting a study to measure in vivo the skin roughness of 25 body sites and validate our technique against currently used silicone replica based methods, which are more time consuming. The device is based on measuring the contrast of laser speckle patterns which are light and dark stochastic interference patterns generated when coherent light interacts with a rough surface. The contrast of images of laser speckle correlates with the roughness of surfaces. Methods: Our study has recruited 40 volunteers (18 males; 22 females; mean age 39.6 6 13.7), who have had 25 body sites measured by our portable device. Body sites were categorized into minimally, intermittently and maximally sun exposed. ANOVA was used to assess for a significant relationship between skin roughness, body site and sun exposure. As well, silicone replicas were created of one body site from each subject and measured with profilometry and compared with the measurements from our device. Results: Statistical analysis showed a significant relationship between body site and measured root-mean-square roughness (P \.001). The values of roughness ranged from 12.1 u 6 1.4 u for the palm, the smoothest site, to 32.4 u 6 1.1 u for the ear lobe, the roughest site. On subgroup analysis, roughness of maximally sun exposed sites (28.0 u 6 0.4 u) was found to be significantly higher than the roughness of intermittently (21.4 u 6 0.4 u) or minimally (21.2 u 6 0.5 u) sun exposed sites. There was a statistically significant correlation between the measurements from our device and the measurements of silicone replicas using profilometry (r ¼ 0.51; P ¼.02). Conclusion: Here we report a data set of in vivo skin roughness of 25 body sites. We found that the roughness measurements from analysis of laser speckle were consistent with previously published results using other assessment methodologies, vary as expected with sun exposure, and correlate directly with measurements of our subjects using profilometry of silicone replicas. Commercial support: None identified. P6887 Persistence of corneodesmosomes and elevation of protease inhibitors LEKTI and SCCA1 in dandruff Bhumika Singh, PhD, Unilever R&D, Bebington, United Kingdom; Clive Harding, Unilever R&D, Bebington, United Kingdom Corneodesmosomes (CDSMs) are the major cohesive component of the stratum corneum (SC) and play a vital role in the water barrier function of this tissue. For normal desquamation to occur, CDSMs must be hydrolyzed by members of the kallikrein family of proteases. The activity of these proteases is in turn controlled by a specific inhibitor: lymphoepithelial Kazal-type related inhibitor (LEKTI). The balance between serine proteases and their cognate inhibitors is essential for SC integrity. In cosmetic, dry skin conditions the hydrolysis of CDSMs is reduced, corneocytes cohesion is increased and desquamation is perturbed leading to the characteristic skin surface flaking. In contrast in dandruff, a condition also associated with severe barrier impairment and perturbed desquamation, it has been reported that this condition is associated with a premature loss of CDSMs. To study the fate of CDSMs in more detail we investigated their distribution in the superficial layers of scalp SC recovered from both healthy individuals and dandruff sufferers. CDSMs were localized by immunohistochemical detection of desmoglein 1 (DSG1). We also investigated the expression of LEKTI and the squamous cell carcinoma antigen 1 (SCCA1), a protease inhibitor associated with perturbed barrier function. In healthy samples, DSG1 distribution was limited to the cellular periphery of adjacent corneocytes within the same plane of SC consistent with the distribution observed on other healthy body sites. In samples of dandruff SC the CDSMs were again readily detectable. Immunofluorescence labelling showed that DSG1 was localized to peripheral/intercellular regions and over the entire surface area, reminiscent of the pattern of abnormal CDSM retention observed in dry skin. Western blot analysis of LEKTI and SCCA1 showed an elevated expression of both proteins in samples of dandruff SC as compared to those from healthy SC. Our findings indicate that, the persistence of nonperipheral CDSMs is a characteristic of the perturbed desquamation seen in dandruff. The observations of increased expression of LEKTI and possibly SCCA1 are consistent with the view that the dandruff condition is characterized by an imbalance in protease/protease inhibitor interaction in the SC. 100% funded by Unilever PLC. AB36 JAM ACAD DERMATOL APRIL 2013

Evidence from gene-expression profiling of the inhibition by niacinamide of both innate and adaptive immune response processes and pathways associated with skin photoaging

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Page 1: Evidence from gene-expression profiling of the inhibition by niacinamide of both innate and adaptive immune response processes and pathways associated with skin photoaging

P6417Down-regulation of type I collagen expression in silibinin-treated humanskin fibroblasts through blocking Smad2/3-dependent signaling pathway:Its potential therapeutic use in the chemoprevention of keloid

Kyu-Suk Lee, MD, PhD, Department of Dermatology, Keimyung University Schoolof Medicine, Daegu, South Korea; Jae-We Cho, MD, PhD, Department ofDermatology , School of Medicine, Keimyung University, Daegu, South Korea;Jun-Il Kwon, MD, Department of Dermatology, School of Medicine, KeimyungUniversity, Daegu, South Korea

Inhibition of the Smad2/3 pathway plays a key role on down-regulation of type Icollagen synthesis, eventually prevention of keloid formation in tissue. In this study,we investigated the effect of silibinin on the proliferation of human skin fibroblasts(HSFs), expressions of type I collagen, matrix metalloproteinase-1, and Smad2/3.Our data show that proliferation rates of fibroblasts were not markedly decreased ina dose- and time-dependent manner of silibinin. Even though silibinin did not exertcytotoxic effect in HSF, the expression of type I collagen was markedly decreased bydose- and time-dependent manner in silibinin-treated HSF. Consistent with, de-creased promoter activity of type I collagen was observed in HSF by silibinintreatment. MMP-1 and MMP-2 expressions were increased in silibinin-treated HSF.Moreover, silibinin-induced down-regulation of type I collagenwere associated withinhibition of Smad2/3 activation in TGF-b1etreated HSF. We further demonstratedthat silibinin attenuated the translocation of Smad2/3 into nucleus in TGF-b1etreated HSF. Taken together, our data indicated that silibinin has the potentialto protect fibrotic change of skin through inducing down-regulation of type Icollagen expression, which is partly mediated by blocking Smad2/3-dependentsignaling pathway in HSF.

AB36

cial support: None identified.

Commer

P6184Evidence from gene-expression profiling of the inhibition by niacinamideof both innate and adaptive immune response processes and pathwaysassociated with skin photoaging

Michael K. Robinson, PhD, The Procter and Gamble Company, Cincinnati, OH,United States; Jay P. Tiesman, PhD, The Procter and Gamble Company,Cincinnati, OH, United States; Kenton D. Juhlin, PhD, The Procter and GambleCompany, Cincinnati, OH, United States; Kevin J. Mills, PhD, The Procter andGamble Company, Cincinnati, OH, United States; Robert L. Binder, PhD, TheProcter and Gamble Company, Cincinnati, OH, United States; RosemarieOsborne, PhD, The Procter and Gamble Company, Cincinnati, OH, UnitedStates; Scott M. Hartman, The Procter and Gamble Company, Cincinnati, OH,United States

Background: Gene expression profiling of photoaged skin revealed strong up-regulation of biologic themes related to immune and inflammatory processes andtheir connection to processes of extracellular matrix, oxido-reductase activity, andprotease activity. This suggests that immune and inflammatory processes contributeto the skin photoaging phenotype. A separate gene expression profiling of skintreated with the known skin benefit agent, niacinamide, showed evidence of asubtle reversal of these or related themes. The purpose of this study was to applyadditional informatics analyses to these datasets to determine the potentialrelevance of innate and adaptive immune response processes and pathways inskin photoaging and the effect on these of niacinamide treatment.

Objective: To determine the potential role of innate and adaptive immune responsesin skin photoaging, and the effects of repeat topical application of niacinamide, viagene expression profiling analysis.

Method: Gene expression datasets were analyzed from 2 studies: (1) a photoagingphenotype study of UV exposed (outer forearm) and UV protected (buttock) skinfrom young (age 18-20) and older (age 60-67) female subjects, and (2) a study offorearm skin (females age 35-55) treated with 3.5% niacinamide-containing orvehicle lotion formulations twice a day for up to 15 days. In both studies, full-thickness skin biopsies were obtained and total RNA was extracted and labeled forAffymetrix GeneChip analysis. The analysis was based on hybridization toAffymetrix microarrays, containing up to 54,613 probe sets. The data weresubjected to rigorous statistical quality control and analysis, gene expressionprofiling, and gene set and subnetwork enrichment analysis. The latter wasspecifically focused on pathways and processes related to innate and adaptiveimmunity.

Results: Significant up-regulation of various immune pathways and processes werenoted in older photoaged versus younger UV protected skin. These included generalprocesses of innate and adaptive immunity and more specific processes such ascellular immunity, T cell function, NK cell function, leukocyte chemotaxis, toll-likereceptor processes, etc. Many of these processes were down-regulated in niacin-amide-treated versus vehicle-treated (or untreated) skin, providing additionalmechanistic insights into the known antiinflammatory effects of this skin benefitagent.

ded by The Procter and Gamble Company.

100% fun

J AM ACAD DERMATOL

P6248In vivo laser speckle measurement of skin roughness versus profilometricmeasurement of silicone replicas

Gurbir Dhadwal, MD, Photomedicine Institute, Department of Dermatology andSkin Science, University of British Columbia, Vancouver, Canada; Bernadette Ortiz-Policarpio, MD, Photomedicine Institute, Department of Dermatology and SkinScience, University of British Columbia, Vancouver, Canada; David McLean, MD,Photomedicine Institute, Department of Dermatology and Skin Science, Universityof British Columbia, Vancouver, Canada; Diana Diao, MD, Photomedicine Institute,Department of Dermatology and Skin Science, University of British Columbia,Vancouver, Canada; Harvey Lui, MD, Photomedicine Institute, Department ofDermatology, UBC; Cancer Control Research and Integrative Oncology, BC CancerAgency, Vancouver, Canada; Liuodmila Tchvialeva, PhD, Photomedicine Institute,Department of Dermatology and Skin Science, University of British Columbia,Vancouver, Canada; Tim Lee, PhD, Photomedicine Institute, Dermatology,University of British Columbia; Cancer Control Research and IntegrativeOncology, BC Cancer; Computing Science, Simon Fraser University, Vancouver,Canada

Background: Roughness is a property used in helping make the diagnosis of manyskin conditions, such as verrucae, actinic keratoses, and skin cancers, and is alsoused to assess the response to treatment in cosmetic dermatology. We developed anovel device and are conducting a study to measure in vivo the skin roughness of 25body sites and validate our technique against currently used silicone replica basedmethods, which are more time consuming. The device is based on measuring thecontrast of laser speckle patterns which are light and dark stochastic interferencepatterns generated when coherent light interacts with a rough surface. The contrastof images of laser speckle correlates with the roughness of surfaces.

Methods: Our study has recruited 40 volunteers (18 males; 22 females; mean age39.6 6 13.7), who have had 25 body sites measured by our portable device. Bodysites were categorized into minimally, intermittently and maximally sun exposed.ANOVA was used to assess for a significant relationship between skin roughness,body site and sun exposure. As well, silicone replicas were created of one body sitefrom each subject and measured with profilometry and compared with themeasurements from our device.

Results: Statistical analysis showed a significant relationship between body site andmeasured root-mean-square roughness (P\.001). The values of roughness rangedfrom 12.1 u 6 1.4 u for the palm, the smoothest site, to 32.4 u 6 1.1 u for the earlobe, the roughest site. On subgroup analysis, roughness of maximally sun exposedsites (28.0 u 6 0.4 u) was found to be significantly higher than the roughness ofintermittently (21.4 u 6 0.4 u) or minimally (21.2 u 6 0.5 u) sun exposed sites.There was a statistically significant correlation between the measurements fromour device and the measurements of silicone replicas using profilometry (r ¼ 0.51;P ¼ .02).

Conclusion: Here we report a data set of in vivo skin roughness of 25 body sites. Wefound that the roughness measurements from analysis of laser speckle wereconsistent with previously published results using other assessment methodologies,vary as expected with sun exposure, and correlate directly with measurements ofour subjects using profilometry of silicone replicas.

cial support: None identified.

Commer

P6887Persistence of corneodesmosomes and elevation of protease inhibitorsLEKTI and SCCA1 in dandruff

Bhumika Singh, PhD, Unilever R&D, Bebington, United Kingdom; Clive Harding,Unilever R&D, Bebington, United Kingdom

Corneodesmosomes (CDSMs) are the major cohesive component of the stratumcorneum (SC) and play a vital role in the water barrier function of this tissue. Fornormal desquamation to occur, CDSMs must be hydrolyzed by members of thekallikrein family of proteases. The activity of these proteases is in turn controlled bya specific inhibitor: lymphoepithelial Kazal-type related inhibitor (LEKTI). Thebalance between serine proteases and their cognate inhibitors is essential for SCintegrity. In cosmetic, dry skin conditions the hydrolysis of CDSMs is reduced,corneocytes cohesion is increased and desquamation is perturbed leading to thecharacteristic skin surface flaking. In contrast in dandruff, a condition alsoassociated with severe barrier impairment and perturbed desquamation, it hasbeen reported that this condition is associated with a premature loss of CDSMs. Tostudy the fate of CDSMs in more detail we investigated their distribution in thesuperficial layers of scalp SC recovered from both healthy individuals and dandruffsufferers. CDSMs were localized by immunohistochemical detection of desmoglein1 (DSG1). We also investigated the expression of LEKTI and the squamous cellcarcinoma antigen 1 (SCCA1), a protease inhibitor associated with perturbed barrierfunction. In healthy samples, DSG1 distributionwas limited to the cellular peripheryof adjacent corneocyteswithin the same plane of SC consistent with the distributionobserved on other healthy body sites. In samples of dandruff SC the CDSMs wereagain readily detectable. Immunofluorescence labelling showed that DSG1 waslocalized to peripheral/intercellular regions and over the entire surface area,reminiscent of the pattern of abnormal CDSM retention observed in dry skin.Western blot analysis of LEKTI and SCCA1 showed an elevated expression of bothproteins in samples of dandruff SC as compared to those from healthy SC. Ourfindings indicate that, the persistence of nonperipheral CDSMs is a characteristic ofthe perturbed desquamation seen in dandruff. The observations of increasedexpression of LEKTI and possibly SCCA1 are consistent with the view that thedandruff condition is characterized by an imbalance in protease/protease inhibitorinteraction in the SC.

ded by Unilever PLC.

100% fun

APRIL 2013