Artículos y publicaciones

American Academy of Dermatology Annual Meeting AAD, Orlando-FL, USA. March 3-7, 2017. Antônio Carlos Vanzo Junior, Liliana Bechelli de Oliveira Torloni, Renan Lage, Andréia Feital da Costa Pereira, Amanda Francielli Pereira, Ana Lúcia Tabarini Alves Pinheiro, Michelle Sabrina da Silva, Gustavo Facchini, Samara Eberlin Nails are structures formed by keratinized squamous cells produced by the matrix that grow on the nail bed. The hardness of the nail plate and its adherence to the underlying bed are key properties that make them protective appendages due to the presence of different types of keratins. Studies have shown the presence of K10 and K14 throughout the stratified epithelium of the nail unit. In this context, filaggrin has the role of stabilizing the network of intermediate filaments of K6 and K16 during hyperproliferative stages and promoting increased epidermal cohesion. With the purpose of maintaining nail health and improving its appearance, the cosmeceutical market studies new strengthening and fortifying formulations that may influence the biological function thereof. Based on the above, the present study evaluates the efficacy of a nail care cosmetic product (NCCP) through preclinical studies for strengthening/repair activity. Human keratinocytes (HaCat) were incubated with three non-cytotoxic concentrations of NCCP for 48 hours for further evaluation of the production of keratins 1, 10, 14, 16, 17, and filaggrin. The in vitro results show that NCCP has a strengthening and nail repair effect by stimulating the production of filaggrin and keratins 1, 10, 14, 16, and 17 in human keratinocytes, favoring nail plate hardness and its adherence to the underlying bed.

Journal of the American Academy of Dermatology, 74(5) Supplement 1, AB27, 2016. DOI: http://dx.doi.org/10.1016/j.jaad.2016.02.110 Costa A, Clerici SP, Facchini G, Pinheiro ALTA, Bonner MY, Arbister J, Eberlin S. Extrinsic aging is attributed to skin changes due to lifestyle and environmental insults. Cigarette smoke contains more than 4,000 toxic compounds including many polycyclic aromatic hydrocarbons, dioxins, and furans; all of them exert negative effects on the skin, contributing to extrinsic skin aging. Cigarette smoke induces the latent and non-functional form of transforming growth factor-β1 (TGF-β1); consequently, cellular responsiveness to TGF-β1 is blocked by this non-functional form, which results in decreased synthesis of extracellular matrix proteins, such as collagen. In this study, we evaluated the effects of honokiol - a natural biphenolic compound derived from the bark of magnolia trees, which presents anti-inflammatory, anti-oxidative, anti-tumour, and neuroprotective properties - through the quantification of collagen and TGF-β1 levels in human skin fragments exposed to cigarette smoke. Human skin fragments were incubated in culture medium and treated with honokiol (10µM and 20µM) and then exposed to cigarette smoke using a cigarette smoke chamber. Total collagen and TGF-β1 levels were measured in supernatants of skin fragments culture through ELISA method. Our results demonstrated that cigarette smoke promotes a significant reduction (51.1%) in collagen synthesis and an increase in TGF-β1 (2.1-fold), compared to the control baseline group. Conversely, honokiol treatment prevented the substantial decline in collagen synthesis in relation to the group exposed to cigarette smoke (8.62% and 29.4% at concentrations of 10µM and 20µM, respectively). Interestingly, incubation of the explants with honokiol plus cigarette smoke led to a further increase in the synthesis of TGF-β1 (86.5% and 19.1% at concentrations of 10µM and 20µM, respectively). Honokiol appears to reverse the collagenolytic effects of smoking-induced TGF-β1 dysfunction.

Arte y Ciencia Cosmética 28(65): 18-24, 2016. ISSN 0122-8072. Pinheiro ALTA, Lizárraga A, Silva MS, Facchini G, Eberlin S. El envejecimiento de la piel representa un proceso insidioso, multifactorial, progresivo y degenerativo, de curso inevitable y prácticamente irreversible. Se lo considera como siendo el resultado de dos procesos sinérgicos de envejecimiento: el proceso intrínseco (o cronológico) y el extrínseco o fotoenvejecimiento. El envejecimiento extrínseco corresponde a los cambios ocurridos en la piel debido al estilo de vida y es influenciado, principalmente, por la radiación ultravioleta (UV), seguido por los productos químicos, tabaquismo, calor y demás insultos ambientales. Los dos procesos aceleran las reacciones sufridas por el tejido cutáneo durante el proceso de envejecimiento, alterando directa o indirectamente el complejo sistema neuro-inmuno-endocrinológico de la piel. Macroscópicamente, el envejecimiento intrínseco acarrea cambios considerables, que incluyen atrofia dérmica, aparición de arrugas, aspereza, pérdida de elasticidad y de la capa lipídica subcutánea. Por otro lado, la apariencia grave de este proceso se debe a los cambios histológicos drásticos representados principalmente por una atrofia profunda de la arquitectura dérmica, con reducción del número y función de los fibroblastos, destrucción de importantes estructuras proteicas, especialmente colágeno y elastina, así como una alteración en la "homeostasis celular proliferativa", resultando en lesiones muchas veces malignas o irreversibles. El fotoenvejecimiento deja la piel áspera, con muchas manchas y arrugas profundas.

IFSCC Magazine 19(2), 69-72, 2016. http://www.ifscc.org/Media/Default/PDFs/IFSCC%20Magazine%200216.pdf Eberlin S, Clerici SP, Eberlin S, Pinheiro ALTA, Pinheiro AS. Skin pigmentation is dependent on both the increased number of melanocytes as the increase on melanogenic enzymes. Exposure to sunlight can increase the production of factors such as melanocyte stimulating hormone and endothelin-1, which trigger the melanization of the skin. In addition to ultraviolet light, studies also show that the infrared radiation and visible light promote changes in skin pigmentation. In this work, we evaluated the consequences of ultraviolet A and B (UVA/UVB), infrared-A (IR-A), and visible light (VL) irradiation on melanin and endothelin-1 synthesis, using an in vitro model of melanocyte and keratinocyte cultures, and an ex vivo model of skin culture. All radiations (UV, IR-A, VL, and association of radiations) produced a significant increase in melanin production when compared to non-irradiated control. UV radiation, VL, and the association of all radiation produced a significant increase in the production of endothelin-1 when compared to non-irradiated control. Skin fragments subjected to all radiation had a higher melanin density in comparison with control fragments. Our results corroborate previous data from the literature and indicated the need for specific photoprotection strategies which are not addressed by the available conventional sunscreens.

Surgical and Cosmetic Dermatology 8(2), 115-20, 2016. DOI: http://dx.doi.org/10.5935/scd1984-8773.20168202 Eberlin S, Facchini G, Eberlin S, Pinheiro ALTA, Silva MS, Pinheiro AS, Costa A. A radiação infravermelha A (IV-A) causa alterações estruturais na pele, similares àquelas provocadas pela exposição prolongada à radiação ultravioleta. A avaliação de eficácia e segurança para produtos cosméticos concentra-se em ensaios in vitro e clínicos. Uma alternativa promissora é a utilização de fragmentos de pele humana provenientes de cirurgias plásticas eletivas, para avaliar os reais benefícios clínicos de um produto aplicado topicamente. O objetivo desta investigação foi correlacionar os efeitos da radiação IV-A em biópsias, fragmentos de pele ex vivo e cultura de fibroblastos humanos, pela quantificação dos mediadores MMP-1, TIMP-1 e GADD45a. Biópsias de 15 voluntárias foram coletadas após aplicações de IV-A durante cinco dias consecutivos. Fragmentos de pele humana provenientes de cirurgia plástica eletiva e cultura de fibroblastos humanos foram expostos à radiação IV-A. Os seguintes mediadores foram mensurados: MMP-1, TIMP-1 e GADD45a para posterior comparação dos resultados. Nos três modelos utilizados, a radiação IV-A induziu aumento de MMP-1, inibiu a síntese de GADD45a e não alterou os valores de TIMP-1. Devido à correlação positiva dos modelos estudados, pode-se sugerir o uso de pele ex vivo como ferramenta plausível e sustentável para suprir diferenças entre conhecimentos gerados a partir de experimentos in vitro e clínicos.

29th IFSCC Congress (International Federation of Societies of Cosmetic Chemists), Orlando, 30 outubro a 02 novembro, 2016. Pinheiro ALTA, Facchini G, Clerici SP, Eberlin S, Pinheiro AS, Eberlin S. For a long time, it was believed that skin photoaging was exclusively attributed to ultraviolet radiation, but lately infrared radiation A (IRA) has attracted interest from the scientific community for inducing similar skin structural changes to those observed by prolonged exposure to ultraviolet radiation. With the advent of the 3Rs policy (Replace, Reduce and Refine), the applied research to the cosmetic market on the understanding of this photoaging provided by IRA was restricted to in vitro and clinical testing. The evaluation of the biological mechanisms using skin biopsies obtained from human volunteers as a test system is a model for understanding the damage triggered by an aggressing agent. However, this procedure can be considered invasive as an ordinary research tool. In this sense, an alternative to fill this gap between in vitro and clinical is the use of skin fragments from elective plastic surgery (ex vivo study). The objective of this study was to correlate the effects of IRA radiation in biopsies, ex vivo skin fragments, and human fibroblasts cultured, through the quantification of MMP-1, TIMP-1, and GADD45a mediators. Biopsies of human skin, as well as the human skin fragments originating from elective plastic surgery, were obtained after approval of the ethics committee and voluntary consent. Together with cultured human fibroblasts (HFF-1), these test systems were incubated for 48 hours at 37°C in a humidified atmosphere with 5% CO 2 . Afterward, they were exposed to a dose of 360 J/cm 2 infrared-A radiation using Hydrosun 750 and HBM1 devices (Hydrosun Medizintechnik GmbH, Müllheim, Germany). After irradiation, test systems were incubated with fresh culture medium and maintained for 24 hours to collect the culture supernatant and quantification of MMP-1, TIMP-1, and GADD45a by enzyme-linked immunosorbent assay. The results demonstrated that in the three models used, IRA radiation induced an increase in MMP-1 and did not alter the values of this protease inhibitor (TIMP-1). A similar effect can be observed with the GADD45a protein, as radiation was able to inhibit the synthesis of this cellular stress sensor in the biopsies, ex vivo fragments, and in cell culture. Therefore, due to the positive correlation of results between the three models studied, it may be suggested that an alternative approach to human biopsies is the use of an ex vivo model, as it is characterized as a plausible and sustainable tool to address the differences between the knowledge generated from in vitro and clinical experiments.

29th IFSCC Congress (International Federation of Societies of Cosmetic Chemists), Orlando, 30 outubro a 02 novembro, 2016. Vanzo-Jr AC, Pereira AFC, Torloni LBO, Lage R, Mercuri M, Viana EAA, Silva SG, Satake CY, Maciente GO, Figueredo TT, Clerici SP, Pinheiro ALTA, Facchini G, Eberlin S. Keratinocytes suffer numerous biochemical reactions during the transition from basal layer to stratum corneum (SC), including synthesis of specific basal (K14) and suprabasal (K10) keratins and cornified envelope-associated proteins. Epidermal differentiation complex (EDC) is constituted through cross-linking of involucrin, loricrin, profilaggrin, among others, on the intracellular surface of the plasma membrane in the upper spinous and granular layers of the epidermis 1-2 . Studies have shown that many factors, such as skin inflammatory processes and prolonged exposure to sunlight, could affect the skin barrier and extracellular matrix, inducing a decrease in the synthesis of the major dermal proteins, collagen and elastin, clinically characterized by wrinkles, rough skin, and loss of water and skin tone. In this study, we evaluated the effects of a moisturizing formulation on the production of envelope proteins filaggrin, involucrin, loricrin, keratin 10 and 14, and extracellular matrix collagen and elastin using an in vitro model of human keratinocytes and fibroblasts. The moisturizing formulation system with a unique, three-dimensional glycol-matrix delivery system that releases moisturizing molecules sequentially into the stratum corneum, resulting in a highly significant improvement of both immediate and long-term skin hydration, containing Glycerin; Enteromorpha Compressa Extract; Hydrolyzed Caesalpinia Spinosa Gum; Caesalpinia Spinosa Gum; Rheum Rhaponticum Root Extract, and Tocopheryl Acetate, was provided by 1MANTECORP SKIN CARE, CECI - Innovation and Consumer Studies Center, Barueri/SP - Brazil. Human dermal fibroblasts and keratinocytes were incubated for 48 hours with three concentrations of the moisturizing formulation previously determined by the XTT method. The levels of collagen and elastin were measured respectively in fibroblast culture supernatant and cell lysate using a precipitation commercial kit (Biocolor, UK). The synthesis of filaggrin, involucrin, loricrin, keratin 10 and keratin 14 was measured in supernatant from keratinocyte culture using a colorimetric competitive enzyme-linked immunosorbent assay - ELISA (Uscn Life Science Inc., Houston, TX, USA). The one-way analysis of variance (ANOVA) test was used followed by Bonferroni post-test with a 5% significance level. Our results demonstrated that incubation of fibroblast cultures with the moisturizing formulation increased the synthesis of extracellular matrix proteins collagen and elastin up to 43.33% and 41.62%, respectively, in relation to the non-treated group. The incubation of keratinocyte cultures with the moisturizing formulation increased levels of involucrin, filaggrin, loricrin, keratin 10, and keratin 14, reaching up to 43.84%, 37.04%, 32.02%, 20.89%, and 18.05%, respectively, in relation to the non-treated control. The water content of the SC and skin surface lipids are important factors in the appearance and function of the skin barrier 3 . Traditional moisturizing ingredients are known to decelerate the loss of skin humidity, increase hydration of the SC, and improve physical and chemical properties of the skin surface, making it moist, smooth, and soft 4 . However, most products claimed for skin moisturizing produce an immediate and transient effect. Thus, optimized skin care cosmetics should be developed to supply a proper moisturizing effect and, at the same time, be capable of balancing the synthesis of skin barrier proteins 5 and protecting against down-regulation of the extracellular matrix. In this study, we evaluated the ability of the moisturizing formulation in the production of proteins involved in stratum corneum and extracellular matrix integrity. The moisturizing formulation increased the levels of all proteins evaluated. Our results indicate a biological activity of this skin care product in the promotion of cellular hydration and skin barrier integrity.

25th EADV (European Academy of Dermatology and Venereology) Congress, Vienna, Austria, 28 September - 2 October, 2016. Torloni LBO, Pereira AFC, Lage R, Vanzo-Jr AC, Mercuri M, Silva SG, Figueredo TT, Pinheiro ALTA, Silva MS, Facchini G, Eberlin S. Melasma is considered the most common and visible pigmentary skin disorder that affects the face and can become a significant aesthetic discomfort. Exposure to UV light and drugs, as well as genetic characteristics, chronic inflammation, and hormonal influences are associated with the appearance of melasma, although the pathogenesis of this skin disorder is not fully understood. Studies have demonstrated a close relationship between melasma and increased expression of vascular markers, such as VEGF (vascular endothelium growth factor) and iNOS (inducible nitric oxide synthase) in the affected areas. VEGF has been implicated in the increased melanocyte activity since the melanocytes exhibit functional receptors for VEGF, whereas iNOS causes the release of NO and hence the stimulation of tyrosinase and transfer of melanin to melanosomes. The aim of this study was to evaluate the antiangiogenic and whitening effects of a depigmentation formulation (BLTX) using an in vitro model of human cell and skin culture. Human fibroblasts, keratinocytes, or melanocytes were incubated with 3 non-cytotoxic concentrations of BLTX and subjected to oxidative stress by UV radiation or inflammatory stress with IL-1α for quantification of melanin, tyrosinase, endothelin-1, PAR-2, VEGF, and iNOS. Cell supernatants and lysates were collected, and mediators were quantified using colorimetric sandwich assay (ELISA). Fragments of human skin, from elective plastic surgery, were treated with BLTX and subjected to histological evaluation with hematoxylin/eosin associated with Fontana-Masson technique for melanin view. A parametric method, the one-way analysis of variance (ANOVA) followed by the Bonferroni test, was used to compare data among all groups. The results demonstrated that BLTX promotes a reduction in VEGF and iNOS protein synthesis in cultured dermal fibroblasts, indicating an antiangiogenic property. In relation to whitening effect, BLTX was able to reduce the production of melanin in both systems, melanocytes and human skin cultures. The depigmenting action was also revealed by decreasing the levels of endothelin-1, PAR-2, and activity of tyrosinase when compared to cultures exposed to UV radiation. The results allow us to infer that BLTX presents an antiangiogenic effect indicating a role in the vascular component of melasma. Furthermore, the whitening property observed reinforces its use in the prevention and treatment of melasma.

9º Congresso Latino-Americano de Órgãos Artificiais e Biomateriais - COLAOB, 13º Congresso da Sociedade Latino Americana de Biomateriais, Órgãos Artificiais e Engenharia de Tecidos - SLABO, Foz do Iguaçu, 24 a 27 de Agosto, 2016. Vitalli LMM, Bastos TC, Facchini G, Eberlin E. Os efeitos bioestimuladores dos raios infravermelho longo (FIR - Far-infrared ray) sobre a proliferação de fibroblastos e osteoblastos têm sido demonstrados em estudos recentes, com destaque para uma potencial contribuição no processo de regeneração de tecidos. Neste sentido, o presente estudo demonstra os resultados obtidos com o desenvolvimento de um compósito polimérico de poliamida 6 emissor de FIR para aplicação em Engenharia de Tecidos e processamento em prototipagem rápida. Para isso, avaliou-se a caracterização das propriedades mecânicas e térmicas através de ensaios de tensão, impacto, TGA e DSC. De modo complementar, pesquisou-se o efeito do compósito sobre a proliferação de células osteoblásticas (linhagem de pré-osteoblastos murinos MC3T3-E1 Subclone 14) através do método de exclusão do azul de Trypan e BrdU. Além disso, seu efeito antimicrobiano foi investigado segundo a norma JIS Z 2801/2010 para as bactérias S. aureus, S. epidermidis e MRSA (Staphylococcus aureus resistente à meticilina). Os ensaios demonstraram que o compósito polimérico de poliamida 6 emissor de FIR possui características compatíveis para o processamento de prototipagem rápida. Ao mesmo tempo, a taxa de proliferação de osteoblastos observada foi significativamente superior para o compósito em comparação com o observado para a placa polimérica sem aditivo, para os dois métodos avaliados. Assim, de acordo com os resultados obtidos, o compósito polimérico contendo aditivos emissores de infravermelho longo se mostrou promissor para aplicação médica (scaffolds para engenharia de tecidos e implantes ósseos).

American Academy of Dermatology 74th Annual Meeting AAD, Washington-DC, USA, March 4-8, 2016. Lage R, Cascais LC, Clerici SP, Eberlin S, Figueredo TT, Mercuri M, Pereira AFC, Pereira CS, Pinheiro ALTA, Torloni LBO, Tubone JY, Vanzo-Jr AC. The pilosebaceous unit is a major target of androgens in human skin. Sebaceous glands are located in the dermis and their ducts end at the terminal portion of the hair follicles. The sebaceous secretion produced by those glands is a complex mixture of lipids containing triglycerides, free fatty acids, cholesterol, and esters. The enzymes 5-alpha-reductase (5-alpha-reductase) and 17-beta-hydroxysteroid dehydrogenase (17-beta-HSD) are directly or indirectly responsible for converting testosterone to dihydrotestosterone (DHT), an androgen which has greater biological activity in the tissues, favoring sebum secretion and acne formation. Besides sebum secretion, the pathogenesis of acne also involves an inflammatory response and proliferation of bacteria aggravating injury. Therefore, oily skin is more likely to develop inflammatory processes such as acne and seborrhea, requiring products to control this oil. In this study, we evaluated the in vitro efficacy of an active ingredient in anti-sebum activity and in the protection of the skin barrier in cultured human fibroblasts and keratinocytes. In fibroblast culture, the active ingredient promotes a significant reduction in the production of 5-alpha-reductase and 17-beta-HSD, in the presence or absence of testosterone. Interestingly, the active ingredient promoted further reductions in the production of 5-alpha-reductase and 17-beta-HSD when incubated concurrently with testosterone and finasteride, indicating an additive or synergistic action in the control of oiliness. In relation to skin barrier properties, the active ingredient promoted significant increases in the synthesis of envelope protein involucrin, keratin 10, and keratin 14, indicating an involvement in maintaining stratum corneum integrity. These results demonstrate significant anti-sebum and anti-androgenic activities of the active ingredient, since the enzymes 5-alpha-reductase and 17-beta-HSD are implicated in the induction of DHT synthesis, stimulating sebum production.

XXII COLAMICQ (Congreso LatinoAmericano de Químicos Cosméticos), Uruguai, 27-29, Outubro, 2015. Vanzo-JR, Antonio Carlos, Pereira, Andreia Feital Costa, Justa, Sérgio Ricardo, Lage, Renan, Mercuri, Maurizio, Cascais, Lúcia Carrito, Silva, Sheila Gomes, Araujo, Roberta Barros, Tubone, Júlia Yamashita, Ferreira, Ana Sofia Barbosa, Moreira, Fernanda, Satake, Cinthia Yuka, Maciente, Giulia Okamoto, Clerici, Stefano Piatto, Facchini, Gustavo, Pinheiro, Ana Lúcia Tabarini Alves, Pinheiro, Adriano Silva, Eberlin, Samara. A derme, bem como as camadas mais profundas da epiderme, são as estruturas mais danificadas durante o processo oxidativo induzido pela radiação solar e outros fatores ambientais. Um evento inevitável é a diminuição na síntese e degradação das principais proteínas dérmicas, colágeno e elastina, além do fator transformador do crescimento beta (TGF-β), essenciais para a sustentação do tecido. Essa diminuição associada ao aumento de enzimas proteolíticas como as metaloproteinases (MMPs) leva a um colapso da matriz extracelular e, consequentemente, aos sinais prematuros do envelhecimento cutâneo. O objetivo deste estudo foi avaliar as propriedades restauradoras e regeneradoras teciduais de um dermocosmético através da quantificação de colágeno total, colágenos tipo I e tipo III, elastina, TGF-β e migração celular, utilizando cultura de fibroblastos humanos. Os resultados demonstraram que o tratamento das culturas celulares com o dermocosmético promoveu um aumento de 18,7% na produção de colágeno total, em relação à condição basal. Em relação aos colágenos tipo I e tipo III, o aumento foi mais pronunciado, sendo que o dermocosmético elevou os níveis dessas proteínas em 1,8 e 2,7 vezes, respectivamente. Embora não significativa, a substância investigacional apresentou uma tendência em aumentar a produção de elastina. A substância investigacional também foi capaz de promover um aumento significativo de 4,1 vezes na síntese de TGF-β. Esses resultados estão em consonância com os obtidos no estudo de regeneração tecidual, onde o dermocosmético promoveu um aumento da migração celular, acelerando o fechamento do scratch. Os resultados obtidos nos permitem inferir que o dermocosmético avaliado exerce um efeito benéfico na prevenção do processo de envelhecimento cutâneo, aumentando a capacidade de retração e o poder tensor da pele, minimizando dessa forma o aparecimento de rugas e acelerando o reparo tecidual.

24th EADV (European Academy of Dermatology and Venereology) Congress, Copenhagen, 7-11 October, 2015. Vanzo ACJR, Pereira AFC, Cascais LC, Mercuri M, Torloni LBO, Lage R, Andrade CC, Yamashita JT, Clerici SP, Eberlin S, Pinheiro ALTA, Pinheiro AS. Extrinsic aging is attributed to changes in the skin due to lifestyle, being influenced mainly by ultraviolet radiation (UV) but also by pollution resulting from smoking, chemicals, heat, and other environmental insults. Solar exposure results in an increased glycation process—formation of AGE (advanced glycation end products), which not only influences the properties of collagen and the extracellular matrix but also affects the interactions of cells with the matrix, resulting in physical changes such as migration, growth, proliferation, differentiation, and gene expression. Heat Shock Proteins (HSP) play an important role in the defense mechanism, decreasing the vulnerability of cells to environmental attack, functioning as a chaperone to protect the structure of procollagen during the unfolding phase by binding to a triple-helix. In this study, we evaluated the preclinical effectiveness of an eye care active compound (ECAC) in protecting against photodamage caused by ultraviolet radiation in cultured human fibroblasts and keratinocytes. We measured the synthesis of HSP-47, AGEs, receptor for AGEs (rAGE), NF-κB, as well as the activity of catalase, superoxide dismutase, and glutathione reductase, using an in vitro model of human fibroblasts and keratinocytes. The active compound decreased the consumption of glutathione reductase, superoxide dismutase, and catalase by 53.3%, 70.1%, and 76.8%, respectively, when compared to the group exposed to UV radiation. We also observed inhibition of DPPH and ABTS radicals at all ECAC concentrations tested. Regarding the synthesis of the inflammatory marker NF-κB, significant reductions were also obtained compared to the only irradiated group. The active compound demonstrated antiglycation activity by decreasing the production of AGEs and HSP-47, as well as increasing receptors for AGEs. The results enable us to infer that ECAC exerts a protective effect against UV radiation through mechanisms involving the capture and neutralization of free radicals, thus sparing the consumption of antioxidant enzymes glutathione reductase, catalase, and superoxide dismutase. A decrease in activation of the HSP-47 chaperone for collagen synthesis protection was also observed, confirming the protective effect of the test substance. Additionally, the compound showed antiglycation activity as it reduced the generation of AGEs induced by UV radiation, as well as increased the amount of unbound receptor for AGE. These findings, associated with the anti-inflammatory effect reported in this study, support the use of this active compound in mitigating the harmful effects of prolonged exposure to UV radiation.

23rd IFSCC Conference (International Federation of Societies of Cosmetic Chemists), Zurich, 21-23 September, 2015. Pereira AFC, Torloni LBO, Lage R, Cascais LC, Andrade CC, Yamashita JT, Moreira F, Clerici SP, Eberlin S, Pinheiro ALTA, Pinheiro AS. About 40-50% of total solar energy is infrared radiation, which penetrates deeply into the human skin, reaching the subcutaneous layer. Studies have emphasized the effect of IR-A on photoaging and skin damage. Physiological doses of IR-A lead to a disturbance of the dermal extracellular matrix by upregulating the collagen-degrading enzyme matrix metalloproteinase-1 (MMP-1) and decreasing antioxidant enzyme activity. As such, infrared induces cytotoxicity, DNA damage, and oxidative stress. Thus, photoprotection of human skin against IR-A must be considered. Even though no specific chemical or physical filters directed against IR-A are available, there is an alternative that can provide broader sunscreen protection using antioxidant ingredients. In this study, we evaluated the in vitro effects of 17 sunscreen formulations containing antioxidant ingredients in the prevention of infrared-A-induced damage in human skin fibroblasts through MMP-1 production. Our results demonstrated that irradiation with infrared A resulted in a significant upregulation of MMP-1 production, up to 47% in relation to non-irradiated controls. All sunscreen formulations produced significant reductions in the release of MMP-1 by human fibroblasts compared to the IR-A group. The results can be expressed as a percentage of protection provided by the test substances against infrared-A radiation, considering the production of MMP-1 at baseline without exposure to the radiation. Under these conditions, sunscreens, when incubated with cultures of fibroblasts, promoted up to 100% protection in the release of MMP-1. These sunscreen formulations are suitable for protection against IR-A-induced MMP-1 upregulation in vitro in human dermal fibroblast cell culture. These effects can be attributed, at least in part, to the antioxidant ability of the active ingredients present in the formulations. Although the precise mechanisms remain to be clarified, the results allow us to infer that the evaluated sunscreens exert a protective effect against the excessive increase in the synthesis of MMP-1 induced by infrared-A radiation - which reaches deeper into the skin - preserving collagen, a fundamental structural component of tissue support, whose damage contributes to the skin aging process.

23rd IFSCC (International Federation of Societies of Cosmetic Chemists) Conference, Zurich, 21-23 September, 2015. Pereira AFC, Torloni LBO, Lage R, Cascais LC, Andrade CC, Yamashita JT, Moreira F, Clerici SP, Eberlin S, Pinheiro ALTA, Pinheiro AS. The generation and activation of reactive oxygen species and other reactive agents can cause damage to cellular DNA, and the immediate repair capacity of such damage is an important cellular mechanism that protects cells and maintains genetic stability, preventing early oncogenesis. Mammalian cells have a defense mechanism designed to maintain genomic integrity and prevent the establishment of permanent damage resulting from genotoxic stress. In this context, the GADD45a protein (growth arrest DNA damage 45 alpha) plays a key role as a cellular stress sensor through interaction with other proteins. In response to stress, GADD45a can promote the control of cell cycle regulation, DNA repair, epigenetic changes, apoptosis, survival, and senescence. The purpose of this study was to evaluate the in vitro effectiveness of 18 sunscreen formulations in protecting against photodamage caused by ultraviolet radiation (UV) in cultured human fibroblasts. Our results showed that UV radiation produced a significant reduction of approximately 60% in the production of GADD45a when compared to the non-irradiated control baseline (P<0.001). One possible explanation for this effect is the increased consumption and degradation of this protein as a result of genotoxic stress, which could result in a transient reduction in the levels of GADD45a in fibroblast cultures. As mentioned, the absence of this protein can lead to genomic instability and deficiencies in the repair of DNA damage. On the other hand, concomitant incubation of cell cultures with photoprotective formulations prevented the marked reduction of this protein caused by UV radiation, promoting up to 100% protection against the decrease in GADD45a. Extrinsic aging is attributed to changes in the skin due to lifestyle, being influenced mainly by ultraviolet radiation (UV), but also by pollution resulting from smoking, chemicals, heat, and other environmental insults. One of the main targets of UV radiation is DNA. The nitrogenous bases may undergo structural changes resulting in pyrimidine dimers that can destabilize the DNA molecule. Under normal exposure conditions, the body has enzyme systems that repair these changes in thymines, but with cumulative and prolonged exposures, this system can fail, and UV radiation can trigger mutations in the DNA structure that lead to the activation of proto-oncogenes. The results presented in this work allow us to infer that cosmetic sunscreens have an important effect in the process of DNA damage repair generated as a result of genotoxic stress from UV radiation by reducing GADD45a consumption and degradation.

23rd IFSCC Conference (International Federation of Societies of Cosmetic Chemists), Zurich, 21-23 September, 2015. Eberlin S, Clerici SP, Eberlin S, Pinheiro ALTA, Pinheiro AS. Skin pigmentation is dependent on both the increased number of melanocytes and the increase in melanogenic enzymes. After exposure of the skin to sunlight, the melanogenesis process begins with the oxidation of the amino acid tyrosine by the enzyme tyrosinase, producing DOPA, which is metabolized to produce melanin. Melanin plays an important role in protecting the skin from the harmful effects of solar radiation, but excessive accumulation of this pigment can become an aesthetic problem. Exposure to sunlight, artificial tanning, pharmaceuticals, and other chemicals, as well as chronic inflammatory processes and hormonal influences, can increase the production of factors such as melanocyte-stimulating hormone and endothelin-1, which trigger the melanization of the skin. In addition to ultraviolet light, studies also show that infrared radiation and visible light promote changes in skin pigmentation. In this work, we evaluated the consequences of ultraviolet A and B (UVA/UVB), infrared-A (IR-A), and visible light (VL) irradiation on melanin and endothelin-1 synthesis, using an in vitro model of melanocyte and keratinocyte cultures, and an ex vivo model of skin culture. All radiation types (UV, IR-A, VL, and Association) produced a significant increase of 8.22%, 5.32%, 5.97%, and 10.56%, respectively, in melanin production when compared to the non-irradiated control (p<0.001), contributing to the process of cutaneous pigmentation. The association of radiation produced a further increase of 4.97% and 4.32%, respectively, in melanin synthesis compared to IR-A and VL radiations applied alone. UV radiation, VL, and the association of all radiation produced a significant increase of 53.27%, 63.81%, and 57.81%, respectively, in the production of endothelin-1 when compared to the non-irradiated control. Despite IR-A showing a trend in increased synthesis of endothelin-1, it did not produce a statistically significant result. Consistent with these results, skin fragments subjected to radiation had a higher melanin density in comparison with control fragments. An increasing number of papers have been published describing the harmful effects of exposure to mainly or exclusively short-wavelength IR (IR-A) radiation in cultures of human skin cells and in human skin biopsies. Similarly, visible light has been identified as not inert radiation, and studies have demonstrated that it increases cutaneous pigmentation, even more than UV radiation. Our results corroborate previous data from the literature and indicate the need for specific photoprotection strategies that are not addressed by conventional sunscreens available.