Artículos y publicaciones

Inflammation & Allergy Drug Targets 13(3), 191-8, 2014. DOI: http://dx.doi.org/10.2174/1871528113666140616112708 Costa A, Eberlin S, Polettini AJ, Pereira AFC, Pereira CS, Ferreira NMC, Dolis E, Torloni LBO. The manifestation of sensitive skin occurs as a consequence of increased permeability of the stratum corneum, as well as the involvement of the neuro-immune-endocrine system. In this study, we evaluated the effects of an active ingredient, SensC, on the production of neuropeptides substance P (SP), enkephalin, and β-endorphin; eicosanoids prostaglandin E2 (PGE2) and leukotriene B4 (LTB4); histamine, TRPV1, and envelope proteins filaggrin and involucrin, using an in vitro model of human cell culture. Our results demonstrated that treatment of keratinocyte cultures with SensC prevented the increase of all evaluated inflammatory mediators induced by interleukin-1 alpha (IL-1α). Similarly, SensC provided a decrease in the synthesis of TRPV1. Regarding the synthesis of envelope proteins, SensC promoted increases in filaggrin and involucrin levels compared to the control group. Considering the absence of appropriate treatment, the availability of ingredients such as SensC, with anti-inflammatory and protective barrier properties, can be a significant tool for preventing neurosensorial symptoms associated with sensitive skin.

Surgical and Cosmetic Dermatology 7(2), 320-4, 2014. http://www.surgicalcosmetic.org.br/detalhe-artigo/355/Estudo-clinico-para-avaliar-a-reducao-da-circunferencia-abdominal-apos-tratamento-com-laser-diodo-de-baixa-frequencia Costa A, Eberlin S, Jorge ACF, Mendonça JA, Kalies AH, Pereira CS. A obesidade é um dos principais problemas de saúde no mundo, devido à sua elevada morbimortalidade. Este estudo avaliou a redução da gordura abdominal em voluntários submetidos à terapia com laser diodo de baixa frequência. Foram avaliados 60 voluntários, entre 18 e 50 anos de idade, com IMC entre 18,5 kg/m² e 30 kg/m² e com gordura abdominal, submetidos ao tratamento realizado duas vezes por semana, durante 31 dias. Avaliou-se a circunferência abdominal em três áreas, com tomadas de imagens fotográficas e ultrassom da área avaliada. Houve redução nas medidas da circunferência abdominal nas áreas avaliadas, com significativa compactação ultrassonográfica dérmica e redução ultrassonográfica da hipoderme. A terapia com laser diodo de baixa frequência apresentou resultados significativos na redução de gordura localizada e medidas de circunferência abdominal.

Journal of the American Academy of Dermatology, 70(5) Supplement 1, AB154, 2014. DOI: http://dx.doi.org/10.1016/j.jaad.2014.01.641 Eberlin S, Silva MS, Polettini AJ, Coletta LCD, Costa A, Queiroz MLS. Recent studies demonstrated that photoaging and skin damage are associated with longer wavelengths, in particular near-infrared radiation. Studies have been conducted to determine the damage caused by infrared-A (IR-A) radiation, particularly the upregulation of MMP-1, using in vitro and/or in vivo investigations. For the detection of MMP-1, authors used several IR simulator devices and determined increases in MMP-1 levels through gene expression, western blotting, and immunohistochemistry techniques. Nevertheless, no information concerning MMP-1 quantification using an ELISA assay had been published. In this study, we validated the optimal incubation time for the release of MMP-1 in culture supernatants by human dermal fibroblasts (HDF) stimulated with IR-A radiation. Our results demonstrated that irradiation with IR-A resulted in a significant upregulation of MMP-1 production, 1.7-fold in relation to the non-irradiated group, only at an incubation time of 72 hours after radiation. After 24 and 48 hours of IR-A radiation, no significant differences were observed when compared to control groups. In this work, we standardized the optimal time for human fibroblast incubation to produce MMP-1 after IR-A radiation. Contrary to previous studies in the literature, where MMP-1 gene expression is observed after 24 hours from IR-A exposure, the production of this protease in supernatants is only detected after 72 hours of incubation.

Journal of the American Academy of Dermatology, 70(5) Supplement 1, AB155, 2014. DOI: http://dx.doi.org/10.1016/j.jaad.2014.01.644 Eberlin S, Pereira AFC, Polettini AJ, Weisz LTM, Mendes C, Lage R, Costa A. Physiological doses of infrared A radiation (IR-A) lead to a disturbance of the dermal extracellular matrix by upregulation of matrix metalloproteinase-1 (MMP-1) and decreased antioxidant enzyme activity. As such, infrared induces cytotoxicity, DNA damage, and oxidative stress. Even though no specific chemical or physical filters directed against IR-A are available, there is an alternative that can provide broader sunscreen protection against photodamage, using antioxidant ingredients. In this study, we evaluated the in vitro effects of two formulations containing the antioxidant ingredients FThe and FCar in the prevention of IR-A-induced damage in human dermal fibroblasts (HDF) through MMP-1 production. Our results demonstrated that irradiation with infrared A resulted in a significant upregulation of MMP-1 production, 1.7-fold in relation to the non-irradiated group. FCar produced a significant reduction in the release of MMP-1 by human fibroblasts compared to the IR-A group. More pronounced effects were observed with FThe, which promoted reductions of up to 1.35-fold in the levels of MMP-1 when compared to the IR-A group. The formulations containing FCar and FThe are suitable for protecting 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 known antioxidant activity of these compounds, although the precise mechanisms remain to be clarified.

23rd EADV Congress (European Academy of Dermatology and Venereology); Amsterdam, October 8-12, 2014. Eberlin S, Polettini AJ, Clerici SP, Eberlin S, Pereira AFC, Lage R, Torloni LBO, Costa A. Evidence suggests that periorbital hyperchromia occurs mainly as a consequence of post-inflammatory hemodynamic congestion, producing a typical bruising aspect on the lower eyelids. The local inflammatory process compromises the microvascular network due to the release of vasoactive mediators, such as nitric oxide (NO), which is synthesized from inducible NO synthase (iNOS) after tissue disturbance. Another common manifestation observed in the periorbital area is the presence of fine wrinkles as a consequence of connective tissue breakdown due to the increased activity of matrix metalloproteinases (MMPs) after cytokine and eicosanoid stimulation. In this study, we evaluated the effects of a cosmetic eye care product (EC) on the production of MMP-1, MMP-9, tissue inhibitor of MMP-1 (TIMP-1) and TIMP-2, collagen, elastin, transforming growth factor (TGF-β1), and iNOS, using an in vitro model of human fibroblasts. UV radiation promoted significant increases in the levels of MMP-1, MMP-9, and iNOS, and decreases in collagen, elastin, TGF-β1, TIMP-1, and TIMP-2 synthesis compared to the control group. Concomitant treatment of cell cultures with EC reduced the levels of MMP-1 and MMP-9 by up to 65% and 72%, respectively, compared to the UV-stressed group. Increased levels of iNOS were also prevented by EC treatment, reaching undetectable levels similar to the baseline control. EC treatment also increased elastin, TGF-β1, and TIMP-1 release in relation to the UV-stressed group (98%, 6%, and 75%, respectively). Although the precise mechanisms related to EC remain to be clarified, our results indicate that the reduction of iNOS levels against deleterious UV radiation may be considered an integral approach to preserving the integrity of the vascular endothelium, providing adequate cutaneous circulation, which is essential for maintaining healthy skin function and is strongly related to skin color. Furthermore, our results indicate a putative effect of EC on extracellular matrix arrangement, strengthening its use in aesthetic eye care.

23rd EADV Congress (European Academy of Dermatology and Venereology); Amsterdam, October 8-12, 2014. Costa A, Eberlin S, Polettini AJ, Jorge ACF, Eberlin S, Pereira AFC, Cascais LC, Dolis E, Frazilho M, Lage R, Torloni LBO. Rosacea is a chronic inflammatory dermatosis characterized by flushing, permanent erythema, telangiectasia, papules, and pustules on the central area of the face, affecting middle-aged patients. Its underlying pathophysiological mechanisms are not completely understood, although the disturbances observed are related to vascular reactivity and inflammatory response. Local production of inflammatory mediators, such as prostaglandins (PG) and histamine (HI), leads to vasodilation and increased vascular permeability due to additional release of VEGF (vascular endothelial growth factor) and inducible nitric oxide synthase (iNOS). In this study, we evaluated the effects of an active ingredient (AI) on the production of PGE2, HI, VEGF, and iNOS, using an in vitro model of human keratinocytes. Incubation of cell cultures with IL-1β promoted significant increases in the synthesis of HI, PGE2, VEGF, and iNOS compared to the control group. Concomitant treatment of cell cultures with AI prevented the increase of all evaluated mediators. In relation to HI and PGE2, AI was able to reduce the levels by up to 24.8% and 47%, respectively, compared to the IL-1β-stressed group. Increased levels of VEGF and iNOS were also prevented, reaching 51.2% and 100%, respectively, in relation to the IL-1β-stressed group. Flushing is one of the most evident signs in Rosacea, and increased blood flow in the superficial dermis is the main factor responsible for this phenomenon. In this work, it was demonstrated that AI exerts vasoprotective and anti-inflammatory effects, suggesting a possible application in skin care product formulations to contribute to the amelioration of Rosacea symptoms.

7º Simpósio de Cosmiatria & Laser da Sociedade Brasileira de Dermatologia, Curitiba - PR, 9-10 de maio, 2014 - Prêmio 1º lugar. Costa, Adilson; Pereira, Caroline S.; Jorge, Adriana C.F.; Polettini, Anelise J.; Eberlin, Samara. O envelhecimento extrínseco é atribuído a mudanças ocorridas na pele devido ao estilo de vida, sendo influenciado, sobretudo, pela radiação ultravioleta (UV), mas também pela poluição, resultante do tabagismo, produtos químicos, calor e demais insultos ambientais. A exposição à radiação UV provoca danos celulares em consequência da produção excessiva de radicais livres e do enfraquecimento dos mecanismos de defesa antioxidantes, como o sistema homeostase redox que consiste na ação das enzimas catalase, superóxido dismutase e glutationa redutase. Da mesma maneira, a fumaça do cigarro contém compostos oxidantes e pró-oxidantes, produzindo radicais livres e aumentando o estresse oxidativo, resultando na formação de hidroperóxidos lipídicos, como o malondialdeído, altamente reativos e instáveis que podem levar à oxidação de lipídeos nas membranas celulares em todo o extrato córneo. A exposição a esses e outros insultos pode resultar na degradação do tecido conectivo com perda da firmeza e elasticidade, glicação do colágeno, enfraquecimento no reparo tecidual e cicatrização de feridas, redução da hidratação, perda da função protetora da barreira cutânea, entre outros. Uma estratégia para o desenvolvimento de fotoprotetores capazes de prevenir o fotodano causado pela radiação UV e pela exposição à poluição envolve a utilização de substâncias antioxidantes que evitam o desequilíbrio redox e mantêm a homeostase celular, contendo a progressão das reações em cadeia que oxidam substratos orgânicos. O objetivo deste estudo foi avaliar a eficácia antioxidante pré-clínica de um complexo de ativos na proteção contra o fotodano causado pela radiação UV e pela fumaça de cigarro através da quantificação das enzimas antioxidantes utilizando um modelo de cultura de células de pele humana. Foram usadas três estratégias para avaliação das propriedades antioxidantes do complexo. A primeira baseou-se na mensuração das enzimas antioxidantes catalase, superóxido dismutase e glutationa redutase, utilizando uma cultura de fibroblastos humanos submetidos à radiação UV. A substância-teste demonstrou um efeito protetor das culturas celulares, poupando o consumo enzimático nos grupos tratados. A segunda parte do trabalho consistiu na determinação da atividade anti-radical livre através da habilidade do complexo de ativos em capturar os radicais DPPH e ABTS. A substância-teste demonstrou uma atividade anti-radical livre através da captura dos radicais ABTS e DPPH. A terceira parte desse estudo foi avaliar a habilidade da substância-teste na proteção contra a peroxidação lipídica causada pela exposição das culturas de fibroblastos humanos à fumaça de cigarro. A exposição à fumaça de cigarro promove um aumento de 34,4% na produção de MDA (malondialdeído) comparado ao controle basal não exposto. Esse resultado demonstra o efeito nocivo da fumaça de cigarro no processo de peroxidação lipídica. Por outro lado, a substância-teste demonstrou um efeito protetor contra a peroxidação lipídica, bloqueando em até 100% a síntese de MDA, quando comparado ao grupo exposto à fumaça de cigarro. Os resultados obtidos nos permitem inferir que esse complexo de ativos pode ser utilizado em produtos cosméticos e dermatológicos, conferindo proteção contra os efeitos nocivos do envelhecimento extrínseco.

72nd Annual Meeting AAD - American Academy of Dermatology, Denver, USA, March 21-25, 2014. Eberlin S, Silva MS, Polettini AJ, Coletta LCD, Costa A, Queiroz MLS. Recent studies demonstrated that photoaging and skin damage are associated with longer wavelengths, in particular near-infrared radiation. Studies have been conducted in order to determine the damage caused by infrared-A (IR-A) radiation, particularly the upregulation of MMP-1, using in vitro and/or in vivo investigations. For the detection of MMP-1, authors used several IR simulator devices and determined increases in MMP-1 levels through gene expression, western blotting, and immunohistochemistry techniques. Nevertheless, no information concerning MMP-1 quantification using ELISA assay has been published. In this study, we validated the optimal incubation time for the release of MMP-1 in culture supernatants by human dermal fibroblasts (HDF) stimulated with IR-A radiation. Our results demonstrated that irradiation with IR-A resulted in a significant upregulation of MMP-1 production, 1.7-fold in relation to the non-irradiated group, only at an incubation time of 72 hours after radiation. After 24 and 48 hours of IR-A radiation, no significant differences were observed when compared to control groups. In this work, we standardized the optimal time for human fibroblast incubation to produce MMP-1 after IR-A radiation. Contrary to previous studies in the literature, where MMP-1 gene expression is observed after 24 hours from IR-A exposure, the production of this protease in supernatants is only detected after 72 hours of incubation.

72nd Annual Meeting AAD - American Academy of Dermatology, Denver, USA, March 21-25, 2014. Eberlin S, Pereira AFC, Polettini AJ, Weisz LTM, Mendes C, Lage R, Costa A. Physiological doses of infrared A radiation (IR-A) lead to a disturbance of the dermal extracellular matrix by upregulation of matrix metalloproteinase-1 (MMP-1) and decreased antioxidant enzyme activity 1 . As such, infrared induces cytotoxicity, DNA damage, and oxidative stress 2 . Even though no specific chemical or physical filters directed against IR-A are available, there is an alternative that can provide broader-spectrum sunscreen and favor protection against photodamage, using antioxidant ingredients 3 , such as Thermus Thermophilus Ferment (FThe) 4 and Carnosine (FCar) 5 . In this study, we evaluated the in vitro effects of two formulations containing the antioxidant ingredients FThe and FCar in the prevention of IR-A-induced damage in human dermal fibroblasts (HDF) through MMP-1 production. HDF were incubated with non-cytotoxic concentrations of FCar and FThe (0.316, 0.100, and 0.0316 mg/mL). Cells were exposed to a non-cytotoxic dose 3 of 360 J/cm 2 IR-A radiation, using a Hydrosun 750T IR-A device, and the irradiance was measured using a Hydrosun HBM1 irradiance measuring device. The levels of MMP-1 were measured after 72 hours in culture supernatant using a commercially available kit. Our results demonstrated that irradiation with infrared A resulted in a significant (P<0.0001, ANOVA-Tukey) upregulation of MMP-1 production (8.37±0.44 ng/mL), 1.7-fold compared to the non-irradiated group (4.91±0.64 ng/mL). FCar at 0.0316 mg/mL produced a significant reduction (P<0.01, ANOVA-Tukey) in the release of MMP-1 by human fibroblasts (6.48±0.66 ng/mL) compared to the IR-A group. More pronounced effects (P<0.001, ANOVA-Tukey) were observed for FThe at 0.100 and 0.0316 mg/mL, promoting reductions of up to 1.35-fold (6.18±0.51 ng/mL and 6.29±0.30 ng/mL, respectively) in the levels of MMP-1 when compared to the IR-A group. The formulations containing FCar and FThe are suitable for protecting 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 known antioxidant activity of these compounds, although the precise mechanisms remain to be clarified.