Antiaging effects of a skin care formulation containing nanoencapsulated antioxidants: A clinical, in vitro, and ex vivo study.

BACKGROUND: The development of effective cosmetic products for the reduction of the signs of skin aging is a complex process which requires an optimized combination of ingredients and specialized systems to deliver the actives to the skin layers. AIM: To evaluate the tolerance and antiaging clinical efficacy of a cosmetic formulation containing a blend of nanoencapsulated antioxidants: ascorbyl palmitate, resveratrol, tocopherol, caffeine, carnosine, and niacinamide. METHODS: Clinical efficacy was determined by subjective and instrumental analyses of collagen synthesis by fluorescence spectroscopy, by three-dimensional imaging analysis of suborbital edema, and by analysis of skin hydration and sebum content by biophysical techniques-Corneometer® and Sebumeter®. RESULTS: The studied formulation was safe and effective for the improvement of skin appearance by increasing collagen synthesis and skin moisturizing and by reducing facial blemishes, swelling, and oiliness. A preclinical exploratory approach using an experimental model of human cell and skin cultures agreed with the observed antiaging effects, identifying mechanisms related to the containment of oxidative stress, reduction of melanin production, increased synthesis of type I procollagen, and regulation of the epidermal cohesion protein filaggrin. CONCLUSIONS: The skin benefits obtained resulted from the combination of the ingredients in the formulation and the nanoencapsulation-based delivery system, which favors the solubility, safety, efficacy, and bioavailability of the preparation to the skin.

Estudo comparativo para avaliação da eficácia cicatrizante de formulações tópicas contendo Triticum aestivum L. (sinônimo Triticum vulgare) em modelo de pele humana nativa / Comparative study to evaluate the wound healing efficacy of topical formulations containing Triticum aestivum L. (Triticum vulgare) in a native human skin model

Introdução: extratos vegetais e ativos derivados de plantas tem sido desenvolvidos com o objetivo de melhorar e potencializar o processo de cicatrização cutânea, dentre eles, o Triticum aestivum L. (sinônimo Triticum vulgare). Objetivo: avaliar o efeito do extrato de grão inteiro (EGTA-PR) e extrato aquoso (EATA-FI) de Triticum aestivum L. na cicatrização cutânea em pele humana ex vivo. Métodos: fragmentos de pele obtidos de cirurgia plástica eletiva foram submetidos a lesões teciduais e tratados com os extratos durante oito dias para avaliação histológica da reepitelização e marcação proteica do fator de crescimento epidérmico (EGF). Resultados: EGTA-PR e EATA-FI aceleraram o processo de reepitelização em cultura de pele humana submetida a lesão tecidual. Adicionalmente, foi observado um aumento da marcação proteica de EGF após o tratamento com EGTA-PR. Conclusão: EGTA-PR apresentou um melhor desempenho na reepitelização quando comparado ao EATA-FI, pois apresentou uma maior marcação proteica para EGF em cultura de pele humana. Da mesma forma, os resultados histológicos mostraram que a redensificação dérmica obtida com o EGTA-PR foi visualmente superior à observada com EATA-FI. Os resultados obtidos são promissores e corroboram as diversas ações biológicas já reportadas na literatura para extrato de Triticum aestivum L. nas etapas da cicatrização tecidual.

Introduction: Plant extracts and actives derived from plants were developed to improve and enhance the skin healing process including Triticum aestivum L. (Triticum vulgare). Purpose: To evaluate the effect of whole grain extract (EGTA-PR) and aqueous extract (EATA-FI) of Triticum aestivum L., on ex vivo skin healing. Methods: Skin fragments obtained from elective plastic surgery were subjected to tissue damage and treated with extracts for eight days for histological evaluation of re-epithelialization and immunofluorescence for epidermal growth factor (EGF). Results: EGTA-PR and EATA-FI accelerated the re-epithelialization process in human skin culture submitted to tissue injury. Additionally, we observed increased EGF protein labeling after treatment with EGTA-PR. Conclusion: EGTA-PR showed a better performance in re-epithelialization when compared to EATA-FI, as it presented a higher protein labeling for EGF in human skin culture. Likewise, the histological results showed that the dermal redensification obtained with EGTA-PR was visually superior to that observed with EATA-FI. The results obtained are promising and corroborate the several biological actions already reported in the literature for Triticum aestivum L. extract in tissue healing stages

Invitro effect of pine bark extract on melanin synthesis, tyrosinase activity, production of endothelin-1, and PPAR in cultured melanocytes exposed to Ultraviolet, Infrared, and Visible light radiation.

BACKGROUND: French maritime pine bark (Pinus pinaster) extract (PBE), the registered trade name of which is Pycnogenol® , has been studied for its depigmenting action due to its antioxidant, anti-inflammatory, and anti-melanogenic activity. However, the mechanisms through which PBE are still not fully clear. OBJECTIVE: Evaluate the impact of PBE on four in vitro parameters closely associated with cutaneous pigmentation, including melanin synthesis, tyrosinase activity, endothelin-1 (ED1), and production of peroxisome proliferator-activated receptor α, δ, and γ (PPAR α, δ, and γ), by studying the modulation of action of ultraviolet radiation A (UVA)/ultraviolet radiation B (UVB), infrared-A (IR-A), visible light (VL), and association of UVA/UVB, IR-A, and VL (ASS). METHODS: Human melanocytes were incubated in a dry extract solution of PBE, exposed to UVA/UVB, IR-A, VL, and ASS for subsequent quantification of melanin, ED1, and PPAR α, δ, and γ. The effects of PBE on inhibition of tyrosinase activity were also performed by monophenolase activity assay. RESULTS: UVA/UVB, IR-A, VL, and ASS radiation caused significant increases in the synthesis of melanin, ED1, and PPAR α, δ, and γ when compared to baseline control. However, PBE significantly reduced the production of melanin, ED1, and PPAR α, δ, and γ, as well as reducing about 66.5% of the tyrosinase activity. CONCLUSIONS: PBE reduces in vitro melanin production by downregulating tyrosinase and reducing pigmentation-related mediators, such as ED1 and PPAR α, δ, and γ, therefore contributing to the inhibition of pathways associated with skin hyperpigmentation.

Ex Vivo Human Skin: An Alternative Test System for Skin Irritation and Corrosion Assays.

Native human skin has been reported in the literature as being an important experimental model for studying skin biology. Studies performed by our group have shown that ex vivo skin, from elective plastic surgery, maintains the biological characteristics of native skin under specific culture conditions. As such, it might be a feasible model for the safety and efficacy testing of topical substances. While Brazil is at the forefront of global regulation implementation, Brazilian researchers are not always able to transfer certain widely used protocols to their laboratories, particularly protocols that involve the use of reconstructed tissues with limited viability, such as those for skin corrosion (OECD TG 431) and irritation testing (OECD TG 439). In this study, we investigated the applicability of the ex vivo skin model to the evaluation of irritation and corrosion potential of a number of proficiency substances described in TG 431 and TG 439. The skin fragments were standardised in size and diameter, and placed into cell culture inserts. The experimental protocol was conducted according to TG 431 and TG 439. The results obtained show that ex vivo skin could represent a promising tool for the evaluation of irritation and corrosion potential of substances (subject to inclusion and exclusion criteria), as recommended by OECD guidelines. While this is a proof-of-concept study, the use of ex vivo skin should be considered for such testing.

Modulation of skin androgenesis and sebum production by a dermocosmetic formulation.

BACKGROUND: Excessive androgenesis in the skin promotes sebaceous hyperproduction which is the onset of acne vulgaris pathogenesis. Free fatty acids and lipid accumulation in the glandular infundibulum culminates in microbiota imbalance, triggering inflammatory response and follicular hyperkeratinization. AIMS: The purpose of this work was to present an alternative cosmetic treatment for acne skin care, focusing on the prevention of sebaceous gland dysregulation. METHODS: Insulin-stimulated human sebocytes were treated with noncytotoxic concentrations of a DTRW cosmetic formulation. After 6 days of incubation, cell lysates were collected for testosterone, 5α-reductase, and dyhidrotestosterone (DHT) quantitation. In parallel, cells were stained with Oil Red O to measure sebum production. RESULTS: Human sebocytes were incubated with insulin to mimic a seborrheic microenvironment with overproduction of intracellular lipids and fatty acids. Concomitant incubation of cell cultures with DRTW was able to promote a 52.97% reduction in intracellular lipid content. The anti-androgenic properties of DRTW had been proved by the reductions of testosterone (↓59.90%), 5α reductase (↓59.34%), and DHT (↓55.98%) levels in sebocyte cultures also stimulated with insulin. CONCLUSION: The results indicate a promising action of DRTW cosmetic formulation in preventing the development of acne lesions by mechanisms involving the modulation of cutaneous androgenesis and consequently the control of sebum overproduction, considered one of the leading causes of acne.

Novel complex of cosmetic ingredients with promising action in preventing hair loss and follicular aging through mechanism involving enrichment of WNT/signaling, mitochondrial activity, and stem cells maintenance.

BACKGROUND: Mechanisms involved in hair metabolism are diverse, and the availability of ingredients that normalize dysfunctions or mitigate the effects of extrinsic stress suffered daily is greatly desired by consumers to improve the aesthetic appearance of hair. AIMS: In this work, we carried out a preclinical exploratory approach to evaluate the effects of a complex of nanoencapsulated active ingredients (AcPi), as well as a cosmetic formulation containing AcPi (ShPi and HtPi) in mechanisms involving hair loss and follicular aging. METHODS: Human hair follicle dermal papilla cells and human scalp culture were treated with AcPi, ShPi, or HtPi and stimulated with UV radiation or testosterone for further measurement of mitochondrial biogenesis, reactive oxygen species (ROS), ß-catenin, dyhidrotestosterone (DHT), collagen XVIIα1 (COL17A1), and cutaneous permeation. RESULTS: Our results demonstrated that AcPi prevents oxidative stress and balances mitochondial activity disturbed by exposure to UV radiation. AcPi also promoted an enrichment of WNT/ß-catenin signaling pathway, stimulating hair growth, and lengthening the anagen phase of hair cycle. ShPi and HtPi were able to prevent hair aging, minimizing the excessive degradation of COL17A1 in hair follicle exposed to UV radiation, in addition to controlling androgenic metabolism by reducing DHT production. CONCLUSION: The integral effects of AcPi have not been completely elucidated; however, these results, associated with clinical evidences, allow us to infer that this ingredient prevents follicular aging, miniaturization, and consequently hair loss by mechanisms involving energetic homeostasis maintenance, antioxidant, and anti-androgenic actions.

The Ex Vivo Skin Model as an Alternative Tool for the Efficacy and Safety Evaluation of Topical Products.

The development of alternative approaches for safety and efficacy testing that avoid the use of animals is a worldwide trend, which relies on the improvement of current models and tools so that they better reproduce human biology. Human skin from elective plastic surgery is a promising experimental model to test the effects of topically applied products. As the structure of native skin is maintained, including cell population (keratinocytes, melanocytes, Langerhans cells and fibroblasts) and dermal matrix (containing collagen, elastin, glycosaminoglycans, etc.), it most closely matches the effects of substances on in vivo human skin. In this review, we present a collection of results that our group has generated over the last years, involving the use of human skin and scalp explants, demonstrating the feasibility of this model. The development of a test system with ex vivo skin explants, of standard size and thickness, and cultured at the air-liquid interface, can provide an important tool for understanding the mechanisms involved in several cutaneous disorders.

Whitening effects of cosmetic formulation in the vascular component of skin pigmentation.

BACKGROUND: Melanin plays an important role in protecting the skin against the harmful effects of solar radiation, but its abnormal accumulation may become an aesthetic problem, such as melasma and age spots. AIMS: 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. METHODS: Human fibroblasts, keratinocytes or melanocytes were treated with 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. 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. RESULTS: 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. CONCLUSION: 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.

Flavonoid-Enriched Plant-Extract-Loaded Emulsion: A Novel Phytocosmetic Sunscreen Formulation with Antioxidant Properties.

The aim of this study was to develop a phytocosmetic sunscreen emulsion with antioxidant effect, containing a blend of flavonoid-enriched plant extracts. In vitro sun protection factor, antioxidant activity, skin irritation, photostability, cutaneous permeation, and retention of flavonoids were evaluated. Thermodynamically stable emulsions were obtained and tested for sensorial analysis after loading the blend of extracts. The selected emulsion was stable when stored at low temperatures (5 C), for which after 120 days the concentration of quercetin and rutin were above their limit of quantification, i.e., 2.8 ± 0.39 µg/mL and 30.39 ± 0.39 µg/mL, respectively. Spreadability, low rupture strength and adhesiveness were shown to be similar to a conventional topical product. Higher brittleness, pseudo-plastic, and viscoelastic behaviors were also recorded for the developed phytocosmetic sunscreen. The product presented a critical wavelength of 387.0 nm and ultraviolet rays A and B (UVA/UVB) rate of 0.78, confirming that the developed formulation shows capacity for UVA/UVB protection, protecting skin against damages caused by Ultraviolet (UV) radiation. Rutin was shown to permeate the skin barrier and was also quantified in the stratum corneum (3.27 ± 1.92 µg/mL) by tape stripping and retention test (114.68 ± 8.70 µg/mL). The developed flavonoid-enriched phytocosmetic was shown to be non-irritant to skin by an in vitro assay. Our results confirm the antioxidant activity, sun protection, and physical properties of the developed phytocosmetic for topical application.

Avaliação pré-clínica dos efeitos profiláticos do extrato de Pinus pinaster (Pycnogenol®) sobre a deposição cutânea de hemossiderina / Pre-clinical evaluation of the profilatic effects of Pinus pinaster extract (Pycnogenol®) on skin hemosiderin deposits

Introdução: A escleroterapia é o método mais utilizado para o tratamento de varizes dos membros inferiores tendo como complicação mais comum o aparecimento de manchas hipercrômicas na região tratada. O Pycnogenol® é conhecido há muito tempo como um flebotônico, anti-inflamatório e despigmentante da pele. Estudos já comprovaram a eficácia deste fármaco na prevenção e no tratamento da hiperpigmentação pós-inflamatória. Objetivo: Avaliar a eficácia do extrato de Pinus pinaster (Pycnogenol®; EPP) na prevenção de depósitos de hemossiderina em cultura de pele humana submetida a estresse inflamatório. Métodos: Fragmentos de pele humana foram estimulados com interleucina 1 alfa (IL-1a) para indução de uma resposta inflamatória e, concomitantemente, tratados com EPP para posterior avaliação histológica e semi-quantificação de hemossiderina. Resultados: A avaliação histológica dos fragmentos de pele expostos à IL-1alfa; revelaram uma densidade de hemossiderina 26,6% maior em comparação ao grupo controle. Por outro lado, os fragmentos de pele incubados concomitantemente com EPP mostraram reduções significativas na deposição de hemossiderina quando comparados ao grupo somente expostos ao microambiente inflamatório. Conclusões: Os resultados apresentados neste estudo apontam para um importante efeito do EPP (Pycnogenol®) na prevenção do acúmulo de hemossiderina originado pelo estresse inflamatório semelhante ao processo pós escleroterapia.

Introduction: Sclerotherapy is the most widely used method for the treatment of varicose veins of the lower limbs. The most common complication is the appearance of hyperchromic spots in the treated region. Pycnogenol® has long been known as a phlebotonic, anti-inflammatory and skin depigmenting drug. Studies have already proven the efficacy of this drug in the prevention and treatment of post inflammatory hyperpigmentation. Objective: To evaluate the effectiveness of Pinus pinaster extract (Pycnogenol®; PPE) in the prevention of hemosiderin deposits in human skin culture submitted to inflammatory stress. Methods: Fragments of human skin were stimulated with interleukin 1 alpha (IL-1 a) to induce an inflammatory response and, concurrently, treated with PPE for further histological evaluation and hemosiderin semi-quantification. Results: The histological evaluation of skin fragments exposed to IL-1 alpha revealed a 26.6% higher hemosiderin density compared with the control group. Moreover, skin fragments incubated concomitantly with PPE showed significant reductions in hemosiderin deposits when compared with the group only exposed to the inflammatory microenvironment. Conclusions: The results presented in this study showed an important effect of PPE (Pycnogenol®) in the prevention of hemosiderin accumulation caused by inflammatory stress similar to the post-sclerotherapy process.

Sterol-standardized phytopharmaceutical from ground cherry: Corticoid-like properties on human keratinocytes and fibroblasts and its effects in a randomized double-blind placebo-controlled clinical trial.

BACKGROUND: Topical corticosteroids have been the most commonly prescribed drugs to treat skin inflammation, but their uses can lead to several adverse effects. Nowadays, new pharmacological strategies have been evaluated to improve dermatologic efficacy and reduce adverse effects, including natural products. OBJECTIVES: The aim of this study was to evaluate and compare the effects of a plant sterol standardized supercritical CO2 phytopharmaceutical of Physalis angulata L. with hydrocortisone on the immune and inflammatory mediators, and skin repair components production. Moreover, we studied effects of both products on the skin microcirculation and temperature in a double-blind placebo-controlled clinical trial. METHODS: Both products were evaluated on the immune (IL-6, IL-10, INF-γ, TNF-α, and IL-1α), inflammatory (COX-2, LOX, PLA2 , PGE2 , LTB4 , histamine, and NF-κB), and repair components (TGF-ß, GM-CSF, collagen, and GAG) production on human keratinocytes and fibroblast in non-stimulated and LPS-stimulated conditions. Indeed, in a randomized double-blind placebo-controlled clinical trial, we evaluated the effects of the both creams on the skin microcirculation and temperature using laser Doppler and infrared thermometer, respectively. RESULTS: Physalis angulata acted on the skin, modulating immune status and inflammatory response producing corticoid-like effects, but different of hydrocortisone, increased skin repair factors. The effects of phytopharmaceutical cream in the clinical trial promoted a better reduction in skin microcirculation and temperature than hydrocortisone. CONCLUSIONS: Taken together, the results indicate that sterol standardized CO2 supercritical preparation of P angulata is a new and innovative phytopharmaceutical with multiple pharmacological effects potentially useful as human skin protective product, particularly against cutaneous inflammatory disorders.

In vitro SPF and Photostability Assays of Emulsion Containing Nanoparticles with Vegetable Extracts Rich in Flavonoids.

The aim of study was to determine the in vitro sun protection factor (SPF) and the photostability profile of a topical formulation composed of nanoparticles loaded with vegetable extracts and to assess its physicochemical properties. Chitosan/tripolyphosphate (TPP) nanoparticles loaded with flavonoids-enriched vegetable extracts (Ginkgo biloba L., Dimorphandra mollis Benth, Ruta graveolens, and Vitis vinifera L.) were produced and characterized for their morphology, mean particle size, zeta potential, and encapsulation efficiency. A final topical formulation was obtained by dispersing chitosan/TPP nanoparticles in an o/w emulsion. Results showed that nanoparticles dispersion exhibited yellowish color, spherical shape, and uniform appearance. Extract-loaded chitosan/TPP nanoparticles showed a mean particle size of 557.11 ± 3.1 nm, polydispersity index of 0.39 ± 0.27, zeta potential of + 11.54 ± 2.1 mV, and encapsulation efficiency of 75.89% of rutin. The recorded texture parameters confirm that the developed formulation is appropriate for skin application. The SPF obtained was 2.3 ± 0.4, with a critical wavelength of 387.0 nm and 0.69 UVA/UVB ratio. The developed formulation exhibited photostability, allowing the release of flavonoids from nanoparticles while retaining rutin into the skin in a higher extension.

Ultraviolet A photosensitivity profile of dexchlorpheniramine maleate and promethazine-based creams: Anti-inflammatory, antihistaminic, and skin barrier protection properties.

BACKGROUND: Unwanted side effects such as dryness, hypersensitivity, and cutaneous photosensitivity are challenge for adherence and therapeutical success for patients using treatments for inflammatory and allergic skin response. AIMS: In this study, we compared the effects of two dermatological formulations, which are used in inflammatory and/or allergic skin conditions: dexchlorpheniramine maleate (DCP; 10 mg/g) and promethazine (PTZ; 20 mg/g). METHODS: We evaluated both formulations for phototoxicity potential, skin irritation, anti-inflammatory and antihistaminic abilities, and skin barrier repair in vitro and ex vivo using the standard OECD test guideline n° 432, the ECVAM protocol n° 78, and cultured skin explants from a healthy patient. Ultraviolet A was chosen as exogenous agent to induce allergic and inflammatory response. RESULTS: Both PTZ and DCP promoted increases in interleukin-1 (IL-1) synthesis in response to ultraviolet A (UVA) radiation compared to control. However, the increase observed with PTZ was significantly greater than the DCP, indicating that the latter has a lower irritant potential. DCP also demonstrated a protective effect on UVA-induced leukotriene B4 and nuclear factor kappa B (NF-κB) synthesis. Conversely, PTZ demonstrates more robust UVA antihistaminic activity. Likewise, PTZ promoted a significantly greater increase in the production of involucrin and keratin 14, both associated with protective skin barrier property. CONCLUSION: In conclusion, these data suggest possible diverging UVA response mechanisms of DCP and PTZ, which gives greater insight into the contrasting photosensitizing potential between DCP and PTZ observed in the patients.

Honokiol protects skin cells against inflammation, collagenolysis, apoptosis, and senescence caused by cigarette smoke damage.

BACKGROUND: Pollution, especially cigarette smoke, is a major cause of skin damage. OBJECTIVES: To assess the effects of the small molecule polyphenol, honokiol, on reversing cigarette smoke-induced damage in vitro to relevant skin cells. METHODS: Keratinocytes (HaCat) cultures were exposed to cigarette smoke and, after 48 hours, IL-1α and IL-8 were measured in cell supernatants. Moreover, TIMP-2 production, apoptosis rate, and senescence ß-galactosidase expression were evaluated in primary human foreskin fibroblasts (HFF-1) cultures. RESULTS: Honokiol at 10 µm reduced IL-1α production by 3.4 folds (P < 0.05) and at 10 and 20 µm reduced IL-8 by 23.9% and 53.1% (P < 0.001), respectively, in HaCat keratinocytes. In HFF-1, honokiol restored TIMP-2 production by 96.9% and 91.9% (P < 0.001), respectively, at 10 and 20 µm, as well as reduced apoptosis by 47.1% (P < 0.001) and 41.3% (P < 0.01), respectively. Finally, honokiol reduced senescence-associated ß-galactosidase expression in HFF-1. CONCLUSION: Honokiol protects both HFF-1 and HaCat against cigarette smoke-induced inflammation, collagenolysis, apoptosis, and senescence.

Metodologia alternativa para o estudo dos efeitos da radiação infravermelha-A sobre a pele humana / Alternative methodology for the study of infrared-A radiation effects on human skin

Introdução: 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 plasticas eletivas, para avaliar os reais beneficios os reais benefícios clínicos de um produto aplicado topicamente. Objetivo: O objetivo desta investigação foi correlacionar os efeitos da radiação IV-A, em biópsias e em fragmentos de pele ex vivo e cultura de fibroblastos humanos, pela quantificação dos mediadores MMP-1, TIMP-1 e GADD45a. Métodos: Coleta de biópsias de 15 voluntárias após aplicações de IV-A durante cinco dias consecutivos. Exposição à radiação IV-A de fragmentos de pele humana provenientes de cirurgia plástica eletiva e cultura de fibroblastos humanos. Mensuração dos mediadores MMP-1, TIMP-1 e GADD45a para posterior comparação dos resultados. 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. Conclusão: 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 clínico.

Introduction: Infrared radiation (IR-A) causes structural changes in the skin, similar to those caused by prolonged exposure to ultraviolet radiation. Evaluation of efficacy and safety of cosmetic products concentrates in in vitro tests and clinical trials. A promising alternative is the use of fragments of human skin from elective cosmetic surgery, to evaluate the actual clinical benefits of a product applied topically. Objective: The objective of this study was to correlate IR-A radiation effects in biopsies and in ex vivo skin fragments and in human fibroblasts culture by quantifying MMP-1, TIMP-1 and GADD45a mediators. Methods: Collection of biopsies from 15 volunteers after IR-A applications for 5 consecutive days. Exposure to IR-A radiation of human skin fragments from elective cosmetic surgery, and human fibroblasts culture. Measurement of MMP-1, TIMP-1 and GADD45a mediators for further comparison of results. Results: In the three models used, the IR-A radiation induced an increase in MMP-1, inhibited the synthesis of GADD45a, and did not changed TIMP-1 values. Conclusion: Due to the positive correlation of the models studied, it may be suggested the use of ex vivo skin as plausible and sustainable tool to overcome differences between knowledge generated from in vitro and clinical experiments.

In vitro effects of infrared A radiation on the synthesis of MMP-1, catalase, superoxide dismutase and GADD45 alpha protein.

Harmful influences in the process of photoaging and skin damage are associated with infrared A (IRA) radiation, such as, disturbance of dermal extracellular matrix by up regulation of matrix metalloproteinase-1 (MMP1). Furthermore, DNA damage, induction of cytotoxicity and oxidative stress by decreasing natural antioxidant ability has been reported after acute exposure to IRA. The present study provides additional evidence that IRA radiation response in human skin fibroblasts produces deleterious effects to the cell, such as accelerating aging and weakening of their antioxidant defense mechanism. Human skin fibroblasts were exposed to a non-cytotoxic dose of IRA radiation and cultured for different periods for further collection of cell-free supernatants and lysates, and quantification of MMP-1, catalase, superoxide dismutase, and GADD45a. Our results corroborate previous published data and strongly indicate a negative impact of IRA radiation on the skin physiological by mechanisms involving reduced endogenous antioxidant enzymatic defense, increased MMP-1 and decreased repair process of DNA by reducing GADD45a protein, in cultured human fibroblasts. From a clinical perspective, IRA radiation acts by mechanisms distinct from those observed in ultraviolet radiation indicating the need for developing and making available cosmetics for skin care with properties beyond protection exerted by traditional sunscreens.

Antiageing Mechanisms of a Standardized Supercritical CO 2 Preparation of Black Jack (Bidens pilosa L.) in Human Fibroblasts and Skin Fragments.

The use of topical retinoids to treat skin disorders and ageing can induce local reactions, while oral retinoids are potent teratogens and produce several unwanted effects. This way, efforts to explore complementary care resources should be supported. Based on this, we evaluate the antiageing effects of a supercritical CO2 extract from Bidens pilosa L. (BPE-CO2A) containing a standardized multicomponent mixture of phytol, linolenic, palmitic, linoleic, and oleic acids. BPE-CO2A was assessed for its effects on human dermal fibroblasts (TGF-ß1 and FGF levels using ELISA; collagen, elastin, and glycosaminoglycan by colorimetric assays, and mRNA expression of RXR, RAR, and EGFr by qRT-PCR) and human skin fragments (RAR, RXR, collagen, elastin, and glycosaminoglycan by immunohistochemical analysis). Levels of extracellular matrix elements, TGF-ß1 and FGF, and EGFr gene expression were significantly increased by BPE-CO2A. The modulation of RXR and RAR was positively demonstrated after the treatment with BPE-CO2A or phytol, a component of BPE-CO2A. The effects produced by BPE-CO2A were similar to or better than those produced by retinol and retinoic acid. The ability to stimulate extracellular matrix elements, increase growth factors, and modulate retinoid and rexinoid receptors provides a basis for the development of preparation containing BPE-CO2A as an antiageing/skin-repair agent.

Neuromodulatory and anti-inflammatory ingredient for sensitive skin: in vitro assessment.

The manifestation of sensitive skin occurs as a consequence of increased permeability of the Stratum corneum, besides the involvement of 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 ß-endorphin; eicosanoids prostaglandin E2 (PGE2) and leukotriene B4 (LTB4); histamine, transient receptor potential vanilloid subfamily member 1 (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). As the same way, SensC provides decrease in the synthesis of TRPV1. Regarding the synthesis of envelope proteins, SensC promoted increases for filaggrin and involucrin levels, when compared to control group. Considering the absence of appropriate treatment, the availability of ingredients, such as SensC, with antiinflammatory and protective barrier properties can be a significant tool for preventing neurosensorial symptoms associated with sensitive skin.

Photoprotective and antioxidant effects of Rhubarb: inhibitory action on tyrosinase and tyrosine kinase activities and TNF-α, IL-1α and α-MSH production in human melanocytes.

BACKGROUND: Exposure to ultraviolet (UV) radiation causes various forms of acute and chronic skin damage, including immunosuppression, inflammation, premature aging and photodamage. Furthermore, it induces the generation of reactive oxygen species, produces proinflammatory cytokines and melanocyte-stimulating hormone (MSH) and increases tyrosinase activity. The aim of this study was to evaluate the potential photoprotective effects of Rheum rhaponticum L. rhizome extract on human UV-stimulated melanocytes. METHODS: The effects of Rheum rhaponticum rhizome extract on tyrosine kinase activity, and on interleukin-1α (IL-1α), tumour necrosis factor α (TNF-α), and α-MSH production in human epidermal melanocytes were evaluated under UV-stimulated and non-stimulated conditions. Antioxidant activity was evaluated by lipid peroxidation and 1,1-dyphenyl-2-picryl-hydrazyl (DPPH) assays, while anti-tyrosinase activity was evaluated by the mushroom tyrosinase method. RESULTS: Rheum rhaponticum L. rhizome extract showed in vitro antioxidant properties against lipid peroxidation, free radical scavenging and anti-tyrosinase activities, and inhibited the production of IL-1α, TNF-α, α-MSH, and tyrosine kinase activity in melanocytes subjected to UV radiation. CONCLUSIONS: These results support the inclusion of Rheum rhaponticum L. rhizome extract into cosmetic, sunscreen and skin care products for the prevention or reduction of photodamage.

Eficácia in vitro e clínica do uso cosmético tópico de fitoestrógenos em pele fotoenvelhecida / In vitro and clinical efficacy of the use of phytoestrogens-based topical cosmetic use in photoaged skin

Introdução: O tratamento do envelhecimento cutâneo representa um desafio clínico. Objetivos: Avaliar a eficácia in vitro e in vivo, e a segurança clínica de cosmético com fitoestrógenos na abordagem do fotoenvelhecimento. Métodos: A etapa in vitro foi realizada pela análise da expressão gênica de fibronectina e pró-colágeno, avaliação da atividade imunomoduladora e análise histoquímica e por imunofluorescência da pele e da junção dermoepidérmica com o produto analisado. No estudo clínico in vivo foi 76 mulheres, foram randomizadas em dois grupos: o Grupo A usou creme contendo complexo de fitoestrógenos e FPS 20 duas vezes ao dia, enquanto o Grupo B usou este mesmo creme associado a outro com função de antienvelhecimento aplicado à noite. O estudo durou 120 dias tendo sido realizadas mensalmente avaliações médicas, da voluntária, ultrassonografia (20MHz), fotografias e biópsias pré e pós-tratamento. Resultados: No estudo in vitro houve aumento na expressão de fibronectina e procolágeno, potencial imunomodulador, representado pelo aumento de IL-1α diminuição de IL-10; melhora da integridade da JDE, aumento da viabilidade e espessura da epiderme, e da síntese de colágeno. in vivo: melhora global subjetiva da aparênciada pele da face; redução de manchas, eritema, poros e porfirina cutânea. O ultrassom e a biópsia revelaram aumento da densidade dérmica (52,7%) e de fibras dérmicas (22,25%), respectivamente. Conclusões: Fitoestrógenos tópicos melhoram a condição geral da pele, avaliada clínicamente, histológicamente e por ultrassonografia; acrescentam-se resultados in vitro de aumento da síntese de fibronectina, prócolágeno e colágeno, melhoria da integridade da junção dermoepidérmica e restauração da resposta imunológica da pele.

Introdução: Introduction: Skin aging is a challenge to treat. Objectives: To evaluate the in vitro and in vivo efficacy and clinical safety of a phytoestrogens-based cosmetic in the management of photoaging. Methods: The in vitro investigation was performed using the analysis of the genic expression of fibronectin and procollagen, evaluation of the immunomodulatory activity (proinflammatory and anti-inflammatory cytokines synthesis) and histochemistry and immunofluorescence analyses of the skin and the dermoepidermal junction. The in vivo investigation ­ performed in 76 women randomized into Group A (phytoestrogens complex cream and SPF 20 twice daily) or Group B (the same product plus a commercially available anti-aging cream applied overnight). The study lasted for 120 days, with physician- and patient-led evaluations, in addition to monthly ultrasound (20 MHz) and photographic analysis. Skin biopsies of the face were performed before and after treatment. Results: The study showed In vitro: increase in the expression of fibronectin, in procollagen, immunomodulator potential, represented by an increase in IL-1α and a decrease in IL-10; improvement in the integrity of the dermoepidermal junction; increase in the viability and thickness of the epidermis; increase in collagen synthesis .In vivo: subjective global improvement of the skin's appearance; reduction in the count and intensity of spots, erythema, skin pores, and cutaneous porphyrin. The ultrasound and biopsy revealed increased dermal density (52.7%) and dermal fibers (22.3%), respectively. Conclusions: The topical use of phytoestrogens-based cosmetics improves the overall condition of the skin.