Cosmetic application of the stem-bark extract of Bertholletia excelsa H.B.K.

OBJECTIVE: The Amazon has a rich biodiversity where many different plant species can be found. This diversity is an important source of bioactive substances, mainly due to the different structural components of their phytometabolites. Research for natural products is a strategy for the development of new agents in therapeutic applications, especially cosmetic applications, that have better pharmacological potential. Within this perspective, the objective of the study was to investigate the cosmetic application (anti-aging potential) of the stem-bark extract of Bertholletia excelsa H.B.K - (SBEBE), popularly known as the Brazil nut tree, here called SBEBE, a noble plant species of the Amazon that is rich in selenium. METHODS: Enzymatic, glycation, proliferation, cell-healing, collagen quantification, toxicity and genotoxicity assays were used. RESULTS: Among the enzymes involved in the extracellular matrix of the skin, SBEBE was able to inhibit only elastase (62.67 ± 3.75) when compared to the standard sivelestat (89.04 ± 0.53), and the extract was also able to inhibit both the oxidative and the non-oxidative pathway. When cell toxicity in fibroblasts (MRC-5) and keratinocytes (HACAT) was evaluated, SBEBE did not present toxicity in 24 h of incubation. After this period, the extract showed average cytotoxicity in 48 and 72 h, but not enough to reach the concentration of 50% of MRC-5 fibroblasts. In the trypan blue assay, the extract promoted fibroblast proliferation in 24, 48 and 72 h of incubation, which was evaluated through exponential cell growth, with emphasis mainly on the lowest concentration with results higher than the standard. When the cell healing capacity was evaluated, in 48 h of exposure to fibroblast, SBEBE was able to induce a cell carpet (cell film) in the cell monolayer scratch assay. CONCLUSIONS: SBEBE stimulated collagen production at all concentrations tested. In the alkaline comet assay, at the lowest concentration, the extract did not induce DNA damage when compared to the reference drug doxorubicin. This study proved that SBEBE extract can be considered an ally in the treatment of skin anti-ageing as a possible biotechnological, phytocosmetic product.

OBJECTIF: L'Amazonie possède une riche biodiversité ou l'on trouve de nombreuses espèces végétales différentes. Cette diversité constitue une source importante de substances bioactives, principalement en raison des différents composants structurels de leurs phytométabolites. La recherche de produits naturels est une stratégie de développement de nouveaux agents à applications thérapeutiques, notamment cosmétiques, présentant un meilleur potentiel pharmacologique. Dans cette perspective, l'objectif de l'étude était d'étudier l'application cosmétique (potentiel anti­âge) de l'extrait d'écorce de tige de Bertholletia excelsa H.B.K ­ (SBEBE), communément connu sous le nom de noix du Brésil, ici appelé SBEBE, un arbre noble, espèce végétale d'Amazonie riche en sélénium. MÉTHODES: Des tests enzymatiques, de glycation, de prolifération, de guérison cellulaire, de quantification du collagène, de toxicité et de génotoxicité ont été utilisés. RÉSULTATS: Parmi les enzymes impliquées dans la matrice extracellulaire de la peau, le SBEBE était capable d'inhiber uniquement l'élastase (62,67 +­ 3,75) par rapport au sivelestat standard (89,04 +­ 0,53), et l'extrait était également capable d'inhiber à la fois la voie oxydative et non­oxydative. Lorsque la toxicité cellulaire dans les fibroblastes (MRC­5) et les kératinocytes (HACAT) a été évaluée, SBEBE n'a présenté aucune toxicité en 24 heures d'incubation. Après cette période, l'extrait a montré une cytotoxicité moyenne en 48 et 72 h, mais pas suffisamment pour atteindre la concentration de 50 % de fibroblastes MRC­5. Dans le test au bleu trypan, l'extrait a favorisé la prolifération des fibroblastes en 24, 48 et 72 heures d'incubation, qui a été évaluée par une croissance cellulaire exponentielle, en mettant l'accent principalement sur la concentration la plus faible avec des résultats supérieurs à la norme. Lorsque la capacité de guérison cellulaire a été évaluée, en 48 heures d'exposition aux fibroblastes, SBEBE a pu induire un tapis cellulaire (film cellulaire) dans le test de grattage de la monocouche cellulaire. CONCLUSIONS: SBEBE a stimulé la production de collagène à toutes les concentrations testées. Dans le test alcalin des comètes, à la concentration la plus faible, l'extrait n'a pas induit de dommages à l'ADN par rapport au médicament de référence, la doxorubicine. Cette étude a prouvé que l'extrait de SBEBE peut être considéré comme un allié dans le traitement anti­âge cutané en tant que possible produit biotechnologique et phytocosmétique.

HPLC-TBARS-EVSC (high-performance liquid chromatography-thiobarbituric acid reactive substances-ex vivo stratum corneum) protocol: Selection of the subjects and approach to present the results.

OBJECTIVE: Nowadays, it is recognized the need for improved safety and efficacy protocols to evaluate the human stratum corneum (SC) and its interaction with topical and cosmetic formulations by minimally or non-invasive methodologies. The aim of our research work was to streamline the HPLC-TBARS-EVSC (high-performance liquid chromatography-thiobarbituric acid reactive substances-ex vivo stratum corneum) methodology, by exploring the results of a group of 18 subjects. METHODS: The study included nine women and nine men aged between 19 and 57 years old with phototypes from II to V. Sites in the forearm of each volunteer were randomly delimited, and the SC was collected by tape stripping. HPLC was used to quantify the MDA-TBA2 (malondialdehyde-thiobarbituric acid) adduct from the tape-stripped SC, irradiated and not by an ultraviolet (UV) simulator chamber. RESULTS: Observing the findings of our present investigation, and the statistical approach applied, the use of the ratio between the treatment site and control would be an adequate strategy to better discriminate and evaluate the results. Additionally, an optimal selection of the volunteers to respond specifically to the purpose of the ex vivo assay also can be considered advantageous. CONCLUSIONS: It seemed that in future studies focusing on the impact of SC UV-induced lipid peroxidation, determined by the HPLC-TBARS-EVSC, the most suitable subjects are females aged less than 35 years old, with phototype II.

OBJECTIF: Aujourd'hui, il est nécessaire d'améliorer les protocoles de sécurité d'emploi et d'efficacité pour évaluer le stratum corneum (SC) humain et son interaction avec les formulations topiques et cosmétiques par des méthodologies peu ou pas invasives. L'objectif de notre travail de recherche était de rationaliser la méthodologie HPLC-TBARS-SCEV, à savoir chromatographie en phase liquide à haute performance (High Performance Liquid Chromatography), substances réactives à l'acide thiobarbiturique (Thiobarbituric Acid Reactive Substances), stratum corneum ex vivo (SCEV), en explorant les résultats d'un groupe de 18 sujets. MÉTHODES: L'étude incluait 9 femmes et 9 hommes âgés de 19 à 57 ans présentant des phototypes II à V. Des sites de l'avant-bras de chaque volontaire ont été délimités de manière aléatoire, et le SC a été recueilli par « tape stripping ¼. La chromatographie en phase liquide à haute performance a été utilisée pour quantifier l'adduit MDA-TBA2 (malondialdéhyde - acide thiobarbiturique) à partir du SC recueilli par « tape stripping ¼, irradié et non irradié par une chambre de simulation à ultraviolets (UV). RÉSULTATS: En observant les résultats de notre recherche actuelle et l'approche statistique appliquée, l'utilisation du rapport entre le site traité et le site contrôle serait une stratégie adéquate pour mieux discriminer et évaluer les résultats. En outre, une sélection optimale des volontaires pour répondre spécifiquement à l'objectif du test ex vivo peut également être considérée comme bénéfique. CONCLUSIONS: Il semble que dans les études futures axées sur l'impact de la peroxydation lipidique induite par UV du SC, déterminé par la méthodologie HPLC-TBARS-SCEV, les sujets les plus appropriés sont les femmes âgées de moins de 35 ans présentant un phototype II.

Skin photoaging in young men: A clinical study by skin imaging techniques.

OBJECTIVE: Differences in skin physiology and lifestyle among people can influence the skin damage caused by solar radiation. Photoprotection habits play an important role to prevent skin photoaging. Thus, the objective of the present study was to evaluate the skin changes resulting from solar exposure in young men by skin imaging techniques. METHODS: Twenty-three male, aged 18-28 years, with and without photoprotection habits participated in the study. Instrumental measurements in terms of dermis thickness and echogenicity (20 mHz ultrasound - DermaScanC® ), morphological and structural skin characterization (Reflectance Confocal Microscopy - VivaScope® 1500) and high-resolution imaging (Visioface® ) were performed in the malar region of the face. RESULTS: Pigmentation disorders, telangiectasia, wrinkles and lower dermis echogenicity were observed in the skin of subjects without photoprotection habits. Reflectance Confocal Microscopy images showed an irregular honeycomb pattern (35%), polycyclic papillae (40%) and coarse collagen fibres (35%), which are related to photoaging. CONCLUSION: Sun exposure without photoprotection results in a loss of dermal echogenicity and changes in the epidermis structure and collagen fibres regardless of chronological age. Thus, the study adds data for a discussion about correct photoprotection habits among young people and can help to increase the use of sunscreens for male public.

OBJECTIFS: Les différences de physiologie cutanée et de mode de vie peuvent influer sur les lésions cutanées causées par le rayonnement solaire. Les habitudes de photoprotection jouent un rôle important dans la prévention du photovieillissement cutané. Dans ce contexte, l'objectif de l'étude était d'évaluer les changements cutanés résultants de l'exposition au soleil chez les jeunes hommes à l'aide de techniques d'imagerie cutanée. MÉTHODES: L'étude a inclus 23 hommes, âgés de 18 à 28 ans, avec et sans habitudes de photoprotection. Des mesures instrumentales d'épaisseur et d'échogénicité du derme (échographie 20 mHz - DermaScanC®), de caractérisation morphologique et structurale de la peau (Reflectance Confocal Microscopy - VivaScope® 1500) et des images haute résolution (Visioface®) ont été réalisées dans la région malaire du visage. RÉSULTATS: Des troubles de la pigmentation, des télangiectasies, des rides et une moindre échogénicité du derme ont été observés sur la peau d'individus sans habitudes de photoprotection. Les images de microscopie confocale à réflectance ont montré un motif en nid d'abeille irrégulier (35%), des papilles polycycliques (40%) et des fibres de collagène épaisses (35%), qui sont liés au photovieillissement. CONCLUSION: L'exposition au soleil sans photoprotection entraîne une perte d'échogénicité cutanée et des modifications de la structure de l'épiderme et des fibres de collagène, quel que soit l'âge chronologique. Enfin, l'étude agrège les données pour une discussion sur les bonnes habitudes de photoprotection chez les jeunes et peut aider à augmenter l'utilisation des écrans solaires par le public masculin.

Effectiveness of a formulation containing peptides and vitamin C in treating signs of facial ageing: three clinical studies.

OBJECTIVE: Vitamin C and peptides are widely used in cosmetic products but there is a paucity of clinical studies showing that the formulations are effective in treating signs of facial ageing. These 3 clinical studies evaluated the effectiveness of an anti-ageing formula containing natural vitamin C (10%), biopeptides (rice and lupin), hyaluronic acid, and Vichy volcanic mineralising water, in amber glass ampoules with no preservatives (Peptide-C ampoules). METHODS: Dansyl chloride fluorescence labelling compared cell turnover for Peptide-C ampoules vs untreated skin in 32 female subjects. Study 2, an open clinical study, evaluated the efficacy on wrinkles of Peptide-C ampoules by investigator clinical scoring based on Dynamical Atlas visual assessment (N = 40) and subject self-assessment questionnaires (N = 47). Study 3, an open clinical study, evaluated wrinkles by instrumental quantification with 3D fringe projection analysis (N = 40) and subject questionnaires (N = 51). RESULTS: The mean cell turnover was faster for skin treated with Peptide-C ampoules compared to untreated skin (17.1 days vs. 19.2 days; P < 0.0001). In study 2, after 28 days application of Peptide-C ampoules, clinical grading of crow's-feet wrinkles, forehead wrinkles and nasolabial folds decreased by 9%, 11% and 5%, respectively (all P < 0.05 vs baseline). Of 47 subjects, 77%, 64% and 79% indicated their skin seemed smoothed out, fine lines were less visible, and skin complexion was more radiant, respectively. In study 3, the number of wrinkles decreased by 11.5% after 29 days application of Peptide-C ampoules vs baseline (P < 0.05) and 65% of subjects responded the fine lines were less visible. CONCLUSION: This formulation of a combination of anti-ageing ingredients in ampoules, allowing a minimalist formula, showed significant results on improving facial wrinkles and radiance.

OBJECTIF: La vitamine C et les peptides sont régulièrement utilisés dans les produits dermocosmétiques mais il existe peu d'études cliniques sur l'efficacité des formulations sur les signes du vieillissement cutané du visage. Trois études cliniques ont évalué l'efficacité d'une formule anti-âge contenant de la vitamine C naturelle (10%), des biopeptides (riz et lupin), de l'acide hyaluronique et de l'eau minéralisante volcanique de Vichy, dans un format d'ampoules en verre ambré, sans conservateur (ampoules Peptide-C). MÉTHODES: Une première étude a comparé par la technique de chlorure de Dansyl le renouvellement cellulaire avec la formulation ampoules Peptide-C et la peau non traitée chez 32 sujets féminins. La seconde étude, en ouvert, a évalué l'efficacité clinique sur les rides des ampoules Peptide-C en se reposant sur les Atlas Dynamiques (N=40) et les questionnaires d'auto-évaluation des sujets (N=47). La troisième étude, ouverte, a évalué les rides par quantification instrumentale avec l'analyse de projection de franges 3D (N=40) et les questionnaires d'autoévaluation des sujets (N=51). RÉSULTATS: Le renouvellement cellulaire était plus rapide pour la peau traitée avec des ampoules de Peptide-C comparées à la peau non traitée (17.1 jours contre 19.2 jours ; p<.0001). Dans l'étude 2, après 28 jours d'application des ampoules Peptide-C, l'évaluation clinique des rides de la patte d'oie, du front et des plis naso-labiaux a montré une amélioration de 9 %, 11 % et 5 %, respectivement (tous p<0,05 vs baseline). Sur 47 sujets, 77%, 64% et 79% ont indiqué que leur peau semblait respectivement lissée, que les ridules étaient moins visibles et que le teint de la peau était plus radieux. Dans l'étude 3, le nombre de rides a diminué de 11,5 % après 29 jours d'application des ampoules Peptide-C par rapport à la baseline (p<0,05) et 65 % des sujets ont répondu que les ridules étaient moins visibles. CONCLUSION: Cette combinaison d'ingrédients anti-âge dans un format d'ampoules, et une formulation minimaliste, a montré des résultats significatifs sur l'amélioration des rides faciales et de l'éclat du teint.

Photoageing-related skin changes in different age groups: a clinical evaluation by biophysical and imaging techniques.

OBJECTIVES: In view of the lack of studies about the morphological and structural changes caused by solar radiation in young people, the aim of the present study was to evaluate the photoageing-related changes in the skin of different age groups by biophysical and imaging techniques. METHODS: Forty-four healthy female subjects were divided into two age groups: Group 1 (G1): 18-35 years old and Group 2 (G2): 40-60 years old. The skin of malar region of the face was evaluated in terms of mechanical properties, disorder in the pigmentation pattern, morphological and structural changes using the Cutometer® , Colorimeter® , Visioface® and Dermascan C® devices and reflectance confocal microscopy (Vivascope® ). RESULTS: The results showed that the main changes in the skin of G1 were related to the pigmentation pattern, the papilla format and depletion of thin collagen fibres. These alterations were also observed in the skin of G2, but with more pronounced effects. CONCLUSION: The knowledge about the skin changes caused by photoageing obtained in this study is very important for the development of dermocosmetic products for more effective treatments particularly focused on this type of skin. Finally, objective characterization of photoageing showed the importance of photoprotective habits since the first years of life in order to retard the appearance of skin changes caused by solar radiation.

OBJECTIFS: Compte tenu du manque d'études sur les modifications structurelles et morphologiques causées par le rayonnement solaire chez les jeunes gens, le but de la présente étude était d'évaluer les changements liés au photovieillissement de la peau de différents groupes d'âges grâce des techniques de biophysique et d'imagerie. MÉTHODES: Quarante-quatre sujets féminins en bonne santé ont été divisés en deux groupes d'âges : Groupe 1 (G1) : 18-35 ans et Groupe 2 (G2) : 40-60 ans. La peau des régions malaires du visage a été évaluée en termes de propriétés mécaniques, troubles du schéma de pigmentation, et changements morphologiques et structurels à l'aide des dispositifs Cutometer®, Colorimeter®, Visioface® et Dermascan C® et de la microscopie confocale par réflectance (Vivascope®). RÉSULTATS: Les résultats ont montré que les principaux changements au niveau de la peau du G1 étaient liés au schéma de pigmentation, au format papillaire et à la déplétion de fines fibres de collagène. Ces altérations ont également été observées au niveau de la peau du G2, mais avec des effets plus prononcés. CONCLUSION: Les connaissances sur les modifications cutanées causées par le photovieillissement acquises grâce à cette étude sont très importantes pour le développement de produits dermo-cosmétiques pour des traitements plus efficaces axés plus particulièrement sur ce type de peau. Et enfin, la caractérisation objective du photovieillissement a montré l'importance des habitudes photoprotectrices depuis les premières années de vie afin de retarder l'apparition de modifications cutanées causées par le rayonnement solaire.

Forearm skin aging: characterization by instrumental measurements.

BACKGROUND: The intrinsic ageing and photoaged skin present biomechanical and morphological differences, which are reflected in the appearance of roughness, superficial and deep wrinkles, atrophy, reduced elasticity, hypo- and hyperpigmentation and actinic keratosis. OBJECTIVE: To evaluate and compare the characteristics of the flexor (with a predominance of intrinsic ageing) and extensor (chronically exposed to sunlight and other environmental factors, with a predominance of photoageing) skin surfaces of the forearms. METHODS: Interventional, prospective, diagnostic study, including 23 females, aged over 60 years. The extensor and flexor faces of forearms were compared in relation to clinical parameters and non-invasive instrumental measurements, such as skin surface, elasticity, hydration as well as dermal thickness and echogenicity. RESULTS: Regarding the water content of the stratum corneum, the flexor face presented an average value higher than the extensor face. The average of measures obtained through images of high-frequency ultrasound demonstrated greater echogenicity of flexor face compared to the extensor face. The measurements of the skin surface showed significant differences between the faces. The roughness and scaliness were lower in the flexor face. Regarding the depth of wrinkles, there was no significant difference between the faces. The average of the measurements was slightly higher in the flexor face, which demonstrated that wrinkles are present in the intrinsic ageing and photoageing. The presence of elastosis and the reduction in elasticity in the clinical aspect of the photoaged face of forearms were according to the results of the non-invasive measurements. CONCLUSION: Meaningful differences in the biophysical characteristics of the extensor and flexor faces of the forearms were detected. Because the non-invasive instrumental measurements correlated with clinical findings, they may represent useful tools to assess efficacy and safety of skin ageing treatments in clinical research.

Elastin structure and its involvement in skin photoageing.

Skin aging is a complex process that may be caused by factors that are intrinsic and extrinsic to the body. Ultraviolet (UV) radiation represents one of the main sources of skin damage over the years and characterizes a process known as photoaging. Among the changes that affect cutaneous tissue with age, the loss of elastic properties caused by changes in elastin production, increased degradation and/or processing produces a substantial impact on tissue esthetics and health. The occurrence of solar elastosis is one of the main markers of cutaneous photoaging and is characterized by disorganized and non-functional deposition of elastic fibers. The occurrence of UV radiation-induced alternative splicing of the elastin gene, which leads to inadequate synthesis of the proteins required for the correct assembly of elastic fibers, is a potential explanation for this phenomenon. Innovative studies have been fundamental for the elucidation of rarely explored photoaging mechanisms and have enabled the identification of effective therapeutic alternatives such as cosmetic products. This review addresses cutaneous photoaging and the changes that affect elastin in this process.

Reliable assessment of forearm photoageing by high-frequency ultrasound: a cross-sectional study.

OBJECTIVE: High-frequency ultrasound is a non-invasive tool used in skin ageing research to assess dermis thickness and echogenicity. This study evaluated the reliability of a range of high-frequency ultrasound parameters and tested their correlation with age and a validated clinical scale for the assessment of forearm skin photoageing; the difference between two body sites according to environmental exposition patterns was also investigated. METHODS: Twenty-three volunteers aged 28-82 years were divided into three groups according to forearm photoageing degree. A 20 MHz ultrasound unit was used to obtain cross-sectional images of the skin by two trained investigators on two different sites: the dorsal forearm (chronically photoexposed skin) and the proximal medial arm (non-photoexposed skin). Several echogenicity parameters were studied for each skin compartment: total dermis, upper dermis and lower dermis, and the ratio between upper and lower dermis. RESULTS: The intraclass correlation coefficient (for complete agreement) between investigators was higher for upper and total dermis echogenicity measures compared with the lower dermis. At the non-photoexposed site, the upper and lower dermis parameter ratio was better correlated with age. At the photoexposed area, total dermis parameters demonstrated higher correlations with clinical score. CONCLUSION: The authors discuss the choice of parameters for forearm photoageing assessment using high-frequency ultrasound.

Evaluation of the impact of urban pollution on the quality of skin: a multicentre study in Mexico.

OBJECTIVE: After pilot and preliminary studies aimed at identifying pertinent biochemical parameters, a multicenter clinical study was performed to evaluate the effect of pollution on human skin. METHODS: The clinical study was performed in collaboration with the 'Centre Régional de lutte contre le cancer de Montpellier' and the 'National Institute of Public Health of Mexico' on 96 subjects in Mexico City (exposed to pollution) and 93 subjects in Cuernavaca (less exposed to pollution). Both biochemical and clinical skin parameters were studied. RESULTS: The study demonstrated significant quantitative and qualitative modifications of parameters related to sebum excretion in Mexico City compared to Cuernavaca one: An increased level of sebum excretion rate, a lower level of vitamin E and squalene in sebum, an increase of lactic acid and a higher erythematous index on the face of the subjects. In the stratum corneum, a significant higher level of carbonylated proteins and a lower level of IL 1α were noticed, as well as a decrease of ATP concentration with a decrease of chymotrysin like activity, without modifications of corneodesmosin content and trypsin like activity. From a clinical point of view, a higher frequency of atopic and urticarial skins, a higher frequency of red dermographism, an important seborrheic status at the forehead level and a lower level of dandruffs were noted in Mexico City population. The analysis taking into account the sex does not modify the observed results. CONCLUSION: The study demonstrated an important impact of polluted environmental conditions on skin quality, evidencing important modifications of superficial biochemical parameters. The cause/effects relationships of these modifications remain, however, to be further assessed by a complementary in vitro/in vivo approaches.

Long-wave infrared radiation reflected by compression stockings in the treatment of cellulite: a clinical double-blind, randomized and controlled study.

BACKGROUND: Cellulite refers to changes in skin relief on the thighs and buttocks of women, with a prevalence of 80-90%, causing dissatisfaction and search for treatment. Etiopathogenesis is multifactorial, as follows: herniation of the hypodermis towards the dermis, facilitated by perpendicular fibrous septa, changes in the dermal extracellular matrix, decreased adiponectin, genetic polymorphism, microcirculation alterations and inflammatory process. There are numerous therapeutic approaches, with little evidence of effectiveness. The long-wave infrared (LWIR) radiation interacts with water, improves microcirculation and stimulates metabolic processes. To date, the use of tissues with potential reflection of LWIR radiation has not been systematically investigated as adjuvant treatment for cellulite. OBJECTIVE: To investigate the efficacy and safety of the treatment of cellulite through the use of compression stockings made with thread reflecting LWIR radiation. PATIENTS AND METHODS: Clinical study of therapeutic intervention, controlled and double-blind, including 30 women, aging from 25 to 40 years, with cellulite of grades II and III on the thighs and buttocks who used compression stockings, "pantyhose" model, made with reflector thread of LWIR radiation, on only one randomized side. Women under other treatments for cellulite and with venous and/or blood insufficiencies were excluded. Evaluation of efficacy by clinical parameters, photographs, Dermatology Life Quality Index (DLQI), cutometry and high frequency ultrasonography and security by observation of adverse events and venous EcoDoppler recordings. RESULTS: DLQI scores showed significant reduction; the two-dimensional high-frequency ultrasonography showed an insignificant increase in dermal echogenicity as well as other efficacy parameters demonstrated no or slight improvement, with no differences between the sides exposed or not to LWIR; and there were no severe adverse events. CONCLUSION: Compression stockings, with or without thread reflector of LWIR, showed slight effects in the appearance of cellulite, but the treatment determined a positive impact on women quality of life.