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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.

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Abstract The combination of avobenzone (AVO) and octyl ρ-methoxycinnamate (OMC) is widely used to ensure broad-spectrum photo-protection because they absorb UVA and UVB, respectively. However, they are thermally and photo unstable because they degrade and undergo photo- tautomerization and trans-cis isomerization, thus reducing their photo-protection efficacy during UV exposure. This study aimed to evaluate the potential use of the antioxidants ferulic acid and resveratrol as stabilizing substances in AVO and OMC mixtures in solution or emulsion. The effects of both antioxidants on the thermal/photo-stability and suppression of the filter singlet state, besides skin permeation, were evaluated. Both antioxidants contributed to preserving OMC and AVO during the thermal stability test, which relates to the maintenance of photo-protection even after storing the formulations at high temperatures. Nevertheless, although resveratrol retained part of the OMC trans isomer and suppressed the AVO singlet state when exposed to UV, no contribution to photo-protection stability was observed, contrary to expectations. Regarding the permeation assay, the addition of both antioxidants was accompanied by a reduction of AVO permeation, while resveratrol increased OMC permeation. Thus, the chemical and physicochemical properties of these antioxidants impacted their efficacy and safety profiles; therefore, further studies are required to establish the real cost-benefit ratio for their use in sunscreens.

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INTRODUCTION: Compared to adults, newborns' skin has a thinner epidermis and stratum corneum with decreased hydration levels, higher transepidermal water loss, and a pH variation between 5.5 and 7.5. These characteristics can predispose to the occurrence of dryness, infections, and dermatological conditions. Water and liquid soap with adequate formulation have shown to be beneficial and safe for newborns' skin. However, studies evaluating the effect of bar soap, products widely used in Brazil and Latin America, are unknown. Therefore, the objective of this study was to compare the effects of liquid and bar soaps on the term newborns' skin. METHODS: This randomized controlled, parallel, single-blind clinical trial was conducted at a public university hospital in São Paulo, Brazil. 100 healthy term newborns with no congenital anomalies, acute diseases, or dermatological conditions were randomized to use liquid soap (experimental group) or bar soap (control group). Skin pH, transepidermal water loss, stratum corneum hydration, sebum content, and skin condition were assessed before and after the first bath, at 48 h, 14 days, and 28 days after birth. These evaluations were performed on the forearm, abdomen, buttocks, and thigh. In addition, the mother's perception of soap use was also evaluated. RESULTS: Data of 100 newborns were analyzed by intention to treat. The rate of retention was 53%. Newborns exposed to the liquid soap presented significantly better skin acidification (p < 0.001) and significantly better stratum corneum hydration (p < 0.001) than the skin of newborns exposed to the bar soap, regardless of the area evaluated. There were no significant differences in transepidermal water loss, sebum content, dryness, erythema, or skin breakdown and the mother's perceptions of the use of the soaps. CONCLUSION: Newborns in the experimental group presented better skin acidification and stratum corneum hydration when compared to newborns in the control group.

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Candelilla wax (CW) and 12-hydroxystearic acid (12HSA) are classic solid-fiber-matrix organogelators. Despite the high number of studies using those ingredients in oily systems, there is scarce literature using a mixture of oil and antioxidants. Vitamin E (VE) is an important candidate for its lipophilicity and several applications on pharmaceutical, cosmetics, and food industries. In this work, we investigated the influences of mixtures between vegetable oil (VO) and VE on the microstructures and rheological properties of CW and 12HSA organogels. A weak gel (G''/G' > 0.1) with a shear-thinning behavior was observed for all samples. The presence of VE impacted the gel strength and the phase transition temperatures in a dose-dependent pattern. Larger and denser packed crystals were seen for 12HSA samples, while smaller and more dispersed structures were obtained for CW organogels. The results obtained in this work allowed the correlation of the structural and mechanical properties of the organogels, which plays an important role in the physical-chemical characteristics of these materials.

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OBJECTIVE: The harmful effects induced by ultraviolet exposition and the significant increment in skin cancer diagnosis confirm the necessity to develop effective and safe sunscreens. Limited efficacy and cutaneous adverse reactions of traditional formulations drove the incorporation of natural extracts into multifunctional sunscreens. Vaccinium myrtillus L. extract (VME), that contains anthocyanins and flavonoids, is a potential candidate for such systems. METHODS: Considering that, we performed in vitro and in vivo tests to evaluate the sun protection factor (SPF), photostability, and safety of sunscreen samples containing VME. RESULTS: As main results, the SPF was reduced in both in vitro and in vivo evaluation in the presence of VME; nonetheless, the samples were photostable and safe. CONCLUSION: Further investigation is required to better understand the unexpected effects of VME over photoprotection, decreasing the SPF value. As a conclusion, even with interesting findings, we highlight the importance of case-by-case investigations to develop multifunctional bioactive sunscreens.

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Background: Glyoxylic acid has emerged as a safe alternative to formol (formaldehyde) use as a hair straightener/relaxer. However, the possible damage to the hair fiber after its application is low known and/or published in the literature. Aims: This work aims to characterize hair locks treated with glyoxylic acid compared to traditional alkaline straighteners such as sodium and guanidine hydroxide and ammonium thioglycolate. Materials and Methods: The morphology of the hair cuticles was observed by scanning electron microscopy. Protein loss was assessed by the Lowry method modified by Peterson and as mechanical properties that were expressed in terms of tensile strength. Results: All products (sodium and guanidine hydroxides and ammonium thioglycolate) caused protein loss of about 2.5 µg/g, except glyoxylic acid that caused the worst damage (3.5 µg/g), in relation to the untreated (virgin) hair (1.12 µg/g), indicating that the chemical treatments can cause hair damage in both cuticles and cortex. The force to break the fibers treated with traditional straighteners based on sodium hydroxide, guanidine hydroxide, and ammonium thioglycolate was statistically the same. Conclusion: The treatment with glyoxylic acid showed rupture tensile statistically equivalent to the alkaline straighteners. The mechanism of action of glyoxylic acid does not appear to be based on breaking and rearrangement of disulfide bridges, but altered them, that influenced the hair strength. However, it is also essential to consider other factors relevant: technical application technique, reaction time, and interval of reapplication of the product, as this can change the pattern of the results obtained.

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BACKGROUND: Essential oils have great interest among the increasing demand for herbal cosmetics in the market. They are natural sources of biologically active ingredients due to the wide application of such compounds as well as their particular chemical composition. Several researches have evaluated the effectiveness of these bioactive ingredients for use in cosmeceuticals, mainly in both hair scalp and shaft hair damage repair. Thus, the amounts and their associations define the properties of these compositions with interest for hair cosmetic use, such as antioxidant, inflammatory, and antimicrobial activities. Because they are complex compounds, their actions on the skin, hair scalp, and shaft are not yet fully understood. AIMS: The purpose of this review is to highlight the relevant researches and findings on essential oils in hair care. METHODS: In order to achieve this objective, the present work comprises an updated bibliographic review related to essential oils used in hair care. RESULTS: It was possible to observe that cosmeceuticals containing essential oils applied to the scalp are preferable for topical activity. Also, it was noticed that there are few reports regarding their use in hair shaft. However, it was found that some oils are used to intensify the brightness and fix the hair color. CONCLUSIONS: This work demonstrated that the use of essential oils in both cosmetic products (industrial application) and those associated with oils carriers (as individual protocols) may lead to satisfactory results in the treatment of some scalp dysfunctions.

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Abstract Development and validation of a simple and fast method of high-performance liquid chromatography with diode array detection (HPLC-DAD) for the simultaneously analysis of rutin, avobenzone, and octyl p-methoxycinnamate is presented. These substances were separated using a Kromasil C18 (250×4.6 mm, 5 μm) column, methanol: water (88:12 v/v) as the mobile phase, and a flow rate of 0.8 mL min−1. The experiment was performed at room temperature and elution was under isocratic conditions. Quantification was performed by external calibration at the wavelength of 325 nm. The validated parameters included linearity, selectivity, precision (repeatability), intermediate precision, accuracy, limit of detection, limit of quantification and robustness. The results of validation were statistically treated using the Action Stat version 3.5.152.34. The selectivity was also evaluated in the presence of two cyclodextrins (2-hydroxypropyl-β-cyclodextrin and β-cyclodextrin sulfobutyl ether sodium). The absence of parallelism between the curves of octyl p-methoxycinnamate in the absence and presence of the β-cyclodextrin sulfobutyl ether sodium in the mobile phase revealed interference from this matrix, thereby indicating the necessity of validating the method in the presence of this, and other matrices. The proposed method was selective, linear, precise, accurate, and robust for the simultaneous determination of rutin, avobenzone, and octyl p-methoxycinnamate.

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Innovative technologies have been designed to improve efficacy and safety of chemical UV filters. Encapsulation can enhance efficacy and reduce transdermal permeation and systemic exposure. The aims of this work were (i) to determine the cutaneous biodistribution of avobenzone (AVO), oxybenzone (OXY), and octyl methoxycinnamate (OMC) incorporated in mesoporous silica SBA-15 and (ii) to perform preclinical (in vitro) and (iii) clinical safety studies to demonstrate their innocuity and to evaluate sun protection factor (SPF) in humans. Skin penetration studies showed that deposition of OXY and AVO in porcine and human skin after application of stick formulation with incorporated filters (stick incorporated filters) was significantly lower than from a marketed (non-encapsulated) stick. Cutaneous deposition and transdermal permeation of OXY in and across human skin were 3.8-and 13.4- fold lower, respectively, after application of stick entrapped filters. Biodistribution results showed that encapsulation in SBA-15 decreased AVO and OXY penetration reaching porcine and human dermis. Greater deposition (and permeation) of OXY in porcine skin than in human skin, pointed to the role of follicular transport. Stick incorporated filters had good biocompatibility in vivo and safety profiles, even under sun-exposed conditions. Entrapment of UV filters improved the SPF by 26% and produced the same SPF profile as a marketed stick. Overall, the results showed that SBA-15 enabled safety and efficacy of UV filters to be increased.

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Carotenoids and coenzyme Q10 are naturally occurring antioxidant compounds that are also found in human skin. These bioactive compounds have been the focus of considerable research due to their antioxidant, anti-inflammatory, and photoprotective properties. In this review, the current state of the art in the encapsulation of carotenoids and coenzyme Q10 in lipid nanoparticles to improve their bioavailability, chemical stability, and skin absorption is discussed. Additionally, the main findings are highlighted on the cytotoxic and photoprotective effects of these systems in the skin.

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Urocanic acid is a chromophore found in the skin that has been identified as an important immunosuppressant and carcinogenesis mediator through its photoisomerization from trans to cis form induced by ultraviolet radiation. Research on analytical methods that explore urocanic acid isomerization is indispensable to fully understand the deleterious effects mediated by this biomarker. In this context, the current relevant analytical methods for determination of these isomers in human samples are summarized in this review. The methods presented here are applicable to human samples collected by noninvasive methods (or minimally invasive), encompassing an array of analytical techniques, including high-performance capillary electrophoresis, confocal Raman spectroscopy, gas chromatography, high-performance liquid chromatography, and mass spectrometry, among others. Developed high-performance liquid chromatography methods have proven to be advantageous, allowing noninvasive collections for in vivo analysis and the confocal Raman, specially, for real-time analysis. Among all these methods, high-performance liquid chromatography is the most investigated one with mass spectrometry or ultraviolet detector, and the mass spectrometry detector being the most studied in the last years, demonstrating high sensitivity, very low detection limits, and accurate identification, especially for clinical investigations.

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BACKGROUND: The growing need to use sunscreens is a concrete reality, and it is associated with the increase in the population's awareness of the ultraviolet (UV) radiation damage. Inorganic UV filters promote the formation of particles/pigments film over the skin surface, reflecting, dispersing, and absorbing the radiation. Investigations of this class of filters demonstrate the emergence of alternative ingredients and new technologies. AIMS AND METHODS: In this review, we presented potential candidates for alternative UV inorganic filters, such as hydroxyapatite, cerium dioxide, and hydrotalcite. RESULTS: Specialized literature identified hydroxyapatite and cerium dioxide as actives with good performances, with a broad spectrum of absorption against UV radiation. Both of them were considered safe against degradation and skin erythema formation. Inorganic compounds as an alternative to circumvent degradation problems of organic UV filters containing PABA were also analyzed, with hydrotalcite having a good performance in improving the performance of classic ingredients, bypassing photoinstability, and improving safety, in addition to preventing skin irritability. CONCLUSION: Further investigations of these ingredients and interactions with sunscreen formulations are necessary to overall explore their safety and efficacy, encouraging future researches for more inorganic UV filters.

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BACKGROUND: Unprotected chronic exposure to ultraviolet (UV) radiation generates many harmful effects to human skin and sunscreens are essential to health, however, traditional products do not provide enough protection against cutaneous oxidative stress, a process amplified by UV radiation. Therefore, the development of multifunctional photoprotective formulations seems to be a more efficacious approach, since these enable the absorption/reflection of UV radiation and maintain the cutaneous homeostasis. OBJECTIVES: In the present study, ferulic acid (FA), a well-known antioxidant, has been combined with two UV filters, bemotrizinol and ethylhexyl triazone, and the safety and efficacy of this formulation has been assessed combining ex vivo and in vivo methods. METHODS: Skin permeation assays were performed by applying the formulation in the volar forearm of participants, after which consecutive samples of the stratum corneum were collected by tape stripping, and the quantification of FA, bemotrizinol and ethylhexyl triazone was performed by high-performance liquid chromatography (HPLC). Also, the FA anti-inflammatory action in combination with the UV filters was probed through a method employing Laser Doppler flowmetry to measure the vasodilatory response to methyl nicotinate topical application. RESULTS: Skin permeation assay was able to characterize the penetration depth of each substance. It should also be noted that a specific HPLC analytical method was developed in this study to enable the rapid simultaneous quantification of the three substances. Results from Laser Doppler flowmetry showed that the FA was able to mitigate the vasodilatory response. CONCLUSIONS: FA proved to be a valuable resource in a multifunction sunscreen, not only providing an increase in the SPF of sunscreens, previously published, but also decreasing the extent of inflammation.

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BACKGROUND: The use of sunscreens is mandatory, especially in countries with high ultraviolet (UV) incidence. In consequence, there has been a growing interest in using compounds from natural sources to develop new multifunctional products that protect human skin from the consequences of UV exposition. Even though there are in vitro methods to determine anti-UV efficacy, it is still required to test photoprotection activity on human skin to validate product performance. AIM AND METHODS: In this review, we summarized all reported clinical studies about sun protection factor (SPF) measurements of sunscreens with natural compounds. We also discussed the probable action mechanism of those actives. RESULTS: Herein, we provided an overview on recent studies concerning photoprotection activity of compounds from natural sources, for example, rutin, ferulic acid, caffeine, shea butter, and plant extracts, mainly presented in sunscreen systems with efficacy clinically established by SPF. CONCLUSION: Our review suggested that even when the in vivo SPF evaluation has inherent difficulties, it is essential to assure the real efficacy of sunscreens. Furthermore, the incorporation of natural compounds could enhance the in vivo SPF values of such sunscreens by different mechanisms. Finally, some compounds derived from natural resources with skin benefits could be used as "green"/natural UV filters that provide broad-spectrum sunscreens with further upgrading of the multifunctional dermocosmetic formulation to enhance aesthetics and even skin health.

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The ascorbyl methylsilanol pectinate (AMP) presents the same functional properties of ascorbic acid (AA). Besides antioxidant and depigmentant activity, the AMP presents silanol in its chemical structure. The aim of this work was to characterize and indentify the AMP alone and in cosmetic formulations. The following techniques were employed: Fourier Transform Infrared Spectrophotometry, particle size distributions, in vitro antioxidant activity with 2.2-diphenyl-1-picrylhydrazyl (DPPH) and Oxigen Radical Absorbance Capacity Assay and High Performace Liquid Chromatography (HPLC) (developed and validated method) for the active ingredient; Microscopy, HPLC and Normal Stability Assay (NSA) for the emulsions. Particle size distributions results showed that the average size of AMP was 1.0 µm and polydispersity index was 0.1. In DPPH assay AA and AMP were statistically the same. The value of ORAC obtained for AMP was 0.74 and for AA in the literature was 0.95. In the NSA the formulations were stable in conditions of 5.0 and 45.0 ± 2.0 ºC for 90 days. Adequate stability at ambient temperature out of reach of light was also observed. Thus, this works presented an acceptable method for quantification of AMP alone and in cosmetic formulations. AMP was an adequate choice for the incorporation in emulsions with antioxidant efficacy.

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Photosynthetic microorganisms convert carbon dioxide and solar radiation into interesting bioactive compounds not yet entirely explored. Several species of microalgae are known to be rich in colored high-valuable components that, although remarkable, are poorly explored as natural sources of pigments for cosmetics. Pigments associated to photosynthetic activity include chlorophyll, ß-carotene, astaxanthin, xanthophylls, and phycobiliproteins, many of which have shown high potential as cosmetic actives due to their antioxidant, immune-enhancing, and anti-inflammatory properties. In the last decade, concern with a young and beautiful appearance has emerged, encouraging many consumers to use anti-aging cosmetics daily. As a result, the cosmetic market has been growing and evolving rapidly to meet consumer expectations. However, due to regular use and the sensitive nature of facial skin, local adverse reactions may often occur, such as irritation, sensitization, or photoreactions, and safety evaluation is mandatory prior to marketing. It is, therefore, understandable that new actives from natural sources, such as microalgae, are perceived as attractive alternatives for consumers who seek ingredients without allergenic potential. Thus, the cosmetic industry has recently started to explore the inclusion of compounds extracted from microalgae and cyanobacteria in innovative formulations. Herein, we revised nontraditional microalgae species for pigment production with cosmetic applications, indicating those that could also be considered potential ingredients for innovative cosmetics. KEY POINTS: • Extraction methods for pigments from photosynthetic microorganisms were compiled. • Innovative cosmeceuticals could be developed with natural pigments. • Safety features of such natural pigments were also described.

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BACKGROUND: Acid straightener products are widely used, and it can cause changes in the hair shaft properties. The pH value of these products established by ANVISA (Brazil's National Health Surveillance Agency) as secure is above 2.0. However, the industries are interested in working at lower pH values in order to increase the straightening effect. Unfortunately, there are a lot of products in the market with pH value under the permitted. OBJECTIVE: Analyze two different pH values (1.0 and 2.0) of acid straightener formulation and the influence of this difference in the hair shaft properties. In order to provide information to professionals as cosmetologists, dermatologists, and hairdressers. METHODS: Combing and colorimetric analyses, tensile strength, differential scanning calorimetry (DSC), environmental scanning electron microscope (ESEM), and tryptophan content. RESULTS: The tresses treated at pH 1.0 had a better result in the straightening capacity, improving the combing test in 59.4%, while those at pH 2.0, only in 33.0% compared with virgin hair. However, the tensile strength, at pH 1.0 decreased by 16.0% and 9.0% to the pH 2.0. In addition, the tryptophan content was lower in the tresses treated with formulation at pH 1.0. The DSC analysis showed impairment in the straightened tresses. The images by ESEM, indicated a possible formation of a film around the fiber. CONCLUSIONS: It was possible to conclude that the pH value interferes in the hair shaft properties. Tresses treated with pH 1.0 had more modifications than tresses treated with pH 2.0.

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BACKGROUND: Hair-straightening treatments may involve the use of thermal devices, which potentially cause damages in the cuticle and cortex of the hair fibers. Particularly, the heat causes denaturation of the α-keratin and the degradation of the components of the hair cortex. OBJECTIVES: The aim of this study was to evaluate the effect of heating on the cuticle and cortex of the Caucasian and Asian virgin hair by using heat iron flat. METHODS: The effects of the heating in the tresses of Caucasian and Asian virgin hair were analyzed by thermogravimetry/differential scanning calorimetry-Fourier transform infrared/mass spectrometry (TG/DSC-MS/FTIR), protein loss analysis (PLA), Fourier transform infrared (FTIR-ATR), and scanning electron microscope (SEM). RESULTS: TG/DSC-MS/FTIR results were: formation of the H2 O between 25-170°C and CO2 , SCO and H2 S from 200°C. Denaturation temperature = 237°C. Protein loss: Asian hair>Caucasian hair. Data presented statistically significant alterations, α = 5, P ≤ 0.05, n = 3. FTIR-ATR: changes in the secondary structural conformation of the protein of hair cuticle. SEM: heat caused damage to hair cuticle. CONCLUSIONS: The results evidenced the importance of the control of temperature in the procedures involving heat. The damage caused by thermal devices showed be dependent of the ethnicity analyzed (Caucasian and Asian hair).

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The Ginoide Hydrolipodystrophy (GHLD), commonly known as cellulite, occurs in 80-90% of the female population after the puberty period and comes from a metabolic modification in the cutaneous adipose tissue. Caffeine has been used in topical formulations due to its lipolytic action. We studied a nanoemulsion (F3) containing caffeine with two surfactants (oleth-3 and oleth-20) by emulsification method by phase inversion temperature inversion (PIT). The polydispersion indices (PDI) showed the reduced deviation of 0.1. The mean droplet size was ~ 40 nm. The evaluated constant of Ostwald, in the refrigerator condition was the most favorable during the stability test. In the In Raman spectroscopy assay, the caffeine bands found in F3 were compatible with those found in the caffeine solution (1337, 652.5 and 558.2 cm-1). There was no interaction of caffeine anhydrous with other ingredients in nanoemulsion. In the in vitro safety assay the result of 1.4 ranked the F3 as slightly irritating. In the natural membrane, cutaneous permeation test (human skin) permeate concentrations did not exceed the saturation concentration of the PBS buffer (48.96 µg/3 mL). The caffeine solution and F3 permeated statistically equal, but the nanoemulsion visually and sensorially improved the caffeine precipitation.

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Unprotected chronic exposure to solar radiation can contribute to premature skin cancer and sunscreens are a key factor to avoid those detrimental effects. Currently, there is a growing interest in the photoprotector and antioxidant potential of bioactive substances, such as rutin, that could increase the sun protection factor (SPF) value and, also, donate multifunctional characteristics to sunscreens. Recent in vitro findings indicated that rutin, when incorporated into sunscreens, can provide antioxidant activity and SPF improvement. However, clinical studies are fundamental to determine this activity, due to the lack of repeatability of in vitro methodology and low correlation with the in vivo data. We aimed at evaluating the clinical safety and in vivo SPF of rutin by comparing sunscreen formulations containing 0.1% (w/w) rutin, 3.0% (w/w) butyl methoxydibenzoylmethane and 8.0% (w/w) octyl dimethyl PABA (2-ethylhexyl 4-(dimethylamino)benzoato) with a similar bioactive-free preparation. Additionally, skin hydration, in vitro SPF and in vitro antioxidant activity of rutin, in association with the ultraviolet (UV) filters, were investigated. The safety profile of the formulations under sun-exposed skin conditions qualified the formulas for clinical efficacy assays. 2,2-Diphenyl-1-picrylhydrazyl (DPPH) test confirmed the antioxidant properties of rutin, revealing around 40% increase in radical scavenging potential when the bioactive compound was present. Rutin in combination with the UV filters robustly elevated the clinical SPF around 70%, when compared with the bioactive-free formulation. To date, this is the first report in the specialized literature of an in vivo SPF measurement of a rutin-containing photoprotective preparation, supporting the claim that rutin is an effective and safe bioactive compound to be used in multifunctional sunscreens.

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