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.

Impact of ferulic acid and resveratrol on the effectiveness and safety of sunscreen

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.

Impact of Acid ("Progressive Brush") and Alkaline Straightening on the Hair Fiber: Differential Effects on the Cuticle and Cortex Properties.

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.

RP-HPLC simultaneous quantification of rutin, avobenzone, and octyl methoxycinnamate in the presence of hydroxypropyl β-cyclodextrin (HPβCD) and sulfobutyl ether β-cyclodextrin (SBEβCD)

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.

Ex vivo penetration analysis and anti-inflammatory efficacy of the association of ferulic acid and UV filters.

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.

The Synergistic Behavior of Antioxidant Phenolic Compounds Obtained from Winemaking Waste's Valorization, Increased the Efficacy of a Sunscreen System.

Grape pomace retains polyphenols in the peels and in the seeds after winemaking, which is indicative of the high valorization potential of this industrial waste. There is strong evidence that phenolics are robust antioxidants and confer photoprotection; thus, it is rational to apply these active compounds from winemaking waste to sunscreens, in order to increase UV protection. Despite the importance of this class of cosmetics to public health, more efficacious strategies are still needed to overcome the problems caused by the photoinstability of some UV filters. The hydroethanolic extract of Vitis vinifera L. grapes was obtained by percolation and then lyophilized. Six formulations were developed: Type I-cosmetic base and UV filters; Type II-cosmetic base and extract; and Type III-cosmetic base, extract and UV filters. Each formulation was prepared in the pHs 5 and 7. The antioxidant activities of the samples were measured by DPPH• and expressed in Trolox® equivalents (TE), and their photostability and in vitro sun protection factor (SPF) were analyzed by diffuse reflectance spectrophotometry. The anti-radical efficiencies observed in the formulations with grape extract were: (II) 590.12 ± 0.01 µmol TE g-1 at pH 5 and 424.51 ± 0.32 µmol TE g-1 at pH 7; (III) 550.88 ± 0.00 µmol TE g-1 at pH 5 and 429.66 ± 0.10 µmol TE g-1, at pH 7, demonstrating that the UV filters, butylmethoxydibenzoyl methane, ethylhexyl methoxycinnamate and ethylhexyl dimethyl 4-aminobenzoic acid had no influence on this effect. The photoprotective efficacy and the photostability of formulation III containing the extract and UV filters at pH 5 suggested that a synergism between the active molecules provided an 81% increase in SPF. Additionally, this was the only sample that maintained a broad spectrum of protection after irradiation. These results confirmed that the grape pomace extract has multifunctional potential for cosmetic use, mainly in sunscreens, granting them superior performance.

Tryptophan and kynurenine determination in human hair by liquid chromatography.

Tryptophan, an amino acid found in hair proteinaceous structure is used as a marker of hair photodegradation. Also, protein loss caused by several chemical/physical treatments can be inferred by tryptophan quantification. Kynurenine is a photo-oxidation product of tryptophan, expected to be detected when hair is exposed mainly to UVB (290-320nm) radiation range. Tryptophan from hair is usually quantified directly as a solid or after alkaline hydrolysis, spectrofluorimetrically. However, these types of measure are not sufficiently specific and present several interfering substances. Thus, this work aimed to propose a quantification method for both tryptophan and kynurenine in hair samples, after alkali hydrolysis process, by using high-performance liquid chromatography (HPLC) with fluorimetric and UV detection. The tryptophan and kynurenine quantification method was developed and validated. Black, white, bleached and dyed (blond and auburn) hair tresses were used in this study. Tryptophan and kynurenine were separated within ∼9min by HPLC. Both black and white virgin hair samples presented similar concentrations of tryptophan, while bleaching caused a reduction in the tryptophan content as well as dyeing process. Unexpectedly, UV/vis radiation did not promote significantly the conversion of tryptophan into its photo-oxidation product and consequently, kynurenine was not detected. Thus, this works presented an acceptable method for quantification of tryptophan and its photooxidation metabolite kynurenine in hair samples. Also, the results indicated that bleaching and dyeing processes promoted protein/amino acids loss but tryptophan is not extensively degraded in human hair by solar radiation.

Hydrolyzed collagen interferes with in vitro photoprotective effectiveness of sunscreens

ABSTRACT The chronological skin aging is a progressive and natural process with genetic and physiological changes. However, ultraviolet (UV) radiation may accelerate the oxidative stress, generating carcinogenesis and photoaging. Natural compounds and their applications are considered a trend in the cosmetic market. The protein-based film-forming compounds play an important role, once it collaborates for the better distribution of sunscreens on the skin. Here we investigated the in vitro photoprotective effectiveness of sunscreens containing the hydrolyzed collagen associated with UVA, UVB and/or inorganic filters. Sunscreens were developed with octocrylene (7.5%), butyl methoxydibenzoylmethane (avobenzone) (3.0%) and/or titanium dioxide (5.0%), associated or not with the hydrolyzed collagen (3.0%). In vitro photoprotective effectiveness was determined in a Labsphere(r) UV2000S by the establishment of the sun protection factor (SPF) and critical wavelength (nm) values. Physicochemical and organoleptic characteristics were also assayed. The hydrolyzed collagen subjectively improved the formulation sensory characteristics. However, this bioactive compound led to a decrease of the SPF values of the photoprotective formulations containing octocrylene alone and octocrylene + butyl methoxydibenzoylmethane + TiO2. This inadequate interaction may be considered during the development of new sunscreens intended to contain protein-based components.

Functional photostability and cutaneous compatibility of bioactive UVA sun care products.

Sunscreens are the most-established approach for photoprotection. The strategy of providing antioxidant properties to sun care products by addition of natural and potent anti-free radical compounds has led to the development of bioactive sunscreens, able to neutralize the harmful effects of ultraviolet (UV) radiation. UVA filters, such as benzophenone-3 (BP) and butyl methoxydibenzoylmethane (BMDBM), can exhibit photodegradation which limits the development of broad spectrum sunscreens. Previous research verified that rutin interacts with filters incorporated in sunscreens. In this work, we focused on the development and evaluation of the efficacy of the sunscreens containing either BP or BMDBM with and without rutin. The addition of rutin to the UVA filters afforded antioxidant properties to the formulations and they were considered safe for human use. Additionally, rutin in combination with either BP or BMDBM increased the antioxidant activity about 40 times when compared with the UVA filters alone. Remarkably, the addition of rutin 0.1% (w/w) to BP 6.0% (w/w) raised the SPF from 24.3±1.53 to 33.3±2.89. In conclusion, these findings demonstrated that the addition of rutin into sunscreens can markedly improve the antioxidant properties of the formulation as well as photostabilize some of the UVA filters.

Influência da rutina na fotoestabilização da avobenzona (filtro UVA) e do ρ-metoxicinamato de octila (filtro UVB) / Influence of rutin in photostabilization of avobenzone (UVA filter) and octyl methoxycinnamate (UVB filter)

Com o intuito de promover proteção de amplo espectro, na maioria dos protetores solares estão associados pelo menos dois filtros orgânicos (UVA e UVB). A combinação da avobenzona (BMBM), filtro UVA, e do p--metoxicinamato de octila (EHMC), filtro UVB, é conhecida e muito utilizada em formulações manipulas e industrializadas, porém apresenta alteração na absorção espectral após exposição à radiação UV. A estratégia empregada com maior frequência para reduzir a instabilidade da combinação é baseada na adição de agentes fotoestabilizadores. A adição de substâncias naturais em formulações fotoprotetoras vem sendo explorada, especialmente o grupo dos flavonoides, como a rutina, que apresenta resultados positivos em relação à eficácia fotoprotetora. O objetivo principal desta pesquisa foi avaliar o potencial da rutina como substância fotoestabilizadora dos filtros EHMC e BMBM. Foram desenvolvidas formulações contendo os dois filtros associados ou não com rutina de acordo planejamento fatorial em três níveis. As formulações foram avaliadas quanto a eficácia fotoprotetora in vitro aplicadas em placas de PMMA e analisadas por espectrofotometria de refletância com esfera de integração antes e após a exposição à radiação UV. As interações moleculares dos filtros com a rutina foram avaliadas por 1H RMN, DSC, TG e análise qualitativa da supressão do estado energético singleto. A adição de rutina nas formulações contendo 5,0% (p/p) de BMBM e 10,0% (p/p) de EHMC promoveu elevação na conservação do FPS in vitro de 53,9% para 65,8 (0,1% de rutina) e 70,8% (1,0% de rutina). As curvas DSC e TG da rutina apresentaram alterações promovidas pela presença dos filtros BMBM e EHMC, indicando interação entre o flavonoide e os filtros. Após dose de 5760 J cm-2 de radiação UV o valor da razão trans/cis para o filtro EHMC em solução adicionado do filtro BMBM foi elevado de 5,5±0,1, sem adição de rutina, para 12,6±0,4, com adição da rutina. A análise qualitativa da supressão do estado singleto indicou que um dos mecanismos envolvidos na fotoestabilização dos filtros BMBM e EHMC é a supressão do estado energético singleto. Os resultados reportados neste estudo indicaram que a adição da rutina em formulações fotoprotetoras representa um caminho simples e efetivo para elevar a fotoestabilidade da combinação dos filtros BMBM e EHMC. A adição da rutina em formulações fotoprotetoras representa uma estratégia promissora, pois aliada a ação fotoestabilizadora, verificada nesse estudo, esse flavonoide possui propriedades antioxidante e quelante de metais que podem colaborar para o desenvolvimento de formulações fotoprotetoras de amplo espectro com aumento da segurança e eficácia

In order to promote broad-spectrum protection, most sunscreens are associated with at least two organic filters (UVA and UVB). The combination of avobenzone (BMBM), UVA filter, and octyl methoxycinnamate (EHMC), UVB filter, is well known and widely used in industrial formulations and pharmaceutical compounding, but shows alteration in spectral absorption after UV radiation exposure. The most commonly used strategy to reduce the instability of the combination is based on the addition of photostabilizer agents. The addition of natural substances in sunscreen formulations has been explored, especially the group of flavonoids such as rutin, which shows positive results regarding photoprotective efficacy. The main objective of this research was to evaluate the potential of rutin as a photostabilizer substance of EHMC and BMBM. Formulations were developed containing the two filters associated or not with rutin, according to factorial design at three levels. The formulations were evaluated for in vitro photoprotective efficacy applied on PMMA plates and analyzed by spectrophotometer with integrating sphere reflectance before and after exposure to UV radiation. Molecular interactions of filters with rutin were evaluated by 1H NMR, DSC, TG and qualitative analysis of the suppression of singlet energy state. The addition of rutin in the formulations containing 5.0 % (w/w) BMBM and 10.0 % (w/w) EHMC promoted an increase in the preservation of in vitro SPF of 53.9% to 65.8 (0.1 % rutin) and 70.8 % (1.0% rutin). The DSC and TG curves of rutin showed changes promoted by the presence of BMBM and EHMC filters, indicating interaction between the flavonoid and filters. After 5760 J cm-2 of UV radiation the value of the trans/cis ratio for the EHMC filter added from the BMBM filter was increased from 5.5 ± 0.1 without addition of rutin, to 12.6 ± 0 4,with the addition of rutin. Qualitative analysis of the suppression of the singlet state indicated that one of the mechanisms involved in the photostabilization BMBM and EHMC filters is suppression of singlet excited state.The results reported in this study indicate that the addition of rutin in sunscreen formulations is a simple and effective way to increase the photostability of the combination of BMBM and EHMC. The addition of rutin in sunscreen formulations represents a promising strategy, for allied with the photostabilization action, observed in this study, this flavonoid has antioxidant and chelating properties of metals that can contribute to the development of broad-spectrum sunscreens formulations with increased safety and efficacy

Stability evaluation of organic Lip Balm

Rising global demand for natural products whose production is harmless to the environment has stimulated the development of natural cosmetics and, within this category, organics (95% organic raw materials). The image of environmentally friendly production is one of the strongest attractions of organic products. Lip balm is a cosmetic product similar to lipstick whose purpose is to prevent lip dryness and protect against adverse environmental factors. The product's characteristics are: resistance to temperature variations, pleasant flavor, innocuousness, smoothness during application, adherence and easy intentional removal. This work involved the development of a lip balm formulated with certified organic raw materials and the execution of stability tests: fusion point determination, evaluation of organoleptic characteristics (color, odor and appearance) and functionality evaluation (spreadability test). The formulation selected after the Preliminary Stability Test was submitted to the Normal Stability Test under the following storage conditions (temperature): Room Temperature (22.0 ± 3.0 ºC), Oven (40.0 ± 2.0 ºC) and Refrigerator (5.0 ± 1.0 ºC), for 90 days. Under the Refrigerator and Room Temperature conditions, spreadability proved adequate, but the surface presented white spots, characterizing the fat bloom, a phenomenon involving the recrystallization of cocoa butter. Storage at 40.0 ± 2.0 ºC (Oven) caused loss of functionality according to the spreadability test, in addition to changes in color, although the aspect was uniform since the fat bloom was not observed (white spots on the surface). The odor remained stable under all conditions as did the melting point, which had a mean of 72.9 ± 1.7 ºC throughout the course of stability testing (90 days).

O aumento da demanda global por produtos naturais, cuja produção não envolva nenhum dano ao meio ambiente, tem estimulado o desenvolvimento de cosméticos naturais e, dentro desta categoria, dos produtos orgânicos (que contêm 95% de matérias-primas orgânicas). O protetor labial é um produto cosmético semelhante ao batom que tem a finalidade de prevenir o ressecamento dos lábios, mantendo a hidratação, e protegê-los contra fatores ambientais adversos. Este trabalho envolveu o desenvolvimento de um protetor labial formulado com matérias-primas orgânicas e avaliação dos parâmetros de estabilidade, como ponto de fusão, características organolépticas e funcionalidade (teste de espalhabilidade). A formulação selecionada após o Estudo de Estabilidade Preliminar foi submetida à Avaliação Normal de Estabilidade, nas seguintes condições de armazenamento (temperatura): Ambiente (22,0 ± 3,0 ºC), Estufa (40,0 ± 2,0 ºC) e Geladeira (5,0 ± 1,0 ºC), por 90 dias. Nas condições de armazenamento em geladeira ou ambiente, a espalhabilidade foi adequada, mas a superfície apresentou pontos esbranquiçados, caracterizando o fenômeno chamado fat bloom, que está relacionado à recristalização da manteiga de cacau. O armazenamento à temperatura elevada (40,0 ± 2,0 ºC) provocou perda de funcionalidade, de acordo com o teste de espalhabilidade, e mudança de cor, apesar do aspecto permanecer uniforme, visto que não foi observado o fenômeno fat bloom. O odor manteve-se estável em todas as condições, assim como o ponto de fusão, com valor médio de 72,9 ºC ± 1,7 ºC durante todo o período de avaliação do teste de estabilidade (90 dias).

Hidrolipodistrofia ginoide: aspectos gerais e metodologias de avaliação da eficácia: [revisão] / Gynoid lipodystrophy: general aspects and methodologies for efficacy evaluation: [review]

A hidrolipodistrofia ginoide (HLDG), popularmente conhecida como celulite, está presente em grande parte da população feminina. Trata-se de umaalteração do relevo cutâneo, envolvendo modificações morfológicas, histoquímicas, bioquímicas e ultraestruturais nos adipócitos, além de alterações na derme e na microcirculação. Existem inúmeras formas de se tratar essa condição, sendo a mais utilizada e de fácil acesso o uso de produtos tópicos contendo princípios ativos com ação vasoprotetora, anti-inflamatória, estimuladores da microcirculação periférica e agentes lipolíticos. Para verificar a eficácia destas formulações, existem metodologias de avaliação envolvendo avaliações instrumentais e clínicas da melhoria do aspecto da celulite. Este artigo teve por objetivo revisar as formas existentes de se avaliar a ação de produtos que atuam na HLDG, considerando os diversos mecanismos de ação, além de revisar aspectos gerais referentes a esta condição.

Gynoid lipodystrophy (HLDG), popularly known as cellulite, is present in a great part of the female population. It is an alteration of the cutaneousrelief involving morphological, histochemical, biochemical and ultrastructural modifications in the adipocytes and also alterations in the dermis andmicrocirculation. There are countless ways of treating this condition, being topic products containing active principles with vessel protective, antiinflammatory actions, peripheral microcirculation stimulators and lipolytic agents the most used and of easiest access. In order to verify the efficacyof these formulations, there are evaluation methodologies involving instrumental and clinical evaluations of the improvement of the aspect of cellulite. Therefore, this article aimed at reviewing the existent ways of evaluating the efficacy of products against gynoid lipodystrophy, taking the different mechanisms of action into account and also at reviewing general aspects concerning this condition.

Comparative study of the stability of free and modified papain incorporated in topical formulations

Papain is an enzyme used in topical formulations as a proteolytic debriding agent for the treatment of open, extensive wounds and burnings. It is also employed as an enhancer for cutaneous permeation of active compounds, chemical peeling and as a progressive depilatory agent. The stability of formulations containing enzymes is not easy. In this research, papain was modified with polyethylene glycol in order to increase the stability of the formulations. The comparative Normal Stability Testing of the topical formulations containing unmodified and modified papain showed that the modified variety presented with a differentiated profile under the adopted temperature conditions (5.0 ± 1.0 °C; 22.0 ± 2.0 °C; 40.0 ± 2.0 °C). The most suitable condition for non-modified papain were 5.0 ± 1.0 °C and, for modified papain, they were 22.0 ± 2.0 °C. These results confirmed the higher stability of modified papain compared to free papain, as well as its potential to be applied in topical formulations.

A papaína é uma enzima utilizada em formulações tópicas como agente proteolítico debridante no tratamento de lesões abertas de grande extensão e queimaduras. É, também, empregada na pele íntegra como agente promotor da permeação cutânea de princípios ativos, peeling químico e como agente depilatório progressivo. A estabilidade de formulações contendo enzimas não é facilmente alcançada. No presente trabalho realizou-se a modificação da enzima com polietilenoglicol, visando maior estabilidade das formulações. A realização do Teste Estabilidade Normal comparativo entre as formulações contendo as formas da enzima não modificada e modificada demonstrou que a última apresentou um perfil de estabilidade diferenciado, nas diferentes condições (5,0 ± 1,0 °C; 22,0 ± 2,0 °C; 40,0 ± 2,0 °C). A condição de 5,0 ± 1,0 °C foi a mais adequada para a formulação contendo papaína não modificada enquanto a 22,0 ± 2,0 °C foi indicada para aquela contendo a forma modificada. Estes resultados confirmaram o aumento da estabilidade da papaína modificada comparada com a livre e seu potencial de aplicação em formulações de uso tópico.

Spectroscopic and thermal characterization of alternative model biomembranes from shed skins of Bothrops jararaca and Spilotis pullatus / Caracterização espectroscópica e térmica de biomembranas modelo alternativo de peles galpão de Bothrops jararaca e Spilotis pullatus

Recently, there has been an interest in the use of shed snake skin as alternative model biomembrane for human stratum corneum. This research work presented as objective the qualitative characterization of alternative model biomembranes from Bothrops jararaca and Spilotis pullatus by FT-Raman, PAS-FTIR and DSC. The employed biophysical techniques permitted the characterization of the biomembranes from shed snake skin of B. jararaca and S. pullatus by the identification of vibrational frequencies and endothermic transitions that are similar to those of the human stratum corneum.

Existe atualmente interesse no uso da muda de pele de cobra como modelos alternativos de biomembranas da pele humana. O presente trabalho apresentou como objetivo a caracterização qualitativa de modelos alternativos de biomembranas provenientes de mudas de pele de cobra da Bothrops jararaca e Spilotis pullatus por espectroscopia Raman (FT-Raman), espectroscopia fotoacústica no infravermelho (PAS-FTIR) e calorimetria exploratória diferencial (DSC). As técnicas biofísicas FT-Raman, PAS-FTIR e DSC permitiram caracterizar qualitativamente os modelos alternativos de biomembranas provenientes das mudas de pele de cobra da B. jararaca e S. pullatus e identificar freqüências vibracionais e transições endotérmicas similares ao estrato córneo humano.

Physical and physicochemical stability evaluation of cosmetic formulations containing soybean extract fermented by Bifidobacterium animalis / Avaliação da estabilidade física e físico-química de formulações cosméticas contendo extrato de soja fermentado por Bifidobacterium animalis

Peel off facial masks, based on polyvinyl alcohol (PVA), are formulations that, after application and drying, form an occlusive film over the face. After removing, they provide cleanness, tensor and moisturizing effects, removing dead cells, residues and other materials deposited on the stratum corneous. The soybean extract fermented by Bifidobacterium animalis has sugars, amino acids, peptides, proteins and free isoflavonoids in high concentrations, when compared to the unfermented extract, providing benefits to the cosmetic formulations like anti-aging effect, moisture, tensor action and emollience. The cosmetic bases of peel off facial masks, added with 5.0 percent w/w of fermented soybean extract, were submitted to Preliminary and Accelerated Stability Studies. Eight (8) preparations were evaluated in several conditions of temperature (-10.0, 5.0, 22.0 and 45.0 ºC) and time (maximum of 15 days), comparing the results with the initial condition (48 h after preparation). The variables observed were: organoleptic characteristics, pH and appearing viscosity value and film drying time. The preparation containing 17.0 percent w/w of PVA and 0.5 percent w/w of guar gum was selected between the eight preparations initially prepared, because it presented the best performance in the stability test, being recommended storage at low temperatures (5.0 ºC).

As máscaras faciais peel off a base de álcool polivinílico (PVA) são formulações que, após a aplicação e secagem, formam um filme oclusivo sobre a face e, após sua remoção, conferem limpeza, ação tensora e hidratação à pele, retirando células mortas do estrato córneo, resíduos e outros materiais depositados. O extrato de soja fermentado por Bifidobacterium animalis possui açúcares, aminoácidos, peptídeos, e alto teor de isoflavonas na forma livre, quando comparado ao leite não fermentado, propiciando benefícios às formulações cosméticas, como ação antienvelhecimento, hidratação, efeito tensor e emoliência. As bases cosméticas de máscaras faciais peel off, acrescidas de extrato de soja fermentado 5,0 por cento p/p, foram submetidas aos ensaios de Estabilidade Preliminar e Acelerada, avaliando-se 8 preparações em diversas condições de temperatura (-10,0; 5,0; 22,0 e 45,0 ºC) e tempo (máximo de 15 dias), em relação à condição inicial (48 h após o preparo). As variáveis observadas envolveram: características organolépticas, valor de pH, viscosidade aparente e tempo de secagem do filme. A preparação contendo 17,0 por cento p/p de PVA e 0,5 por cento p/p de goma guar foi a selecionada dentre as oito preparações elaboradas inicialmente, por ter apresentado melhor desempenho no teste de estabilidade, sendo recomendado o armazenamento em temperatura reduzida (5,0 ºC).

Hair fiber characteristics and methods to evaluate hair physical and mechanical properties / Características da fibra do cabelo e métodos para avaliar o cabelo propriedades físicas e mecânicas

The hair thread is a natural fiber formed by keratin, a protein containing high concentration of sulfur coming from the amino acid cystine. The main physical proprieties of the hair depend mostly on its geometry; the physical and mechanical properties of hair involve characteristics to improve: elasticity, smoothness, volume, shine, and softness due to both the significant adherence of the cuticle scales and the movement control (malleability), as well as the easiness of combing, since they reduce the fibers static electricity. The evaluation of these effects on hair may be carried out by several methods, as: optical and electron microscopy, mechanical resistance measuring, shine evaluation and optical coherence tomography (OCT).

O cabelo é uma fibra natural formada por queratina, uma proteína composta por teor elevado de enxofre proveniente da cistina. As propriedades principais do cabelo dependem de sua geometria, estrutura física. Características físicas e mecânicas das fibras capilares envolvem propriedades que melhoram: elasticidade, maciez, volume, maleabilidade, facilidade para o ato de pentear e brilho. A avaliação de tais propriedades do cabelo pode ser obtida por métodos diversos, como: microscopia óptica e eletrônica, mensuração da resistência mecânica, determinação do brilho e tomografia por coerência óptica (OCT).

Influence of urea, isopropanol, and propylene glycol on rutin in vitro release from cosmetic semisolid systems estimated by factorial design.

Rutin, one of the major flavonoids found in an assortment of plants, was reported to act as a sun protection factor booster with high anti-UVA defense, antioxidant, antiaging, and anticellulite, by improvement of the cutaneous microcirculation. This research work aimed at evaluating the rutin in vitro release from semisolid systems, in vertical diffusion cells, containing urea, isopropanol and propylene glycol, associated or not, according to the factorial design with two levels with center point. Urea (alone and in association with isopropanol and propylene glycol) and isopropanol (alone and in association with propylene glycol) influenced significant and negatively rutin liberation in diverse parameters: flux (microg/cm(2).h); apparent permeability coefficient (cm/h); rutin amount released (microg/cm(2)); and liberation enhancement factor. In accordance with the results, the presence of propylene glycol 5.0% (wt/wt) presented statistically favorable to promote rutin release from this semisolid system with flux = 105.12 +/- 8.59 microg/cm(2).h; apparent permeability coefficient = 7.01 +/- 0.572 cm/h; rutin amount released = 648.80 +/- 53.01 microg/cm(2); and liberation enhancement factor = 1.21 +/- 0.07.

Evaluation of in vitro percutaneous enhancement effect of papain and pequi oil on diclofenac sodium permeation through human skin / Avaliação in vitro da papaína e do óleo de pequi como promotores de permeação cutânea para diclofenaco de sódio em pele humana

The purpose of this research was to determine the potential of papain and pequi oil as penetration enhancers for diclofenac sodium (DS) across human skin in vitro. The permeation studies were conducted with vertical diffusion cells. The enhancers were associated or not in gels in different concentrations. In vitro studies reveled that papain 0.2 percent (w/v) presented an elevated enhancer property for diclofenac sodium (J = 0.3369 mg/cm²x h). Pequi oil 10 percent (w/v) generated a reduced flux value (J = 0.1848 mg/cm²x h) and a combination of both enhancers presented a medium value of J = 0.2187 mg/cm²x h. Papain was found to be better enhancer than pequi oil.

O objetivo desta pesquisa foi determinar in vitro o potencial da papaína e do óleo de pequi como promotores de penetração cutânea para o diclofenaco de sódio (DS) através de pele humana. Os estudos de penetração foram conduzidos em células de difusão vertical. Os promotores foram associados ou não em géis em concentrações distintas. A avaliação in vitro revelou que a papaína 0,2 por cento p/p apresentou propriedade promotora maior para o diclofenaco de sódio (J = 0,3369 mg/cm²x h). O óleo de pequi 10,0 por cento p/v promoveu redução do fluxo (J = 0,1848 mg/cm²x h) e a combinação de ambos os promotores apresentou valor mediano de fluxo de J = 0,2187 mg/cm²x h. A partir dos resultados, verificou-se que a papaína exerceu ação promotora de penetração cutânea melhor que o óleo de pequi.

Estabilidade e estudo de penetração cutânea in vitro da rutina veiculada em uma emulsão cosmética através de um modelo de biomembrana alternativo / Stability and in vitro penetration study of rutin incorporated in a cosmetic emulsion through an alternative model biomembrane

A rutina é empregada como antioxidante e na prevenção da fragilidade capilar. Estudos de penetração in vitro através da pele humana seria a situação ideal, entretanto, há dificuldades de sua obtenção e manutenção de sua viabilidade. Entre os demais modelos de membrana, a muda de pele de cobra se apresenta como estrato córneo puro, fornecendo barreira similar ao humano e é obtida sem a morte do animal. Os objetivos desta pesquisa foram desenvolver e avaliar a estabilidade de uma emulsão cosmética contendo rutina e, como promotor de penetração cutânea, o propilenoglicol; e avaliar a penetração e a retenção cutânea in vitro da referida substância ativa da formulação, empregando um modelo de biomembrana alternativo. A emulsão foi desenvolvida com rutina e propilenoglicol, ambos a 5,0 por cento p/p. Quantificou-se a rutina das emulsões por espectrofotometria a 361,0 nm, método previamente validado. A penetração e retenção cutânea in vitro foram realizadas em células de difusão vertical com muda de pele de cobra de Crotalus durissus, como modelo de biomembrana alternativo, e água destilada e álcool etílico absoluto 99,5 por cento (1:1), como fluido receptor. O experimento foi conduzido em um período de seis horas, a 37,0 ± 0,5 ºC e agitação constante de 300 rpm. Empregou-se o método espectrofotométrico validado a 410,0 nm para a quantificação da rutina após penetração e retenção cutânea. A emulsão não promoveu a penetração cutânea da rutina através da muda de pele de C. durissus, retendo 0,931 ± 0,0391 mg de rutina/mg de muda de pele de cobra. Nas condições de armazenamento a 25,0 ± 2,0 ºC; 5,0 ± 0,5 ºC e 45,0 ± 0,5 ºC, a emulsão apresentou-se quimicamente estável durante 30 dias. De acordo com os resultados, a emulsão não favoreceu a penetração cutânea da rutina, mas apenas sua retenção no estrato córneo de C. durissus, condição considerada estável no período de 30 dias.

Rutin is employed as antioxidant and to prevent the capillary fragility and, when incorporated in cosmetic emulsions, it must target the action site. In vitro cutaneous penetration studies through human skin is the ideal situation, however, there are difficulties to obtain and to maintain this tissue viability. Among the membrane models, shed snake skin presents itself as pure stratum corneum, providing barrier function similar to human and it is obtained without the animal sacrifice. The objectives of this research were the development and stability evaluation of a cosmetic emulsion containing rutin and propylene glycol (penetration enhancer) and the evaluation of rutin in vitro cutaneous penetration and retention from the emulsion, employing an alternative model biomembrane. Emulsion was developed with rutin and propylene glycol, both at 5.0 percent w/w. Active substance presented on the formulation was quantified by a validated spectrophotometric method at 361.0 nm. Rutin cutaneous penetration and retention was performed in vertical diffusion cells with shed snake skin of Crotalus durissus, as alternative model biomembrane, and distilled water and ethanol 99.5 percent (1:1), as receptor fluid. The experiment was conducted for six hours, at 37.0 ± 0.5 ºC with constant stirring of 300 rpm. Spectrophotometry at 410.0 nm, previously validated, determined the active substance after cutaneous penetration/retention. Emulsion did not promote rutin cutaneous penetration through C. durissus skin, retaining 0.931 ± 0.0391 mg rutin/mg shed snake skin. The referred formulation was chemically stable for 30 days after stored at 25.0 ± 2.0 ºC, 5.0 ± 0.5 ºC and 45.0 ± 0.5 ºC. In conclusion, it has not been verified the active cutaneous penetration through the model biomembrane, but only its retention on the Crotalus durissus stratum corneum, condition considered stable for 30 days.

In vitro safety assessment of papain on human skin: A qualitative Light and Transmission Electron Microscopy (TEM) study / Avaliação in vitro da segurança de uso da papaína em pele humana: Estudo qualitativo por microscopia de luz e eletrônica de transmissão (MET)

Papain is a thiol proteolytic enzyme widely used in dermatology that found applications in wound treatment. Recently, papain was also used as absorption enhancer which can modify the peptide/protein material in the bilayer domain. We investigated papain safety using human skin that was exposed to papain in vitro at different times: 4, 24 and 48 hours. The samples were examined using Light and Transmission Electron Microscopy (TEM) to study of the mechanisms involved in enhancer-skin interaction. After 24 hours, changes occurred in corneosomes. However, samples of 48 hours did not show major changes in agreement with the control. These findings indicated that papain could be used safely onto the skin.

Papaína é uma enzima proteolítica amplamente utilizada na dermatologia para o tratamento de feridas. Atualmente, a papaína também tem sido empregada como promotor de absorção cutânea passível de modificar os domínios protéicos da epiderme. Nesta pesquisa investigou-se in vitro a segurança da papaína, utilizando pele humana exposta a enzima em diferentes períodos de tempo de contato: 4, 24 e 48 horas. As amostras foram avaliadas por Microscopia de Luz e Eletrônica de Transmissão (MET), técnicas que podem ser utilizadas no estudo dos mecanismos envolvidos na interação de promotores de absorção cutânea e a pele. Após 24 horas de contato entre a pele a solução de papaína, mudanças ocorreram nos corneossomos, no entanto, as amostras em contato por 24 horas não evidenciaram alterações relevantes comparadas com o controle. Os resultados indicaram que a papaína poderia ser seguramente aplicada sobre a pele.