Rosacea Topical Treatment and Care: From Traditional to New Drug Delivery Systems.

Rosacea is a multifactorial chronic inflammatory dermatosis characterized by flushing, nontransient erythema, papules and pustules, telangiectasia, and phymatous alterations accompanied by itching, burning, or stinging, the pathophysiology of which is not yet fully understood. Conventional topical treatments usually show limited efficacy due to the physical barrier property of the skin that hinders skin penetration of the active ingredients, thereby hampering proper drug skin delivery and the respective therapeutic or cosmetic effects. New advances regarding the physiopathological understanding of the disease and the underlying mechanisms suggest the potential of new active ingredients as promising therapeutic and cosmetic approaches to this dermatosis. Additionally, the development of new drug delivery systems for skin delivery, particularly the potential of nanoparticles for the topical treatment and care of rosacea, has been described. Emphasis has been placed on their reduced nanometric size, which contributes to a significant improvement in the attainment of targeted skin drug delivery. In addition to the exposition of the known pathophysiology, epidemiology, diagnosis, and preventive measures, this Review covers the topical approaches used in the control of rosacea, including skin care, cosmetics, and topical therapies, as well as the future perspectives on these strategies.

In vitro performance of free and encapsulated bromelain.

For centuries, bromelain has been used to treat a range of ailments, even though its mechanism of action is not fully understood. Its therapeutic benefits include enzymatic debridement of the necrotic tissues of ulcers and burn wounds, besides anti-inflammatory, anti-tumor, and antioxidant properties. However, the protease is unstable and susceptible to self-hydrolysis over time. To overcome the stability issues of bromelain, a previous study formulated chitosan-bromelain nanoparticles (C-B-NP). We evaluated the optimized nanoformulation for in vitro antioxidant, cell antiproliferative activities and cell migration/proliferation in the scratch assay, comparing it with free bromelain. The antioxidant activity of free bromelain was concentration and time-dependent; after encapsulation, the activity level dropped, probably due to the slow release of protein from the nanoparticles. In vitro antiproliferative activity was observed in six tumor cell lines for free protein after 48 h of treatment (glioma, breast, ovarian, prostate, colon adenocarcinoma and chronic myeloid leukemia), but not for keratinocyte cells, enabling its use as an active topical treatment. In turn, C-B-NP only inhibited one cell line (chronic myeloid leukemia) and required higher concentrations for inhibition. After 144 h treatment of glioma cells with C-B-NP, growth inhibition was equivalent to that promoted by the free protein. This last result confirmed the delayed-release kinetics of the optimized formulation and bromelain integrity. Finally, a scratch assay with keratinocyte cells showed that C-B-NP achieved more than 90% wound retraction after 24 h, compared to no retraction with the free bromelain. Therefore, nanoencapsulation of bromelain with chitosan conferred physical protection, delayed release, and wound retraction activity to the formulation, properties that favor topical formulations with a modified release. In addition, the promising results with the glioma cell line point to further studies of C-B-NP for anti-tumor treatments.

In vitro activity and formulation of a flavonoid-containing cashew pulp extract for the topical treatment of acne and the protection of skin against premature aging.

The cashew nut is an important product in Brazil, both for consumption and export, with the pulp of the cashew fruit being considered a by-product despite its high flavonoid content. In this study, the use of cashew pulp extract as a treatment for acne and in the prevention of early skin damage was investigated. Its flavonoid content was determined using spectrophotometric identification, and its effects on cell and bacterial viability, the migration of keratinocytes, and antioxidant activity in vitro were evaluated. Furthermore, it was incorporated into an emulsion for topical administration, and the physical-chemical stability parameters of the formulation were determined. The cashew pulp contained flavonoids with healing and antioxidant activity, and was not toxic to keratinocyte cells in a viability test. The flavonoid-rich formulation was stable, indicating that this is a promising formulation for use in the treatment of acne and protection of skin against premature damage.[Figure: see text].

In vitro solar protection factor, antioxidant activity, and stability of a topical formulation containing Benitaka grape (Vitis vinifera L.) peel extract.

A flavonoid enriched extract (FE) was obtained from grape peels, and in vitro SPF, antioxidant activity, and effects on cell viability of this extract were tested with the intent to develop a cosmetic product. A formulation was developed with the FE, and the stability of this mixture was evaluated in terms of pH, density, viscosity, and SPF (90-days). FE showed no cytotoxicity to human keratinocytes and an in vitro SPF of 18.56 (UV-spectrophotometry). Further, FE showed a UVA protection factor of 3.17 ± 0.2, a critical wavelength of 318.0 ± 0.1 and a UVA/UVB of 0.9. Antioxidant activity assays resulted in 92.08% and 86.85% of activity against DPPH and ABTS (IC50 = 296.90 ± 1.2 µg/mL and 643.13 ± 0.9 µg/mL), respectively. Finally, SPF of formulation with FE was 12.45. Results from the in vitro SPF and product stability tests (especially storage under refrigeration), indicate that FE is a promising compound for use as an innovative sunscreen formulation.

Evaluation of In Vitro Solar Protection Factor (SPF), Antioxidant Activity, and Cell Viability of Mixed Vegetable Extracts from Dirmophandra mollis Benth, Ginkgo biloba L., Ruta graveolens L., and Vitis vinífera L.

The aim of this study was to validate a HPLC method for the assay of flavonoids in extracts obtained from natural sources, i.e., from Dirmophandra mollis Benth, Ginkgo biloba L., Ruta graveolens L., and Vitis vinífera L. The potential sun protecting effect, antioxidant activity, and cell viability of the extracts were also determined. Individual extracts (obtained from each individual species) and a mixed extract (containing the four extracts) were analyzed by the validated HPLC method for the identification of flavonoids and quantification of rutin and quercetin. An in vitro cell viability study was carried out using the neutral red method. The in vitro sun protection factor was determined by spectral transmittance and in vitro antioxidant efficacy was evaluated against DPPH, ABTS, and AAPH radicals. The HPLC method used for the identification and quantification of flavonoids in extracts exhibited linearity, precision, accuracy, and robustness. Detection and quantification limits were, respectively, 2.881 ± 0.9 µg·mL-1 and 0.864 ± 0.9 µg·mL-1 for quercetin, and 30.09 ± 1 µg·mL-1 and 9.027 ± 1.1 µg·mL-1 for rutin. All extracts did not affect cell viability at the evaluated concentration range and exhibited a sun protection effect and antioxidant activity. Among the evaluated extracts, Ginkgo biloba L. and the mixed extract depicted the most expressive antioxidant activity. The mixed extract exhibited sunscreen protection against ultraviolet A (UVA) and ultraviolet B (UVB) and a critical wavelength of 372.7 ± 0.1. Our results translate the enhanced flavonoids' composition of the mixed extract, which may be a potential alternative over sunscreens and antioxidants in pharmaceutic/cosmetic formulations.

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

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

Effect of Polysaccharide Sources on the Physicochemical Properties of Bromelain-Chitosan Nanoparticles.

Bromelain, a set of proteolytic enzymes potential pharmaceutical applications, was encapsulated in chitosan nanoparticles to enhance enzyme stability, and the effect of different chitosan sources was evaluated. Chitosan types (i.e., low molecular weight chitosan, chitosan oligosaccharide lactate, and chitosan from shrimp shells) produced nanoparticles with different physicochemical properties, however in all cases, particle size and zeta potential decreased, and polydispersity index increased after bromelain addition. Bromelain encapsulation was higher than 84% and 79% for protein content and enzymatic activity, respectively, with low molecular weight chitosan presenting the highest encapsulation efficiency. Nanoparticle suspension was also tested for accelerated stability and rheological behavior. For the chitosan-bromelain nanoparticles, an instability index below 0.3 was recorded and, in general, the loading of bromelain in chitosan nanoparticles decreased the cohesiveness of the final suspension.

In vitro antioxidant activity and solar protection factor of blackberry and raspberry extracts in topical formulation.

BACKGROUND: Berries are known for their antioxidant activity due to the presence of flavonoids. Antioxidants' usage guarantees skin protection against free radicals and, flavonoids, especially, can act as sunscreen. The aim of this paper was to evaluate solar protection factor (SPF) and antioxidant activity in vitro of blackberry and raspberry extracts incorporated in topical formulation and study their stability. RESULTS: Raspberry presented 29.93 mg of anthocyanins/100 g of fruit and blackberry 65.58 mg of anthocyanins/100 g of fruit. In vitro solar protection factor was 54.57 to blackberry and 37.32 to raspberry. When incorporated in O/W emulsions, final formulations showed light pink color, creamy aspect, and typical fruity odor. Formulations submitted to indirect light and stove showed odor and color alterations. CONCLUSIONS: Based on the results, formulations containing extracts should be kept under refrigeration in opaque package to ensure stability due to the presence of flavonoids. In addition, blackberry and raspberry extracts are potential natural alternatives to be used as sunscreen and to prevent skin aging.

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

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

Vitamin C in Acerola and Red Plum Extracts: Quantification via HPLC, in Vitro Antioxidant Activity, and Stability of their Gel and Emulsion Formulations.

BACKGROUND: The fruits acerola and red plum are known to be good sources of antioxidants, particularly vitamin C. Antioxidants are compounds that protect organisms from biomolecular damage, such as accelerated aging, caused by free radicals. OBJECTIVE: The objective of this study was to extract vitamin C from acerola and red plum, incorporate these extracts into different topical formulations, and evaluate the physicochemical stabilities of these formulations under stress conditions. METHODS: Vitamin C was extracted from acerola and red plum via dynamic maceration for 2 h at 50 ± 2°C and was quantified via HPLC. In vitro antioxidant activities were evaluated using DPPH assays. The extracts were then incorporated into emulsion and gel formulations in two types of packaging, and stability studies were carried out. RESULTS: Red plum and acerola extracts were orange and red and contained vitamin C concentrations of 2732.70 ± 93.01 mg/100 g and 2.60 ± 1.2 mg/100 g, respectively. In vitro antioxidant activity resulted in over 90.0% inhibition of free radicals at 0.01 mL/mL acerola extract and 0.1 mL/mL red plum extract. In the stability study, pH values decreased for both acerola formulations when stored in the oven or in transparent glass containers. Formulations containing red plum extract were stable under all conditions. Acerola extracts contained a higher concentration of vitamin C than red plum extracts. Both extracts possessed antioxidant activity, although the acerola-based formulation was unstable when stored at high temperatures or in transparent glass containers. HIGHLIGHTS: Extracts from red plum and acerola contained vitamin C; antioxidant activity of the extracts resulted in over 90.0% inhibition of free radicals. Formulations containing red plum were stable under all tested conditions, and formulations containing acerola were unstable when stored in the oven or in transparent glass containers.

Bromelain Loading and Release from a Hydrogel Formulated Using Alginate and Arabic Gum.

An ideal wound dressing ensures a moist environment around the wound area and absorbs exudates from the wound surface. Topical application of bromelain to incised wounds has been shown to reprogram the wound microenvironment to promote effective tissue repair. Combining the characteristics of hydrogels and bromelain is therefore of great interest. Herein, we describe the development of a hydrogel, formulated using alginate and Arabic gum, for bromelain loading and release. The hydrogel formulation was evaluated using response surface methodology, considering the pH value and the concentration of alginate and Arabic gum. Bromelain loading and release were evaluated based on passive diffusion. Differential scanning calorimetry and Fourier transform infrared spectroscopy were performed to confirm bromelain immobilization in the hydrogel. The final hydrogel formulation had a swelling ratio of 227 % and incorporated 19 % of bromelain from a bromelain solution. Bromelain immobilization in the hydrogel was the result of hydrogen bond formation and was optimal at 4 °C after 4 h of contact. This evidence suggests that bromelain entrapment into a hydrogel is a promising strategy for the development of wound dressings that support the debridement of burns and wounds.

Photostability study of commercial sunscreens submitted to artificial UV irradiation and/or fluorescent radiation.

Sunscreens contain molecules with the ability to absorb and/or reflect UVA (ultraviolet A) and UVB (ultraviolet B) radiation, thereby preventing this radiation from reaching the epidermis or dermis. Their photo stabilities after exposure to UV radiation are well known and described, but there is little data on the stability of these filters after fluorescent indoors light radiation, such as from light emitted by commercial lamps present in homes and offices. Those lamps can expose people to varying levels of UVB, UVA, visible light, and IR (infrared). This study assesses the photostability of four different commercial products containing chemical sun filters after artificial UV and fluorescent irradiation, correlating the UVB and UVA absorption efficiencies of each product against the different types of radiation. The tested products were applied on a plate of polymethylmethacrylate (PMMA) and irradiated by a solar simulator with specific filters for UVA and UVB and a commercial fluorescent light source. According to the results, three formulations did not show photostability, suffering significant changes in their UV absorption spectra, and one of the selected formulations can be considered photostable. This reinforces the importance of conducting stability studies for sunscreen formulations in different conditions, including under artificial (indoor) light exposure.

Antioxidant Activity and Validation of Quantification Method for Lycopene Extracted from Tomato.

Lycopene is a carotenoid found in tomatoes with potent antioxidant activity. The aim of the study was to obtain an extract containing lycopene from four types of tomatoes, validate a quantification method for the extracts by HPLC, and assess its antioxidant activity. Results revealed that the tomatoes analyzed contained lycopene and antioxidant activity. Salad tomato presented the highest concentration of this carotenoid and antioxidant activity. The quantification method exhibited linearity with a correlation coefficient of 0.9992. Tests for the assessment of precision, accuracy, and robustness achieved coefficients with variation of less than 5%. The LOD and LOQ were 0.0012 and 0.0039 µg/mL, respectively. Salad tomato can be used as a source of lycopene for the development of topical formulations, and based on performed tests, the chosen method for the identification and quantification of lycopene was considered to be linear, precise, exact, selective, and robust.

Development and evaluation of an emulsion containing lycopene for combating acceleration of skin aging

Lycopene, a carotenoid and potent antioxidant is found in large quantities in tomatoes. Lycopene combats diseases, such as cardiovascular disease and different types of cancer, including prostate cancer. However, its topical use in emulsion form for the combat of skin aging is under-explored. The aim of the present study was to develop an emulsion containing lycopene extracted from salad tomatoes and evaluate its cytotoxicity, stability, rheological behavior, antioxidant activity and phytocosmetic permeation. The developed cosmetic comprised an oil phase made up of shea derivatives and was evaluated in terms of its physiochemical stability, spreadability, thermal analysis, rheological behavior, microbiological quality, cytotoxicity, antioxidant activity, cutaneous permeation and retention. The results demonstrate that this phytocosmetic is stable, exhibits satisfactory rheological behavior for a topical formula and is a promising product for combating skin aging.

Licopeno é um carotenóide com potente atividade antioxidante encontrado em grande quantidade no tomate e usado no combate a diversas doenças como doenças cardiovasculares e diferentes tipos de cânceres, incluindo o câncer de próstata. O objetivo desse trabalho foi desenvolver uma emulsão contendo extrato de licopeno obtido do tomate salada e avaliar a citotoxicidade do extrato, a estabilidade, o comportamento reológico, atividade antioxidante e permeação do fitocosmético. O cosmético foi desenvolvido utilizando fase oleosa contendo derivados de Karité e submetido à avaliação da estabilidade físico-química, espalhabilidade, análise térmica, comportamento reológico, qualidade microbiológica, citotoxicidade, atividade antioxidante e testes de permeação e retenção cutânea. Os resultados demonstraram que o fitocosmético é estável, apresenta comportamento reológico desejável para uma formulação tópica e é um produto promissor para ser utilizado no combate à aceleração do envelhecimento cutâneo.

Design of antiseptic formulations containing extract of Plinia cauliflora / Concepção de formulações anti-sépticas que contenham extrato de Plinia cauliflora

The leaves of the Brazilian species Plinia cauliflora were used to obtain active hydroalcoholic extract and fractions enabling the development of efficient antiseptic pharmaceutical formulations. A chemical composition of 70 percent ethanol extract, aqueous and ethyl acetate fractions was analyzed by thin-layer chromatography and for phenol content. Antimicrobial activity was tested against Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, Lactobacillus acidophilus and Candida albicans by the agar diffusion method and the minimum inhibitory concentration was assayed by broth microdilution. Extract microbiological quality was tested to avoid contamination in the formulations. A mouthwash and a topical cream containing the extract were developed and antiseptic activity was assessed by agar diffusion. Sensory and physicochemical stability of the formulations were assayed. Chromatography indicated the presence of terpenes, flavonoids and tannins in the extract and fractions and total phenol content were found to be high. The plant samples were active against all the microorganisms tested, except for Lactobacillus acidophilus. Both topical formulations showed antiseptic activity and stability. Thus, these may be used as antimicrobials in skin infections, but would be more useful in the treatment of candidiasis.

As folhas da espécie brasileira Plinia cauliflora foram utilizadas a fim de se obter um extrato hidroalcoólico e frações ativas proporcionando o desenvolvimento de eficazes formulações farmacêuticas antissépticas. A composição química do extrato etanólico 70 por cento, fração aquosa e acetato de etila foi analisada por cromatografia em camada delgada e teor de fenóis. A atividade antimicrobiana foi testada frente a Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, Lactobacillus acidophilus e Candida albicans por difusão em ágar e a concentração inibitória mínima foi determinada por microdiluição. A qualidade microbiológica do extrato foi avaliada para evitar a contaminação das formulações. Foram desenvolvidos um enxaguatório bucal e um creme tópico contendo o extrato sendo que a atividade antisséptica foi ensaiada por difusão em ágar. A estabilidade sensorial e físico-química foram testadas. A cromatografia indicou a presença de terpenos, flavonóides e taninos no extrato e frações, observando-se alto teor de fenóis totais. As amostras vegetais foram ativas contra todos os micro-organismos testados, exceto Lactobacillus acidophilus. Ambas as formulações apresentaram atividade antisséptica e estabilidade. Desta forma, infere-se que as formulações desenvolvidas podem ser utilizadas como antissépticas em infecções de pele, podendo ser mais eficazes no tratamento de candidíase.