RESUMO
In this study, we developed a water-in-oil microemulsion containing vitamin A (retinol) and vitamin E (α-tocopherol), which serves as a multifunctional nanosystem that co-delivers antioxidants and displayed additive effect against acute skin inflammation. Microemulsion (ME) was prepared by mixing a surfactant blend (Tween 80 and propylene glycol, 5:1) with isopropyl myristate and water (ratio of 50:40:10, respectively). Vitamin A (0.05% w/w concentration) and/or vitamin E (0.1% w/w concentration) were incorporated into the surfactant mixture of ME by stirring with a magnetic stirrer for 30 min. This multifunctional ME displayed physical stability, with low cytotoxicity in 3T3 cell line, as well as cellular internalization into the cytosol. In vivo treatments using ME delivering α-tocopherol reduced dermal expression of TNF-α by 1.3-fold (pâ¯<â¯0.01), when compared to unloaded ME treatment group. When retinol was added into the ME containing α-tocopherol, it further reduced TNF-α expression by 2-fold (pâ¯<â¯0.001), suggesting the additive effect of vitamin E and vitamin A in the treatment against skin inflammation. In conclusion, we successfully developed the use of water-in-oil ME to pack both vitamin E and vitamin A, and demonstrated for the first time its anti-inflammatory potential when applied topically to TPA-induced inflamed skin.
Assuntos
Sistemas de Liberação de Medicamentos , Inflamação/tratamento farmacológico , Inflamação/patologia , Pele/patologia , Vitamina A/administração & dosagem , Vitamina E/administração & dosagem , Doença Aguda , Administração Tópica , Animais , Varredura Diferencial de Calorimetria , Sobrevivência Celular/efeitos dos fármacos , Emulsões , Células HaCaT , Humanos , Camundongos , Camundongos Pelados , Células NIH 3T3 , Pele/efeitos dos fármacos , Suínos , Vitamina A/farmacologia , Vitamina A/uso terapêutico , Vitamina E/farmacologia , Vitamina E/uso terapêuticoRESUMO
Since psoriasis is an immuno-mediated skin disease, long-term therapies are necessary for its treatment. In clinical investigations, tacrolimus (TAC), a macrolide immunosuppressive inhibitor of calcineurin, arises as an alternative for the treatment of psoriasis, acting in some cytokines involved in the pathogenesis of the disease. Here, we aim to study the psoriasis treatment with TAC and siRNA for one of most cytokines expressed in psoriasis, the TNF-α. A multifunctional nanostructure lipid carrier (NLC) was developed to co-delivery TAC and siRNA. Results showed that the particle size and zeta potential were around 230 nm and + 10 mV, respectively. The release study demonstrated a controlled release of TAC, and the permeation and retention profile in the skin tissue show to be promising for topical application. The cell viability and uptake in murine fibroblast presented low toxicity associated to uptake of NLC in 4 h, and finally, the in vivo animal model demonstrates the efficiency of the NLC multifunctional, exhibiting a reduction of the cytokine TNF-α expression about 7-fold and presenting a synergic effect between the TAC and TNF-α siRNA. The developed system was successfully to treat in vivo psoriatic animal model induced by imiquimod and the synergic combination was reported here for the first time. Graphical abstract.
Assuntos
Imiquimode/efeitos adversos , Psoríase/tratamento farmacológico , RNA Interferente Pequeno/administração & dosagem , Tacrolimo/administração & dosagem , Fator de Necrose Tumoral alfa/genética , Administração Cutânea , Animais , Preparações de Ação Retardada , Modelos Animais de Doenças , Regulação para Baixo , Sinergismo Farmacológico , Feminino , Lipossomos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Células NIH 3T3 , Nanopartículas , Tamanho da Partícula , Psoríase/induzido quimicamente , Psoríase/genética , RNA Interferente Pequeno/farmacologia , Tacrolimo/farmacologia , Fator de Necrose Tumoral alfa/antagonistas & inibidoresRESUMO
Celecoxib (CXB) is a widely used anti-inflammatory drug that also acts as a chemopreventive agent against several types of cancer, including skin cancer. As the long-term oral administration of CXB has been associated with severe side effects, the skin delivery of this drug represents a promising alternative for the treatment of skin inflammatory conditions and chemoprevention of skin cancer. We prepared and characterized liquid crystalline systems based on glyceryl monooleate and water containing penetration enhancers which were primarily designed to promote skin delivery of CXB. Analysis of their phase behavior revealed the formation of cubic and hexagonal phases depending on the systems' composition. The systems' structure and composition markedly affected the in vitro CXB release profile. Oleic acid reduced CXB release rate, but association oleic acid/propylene glycol increased the drug release rate. The developed systems significantly reduced inflammation in an aerosil-induced rat paw edema model. The systems' composition and liquid crystalline structure influenced their anti-inflammatory potency. Cubic phase systems containing oleic acid/propylene glycol association reduced edema in a sustained manner, indicating that they modulate CXB release and permeation. Our findings demonstrate that the developed liquid crystalline systems are potential carriers for the skin delivery of CXB.
Assuntos
Celecoxib/química , Glicerídeos/química , Cristais Líquidos/química , Pele/metabolismo , Administração Cutânea , Animais , Anti-Inflamatórios/química , Anti-Inflamatórios/farmacologia , Celecoxib/farmacologia , Química Farmacêutica/métodos , Portadores de Fármacos/química , Sistemas de Liberação de Medicamentos/métodos , Liberação Controlada de Fármacos/efeitos dos fármacos , Edema/tratamento farmacológico , Inflamação/tratamento farmacológico , Masculino , Ácido Oleico/química , Permeabilidade/efeitos dos fármacos , Propilenoglicol/química , Ratos , Ratos Wistar , Absorção Cutânea/efeitos dos fármacos , Absorção Cutânea/fisiologia , Solubilidade/efeitos dos fármacos , Água/químicaRESUMO
We have designed a microemulsion (ME) containing Ketoprofen (KET) for anti-inflammatory effect evaluated using the rat paw edema model. The ME was prepared by adding propylene glycol (PG) loaded with 1% KET/water (3:1, w/w), to a mixture of sorbitan monooleate and polysorbate 80 (47.0%) at 3:1 (w/w) and canola oil (38.0%). The physicochemical characterization of KET-loaded ME involved particle size and zeta potential determination, entrapment efficiency, calorimetric analysis, and in vitro drug release. The in vivo anti-inflammatory study employed male Wistar rats. Measurement of the foot volume was performed using a caliper immediately before and 2, 4, and 6 h after injection of Aerosil. KET-loaded ME showed particle size around 20 nm, with zeta potential at -16 mV and entrapment efficiency at 70%. Moreover, KET was converted to the amorphous state when loaded in the formulation and it was shown that the drug was slowly released from the ME. Finally, the in vivo biological activity was similar to that of the commercial gel, but ME better controlled edema at 4 h. These results demonstrated that the ME formulation is an alternative strategy for improving KET skin permeation for anti-inflammatory effect. Furthermore, our findings are promising considering that the developed ME was loaded with only 1% KET, and the formulation was able to keep a similar release profile and in vivo effect compared to the commercial gel with 2.5% KET. Therefore, the KET-loaded developed herein ME is likely to have a decreased side effect compared with that of the commercial gel, but both presented the same efficacy.
Assuntos
Anti-Inflamatórios não Esteroides/administração & dosagem , Cetoprofeno/administração & dosagem , Pele/metabolismo , Animais , Liberação Controlada de Fármacos , Edema/tratamento farmacológico , Emulsões/química , Cetoprofeno/química , Cetoprofeno/farmacocinética , Cetoprofeno/farmacologia , Masculino , Tamanho da Partícula , Ratos , Ratos Wistar , Absorção CutâneaRESUMO
Introdução: O uso de animais em pesquisa científica vem sendo ponto de discussão na última década, baseado na teoria dos 3Rs - Reduction, Refinement and Replacement. Objetivo: Avaliar o uso da pele de cobra brasileira (Crotalus durissus terrificus) como membrana biológica para ensaios in vitro de permeação cutânea, usando nicotina em adesivos transdérmicos como medicamento modelo. Há muitas similaridades nas taxas de permeação entre o estrato córneo humano e a pele de cobra, a qual sugere o uso da pele de cobra como um modelo de barreira nos ensaios de permeação através do estrato córneo. A pele de cobra pode vir a ser um substituto em potencial para o uso de animais de laboratórios, reduzindo o número de animais utilizados nas pesquisas, a dor e desconforto dos animais durante os experimentos. É importante procurar alternativas para substituição dos animais tanto em ensaios laboratoriais in vitro como nos ensaios in vivo. Material e Método: Experimento de laboratório. Ensaio in vitro de permeação cutânea utilizando pele de cobra pré-hidratada em água durante 12, 24 e 48 horas...
Introduction: Based on the theory of the 3Rs - Reduction, Refinement and Replacement, the use of animals in scientific research has been focused in the past decade. Objective: To assess the use of Brazilian snake (Crotalus durissus terrificus) skin as a biological membrane for in vitro skin permeation studies using nicotine transdermal patches as a drug model. There are many similarities in the permeation rates between the snake skin and the human stratum corneum, which suggest the use of this tissue as a barrier model to test drug permeation through the stratum corneum. The use of snake skin may become a potential replacement for the use of laboratory animals, reducing this way the number of animals used in research, as well as the pain and discomfort of animals during the experiments. It is important to seek alternatives in order to replace these animals both in vitro and in vivo laboratory assays. Materials and Methods: Laboratory Experiment During in vitro skin permeation studies using pre-treated with water hydration for 12, 24 and 48 hours snake skin, nicotine was used as a model drug, as well as a vertical diffusion cell as apparatus. The nicotine values permeated through snake skin were determined by High Performance Liquid Chromatography, being...
Introducción: El uso de animales en la investigación científica está siendo tema de debate en la última década, basado en la teoría de las 3Rs - Reducción, Refinamiento y Reemplazo. Objetivo: Evaluar el uso de la piel de serpiente brasileña (Crotalus durissus terrificus) como membrana biológica para las pruebas in vitro de la permeabilidad cutánea, el uso de la nicotina en parches transdérmicos como modelo de producto medicinal. Existen muchas similitudes entre los índices de permeabilidad en el estrato córneo y la serpiente humana, lo que sugiere el uso como modelo de barrera en las pruebas de la infiltración a través del estrato córneo. La serpiente puede ser un sustituto potencial para el uso de animales de laboratorio, reduciendo el número de animales utilizados en la investigación, el dolor y el sufrimiento de los animales durante los experimentos. Es importante buscar alternativas para animales de reemplazo tanto en pruebas de laboratorio ensayos in vivo e in vitro. Material y Método: Experimento de laboratorio - En ensayos de permeación in vitro utilizando piel de serpiente pre-tratados con hidratación en agua durante 12, 24 y 48 horas, la nicotina fue utilizado como um modelo de droga y célula de difusión vertical como lo aparato. Los valores de nicotina permeada por la piel de serpiente...
Assuntos
Animais , Animais de Laboratório , Crotalus cascavella , Técnicas In Vitro/métodosRESUMO
PDT has been used in the treatment of malignant brain tumors for the last 2 decades. It is based on the interaction of a photosensitizer (PS) and light of an appropriate wavelength, with generation of oxygen species, mainly singlet oxygen. Brain is particularly susceptible to oxidative stress; therefore the study of PDT effects on cerebral mitochondria might provide mechanistic insights into the action of the therapy, contributing to its optimization. In the present study, we addressed the mitochondrial toxicity of the second generation PS, zinc phthalocyanine tetrasulfonate (ZnPcS4), on rat brain isolated mitochondria, by investigating both intrinsic toxicity and photodynamic action. At higher concentrations (15 and 25 microM/mg protein) ZnPcS4 caused (a) inhibition of state-3 respiration and (b) decrease of RCR and ADP/O. Electrochemical potential, state-4 respiration and Ca2+ retention capacity were not affected. Cytochrome c release was not observed. Coupled with 600 or 1800 mJ/cm2 laser irradiation, ZnPcS4 (5 microM/mg protein) caused more intense effects on state 3, RCR and ADP/O; moreover state-4 respiration and membrane potential were affected. Besides that, Ca2+ and cytochrome c release were induced. Cyclosporine A (CsA) decreased Ca2+ release and ameliorated the electrochemical potential, suggesting that membrane permeability transition (MPT) might be involved in the photodynamic effect. The low intrinsic toxicity and the high photodynamic effect on rat brain mitochondria induced by ZnPcS4, allied to its improved photophysical properties, might indicate its potential for the treatment of malignant brain tumors.