Dispersões sólidas de trans-resveratrol incorporadas em membranas de polivinilpirrolidona para aplicação cutânea: desenvolvimento, caracterização e avaliação biológica in vitro e in vivo
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Data
2021-11-19
Autores
Riccio, Bruno Vincenzo Fiod
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Editor
Universidade Estadual Paulista (Unesp)
Resumo
A inflamação cutânea se relaciona à síntese de mediadores inflamatórios que quando em quantidades exacerbadas e produzidos cronicamente podem gerar doenças como a dermatite atópica. O trans-resveratrol demonstra ser um efetivo agente anti-inflamatório devido à redução ou inibição de mediadores como histamina, citocinas e a ciclooxigenase-2. O desenvolvimento de dispersões sólidas de trans-resveratrol pode melhorar a sua solubilidade. De forma a viabilizar a administração cutânea das dispersões sólidas, membranas de polivinilpirrolidona (PVP) podem ser interessantes, pois se trata de um polímero capaz de evitar a recristalização do trans-resveratrol. O estudo objetivou desenvolver, caracterizar e avaliar a citotoxicidade in vitro e a atividade anti-inflamatória in vivo de dispersões sólidas incorporadas em membranas de PVP para a administração cutânea, sendo algo inédito do ponto de vista tecnológico. Foram desenvolvidas dispersões sólidas contendo trans-resveratrol constituídas por quitosana e tensoativo (Poloxamer® 407 ou TPGS), bem como duas formulações sem tensoativos, as quais foram caracterizadas empregando análises morfológicas e físico-químicas. Foi desenvolvido um método analítico por cromatografia líquida de alta eficiência (CLAE) para a quantificação do trans-resveratrol. Após a escolha das formulações de melhor desempenho, duas foram incorporadas em membranas poliméricas de PVP, sendo feitas também outras duas sem fármaco como controle. As membranas foram caracterizadas quanto às suas propriedades de barreira e físico-químicas, bem como análises in vitro e in vivo. De acordo com os resultados, foi possível atingir até 56% de rendimento na produção das dispersões sólidas de trans-resveratrol empregando secagem por aspersão (spray dryer), sendo que dentre as formulações quase todas obtiveram ângulo de repouso excelente, bem como através da difração de raios-X (DRX) foi possível constatar que as dispersões sólidas conseguiram amorfizar o trans-resveratrol com sucesso. As dispersões sólidas também aumentaram a solubilidade do fármaco em água, com algumas formulações dobrando a sua solubilidade. As membranas conseguiram manter o trans-resveratrol sem recristalizar por mais de dois meses, conforme evidenciado pelo DRX, além de apresentarem bioadesão, favorável para a administração cutânea. De acordo com o ensaio de liberação in vitro, as membranas obtidas não liberaram totalmente o fármaco em 24 h, obtendo valores de 82,27±2,60 a 92,81±2,50%, sendo assim um perfil de liberação prolongado. Ensaios de retenção in vitro evidenciaram que as quantidades de trans-resveratrol retidas nas camadas da derme e epiderme em 24 h variaram de 42,88 a 53,28%, estando disponíveis para promover o efeito farmacológico. Observou-se também que as citotoxicidades das formulações não foram expressivas. Por fim, no ensaio in vivo, foi possível observar que as formulações apresentaram efeito anti-inflamatório favorável ao tratamento farmacológico proposto, obtendo-se a redução de até 66% no modelo de inflamação de orelha de camundongo. Os resultados demonstram que o sistema desenvolvido é capaz de ser utilizado como agente anti-inflamatório para o tratamento da dermatite atópica.
Cutaneous inflammation is related to the synthesis of inflammatory mediators that, when in exacerbated amounts and chronically produced, can generate diseases such as atopic dermatitis. Trans-resveratrol proves to be an effective anti-inflammatory agent due to the reduction or inhibition of mediators such as histamine, cytokines and cyclooxygenase-2. The development of solid dispersions containing trans-resveratrol can improve its solubility. In order to enable the cutaneous administration of solid dispersions, polyvinylpyrrolidone (PVP) membranes can be interesting, since it is a polymer capable of preventing the recrystallization of trans-resveratrol, being something unprecedented from a technological point of view. The study aimed to develop, characterize and evaluate the in vitro cytotoxicity and in vivo anti-inflammatory activity of solid dispersions incorporated in PVP membranes for cutaneous administration. Solid dispersions containing trans-resveratrol constituted by chitosan and surfactant (Poloxamer® 407 or TPGS) were developed, as well as two formulations without surfactants, which were characterized using morphological and physicochemical analyses. An analytical method by high performance liquid chromatography (HPLC) was developed for the quantification of trans-resveratrol. After choosing the best performing formulations, two were incorporated into polymeric PVP membranes and two others were also made without the drug as a control. The membranes were characterized regarding their barrier and physicochemical properties, as well as in vitro and in vivo analyses. According to the results, it was possible to reach up to 56% yield in the production of solid dispersions containing trans-resveratrol using spray drying. According to the X-ray diffraction (XRD) it was possible to verify that the solid dispersions were able to amorphize the trans-resveratrol successfully. Solid dispersions also increased the drug's solubility in water, with some formulations doubling its solubility.The membranes managed to maintain trans-resveratrol without recrystallizing for more than two months, as evidenced by the XRD, in addition to presenting bioadhesion, favorable for cutaneous administration. According to the in vitro release assay, the membranes obtained did not fully release the drug in 24 h, obtaining values from 82.27±2.60 to 92.81±2.50%, thus being a profile of extended release. The in vitro retention assays showed that the amounts of trans-resveratrol retained in the dermis and epidermis layers within 24 h ranged from 42.88 to 53.28%, being available to promote the pharmacological effect. It was also observed that the cytotoxicities of the formulations were not expressive. Finally, in the in vivo test, it was possible to observe that the formulations presented an anti-inflammatory effect favorable to the proposed pharmacological treatment, obtaining a reduction of up to 66% in mouse ear inflammation model. The results demonstrate that the developed system is capable of being used as an anti-inflammatory agent for the treatment of atopic dermatitis.
Cutaneous inflammation is related to the synthesis of inflammatory mediators that, when in exacerbated amounts and chronically produced, can generate diseases such as atopic dermatitis. Trans-resveratrol proves to be an effective anti-inflammatory agent due to the reduction or inhibition of mediators such as histamine, cytokines and cyclooxygenase-2. The development of solid dispersions containing trans-resveratrol can improve its solubility. In order to enable the cutaneous administration of solid dispersions, polyvinylpyrrolidone (PVP) membranes can be interesting, since it is a polymer capable of preventing the recrystallization of trans-resveratrol, being something unprecedented from a technological point of view. The study aimed to develop, characterize and evaluate the in vitro cytotoxicity and in vivo anti-inflammatory activity of solid dispersions incorporated in PVP membranes for cutaneous administration. Solid dispersions containing trans-resveratrol constituted by chitosan and surfactant (Poloxamer® 407 or TPGS) were developed, as well as two formulations without surfactants, which were characterized using morphological and physicochemical analyses. An analytical method by high performance liquid chromatography (HPLC) was developed for the quantification of trans-resveratrol. After choosing the best performing formulations, two were incorporated into polymeric PVP membranes and two others were also made without the drug as a control. The membranes were characterized regarding their barrier and physicochemical properties, as well as in vitro and in vivo analyses. According to the results, it was possible to reach up to 56% yield in the production of solid dispersions containing trans-resveratrol using spray drying. According to the X-ray diffraction (XRD) it was possible to verify that the solid dispersions were able to amorphize the trans-resveratrol successfully. Solid dispersions also increased the drug's solubility in water, with some formulations doubling its solubility.The membranes managed to maintain trans-resveratrol without recrystallizing for more than two months, as evidenced by the XRD, in addition to presenting bioadhesion, favorable for cutaneous administration. According to the in vitro release assay, the membranes obtained did not fully release the drug in 24 h, obtaining values from 82.27±2.60 to 92.81±2.50%, thus being a profile of extended release. The in vitro retention assays showed that the amounts of trans-resveratrol retained in the dermis and epidermis layers within 24 h ranged from 42.88 to 53.28%, being available to promote the pharmacological effect. It was also observed that the cytotoxicities of the formulations were not expressive. Finally, in the in vivo test, it was possible to observe that the formulations presented an anti-inflammatory effect favorable to the proposed pharmacological treatment, obtaining a reduction of up to 66% in mouse ear inflammation model. The results demonstrate that the developed system is capable of being used as an anti-inflammatory agent for the treatment of atopic dermatitis.
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Palavras-chave
Amorfização, Dermatite atópica, Cristalinidade, Permeação Cutânea, Solubilidade