Estudo comparativo do perfil de retenção e permeação cutânea in vitro e ex vivo de peróxido de benzoíla e avaliação da viabilidade de diferentes membranas biológicas em relação ao tempo de armazenamento
Data
2024-07-29
Autores
Brighenti, Murilo de Souza 
Orientadores
Silva, Vânia Rodrigues Leite e
Tipo
Dissertação de mestrado
Título da Revista
ISSN da Revista
Título de Volume
Resumo
A acne vulgar pode ser definida como uma dermatite inflamatória crônica. Peróxido de benzoíla é um dos agentes tópicos mais prescritos para esse tratamento. Em contato com a pele, ocorre a rápida metabolização de peróxido de benzoíla em ácido benzóico e espécies reativas de oxigênio, estas capazes de oxidar proteínas bacterianas presentes na epiderme, reduzindo consideravelmente a carga bacteriana das espécies Cutibacterium e Staphylococcus. Ensaios de liberação in vitro, podem ser utilizados como uma ferramenta importante para avaliar, de maneira prévia, o comportamento físico-químico e a performance de formulações semissólidas e ensaios de permeação cutânea ex vivo são amplamente utilizados para avaliação do perfil de segurança e eficácia de um medicamento tópico, a partir da utilização de membrana biológica provenientes de humanos e de animais. O objetivo deste trabalho foi avaliar e comparar os perfis de liberação in vitro e o perfis de retenção e permeação cutânea ex vivo de formulações semissólidas contendo peróxido de benzoíla, adquiridas no mercado farmacêutico nacional. Os estudos de retenção e permeação cutânea ex vivo foram conduzidos utilizando pele suína e pele humana, a fim de comparar os perfis de retenção e permeação de ambas as formulações avaliadas utilizando-se duas fontes de membrana biológica distintas. A partir dos ensaios de liberação in vitro foi possível evidenciar diferenças físico-químicas entre as formulações avaliadas. A formulação semissólida A apresentou taxa de liberação média de 952,86 ± 24,88 mg/cm²/h½, enquanto as formulações semissólidas B e C apresentaram taxa de liberação média de 505,59 ± 51,21 e 448,64 ± 20,57 mg/cm²/h½, respectivamente. Após a condução dos ensaios de retenção e permeação cutânea ex vivo, utilizando pele de suína e pele humana como membrana biológica foi possível evidenciar que não há diferença estatisticamente relevante entre os resultados obtidos para as três formulações semissólidas avaliadas em relação aos parâmetros: fluxo de permeação, massa total permeada e lag time. Com relação ao perfil de retenção cutânea, a formulação A apresentou uma maior capacidade de retenção cutânea, quando comparada as formulações semissólidas B e C. Além disso, por meio de ensaios de validação analítica, comprovou-se que as metodologias analíticas desenvolvidas e empregadas para a quantificação das amostras provenientes dos estudos de liberação, retenção e permeação cutânea ex vivo são apropriadas para os propósitos pretendidos. Avaliou-se os potenciais efeitos no perfil de retenção e permeação de peróxido de benzoíla após submeter as membranas biológicas à –20 °C em diferentes tempos. Os resultados indicam que a pele suína apresentou alterações significativas após o período de 3 meses, enquanto a pele humana somente após 6 meses.
Acne vulgaris can be defined as a chronic inflammatory dermatitis. Benzoyl peroxide is one of the most prescribed topical agents for this treatment. Upon contact with the skin, benzoyl peroxide is rapidly metabolized into benzoic acid and reactive oxygen species, capable of oxidizing bacterial proteins present in the epidermis, significantly reducing the bacterial load of Cutibacterium and Staphylococcus species. Due to the widespread use of benzoyl peroxide in topical formulations, an evaluation related to the safety of these formulations is necessary. In vitro release assays, which use only synthetic membranes, can be used as an important tool to assess, in advance, the quality, physicochemical behavior, and performance of semi-solid formulations that are in development or already available on the market. Additionally, ex vivo cutaneous permeation assays are widely used to evaluate the safety and efficacy profile of a topical medication, using biological membranes from humans and animals. The objective of this work was to evaluate and compare the in vitro release profiles and ex vivo cutaneous retention and permeation profiles of three semi-solid formulations containing 5% benzoyl peroxide, acquired in the national pharmaceutical market. Ex vivo retention and permeation studies were conducted using porcine ear skin and human skin to compare the retention and permeation profiles of both formulations using two different sources of biological membrane. From the in vitro release assays, it was possible to highlight physicochemical differences between the semi-solid formulations containing 5% benzoyl peroxide evaluated. Semi-solid formulation A showed an average release rate of 952.86 ± 24.88 mg/cm²/h½, while semi-solid formulations B and C showed an average release rate of 505.59 ± 51.21 and 448.64 ± 20.57 mg/cm²/h½, respectively. After conducting ex vivo retention and permeation assays using porcine ear skin and human skin as biological membrane and statistical evaluation using the ANOVA - Tukey test model, it was possible to show that there is no statistically significant difference between the results obtained for the three semi-solid formulations evaluated in relation to the parameters: permeation flux, total permeated mass, and lag time. Regarding the cutaneous retention profile, formulation A showed a higher cutaneous retention capacity when compared to semi-solid formulations B and C. Additionally, through analytical validation assays, it was confirmed that the analytical methodologies developed and employed for the quantification of samples from in vitro release, retention, and ex vivo cutaneous permeation studies are appropriate for the intended purposes. The potential effects on the retention and permeation profile of benzoyl peroxide were evaluated after subjecting biological membranes to -20 °C for different durations. The results indicate that porcine skin showed significant changes after a 3-month period, while human skin only showed changes after 6 months.
Acne vulgaris can be defined as a chronic inflammatory dermatitis. Benzoyl peroxide is one of the most prescribed topical agents for this treatment. Upon contact with the skin, benzoyl peroxide is rapidly metabolized into benzoic acid and reactive oxygen species, capable of oxidizing bacterial proteins present in the epidermis, significantly reducing the bacterial load of Cutibacterium and Staphylococcus species. Due to the widespread use of benzoyl peroxide in topical formulations, an evaluation related to the safety of these formulations is necessary. In vitro release assays, which use only synthetic membranes, can be used as an important tool to assess, in advance, the quality, physicochemical behavior, and performance of semi-solid formulations that are in development or already available on the market. Additionally, ex vivo cutaneous permeation assays are widely used to evaluate the safety and efficacy profile of a topical medication, using biological membranes from humans and animals. The objective of this work was to evaluate and compare the in vitro release profiles and ex vivo cutaneous retention and permeation profiles of three semi-solid formulations containing 5% benzoyl peroxide, acquired in the national pharmaceutical market. Ex vivo retention and permeation studies were conducted using porcine ear skin and human skin to compare the retention and permeation profiles of both formulations using two different sources of biological membrane. From the in vitro release assays, it was possible to highlight physicochemical differences between the semi-solid formulations containing 5% benzoyl peroxide evaluated. Semi-solid formulation A showed an average release rate of 952.86 ± 24.88 mg/cm²/h½, while semi-solid formulations B and C showed an average release rate of 505.59 ± 51.21 and 448.64 ± 20.57 mg/cm²/h½, respectively. After conducting ex vivo retention and permeation assays using porcine ear skin and human skin as biological membrane and statistical evaluation using the ANOVA - Tukey test model, it was possible to show that there is no statistically significant difference between the results obtained for the three semi-solid formulations evaluated in relation to the parameters: permeation flux, total permeated mass, and lag time. Regarding the cutaneous retention profile, formulation A showed a higher cutaneous retention capacity when compared to semi-solid formulations B and C. Additionally, through analytical validation assays, it was confirmed that the analytical methodologies developed and employed for the quantification of samples from in vitro release, retention, and ex vivo cutaneous permeation studies are appropriate for the intended purposes. The potential effects on the retention and permeation profile of benzoyl peroxide were evaluated after subjecting biological membranes to -20 °C for different durations. The results indicate that porcine skin showed significant changes after a 3-month period, while human skin only showed changes after 6 months.