Novas perspectivas de tratamentos para a acne através de pos-bióticos e ácido salicílico nanoencapsulado para o controle da microbiota da pele
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Data
2023-03-30
Autores
Miranda, Rute Gonçalves [UNIFESP]
Orientadores
Silva, Vânia Rodrigues Leite e [UNIFESP]
Tipo
Dissertação de mestrado
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Resumo
Por muito tempo a acne foi definida pela proliferação do Cutibacterium acnes (C.acnes), mas recentemente discute-se a hipótese de que uma possível disbiose entre os subgrupos do C.acnes é que pode levar as lesões aceicas. O presente estudo tem o objetivo de analisar a influência de três estratégias no desenvolvimento de C.acnes com intuito de restabelecer o equilíbrio da microbiota da pele acneica que são: fornecimento do pós-biótico Lactobacillus acidophylus (La), de lipase adsorvida em sílica mesoporosa (SBA-15) e administração de ácido salicílico (AS) em nanopartículas lipídicas. Para isso, foi obtida a formulação de carreadores lipídicos nanoestruturados contendo AS, sendo caracterizado quanto ao teor de AS e liberação frente ao pH por UV-VIS, e análise do tamanho de partícula por difração a laser (DL); o complexo de lipase incorporada na sílica SBA-15 (LIP-SIL), e em sequência caracterizada quanto ao teor de lipase por espectrofotometria, atividade enzimática e conservação da atividade por 30 dias; os La foram cultivados em BHI Caldo para a construção da curva de crescimento e os microorganismos foram lavados e ultrassonicados para obtenção dos lisados como pós-bióticos; da mesma forma foi realizado para obtenção e padronização do C.acnes para utilização no teste in vitro de desafio do microrganismo frente a cada uma das estratégias apontadas. Os valores de absorbância em leitor de placas revelaram o crescimento do C.acnes nas condições empregadas. As nanopartículas lipídicas contendo AS apresentaram teor de recuperação de fármaco de 90,6% p/V, já a análise de tamanho revelou cerca de 90 – 100% das partículas lipídicas com tamanho menor que 500 nm, indicando a capacidade das partículas atuarem no interior dos folículos pilosos. Os ensaios de liberação demonstraram que as nanopartículas foram capazes de liberar totalmente o conteúdo de AS para valores de pH acima do pKa do fármaco em um período de 8h. Foi incorporada à sílica 44%p/p de lipase com atividade registrada de 60% (T0), sendo mantida atividade de 66,37% após 30 dias. O cultivo de C.acnes e La demostraram melhor crescimento em 72h. Os resultados de crescimento do C.acnes frente as estratégias terapêuticas demostraram que todas contribuíram para inibição do crescimento do C.acnes após 24 e 48h de exposição, sendo capazes de interferir no seu desenvolvimento. Conclui-se que as estratégias terapêuticas propostas têm potencial para aplicação em formulações cosméticas para o tratamento da acne.
For a long time, acne was defined by the external perspectives of Cutibacterium acnes (C.acnes), but recently the hypothesis has been discussed that a possible dysbiosis between C.acnes subgroups is what can lead to acneic lesions. The present study aims to analyze the influence of three strategies on the development of C.acnes with the aim of restoring the balance of the microbiota of acneic skin, which are: supply of the post-biotic Lactobacillus acidophylus (La), of lipase adsorbed on mesoporous sílica (SBA-15) and administration of salicylic acid (SA) in lipid nanoparticles. For this, it was subjected to a formulation of nanostructured lipid carriers containing AS, characterized in terms of AS content and released against pH by UV-VIS, and particle size analysis by laser diffraction (DL); the lipase complex created on SBA-15 silica (LIP-SIL), and in a sequence identified in terms of lipase content by spectrophotometry, enzymatic activity and conservation of activity for 30 days; the La were cultivated in BHI Caldo for the construction of the growth curve and the microorganisms were washed and ultrasonicated to obtain the lysates as postbiotics; in the same way, it was carried out to obtain and standardize the C.acnes for use in the in vitro challenge test of the microorganism against each of the indicated strategies. The absorbance values in a plate reader revealed the growth of C.acnes in the tasks employed. The lipid nanoparticles containing AS showed drug recovery content of 90.6% w/v, while the size analysis revealed about 90 - 100% of the lipid particles with a size smaller than 500 nm, indicating the ability of the particles to act inside of the hair follicles. The release assays showed that the nanoparticles were able to completely release the AS content at pH values above the pKa of the drug in a period of 8h. Silica was created with 44%w/w of lipase with a registered activity of 60% (T0), with an activity of 66.37% maintained after 30 days. The culture of C.acnes and La showed better growth in 72h. The results of growth of C.acnes compared to the therapeutic strategies showed that all directed towards the growth of C.acnes after 24 and 48 hours of exposure, being capable of interfering in its development. It is concluded that the proposed therapeutic strategies have potential for application in cosmetic formulations for the treatment of acne.
For a long time, acne was defined by the external perspectives of Cutibacterium acnes (C.acnes), but recently the hypothesis has been discussed that a possible dysbiosis between C.acnes subgroups is what can lead to acneic lesions. The present study aims to analyze the influence of three strategies on the development of C.acnes with the aim of restoring the balance of the microbiota of acneic skin, which are: supply of the post-biotic Lactobacillus acidophylus (La), of lipase adsorbed on mesoporous sílica (SBA-15) and administration of salicylic acid (SA) in lipid nanoparticles. For this, it was subjected to a formulation of nanostructured lipid carriers containing AS, characterized in terms of AS content and released against pH by UV-VIS, and particle size analysis by laser diffraction (DL); the lipase complex created on SBA-15 silica (LIP-SIL), and in a sequence identified in terms of lipase content by spectrophotometry, enzymatic activity and conservation of activity for 30 days; the La were cultivated in BHI Caldo for the construction of the growth curve and the microorganisms were washed and ultrasonicated to obtain the lysates as postbiotics; in the same way, it was carried out to obtain and standardize the C.acnes for use in the in vitro challenge test of the microorganism against each of the indicated strategies. The absorbance values in a plate reader revealed the growth of C.acnes in the tasks employed. The lipid nanoparticles containing AS showed drug recovery content of 90.6% w/v, while the size analysis revealed about 90 - 100% of the lipid particles with a size smaller than 500 nm, indicating the ability of the particles to act inside of the hair follicles. The release assays showed that the nanoparticles were able to completely release the AS content at pH values above the pKa of the drug in a period of 8h. Silica was created with 44%w/w of lipase with a registered activity of 60% (T0), with an activity of 66.37% maintained after 30 days. The culture of C.acnes and La showed better growth in 72h. The results of growth of C.acnes compared to the therapeutic strategies showed that all directed towards the growth of C.acnes after 24 and 48 hours of exposure, being capable of interfering in its development. It is concluded that the proposed therapeutic strategies have potential for application in cosmetic formulations for the treatment of acne.
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Citação
MIRANDA, Rute Gonçalves. Novas perspectivas de tratamentos para a acne através de pos-bióticos e ácido salicílico nanoencapsulado para o controle da microbiota da pele. 2023. 161 f. Dissertação (Mestrado em Medicina Translacional) - Escola Paulista de Medicina, Universidade Federal de São Paulo (UNIFESP). São Paulo, 2023.