Impacto na migração de queratinócitos por modulação das enzimas LOX : aplicabilidade em formulações tópicas cutâneas
Miniatura indisponível
Data
2023
Título da revista
ISSN da revista
Título do Volume
Editora
Resumo
Uma ferida cutânea é definida por qualquer alteração na integridade das camadas da pele. Estima-se que 2 milhões de europeus sofrem de lesões cutâneas sendo um importante fator influenciador da qualidade de vida do paciente. Aquando da disrupção da pele, o sistema tegumentar possui capacidade para se autorregenerar, processo esse designado de cicatrização de lesões cutâneas. Constitui um processo vital e fundamental que tem como principal objetivo a restauração da integridade e função normal da pele, onde estão envolvidos inúmeros componentes celulares como por exemplo, as enzimas LOX. Estas catalisam reações de reticulação de colagénio e elastina no tecido conjuntivo, permitindo a ligação cruzada entre estes dois compostos da matriz extracelular da pele, facilitando o processo de migração celular no processo de cicatrização de feridas. De modo a otimizar tratamentos para maximizar a cura das feridas cutâneas, a presente dissertação teve como principal objetivo estudar o impacto de compostos selecionados, com atividade inibidora das enzimas LOX, na migração coletiva de queratinócitos, e posterior desenvolvimento de nano formulações para aplicação cutânea, tendo em vista a otimização do processo de cicatrização de feridas cutâneas, contribuindo para melhoria de problemas relacionados com estas e podendo aumentar a qualidade de vida do paciente. Usando um ensaio in vitro, o efeito dos compostos inibidores das enzimas LOX BAPN, EPCU e PN1 na migração coletiva de células HaCaT foi avaliado. Ao contrário do que seria de esperar, tendo em conta o efeito deste tipo de compostos noutras linhas celulares, observou-se que dois dos compostos aumentaram significativamente a migração celular coletiva, enquanto outro composto não teve qualquer impacto migração celular. Posteriormente, foram produzidas e desenvolvidas nano formulações compostas por nanopartículas lipídicas solidas contendo BAPN e avaliou-se a sua estabilidade ao longo do tempo assim como o seu impacto aquando da sua aplicação cutânea. De uma forma geral, as nano formulações desenvolvidas apresentaram valores de pH, diâmetro hidrodinâmico e índice de polidispersão estáveis ao longo dos 30 dias de estudo. Os resultados do estudo in vivo indiciam uma boa compatibilidade cutânea das nanoformulacões, mas não foram observadas diferenças relativamente ao controlo na sua eficácia, pelo que serão necessários estudos futuros com um painel mais alargado de voluntários. Palavras-chave: enzimas LOX, cicatrização de feridas, ensaio de risco, migração celular coletiva, nanotecnologia, aplicação cutânea
A skin wound is defined by any alteration in the integrity of the skin layers. It is estimated that 2 million Europeans suffer from skin lesions, which is an important factor influencing the patient's quality of life. When the skin is disrupted, the integumentary system has the capacity to self-regenerate, a process called healing of skin lesions. It constitutes a vital and fundamental process whose main objective is to restore the integrity and normal function of the skin, where numerous cellular components are involved, such as the LOX enzymes. These catalyze cross-linking reactions of collagen and elastin in connective tissue, thus allowing cross-linking between these two compounds in the extracellular matrix of the skin, facilitating the process of cell migration in the wound healing process. In order to optimize treatments to maximize the healing of cutaneous wounds, the present dissertation had as main objective to study the impact of selected compounds, with inhibitory activity of LOX enzymes, in the collective migration of keratinocytes, and later development of nanoformulations for cutaneous application, having with a view to optimizing the healing process of cutaneous wounds, contributing to the improvement of problems related to these and possibly increasing the patient's quality of life. Using an in vitro assay, the effect of LOX BAPN, EPCU-5 and PN1 inhibitor compounds on collective migration of HaCaT cells was evaluated. Contrary to what would be expected considering the effect of this type of compounds on other cell lines, it was observed that two of the compounds significantly increased collective cell migration, while another compound did not have any impact on cell migration. Subsequently, nanoformulations composed of solid lipid nanoparticles containing BAPN were produced and developed and their stability over time was evaluated, as well as their impact when applied to the skin. In general, the developed nanoformulations showed stable pH, hydrodynamic diameter and polydispersion index values over the 30 days of the study. The results of the in vivo study indicate a good cutaneous compatibility of the nanoformulations, but no differences were observed regarding the control in its effectiveness, so future studies with a larger panel of volunteers will be necessary. Keywords: LOX enzymes, wound healing, risk assay, collective cell migration, nanotechnology, skin application
A skin wound is defined by any alteration in the integrity of the skin layers. It is estimated that 2 million Europeans suffer from skin lesions, which is an important factor influencing the patient's quality of life. When the skin is disrupted, the integumentary system has the capacity to self-regenerate, a process called healing of skin lesions. It constitutes a vital and fundamental process whose main objective is to restore the integrity and normal function of the skin, where numerous cellular components are involved, such as the LOX enzymes. These catalyze cross-linking reactions of collagen and elastin in connective tissue, thus allowing cross-linking between these two compounds in the extracellular matrix of the skin, facilitating the process of cell migration in the wound healing process. In order to optimize treatments to maximize the healing of cutaneous wounds, the present dissertation had as main objective to study the impact of selected compounds, with inhibitory activity of LOX enzymes, in the collective migration of keratinocytes, and later development of nanoformulations for cutaneous application, having with a view to optimizing the healing process of cutaneous wounds, contributing to the improvement of problems related to these and possibly increasing the patient's quality of life. Using an in vitro assay, the effect of LOX BAPN, EPCU-5 and PN1 inhibitor compounds on collective migration of HaCaT cells was evaluated. Contrary to what would be expected considering the effect of this type of compounds on other cell lines, it was observed that two of the compounds significantly increased collective cell migration, while another compound did not have any impact on cell migration. Subsequently, nanoformulations composed of solid lipid nanoparticles containing BAPN were produced and developed and their stability over time was evaluated, as well as their impact when applied to the skin. In general, the developed nanoformulations showed stable pH, hydrodynamic diameter and polydispersion index values over the 30 days of the study. The results of the in vivo study indicate a good cutaneous compatibility of the nanoformulations, but no differences were observed regarding the control in its effectiveness, so future studies with a larger panel of volunteers will be necessary. Keywords: LOX enzymes, wound healing, risk assay, collective cell migration, nanotechnology, skin application
Descrição
Orientação: Nuno Saraiva
Palavras-chave
MESTRADO INTEGRADO EM CIÊNCIAS FARMACÊUTICAS, CIÊNCIAS FARMACÊUTICAS, PHARMACEUTICAL SCIENCES, CICATRIZAÇÃO DE FERIDAS, WOUND HEALING, NANOTECNOLOGIA, NANOTECHNOLOGY, ADMINISTRAÇÃO CUTÂNEA, CUTANEOUS ADMINISTRATION