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Authors
Abstract(s)
A presente dissertação de mestrado encontra-se dividida em três capítulos, sendo o
primeiro capítulo referente à componente de Investigação, o segundo capítulo ao estágio
curricular em Farmácia Hospitalar e, por último, o terceiro capítulo relativo ao estágio
curricular em Farmácia Comunitária.
O primeiro capítulo, intitulado “Avaliação da utilização de probióticos no tratamento
complementar da acne vulgaris”, baseia-se na investigação desenvolvida no Centro de
Investigação em Ciências da Saúde da Universidade da Beira Interior (CICS-UBI), sob a
orientação da Professora Doutora Ana Palmeira de Oliveira, da Mestre Ana Sofia Oliveira
e do Mestre Carlos Gaspar. A acne vulgaris é uma doença de pele comum que afeta
principalmente os adolescentes. Formas severas de acne podem causar desfiguração e
cicatrizes, resultando em baixa autoestima, dificuldade na interação social, e angústia
psicológica. Esta condição afeta a unidade pilossebácea da pele e tem uma natureza
multifatorial que envolve a interação de quatro fatores fisiopatológicos principais,
nomeadamente: aumento da produção de sebo induzida por hormonas; aumento da
queratinização; processos imunológicos e inflamação; e aumento da proliferação
bacteriana de Cutibacterium acnes no folículo piloso. Cutibacterium acnes é uma
bactéria gram-positiva anaeróbia que coloniza e vive na pele humana, especialmente em
folículos pilossebáceos. Embora esta bactéria seja classificada como simbiótica, pode agir
como um agente patogénico oportunista e muitas vezes causar infeções. Uma panóplia
de tratamentos está disponível para a acne vulgaris. No entanto não existe uma cura
definitiva para esta condição e, além disso, alguns dos tratamentos disponíveis
apresentam efeitos adversos significativos. Portanto, existe a necessidade de desenvolver
novas opções de tratamento que atuem de forma segura e eficaz no tratamento ou
controlo da acne. Os probióticos são microrganismos vivos que, quando administrados
em quantidades adequadas, podem conferir um benefício para a saúde do hospedeiro.
Considerando o Cutibacterium acnes como o principal agente patogénico na acne, os
probióticos parecem demonstrar benefícios diretos e indiretos no tratamento da acne: ao
inibirem diretamente o crescimento de Cutibacterium acnes e diminuírem a resposta
inflamatória; e indiretamente, reduzindo alguns dos efeitos secundários dos tratamentos
clássicos, nomeadamente a resistência bacteriana, apresentando-se por isso como uma
alternativa promissora à utilização de antibióticos. O objetivo do presente trabalho foi
estudar o perfil probiótico de diferentes bactérias ácido-lácticas com o intuito de avaliar
o seu potencial como alternativa ou adjuvante às terapêuticas convencionais utilizadas no tratamento da acne vulgaris. Para isso, a atividade antimicrobiana de sobrenadantes
produzidos por estirpes isoladas da pele de voluntários saudáveis e de probióticos
comerciais foi determinada através de dois métodos distintos: o método de microdiluição
em caldo contra o principal microrganismo patogénico da acne vulgaris, Cutibacterium
acnes; e pelo método de difusão agar contra Cutibacterium acnes e duas outras bactérias
envolvidas no desenvolvimento da doença, nomeadamente Staphylococcus aureus e
Staphylococcus epidermidis. A atividade das estirpes em estudo foi determinada
adicionalmente através do ensaio preliminar de competição direta contra Cutibacterium
acnes, Staphylococcus aureus e Staphylococcus epidermidis. A deteção de bacteriófagos
e a determinação de unidades formadores de placas foi realizada através do ensaio de
difusão em placa após verificação de um perfil de atividade antimicrobiana distinto, para
uma das estirpes. Verificou-se que os sobrenadantes produzidos pelas estirpes em estudo
reduzem ou inibem o crescimento dos microrganismos indicadores (Cutibacterium
acnes, Staphylococcus aureus e Staphylococcus epidermidis) e que os sobrenadantes
produzidos pelas estirpes probióticas têm mais atividade do que os sobrenadantes
produzidos pelas estirpes da pele. Em relação às estirpes em estudo, verificou-se que há
um aumento de atividade das estirpes isoladas da pele e uma diminuição de atividade
das estirpes isoladas de probióticos quando estas são comparadas com os seus respetivos
sobrenadantes. O bacteriófago isolado mostra atividade e seletividade contra
Cutibacterium acnes. Estes resultados mostraram que as estirpes isoladas de probióticos
comerciais e o bacteriófago isolado da estirpe bacteriana OF2 têm o potencial de serem
empregues como alternativa ou adjuvante às terapêuticas convencionais utilizadas no
tratamento da acne vulgaris. São necessários estudos futuros para compreender o
mecanismo pelo qual estas estirpes interferem na patogénese da acne e afim de avaliar a
sua segurança com vista a uma futura aplicação na pele.
O segundo capítulo relata a minha experiência no estágio em Farmácia Hospitalar,
decorrido entre 7 de fevereiro e 25 de março de 2022, no Centro Hospitalar Universitário
Cova da Beira, na Covilhã, sob a orientação da Dr.ª Maria Olímpia Fonseca.
O terceiro capítulo descreve a experiência adquirida e atividades desenvolvidas ao longo
do estágio em Farmácia Comunitária, que decorreu entre 28 de março a 28 de junho de
2022, na Farmácia São Cosme, na Covilhã, sob a orientação do Dr. Carlos Tavares.
This dissertation is divided into three chapters, the first chapter referring to the Research component, the second chapter to the curricular internship in Hospital Pharmacy and, finally, the third chapter concerning the curricular internship in Community Pharmacy. The first chapter entitled “Assessment of the use of probiotics in the complementary treatment of acne vulgaris”, is based on the research carried out at the Health Sciences Research Center of the University of Beira Interior (CICS-UBI), under the supervision of Professor Ana Palmeira de Oliveira, Dr. Ana Sofia Oliveira, and Dr. Carlos Gaspar. Acne vulgaris is a multifactorial skin condition that affects mainly teenagers. Severe forms of acne can cause disfigurement and scarring, resulting in low self-esteem, difficulty in social interaction, and psychological distress. This condition affects the pilosebaceous unit of the skin and has a multifactorial nature that involves the interaction of four main pathophysiological hallmarks, namely: increased sebum production induced by hormones; increased keratinization; immune processes and inflammation; and increased bacterial proliferation of Cutibacterium acnes in the hair follicle. Cutibacterium acnes is an anaerobic gram-positive bacterium that colonizes the human skin, especially in pilosebaceous follicles. Although this bacterium is classified as symbiotic, it can act as an opportunistic pathogen and often cause infection. A plethora of treatments are available for acne vulgaris. However, there is no definitive cure for this condition and, additionally, some of the available treatments can cause significant adverse effects. Therefore, there is a need to develop new treatment options that act safely and effectively in the treatment of acne. Probiotics are live microorganisms that, when administered in adequate amounts, may confer a health benefit on the host. Considering the bacterium Cutibacterium acnes as one of the main pathogenic agent in acne, probiotics seem to demonstrate direct and indirect benefits in the treatment of acne: by directly inhibiting the growth of Cutibacterium acnes and decreasing the inflammatory response; and indirectly, by reducing some of the side effects associated with classic treatments, namely bacterial resistance, thus presenting itself as a promising alternative to the use of antibiotics. The objective of the present work was to study the probiotic profile of different lactic acid bacteria to evaluate their potential as an alternative or adjuvant to conventional therapies used in the treatment of acne vulgaris. The antimicrobial activity of supernatants produced by strains isolated from the skin of healthy volunteers and commercial probiotics was determined using two different methods: the broth microdilution method against the main pathogenic microorganism of acne vulgaris, Cutibacterium acnes; and the agar diffusion method against Cutibacterium acnes and two other bacteria involved in the development of the disease, namely Staphylococcus aureus and Staphylococcus epidermidis. The activity of the strains under study was additionally determined through the preliminary assay of direct competition against Cutibacterium acnes, Staphylococcus aureus and Staphylococcus epidermidis. Detection of bacteriophages and determination of plaque forming units was performed using the plaque diffusion assay after verifying a distinct antimicrobial activity profile for one of the strains. It was found that the supernatants produced by the strains under study reduce or inhibit the growth of indicator microorganisms (Cutibacterium acnes, Staphylococcus aureus and Staphylococcus epidermidis) and that the supernatants produced by the probiotic strains were more active than the supernatants produced by the skin strains. Regarding the strains under study, it was found that there is an increase in activity of strains isolated from the skin and a decrease in activity of strains isolated from probiotics when their activity is compared to the one produced by their respective supernatants. The isolated bacteriophage shows activity and selectivity against Cutibacterium acnes. These results showed that the isolated strains of commercial probiotics and the bacteriophage isolated from the bacterial strain OF2 have the potential to be used as an alternative or adjuvant to conventional therapies used in the treatment of acne vulgaris. Future studies are needed to understand the mechanism by which these strains interfere with the pathogenesis of acne and to assess their safety concerning a future skin application. The second chapter reports my experience in the internship in Hospital Pharmacy, which took place between February 7th and March 25th, 2022, at Centro Hospitalar Universitário Cova da Beira, in Covilhã, under the guidance of Dr. Maria Olímpia Fonseca. The third chapter describes the experience gained and activities developed during the internship in Community Pharmacy, which took place from March 28th and June 28th, 2022, at Farmácia São Cosme, in Covilhã, under the guidance of Dr. Carlos Tavares.
This dissertation is divided into three chapters, the first chapter referring to the Research component, the second chapter to the curricular internship in Hospital Pharmacy and, finally, the third chapter concerning the curricular internship in Community Pharmacy. The first chapter entitled “Assessment of the use of probiotics in the complementary treatment of acne vulgaris”, is based on the research carried out at the Health Sciences Research Center of the University of Beira Interior (CICS-UBI), under the supervision of Professor Ana Palmeira de Oliveira, Dr. Ana Sofia Oliveira, and Dr. Carlos Gaspar. Acne vulgaris is a multifactorial skin condition that affects mainly teenagers. Severe forms of acne can cause disfigurement and scarring, resulting in low self-esteem, difficulty in social interaction, and psychological distress. This condition affects the pilosebaceous unit of the skin and has a multifactorial nature that involves the interaction of four main pathophysiological hallmarks, namely: increased sebum production induced by hormones; increased keratinization; immune processes and inflammation; and increased bacterial proliferation of Cutibacterium acnes in the hair follicle. Cutibacterium acnes is an anaerobic gram-positive bacterium that colonizes the human skin, especially in pilosebaceous follicles. Although this bacterium is classified as symbiotic, it can act as an opportunistic pathogen and often cause infection. A plethora of treatments are available for acne vulgaris. However, there is no definitive cure for this condition and, additionally, some of the available treatments can cause significant adverse effects. Therefore, there is a need to develop new treatment options that act safely and effectively in the treatment of acne. Probiotics are live microorganisms that, when administered in adequate amounts, may confer a health benefit on the host. Considering the bacterium Cutibacterium acnes as one of the main pathogenic agent in acne, probiotics seem to demonstrate direct and indirect benefits in the treatment of acne: by directly inhibiting the growth of Cutibacterium acnes and decreasing the inflammatory response; and indirectly, by reducing some of the side effects associated with classic treatments, namely bacterial resistance, thus presenting itself as a promising alternative to the use of antibiotics. The objective of the present work was to study the probiotic profile of different lactic acid bacteria to evaluate their potential as an alternative or adjuvant to conventional therapies used in the treatment of acne vulgaris. The antimicrobial activity of supernatants produced by strains isolated from the skin of healthy volunteers and commercial probiotics was determined using two different methods: the broth microdilution method against the main pathogenic microorganism of acne vulgaris, Cutibacterium acnes; and the agar diffusion method against Cutibacterium acnes and two other bacteria involved in the development of the disease, namely Staphylococcus aureus and Staphylococcus epidermidis. The activity of the strains under study was additionally determined through the preliminary assay of direct competition against Cutibacterium acnes, Staphylococcus aureus and Staphylococcus epidermidis. Detection of bacteriophages and determination of plaque forming units was performed using the plaque diffusion assay after verifying a distinct antimicrobial activity profile for one of the strains. It was found that the supernatants produced by the strains under study reduce or inhibit the growth of indicator microorganisms (Cutibacterium acnes, Staphylococcus aureus and Staphylococcus epidermidis) and that the supernatants produced by the probiotic strains were more active than the supernatants produced by the skin strains. Regarding the strains under study, it was found that there is an increase in activity of strains isolated from the skin and a decrease in activity of strains isolated from probiotics when their activity is compared to the one produced by their respective supernatants. The isolated bacteriophage shows activity and selectivity against Cutibacterium acnes. These results showed that the isolated strains of commercial probiotics and the bacteriophage isolated from the bacterial strain OF2 have the potential to be used as an alternative or adjuvant to conventional therapies used in the treatment of acne vulgaris. Future studies are needed to understand the mechanism by which these strains interfere with the pathogenesis of acne and to assess their safety concerning a future skin application. The second chapter reports my experience in the internship in Hospital Pharmacy, which took place between February 7th and March 25th, 2022, at Centro Hospitalar Universitário Cova da Beira, in Covilhã, under the guidance of Dr. Maria Olímpia Fonseca. The third chapter describes the experience gained and activities developed during the internship in Community Pharmacy, which took place from March 28th and June 28th, 2022, at Farmácia São Cosme, in Covilhã, under the guidance of Dr. Carlos Tavares.
Description
Keywords
Acne Vulgaris Atividade Antimicrobiana Bactérias Ácido-Lácticas Bacteriófagos Cutibacterium Acnes Farmácia Comunitária Farmácia Hospitalar Pele Probióticos Terapia Fágica