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Estudo piloto sobre o microbioma e resistoma da peri-implantite através de uma abordagem metagenómica
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Abstract(s)
Objetivo: Com a crescente colocação de implantes dentários, as doenças periimplantares, das quais se destaca a peri-implantite (PI), são uma complicação cada vez mais prevalente. Os objetivos deste trabalho experimental foram sobretudo metodológicos 1) seleção e recrutamento de pacientes, 2) recolha de amostras de saliva e de biofilme subgengival, 3) extração do DNA microbiano, 4) envio das amostras para sequenciação metagenómica de shotgun, 5) análise da composição taxonómica e identificação de genes de resistência aos antibióticos (ARGs) de algumas amostras sequenciadas. A concretização deste plano abrirá caminho para um conhecimento mais aprofundado dos perfis taxonómicos, funcionais e da prevalência de ARGs das comunidades microbianas presentes em tecidos peri-implantares saudáveis e com PI.
Metodologia: Foram recrutados pacientes com implantes saudáveis (Grupo HI) e pacientes com coocorrência de implantes com peri-implantite e saudáveis (Grupo PI) na Clínica Dentária Egas Moniz, tendo em conta determinados critérios, após aprovação pela Comissão de Ética Egas Moniz. Amostras de saliva não estimulada, de biofilme subgengival (com recurso a periopapers) foram recolhidas, e usadas para extração do
DNA microbiano total. Estas amostras foram submetidas a sequenciação metagenómica de shotgun, realizada através de tecnologias Illumina.
Resultados: Dos 55 pacientes recrutados, 10 foram usados nos ensaios preliminares e 38 pacientes (20 pertencentes ao grupo HI e 18 pertencentes ao grupo PI) proporcionaram a obtenção de amostras de DNA microbiano para prosseguir para a sequenciação metagenómica. Ambos os grupos de estudo revelaram homogeneidade quanto às características demográficas e clínicas. O estudo da composição taxonómica (filo, género e espécie) e a deteção de ARGs foram feitos apenas nas amostras de dois pacientes (10 e 21).
Conclusão: Os objetivos propostos foram atingidos, uma vez que se cumpriu o plano experimental antecipado. A análise metagenómica de amostras de apenas dois pacientes é limitada e impede o alcançar de conclusões significativas e abrangentes.
Aim: With the increase in the placement of dental implants, peri-implant diseases, of which peri-implantitis (PI) stands out, are an increasingly prevalent complication. The objectives of this experimental work were essentially methodological: 1) selection and recruitment of patients, 2) collection of saliva and subgingival biofilm samples, 3) extraction of microbial DNA, 4) sending the samples for shotgun metagenomic sequencing, 5) analysis of the taxonomic composition and identification of antibiotic resistance genes (ARGs) of some sequenced samples. The achievement of this plan will pave the way for in-depth knowledge of the taxonomic and functional profiles and the prevalence of ARGs of the microbial communities present in healthy and PI peri-implant tissues. Methodology: Patients with healthy implants (Group HI) and patients with co-occurring PI-affected and healthy implants (Group PI) were recruited from the Egas Moniz Dental Clinic, considering certain criteria, and after approval by the Egas Moniz Ethics Committee. Samples of unstimulated saliva and subgingival biofilm (using periopapers) were collected and used to extract total microbial DNA. These samples were subjected to shotgun metagenomic sequencing, using Illumina technologies. Results: Of the 55 recruited patients, 10 were used in the preliminary assays and 38 patients (20 belonging to the HI group and 18 belonging to the PI group) provided microbial DNA samples to proceed to metagenomic sequencing. Both study groups showed homogeneity in terms of demographic and clinical characteristics. The study of the taxonomic composition (phylum, genus and species) and the detection of ARGs were only carried out on the samples of two patients (10 and 21). Conclusion: The proposed objectives were achieved since the anticipated experimental plan was fulfilled. The metagenomic analysis of samples from only two patients is limited and prevents significant and comprehensive conclusions from being reached.
Aim: With the increase in the placement of dental implants, peri-implant diseases, of which peri-implantitis (PI) stands out, are an increasingly prevalent complication. The objectives of this experimental work were essentially methodological: 1) selection and recruitment of patients, 2) collection of saliva and subgingival biofilm samples, 3) extraction of microbial DNA, 4) sending the samples for shotgun metagenomic sequencing, 5) analysis of the taxonomic composition and identification of antibiotic resistance genes (ARGs) of some sequenced samples. The achievement of this plan will pave the way for in-depth knowledge of the taxonomic and functional profiles and the prevalence of ARGs of the microbial communities present in healthy and PI peri-implant tissues. Methodology: Patients with healthy implants (Group HI) and patients with co-occurring PI-affected and healthy implants (Group PI) were recruited from the Egas Moniz Dental Clinic, considering certain criteria, and after approval by the Egas Moniz Ethics Committee. Samples of unstimulated saliva and subgingival biofilm (using periopapers) were collected and used to extract total microbial DNA. These samples were subjected to shotgun metagenomic sequencing, using Illumina technologies. Results: Of the 55 recruited patients, 10 were used in the preliminary assays and 38 patients (20 belonging to the HI group and 18 belonging to the PI group) provided microbial DNA samples to proceed to metagenomic sequencing. Both study groups showed homogeneity in terms of demographic and clinical characteristics. The study of the taxonomic composition (phylum, genus and species) and the detection of ARGs were only carried out on the samples of two patients (10 and 21). Conclusion: The proposed objectives were achieved since the anticipated experimental plan was fulfilled. The metagenomic analysis of samples from only two patients is limited and prevents significant and comprehensive conclusions from being reached.
Description
Dissertação para obtenção do grau de Mestre no Instituto Universitário Egas Moniz
Keywords
Implante saudável Peri-implantite Microbioma Resistoma