Microbial composition and diversity in intraradicular biofilm formed in situ: New concepts based on next-generation sequencing.

IF 2.8 3区 医学 Q1 DENTISTRY, ORAL SURGERY & MEDICINE Molecular Oral Microbiology Pub Date : 2024-10-01 Epub Date: 2024-03-18 DOI:10.1111/omi.12462
Felipe Barros Matoso, Francisco Montagner, Fabiana Soares Grecca, Pabulo Henrique Rampelotto, Patrícia Maria Poli Kopper
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Abstract

This study aimed to characterize the taxonomic composition of intraradicular multispecies biofilms (IMBs) formed in situ in a model to reproduce clinical conditions. Twelve palatal roots of maxillary molars had its canals prepared. Two roots were randomly selected to sterility control. Ten intraoral prosthetic appliances with lateral slots were fabricated. The roots were positioned in the slots with the canal access open to the oral cavity. Eight volunteers wore the appliance for 21 days, and two wore it at two different time points. One root from each appliance was removed and stored at -20°C until DNA extraction and sequencing (n = 10). Biofilm was analyzed using next-generation sequencing and bioinformatics. The V4 hyper-variable region of the 16SrRNA gene was amplified and sequenced. For data analyses, the mothur pipeline was used for 16SrRNA processing, and subsequent analyses of the sequence dataset were performed in R using the MicrobiomeAnalyst R package. The taxonomy-based analysis of bacterial communities identified 562 operational taxonomic units (OTUs), which belonged to 93 genera, 44 families, and 8 phyla. Bacterial colonization was different for each biofilm, and samples did not have the same group of bacteria. Alpha and beta diversity analysis revealed some general patterns of sample clustering. A core microbiome of prevalent OTUs and genera was identified. IMBs were heterogeneous when analyzed individually, but some diversity patterns were found after sample clustering. The experimental model seemed to reproduce the actual biofilm composition in endodontic infections, which suggests that it may be used to evaluate disinfection protocols.

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原位形成的关节内生物膜中的微生物组成和多样性:基于新一代测序的新概念。
本研究旨在重现临床条件下在原位形成的牙槽内多物种生物膜(IMBs)的分类组成特征。研究人员制备了 12 个上颌磨牙的腭根。随机选取两个牙根进行无菌控制。制作了十个带有侧槽的口内修复装置。牙根被放置在槽沟中,牙槽通向口腔。八名志愿者佩戴义齿 21 天,两名志愿者在两个不同的时间点佩戴义齿。从每个义齿上取下一个牙根,保存在零下 20°C 的温度下,直到 DNA 提取和测序(n = 10)。使用新一代测序和生物信息学分析了生物膜。对 16SrRNA 基因的 V4 超变区进行了扩增和测序。在数据分析中,使用了mothur管道进行16SrRNA处理,并使用MicrobiomeAnalyst R软件包在R语言中对序列数据集进行了后续分析。基于分类学的细菌群落分析确定了 562 个操作分类单元(OTUs),它们隶属于 93 属、44 科和 8 门。每个生物膜的细菌定植情况不同,样本中的细菌群也不尽相同。阿尔法和贝塔多样性分析揭示了样本聚类的一些一般模式。确定了一个由主要 OTU 和菌属组成的核心微生物组。单独分析时,IMBs 是异质的,但样本聚类后发现了一些多样性模式。该实验模型似乎再现了牙髓感染中实际的生物膜组成,这表明它可用于评估消毒方案。
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来源期刊
Molecular Oral Microbiology
Molecular Oral Microbiology DENTISTRY, ORAL SURGERY & MEDICINE-MICROBIOLOGY
CiteScore
6.50
自引率
5.40%
发文量
46
审稿时长
>12 weeks
期刊介绍: Molecular Oral Microbiology publishes high quality research papers and reviews on fundamental or applied molecular studies of microorganisms of the oral cavity and respiratory tract, host-microbe interactions, cellular microbiology, molecular ecology, and immunological studies of oral and respiratory tract infections. Papers describing work in virology, or in immunology unrelated to microbial colonization or infection, will not be acceptable. Studies of the prevalence of organisms or of antimicrobials agents also are not within the scope of the journal. The journal does not publish Short Communications or Letters to the Editor. Molecular Oral Microbiology is published bimonthly.
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