Effects of Sequential Antimicrobial Phases on Root Canal Microbiome Dynamics in Two-Visit Treatment of Primary Apical Periodontitis: A Longitudinal Experimental Study

Background: Effective management of primary apical periodontitis depends on understanding the dynamic interactions within the root canal microbiome. This study aimed to investigate the effect of sequential antimicrobial phases on the root canal microbiome during a two-visit treatment approach, with...

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Main Authors: Bertan Kesim, Seda Tezcan Ülger, Gönül Aslan, Yakup Üstün, Ayşe Tuğba Avcı, Mustafa Öner Küçük
Format: Article
Language:English
Published: MDPI AG 2024-12-01
Series:Life
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Online Access:https://www.mdpi.com/2075-1729/14/12/1696
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author Bertan Kesim
Seda Tezcan Ülger
Gönül Aslan
Yakup Üstün
Ayşe Tuğba Avcı
Mustafa Öner Küçük
author_facet Bertan Kesim
Seda Tezcan Ülger
Gönül Aslan
Yakup Üstün
Ayşe Tuğba Avcı
Mustafa Öner Küçük
author_sort Bertan Kesim
collection DOAJ
description Background: Effective management of primary apical periodontitis depends on understanding the dynamic interactions within the root canal microbiome. This study aimed to investigate the effect of sequential antimicrobial phases on the root canal microbiome during a two-visit treatment approach, with a focus on calcium hydroxide medication. Methods: Samples were collected from three teeth across four treatment phases: initial infection (S1), after chemomechanical preparation (S2), after intracanal medication (S3), and after a final flush (S4). DNA was extracted, and the V3–V4 regions of the 16S rRNA gene were sequenced using Illumina MiSeq. Sequencing data were analyzed with QIIME 2, and differentially abundant taxa were identified using linear discriminant analysis effect size (LEfSe). Results: While microbial community composition did not differ significantly between phases, the <i>Firmicutes</i>/<i>Bacteroidetes</i> ratio decreased after the antimicrobial stages. LEfSe analysis revealed higher abundances of <i>Lactobacillales</i>, <i>Arthrobacter</i>, and <i>Veillonella</i> in the untreated (CMP) group. <i>Bifidobacterium longum</i> was relatively more abundant in the intracanal medication (ICM) phase, and <i>Dorea formicigenerans</i> was more abundant in the final-flush (FF) phase. Conclusions: Although calcium hydroxide treatment did not induce statistically significant changes in overall root canal microbial composition, trends such as a reduction in the <i>Firmicutes</i>/<i>Bacteroidetes</i> ratio and a relative increase in <i>Bifidobacterium longum</i> numbers suggest potential ecological shifts. The observed relative increase in <i>Bifidobacterium longum</i> numbers may represent a hypothesis-driven observation reflecting indirect ecological effects rather than direct pH modulation. While visual patterns (e.g., PCA clustering) were observed, they lacked statistical support. Further studies with larger sample sizes are needed to validate these observations and assess the potential role of beneficial bacteria in root canal treatments.
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spelling doaj-art-aa6c72e6ded14739aab1c47c60da76c52024-12-27T14:36:20ZengMDPI AGLife2075-17292024-12-011412169610.3390/life14121696Effects of Sequential Antimicrobial Phases on Root Canal Microbiome Dynamics in Two-Visit Treatment of Primary Apical Periodontitis: A Longitudinal Experimental StudyBertan Kesim0Seda Tezcan Ülger1Gönül Aslan2Yakup Üstün3Ayşe Tuğba Avcı4Mustafa Öner Küçük5Department of Endodontics, Faculty of Dentistry, Nuh Naci Yazgan University, Kayseri 38170, TurkeyDepartment of Medical Microbiology, Faculty of Medicine, Mersin University, Mersin 33343, TurkeyDepartment of Medical Microbiology, Faculty of Medicine, Mersin University, Mersin 33343, TurkeyDepartment of Endodontics, Faculty of Dentistry, Erciyes University, Kayseri 38039, TurkeyDepartment of Endodontics, Faculty of Dentistry, Erciyes University, Kayseri 38039, TurkeyDepartment of Microbiology, Faculty of Dentistry, Nuh Naci Yazgan University, Kayseri 38170, TurkeyBackground: Effective management of primary apical periodontitis depends on understanding the dynamic interactions within the root canal microbiome. This study aimed to investigate the effect of sequential antimicrobial phases on the root canal microbiome during a two-visit treatment approach, with a focus on calcium hydroxide medication. Methods: Samples were collected from three teeth across four treatment phases: initial infection (S1), after chemomechanical preparation (S2), after intracanal medication (S3), and after a final flush (S4). DNA was extracted, and the V3–V4 regions of the 16S rRNA gene were sequenced using Illumina MiSeq. Sequencing data were analyzed with QIIME 2, and differentially abundant taxa were identified using linear discriminant analysis effect size (LEfSe). Results: While microbial community composition did not differ significantly between phases, the <i>Firmicutes</i>/<i>Bacteroidetes</i> ratio decreased after the antimicrobial stages. LEfSe analysis revealed higher abundances of <i>Lactobacillales</i>, <i>Arthrobacter</i>, and <i>Veillonella</i> in the untreated (CMP) group. <i>Bifidobacterium longum</i> was relatively more abundant in the intracanal medication (ICM) phase, and <i>Dorea formicigenerans</i> was more abundant in the final-flush (FF) phase. Conclusions: Although calcium hydroxide treatment did not induce statistically significant changes in overall root canal microbial composition, trends such as a reduction in the <i>Firmicutes</i>/<i>Bacteroidetes</i> ratio and a relative increase in <i>Bifidobacterium longum</i> numbers suggest potential ecological shifts. The observed relative increase in <i>Bifidobacterium longum</i> numbers may represent a hypothesis-driven observation reflecting indirect ecological effects rather than direct pH modulation. While visual patterns (e.g., PCA clustering) were observed, they lacked statistical support. Further studies with larger sample sizes are needed to validate these observations and assess the potential role of beneficial bacteria in root canal treatments.https://www.mdpi.com/2075-1729/14/12/169616S ribosomal RNA sequencingcalcium hydroxideantimicrobial proceduresmicrobiome modulation2-visit root canal treatment<i>Bifidobacterium longum</i>
spellingShingle Bertan Kesim
Seda Tezcan Ülger
Gönül Aslan
Yakup Üstün
Ayşe Tuğba Avcı
Mustafa Öner Küçük
Effects of Sequential Antimicrobial Phases on Root Canal Microbiome Dynamics in Two-Visit Treatment of Primary Apical Periodontitis: A Longitudinal Experimental Study
Life
16S ribosomal RNA sequencing
calcium hydroxide
antimicrobial procedures
microbiome modulation
2-visit root canal treatment
<i>Bifidobacterium longum</i>
title Effects of Sequential Antimicrobial Phases on Root Canal Microbiome Dynamics in Two-Visit Treatment of Primary Apical Periodontitis: A Longitudinal Experimental Study
title_full Effects of Sequential Antimicrobial Phases on Root Canal Microbiome Dynamics in Two-Visit Treatment of Primary Apical Periodontitis: A Longitudinal Experimental Study
title_fullStr Effects of Sequential Antimicrobial Phases on Root Canal Microbiome Dynamics in Two-Visit Treatment of Primary Apical Periodontitis: A Longitudinal Experimental Study
title_full_unstemmed Effects of Sequential Antimicrobial Phases on Root Canal Microbiome Dynamics in Two-Visit Treatment of Primary Apical Periodontitis: A Longitudinal Experimental Study
title_short Effects of Sequential Antimicrobial Phases on Root Canal Microbiome Dynamics in Two-Visit Treatment of Primary Apical Periodontitis: A Longitudinal Experimental Study
title_sort effects of sequential antimicrobial phases on root canal microbiome dynamics in two visit treatment of primary apical periodontitis a longitudinal experimental study
topic 16S ribosomal RNA sequencing
calcium hydroxide
antimicrobial procedures
microbiome modulation
2-visit root canal treatment
<i>Bifidobacterium longum</i>
url https://www.mdpi.com/2075-1729/14/12/1696
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