Advertisement

Impact of an Enhanced Infection Control Protocol on the Microbial Community Profile in Molar Root Canal Treatment: An in Vivo NGS Molecular Study

  • Shatha Zahran
    Correspondence
    Address requests for reprints to Dr Shatha Zahran, BDS, MSD, PhD, Faculty of Dentistry, Oral & Craniofacial Sciences, King’s College London, London, United Kingdom.
    Affiliations
    Department of Endodontics, Faculty of Dentistry, King Abdulaziz University, Jeddah, Saudi Arabia

    Department of Endodontics, Centre for Oral, Clinical and Translational Sciences, Faculty of Dentistry, Oral & Craniofacial Sciences, King’s College London, London, United Kingdom
    Search for articles by this author
  • Francesco Mannocci
    Affiliations
    Department of Endodontics, Centre for Oral, Clinical and Translational Sciences, Faculty of Dentistry, Oral & Craniofacial Sciences, King’s College London, London, United Kingdom
    Search for articles by this author
  • Garrit Koller
    Affiliations
    Department of Endodontics, Centre for Oral, Clinical and Translational Sciences, Faculty of Dentistry, Oral & Craniofacial Sciences, King’s College London, London, United Kingdom

    Centre for Host Microbiome Interactions, Faculty of Dentistry, Oral & Craniofacial Sciences, King’s College London, London, United Kingdom

    LCN – London Centre for Nanotechnology, London, United Kingdom
    Search for articles by this author
Published:September 07, 2022DOI:https://doi.org/10.1016/j.joen.2022.08.008

      Abstract

      Introduction

      Recent findings demonstrated that 1-year cone-beam computed tomography–based outcomes of molar root canal treatment were improved through an enhanced infection protocol (EnP), when compared with a current best-practice standard infection control protocol (StP). The EnP comprised measures to reduce iatrogenic contamination from direct and indirect contact surfaces, including the replacement of the rubber dams, gloves, files, all instruments, and surface barriers before root canal obturation. The aim of this study was to investigate the effect of such an EnP on resident microbiome present after chemomechanical instrumentation and the protocol ability in reducing iatrogenic contamination in molar teeth during root canal treatment.

      Methods

      Molar teeth were block-randomized to receive treatment under EnP or StP. To compare the differential effect of the protocol on the identity of bacteria present, 150 matched DNA extracts from 75 molar teeth samples (StP, n = 39; EnP, n = 36) were evaluated. Samples were taken before (S1) and after (S2) chemomechanical preparation and were subjected to next-generation sequencing of the V3-V4 region of the 16S rRNA gene before bioinformatical identification using the HOMD oral microbiome database and downstream taxonomic processing, providing measures of richness and diversity of bacteria and significant bacterial taxa during chemomechanical instrumentation and the effect of the 2 treatment groups.

      Results

      Eighty-eight microbial taxa were significantly more abundant in StP S2 samples, including endodontically relevant contaminants taxa as Actinomyces, Cutibacterium, and Haemophilus. The S2 samples demonstrated fewer residual bacterial species in the EnP group, with 26.8 observed species compared with 38.3 in the StP. Reduced diversity and richness measures were noted in the EnP pre-obturation samples compared with the StP in OTU, Chao1, and ACE indices (P ≤ .05). Differential microbial identities between S1 and S2 samples and protocols demonstrated that the previously observed increased effectiveness of the EnP protocol was likely to prevent recontamination or de novo contamination of the root canal space during treatment.

      Conclusions

      The implemented EnP resulted in a specific reduction of microbial taxa often associated with recontamination or iatrogenic contamination, suggesting the basis for improved infection control measures during root canal treatment.

      Key Words

      To read this article in full you will need to make a payment

      Purchase one-time access:

      Academic & Personal: 24 hour online accessCorporate R&D Professionals: 24 hour online access
      One-time access price info
      • For academic or personal research use, select 'Academic and Personal'
      • For corporate R&D use, select 'Corporate R&D Professionals'

      Subscribe:

      Subscribe to Journal of Endodontics
      Already a print subscriber? Claim online access
      Already an online subscriber? Sign in
      Institutional Access: Sign in to ScienceDirect

      References

        • Schirrmeister J.F.
        • Liebenow A.L.
        • Braun G.
        • et al.
        Detection and eradication of microorganisms in root-filled teeth associated with periradicular lesions: an in vivo study.
        J Endod. 2007; 33: 536-540
      1. Siqueira J, Rôças I. Microbiology of endodontic infections. In: Rotstein I, Ingle JI, eds. Ingle’s Endodontics 7. Raleigh, NC: PMPH; 612–620.

        • Niazi S.A.
        • Vincer L.
        • Mannocci F.
        Glove contamination during endodontic treatment is one of the sources of nosocomial endodontic propionibacterium acnes infections.
        J Endod. 2016; 42: 1202-1211
        • Saeed M.
        • Koller G.
        • Niazi S.
        • et al.
        Bacterial contamination of endodontic materials before and after clinical storage.
        J Endod. 2017; 43: 1852-1856
        • Rorslett Hardersen L.
        • Enersen M.
        • Kristoffersen A.K.
        • et al.
        Maintenance of the aseptic working field during endodontic treatment.
        Acta Odontol Scand. 2019; 77: 1-6
        • Zahran S.
        • Mannocci F.
        • Koller G.
        Assessing the iatrogenic contribution to contamination during root canal treatment.
        J Endod. 2022; 48: 479-486
        • Byrd A.L.
        • Belkaid Y.
        • Segre J.A.
        The human skin microbiome.
        Nature Reviews Microbiology. 2018; 16: 143-155
        • Wong J.
        • Manoil D.
        • Näsman P.
        • et al.
        Microbiological aspects of root canal infections and disinfection strategies: an update review on the current knowledge and challenges.
        Frontiers in Oral Health. 2021; 2: 1-19
        • Lazarevic V.
        • Whiteson K.
        • Huse S.
        • et al.
        Metagenomic study of the oral microbiota by Illumina high-throughput sequencing.
        J Microbiol Methods. 2009; 79: 266-271
        • Siqueira Jr., J.F.
        • Alves F.R.
        • Rôças I.N.
        Pyrosequencing analysis of the apical root canal microbiota.
        J Endod. 2011; 37: 1499-1503
        • Manoil D.
        • Al-Manei K.
        • Belibasakis G.N.
        A systematic review of the root canal microbiota associated with apical periodontitis: lessons from next-generation sequencing.
        Proteomics Clin Appl. 2020; : e1900060
        • Hong B.-Y.
        • Lee T.-K.
        • Lim S.-M.
        • et al.
        Microbial analysis in primary and persistent endodontic infections by using pyrosequencing.
        J Endod. 2013; 39: 1136-1140
        • Keskin C.
        • Demiryürek E.Ö.
        • Onuk E.E.
        Pyrosequencing analysis of cryogenically ground samples from primary and secondary/persistent endodontic infections.
        J Endod. 2017; 43: 1309-1316
        • Anderson A.C.
        • Al-Ahmad A.
        • Elamin F.
        • et al.
        Comparison of the bacterial composition and structure in symptomatic and asymptomatic endodontic infections associated with root-filled teeth using pyrosequencing.
        PloS One. 2013; 8: e84960
        • Siqueira Jr., J.F.
        • Antunes H.S.
        • Rôças I.N.
        • et al.
        Microbiome in the apical root canal system of teeth with post-treatment apical periodontitis.
        PloS One. 2016; 11: e0162887
        • Zahran S.
        • Patel S.
        • Koller G.
        • Mannocci F.
        The impact of an enhanced infection control protocol on molar root canal treatment outcome: a randomized clinical trial.
        Int Endod J. 2021;
        • European Society of Endodontology
        Quality guidelines for endodontic treatment: consensus report of the European Society of Endodontology.
        Int Endod J. 2006; 39: 921-930
        • Zahran S.
        • Witherden E.
        • Mannocci F.
        • Koller G.
        Characterization of root canal microbiota in teeth diagnosed with irreversible pulpitis.
        J Endod. 2021; 47: 415-423
        • Weber N.
        • Liou D.
        • Dommer J.
        • et al.
        Nephele: a cloud platform for simplified, standardized and reproducible microbiome data analysis.
        Bioinformatics. 2018; 34: 1411-1413
        • Caporaso J.G.
        • Kuczynski J.
        • Stombaugh J.
        • et al.
        QIIME allows analysis of high-throughput community sequencing data.
        Nat Methods. 2010; 7: 335-336
        • Kopylova E.
        • Noe L.
        • Touzet H.
        SortMeRNA: fast and accurate filtering of ribosomal RNAs in metatranscriptomic data.
        Bioinformatics. 2012; 28: 3211-3217
        • Chen T.
        • Yu W.H.
        • Izard J.
        • et al.
        The Human Oral Microbiome Database: a web accessible resource for investigating oral microbe taxonomic and genomic information.
        Database, Oxford2010
        • Kim B.R.
        • Shin J.
        • Guevarra R.
        • et al.
        Deciphering diversity indices for a better understanding of microbial communities.
        J Microbiol Biotechnol. 2017; 27: 2089-2093
        • Sunde P.T.
        • Tronstad L.
        • Eribe E.R.
        • et al.
        Assessment of periradicular microbiota by DNA-DNA hybridization.
        Endod Dent Traumatol. 2000; 16: 191-196
        • Sjögren U.
        • Figdor D.
        • Persson S.
        Influence of infection at the time of root filling on the outcome of endodontic treatment of teeth with apical periodontitis.
        Int Endod J. 1997; 30: 297-306
        • Siqueira Jr., J.F.
        • Rôças I.N.
        • Souto R.
        • et al.
        Checkerboard DNA-DNA hybridization analysis of endodontic infections.
        Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 2000; 89: 744-748
        • Brook I.
        • Edith H.F.
        Infections caused by Propionibacterium species.
        Reviews of Infectious Diseases. 1991; 13: 819-822
        • Nardello L.C.L.
        • Vilela B.G.
        • Fernandes F.S.
        • et al.
        Analysis of active bacteria persisting after chemomechanical procedures: an RNA- and DNA-based molecular study.
        J Endod. 2020; 46: 1570-1576
        • Gomes B.P.
        • Vianna M.E.
        • Matsumoto C.U.
        • et al.
        Disinfection of gutta-percha cones with chlorhexidine and sodium hypochlorite.
        Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 2005; 100: 512-517
        • Kunin V.
        • Engelbrektson A.
        • Ochman H.
        • Hugenholtz P.
        Wrinkles in the rare biosphere: pyrosequencing errors can lead to artificial inflation of diversity estimates.
        Environ Microbiol. 2010; 12: 118-123
        • van der Horst J.
        • Buijs M.J.
        • Laine M.L.
        • et al.
        Sterile paper points as a bacterial DNA-contamination source in microbiome profiles of clinical samples.
        J Dent. 2013; 41: 1297-1301
        • Park O.-J.
        • Jeong M.-H.
        • Lee E.-H.
        • et al.
        A pilot study of chronological microbiota changes in a rat apical periodontitis model.
        Microorganisms. 2020; 8: 1174
        • Weyrich L.S.
        • Farrer A.G.
        • Eisenhofer R.
        • et al.
        Laboratory contamination over time during low-biomass sample analysis.
        Molecular Ecology Resources. 2019; 19: 982-996