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      Chemical and mechanical influence of root canal irrigation on biofilm removal from lateral morphological features of simulated root canals, dentine discs and dentinal tubules

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          Abstract

          Aim

          To investigate the anti‐biofilm efficacy of irrigation using a simulated root canal model, the chemical effect of irrigants against biofilms grown on dentine discs and their impact on biofilm viscoelasticity, the efficacy of the irrigants in decontaminating infected dentinal tubules and the capacity of bacteria to regrow.

          Methodology

          Biofilm removal, viscoelastic analysis of remaining biofilms and bacterial viability were evaluated using a simulated root canal model with lateral morphological features, dentine discs and a dentinal tubule model, respectively. Experiments were conducted using a two‐phase irrigation protocol. Phase 1: a modified salt solution (RISA) and sodium hypochlorite (NaOCl) were used at a low flow rate to evaluate the chemical action of the irrigants. Ultrasonic activation (US) of a chemically inert solution (buffer) was used to evaluate the mechanical efficacy of irrigation. Phase 2: a final irrigation with buffer at a high flow rate was performed for all groups. Optical coherence tomography (OCT), low load compression testing (LLCT) and confocal scanning laser microscopy analysis were used in the different models. One‐way analysis of variance ( anova) was performed for the OCT and LLCT analysis, whilst Kruskal–Wallis and Wilcoxon ranked tests for the dentinal tubule model.

          Results

          US and high flow rate removed significantly more biofilm from the artificial lateral canal. For biofilm removal from the artificial isthmus, no significant differences were found between the groups. Within‐group analysis revealed significant differences between the steps of the experiment, with the exception of NaOCl. For the dentine discs, no significant differences regarding biofilm removal and viscoelasticity were detected. In the dentinal tubule model, NaOCl exhibited the greatest anti‐biofilm efficacy.

          Conclusions

          The mechanical effect of irrigation is important for biofilm removal. An extra high flow irrigation rate resulted in greater biofilm removal than US in the artificial isthmus. The mechanical effect of US seemed to be more effective when the surface contact biofilm–irrigant was small. After the irrigation procedures, the remaining biofilm could survive after a 5‐day period. RISA and NaOCl seemed to alter post‐treatment remaining biofilms.

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          Most cited references74

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          A Threshold Selection Method from Gray-Level Histograms

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            The biofilm matrix.

            The microorganisms in biofilms live in a self-produced matrix of hydrated extracellular polymeric substances (EPS) that form their immediate environment. EPS are mainly polysaccharides, proteins, nucleic acids and lipids; they provide the mechanical stability of biofilms, mediate their adhesion to surfaces and form a cohesive, three-dimensional polymer network that interconnects and transiently immobilizes biofilm cells. In addition, the biofilm matrix acts as an external digestive system by keeping extracellular enzymes close to the cells, enabling them to metabolize dissolved, colloidal and solid biopolymers. Here we describe the functions, properties and constituents of the EPS matrix that make biofilms the most successful forms of life on earth.
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              Physiological heterogeneity in biofilms.

              Biofilms contain bacterial cells that are in a wide range of physiological states. Within a biofilm population, cells with diverse genotypes and phenotypes that express distinct metabolic pathways, stress responses and other specific biological activities are juxtaposed. The mechanisms that contribute to this genetic and physiological heterogeneity include microscale chemical gradients, adaptation to local environmental conditions, stochastic gene expression and the genotypic variation that occurs through mutation and selection. Here, we discuss the processes that generate chemical gradients in biofilms, the genetic and physiological responses of the bacteria as they adapt to these gradients and the techniques that can be used to visualize and measure the microscale physiological heterogeneities of bacteria in biofilms.
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                Author and article information

                Contributors
                l.w.m.van.der.sluis@umcg.nl
                Journal
                Int Endod J
                Int Endod J
                10.1111/(ISSN)1365-2591
                IEJ
                International Endodontic Journal
                John Wiley and Sons Inc. (Hoboken )
                0143-2885
                1365-2591
                19 November 2020
                January 2021
                : 54
                : 1 ( doiID: 10.1111/iej.v54.1 )
                : 112-129
                Affiliations
                [ 1 ] Department of Dentistry, Endodontics and Dental Materials Bauru School of Dentistry University of São Paulo Bauru Brazil
                [ 2 ] Center for Dentistry and Oral Hygiene University Medical Center Groningen University of Groningen Groningen The Netherlands
                [ 3 ] Department of Biomedical Engineering University Medical Center Groningen University of Groningen Groningen The Netherlands
                [ 4 ] Department of Orthodontics University Medical Center Groningen University of Groningen Groningen The Netherlands
                Author notes
                [*] [* ] Correspondence: Luc van der Sluis, Center for Dentistry and Oral Hygiene, University Medical Center Groningen, University of Groningen, 9712 CP, Groningen, The Netherlands (Tel.: + 31 50 3616216; e‐mail: l.w.m.van.der.sluis@ 123456umcg.nl ).

                Author information
                https://orcid.org/0000-0002-3390-9371
                https://orcid.org/0000-0002-3449-231X
                https://orcid.org/0000-0002-8342-8939
                https://orcid.org/0000-0002-1238-2160
                Article
                IEJ13399
                10.1111/iej.13399
                7839520
                32880989
                e8f87a29-313c-4c0a-9411-03e69634c42b
                © 2020 The Authors. International Endodontic Journal published by John Wiley & Sons Ltd on behalf of British Endodontic Society

                This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

                History
                : 20 December 2019
                : 26 August 2020
                Page count
                Figures: 7, Tables: 2, Pages: 19, Words: 11224
                Funding
                Funded by: European Society of Endodontology
                Funded by: Abel Tasman Talent Program
                Funded by: Coordenação de Aperfeiçoamento de Pessoal de Nível Superior , open-funder-registry 10.13039/501100002322;
                Funded by: Fundação de Amparo à Pesquisa do Estado de São Paulo , open-funder-registry 10.13039/501100001807;
                Award ID: 2010/20196‐3
                Categories
                Original Article
                Basic Research – Biological
                Custom metadata
                2.0
                January 2021
                Converter:WILEY_ML3GV2_TO_JATSPMC version:5.9.6 mode:remove_FC converted:27.01.2021

                Dentistry
                biofilm,confocal laser scanning microscopy,optical coherence tomography,polysaccharides,risa,sodium hypochlorite,ultrasound

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