Antimicrobial Efficacy of Different Irrigant Solutions Using a Novel Biofilm Model: An In Vitro Confocal Laser Scanning Microscopy Experiment.

AIM: To determine the ability of different irrigation solutions to biomechanically remove Enterococcus faecalis biofilm from a novel artificial root canal model during chemomechanical preparation. METHODS: High resolution micro-computer-tomography scans of a mandibular molar's mesial root were used to produce 50 identical 3D-printed resin root canal models. These were cultured with E.faecalis over seven days to generate biofilm and subjected to chemomechanical preparation using: saline; 17% ethylenediaminetetraacetic acid (EDTA) or 2% sodium hypochlorite (NaOCl) alongside positive/negative controls (n = 10). Canals were prepared to 40/.06 taper, with 1 mL irrigation between instruments, followed by 5 mL penultimate rinse, 30 s ultrasonic activation and 5 mL final rinse. Residual biofilm volume (pixels) was determined following immunofluorescent staining and confocal-laser-scanning-microscopy imaging. Statistical comparisons were made using Kruskal-Wallis with post-hoc Dunn's tests (α ⟨0.05). RESULTS: In all canal thirds, the greatest biofilm removal was observed with NaOCl, followed by EDTA and saline. The latter had significantly higher E.faecalis counts than NaOCl and EDTA (P ⟨0.01). However, no statistical differences were found between EDTA and NaOCl or saline and positive controls (P ⟩0.05). CONCLUSIONS: Within limitations of this model, 17% EDTA was found to be as effective as 2% NaOCl at eradicating E.faecalis biofilm following chemomechanical preparation. Further investigations with multi-species biofilms are encouraged.

The presence of smear layer affects the antimicrobial action of root canal sealers.

AIM: To assess the chemical and microstructural characteristics of dentine after the use of two irrigation protocols and correlate this with the antimicrobial properties of hydraulic calcium silicate cement (HCSC) sealers and changes to the dentine structure/chemistry after sealer placement. METHODOLOGY: Two irrigation protocols - Protocol A using 2% NaOCl used 5 mL/5 min and Protocol B with 2% NaOCl (5 mL/5 min) followed by 17% EDTA (5 mL/3 min) - were used to prepare dentine. The chemical and microstructural changes following irrigation were assessed by scanning electron microscopy (SEM), energy dispersive spectroscopy (EDS) and Fourier transform infrared (FT-IR) spectroscopy (n = 5) on dentine obtained from the mid-root and coronal parts of extracted human teeth. Four sealers (AH Plus, BioRoot, MTA Fillapex, TotalFill) were characterized by SEM/EDS (n = 3). The ability of the sealers to eradicate intratubular Enterococcus faecalis biofilms was assessed by live/dead dye and confocal laser scanning microscopy to measure the percentage of living cells. The effect of combined irrigation and root filling on dentine was assessed by SEM and EDS analysis (n = 5). Statistical analysis was undertaken using one-way anova and a number of post hoc tests to detect intergroup differences. The F-test was used for comparison of variances in the microbiology testing. RESULTS: The use of NaOCl alone left the smear layer intact, with traces of chlorine remaining on dentine. The use of BioRoot sealer restored the calcium levels of dentine which had been depleted by the irrigation with EDTA. BioRoot exhibited antimicrobial properties against intratubular bacteria even in the presence of smear layer (Protocol A). Smear layer removal improved the bactericidal effect of all sealers and Ca2+ leaching. The use of a chelating agent was important for the intratubular sealer penetration for AH Plus but not the other sealers. CONCLUSION: The removal of smear layer was necessary for penetration of AH Plus into the dentinal tubules. BioRoot was a more effective sealer in reducing the bacterial load in the dentinal tubules than the other materials tested and the presence of smear layer did not affect its activity.

Investigation of microbial profile, levels of endotoxin and lipoteichoic acid in teeth with symptomatic irreversible pulpitis: a clinical study.

AIM: To investigate the microbial profile, and levels of endotoxin (LPS) and lipoteichoic acid (LTA), in infected dentine (ID) and root canals (RC) at different phases of root canal treatment in teeth with symptomatic irreversible pulpitis. METHODOLOGY: Ten volunteers were included, and samples were collected from infected dentine (ID) and the root canal lumen (RC) using sterile excavators and paper points, respectively. RC samples were taken before (S1) and after (S2) chemo-mechanical canal preparation (CMP), and after intracanal medication (ICM; S3). Checkerboard DNA-DNA hybridization was used for microbial analysis. The levels of LPS and LTA were evaluated using the limulus amebocyte lysate assay and ELISA, respectively. Shapiro-Wilk's test was used to verify data normality. Friedman's test was used to evaluate statistical differences using checkerboard DNA-DNA hybridization in the ID and RC at the different phases of the RC treatment. Post hoc Dunn's multiple comparison test was used to verify significant differences recorded at the different time-points. The levels of LPS and LTA were analysed statistically by using repeated measures anova and Tukey's post hoc test to evaluate differences in both sites. The significance level was set at 5% (P < 0.05). RESULTS: A total of 40 DNA probes were used for microbial investigation of ID and RC samples using checkerboard DNA-DNA hybridization. The levels and complexity of bacteria were similar in the ID and initial RC samples. The levels of LPS and LTA in ID were significantly higher than the initial RC samples (S1; P < 0.05). Canal preparation was effective in significantly decreasing the levels of bacteria, LPS and LTA (P < 0.05). ICM did not provide additional reduction in the levels of bacteria and LPS (P > 0.05). However, a significant reduction in the levels of LTA was observed after ICM (P < 0.05). CONCLUSION: The microbial profile of infected dentine and root canals of teeth with irreversible pulpitis was complex, harbouring different species including Gram-positive and Gram-negative, cocci and bacilli, and facultative and strict anaerobes. Root canal preparation was effective in reducing the levels of bacteria, LPS and LTA from the root canals of teeth with pulpitis.

Comparison of two case difficulty assessment methods on cohorts of undergraduate dental students - a multi-centre study.

AIM: To compare the educational benefits and user friendliness of two anonymized endodontic case difficulty assessment (CDA) methods. METHODOLOGY: A cohort (n = 206) of fourth-year undergraduate dental students were recruited from four different Dental Schools and divided randomly into two groups (Group A and B). The participants assessed six test endodontic cases using anonymized versions of the American Association of Endodontists (AAE) case difficulty assessment form (AAE Endodontic Case Difficulty Assessment Form and Guidelines, 2006) and EndoApp, a web-based CDA tool. Group A (n = 107) used the AAE form for assessment of the first three cases, followed by EndoApp for the latter. Group B (n = 99) used EndoApp for the initial three cases and switched to the AAE form for the remainder. Data were collected online and analysed to assess participants' knowledge reinforcement and agreement with the recommendation generated. Statistical analysis was performed using the two-way mixed model anova, Cohen's Kappa (κ) and independent t-tests, with the levels of significance set at P < 0.05. Additionally, participants' feedback and preference for CDA was also gathered. RESULTS: There was a significant increase in knowledge reinforcement for the AAE form and EndoApp (P = 0.001) after assessment of the first three test cases. However, this increase was not significant (P = 0.842) between the CDA methods. Overall, the AAE form and EndoApp had slight (κ = 0.176, P < 0.001) and substantial (κ = 0.668, P < 0.001) levels of agreement, respectively, and the difference was statistically significant (P < 0.001). Participants' feedback on user friendliness favoured EndoApp for all parameters measured. EndoApp was preferred by 65% of the cohort, whereas only 11% chose the AAE form for CDA. CONCLUSIONS: Both the AAE form and EndoApp were beneficial for dental education. EndoApp was reliable in helping with decisions to treat or refer, and combined with user friendliness, it was the preferred choice for CDA.

The influence of irrigant activation, concentration and contact time on sodium hypochlorite penetration into root dentine: an ex vivo experiment.

AIM: To establish whether irrigant activation techniques, namely manual dynamic activation (MDA), passive ultrasonic irrigation (PUI) and sonic irrigation (SI), improve the tubular penetration of sodium hypochlorite (NaOCl) into root dentine when compared with conventional needle irrigation (CNI). Secondly, investigate if increasing NaOCl concentration and/or contact time improves the performance of these techniques. METHODOLOGY: A total of 83 extracted human maxillary permanent canines were decoronated to 15 mm, and root canals prepared to a size 40, .10 taper. Root dentine was stained with crystal violet for 72 h and embedded in silicone. Eighty specimens were randomly distributed into 16 groups (n = 5) according to the irrigant activation technique, NaOCl concentration (2%; 5.25%) and irrigant contact time (10 min; 20 min). All activation techniques were used for 60 s in the last minute of irrigation. Additionally, three teeth were not exposed to NaOCl to confirm adequate dentine staining had occurred (i.e. negative control). All specimens were subsequently dissected, observed under a light microscope and NaOCl penetration depth (µm) determined by measuring the average width of bleached dentine using ImageJ software. Statistical comparisons were made with paired and unpaired t-tests, anovas followed by post hoc Tukey's and Dunnett's tests, and a general linear model (α < 0.05). RESULTS: Overall, NaOCl penetration ranged from 38.8 to 411.0 µm with MDA, PUI and SI consistently resulting in significantly greater tubular infiltration than CNI (P < 0.05). The deepest measurements in the coronal, middle and apical segments were all recorded in the MDA; 5.25%; 20 min group and the least in the CNI; 2%; 10 min group. Increasing either irrigant concentration or contact time resulted in significantly greater NaOCl penetration depths for all techniques and segments of the canal (P < 0.05). However, when irrigant concentration and contact time were increased together, a significant interaction effect between these two independent variables was observed on overall NaOCl penetration (P < 0.05). CONCLUSIONS: Agitating irrigants with MDA, PUI or SI, as well as using greater irrigant concentrations or contact times, potentiated NaOCl penetration into root dentine. However, longer durations of NaOCl exposure at lower concentrations resulted in similar depths of tubular penetration as those achieved at higher concentrations.

Management of deep caries and the exposed pulp.

Caries prevalence remains high throughout the world, with the burden of disease increasingly affecting older and socially disadvantaged groups in Western cultures. If left untreated, caries will advance through dentine stimulating pulpitis and eventually pulp infection and necrosis; however, if conservatively managed, pulpal recovery occurs even in deep carious lesions. Traditionally, deep caries management was destructive with nonselective (complete) removal of all carious dentine; however, the promotion of minimally invasive biologically based treatment strategies has been advocated for selective (partial) caries removal and a reduced risk of pulp exposure. Selective caries removal strategies can be one-visit as indirect pulp treatment or two-visit using a stepwise approach. Management strategies for the treatment of the cariously exposed pulp are also shifting with avoidance of pulpectomy and the re-emergence of vital pulp treatment (VPT) techniques such as partial and complete pulpotomy. These changes stem from an improved understanding of the pulp-dentine complex's defensive and reparative response to irritation, with harnessing the release of bioactive dentine matrix components and careful handling of the damaged tissue considered critical. Notably, the development of new pulp capping materials such as mineral trioxide aggregate, which although not an ideal material, has resulted in more predictable treatments from both a histological and a clinical perspective. Unfortunately, the changes in management are only supported by relatively weak evidence with case series, cohort studies and preliminary studies containing low patient numbers forming the bulk of the evidence. As a result, critical questions related to the superiority of one caries removal technique over another, the best pulp capping biomaterial or whether pulp exposure is a negative prognostic factor remain unanswered. There is an urgent need to promote minimally invasive treatment strategies in Operative Dentistry and Endodontology; however, the development of accurate diagnostic tools, evidence-based management strategies and education in management of the exposed pulp are critical in the future.

A systematic review of methods used to sample and analyse periradicular tissue fluid during root canal treatment.

AIM: The primary aim was to identify techniques used to sample and analyse periradicular tissue fluid (PTF) in permanent teeth diagnosed with apical disease during root canal treatment. Secondly, to identify the types of inflammatory mediators studied using this approach. METHODOLOGY: Data Sources: PubMed, EMBASE, Cochrane Library, Science Direct, Web of Science and OpenGrey. Eligibility Criteria: Clinical studies published until 1 June 2018 which utilized orthograde techniques to sample and analyse PTF were included. Cell culture, laboratory or animal studies and those concerned with investigating inflammatory mediator activity from within healthy or diseased pulp tissue, and not periradicular tissues, were excluded. Study appraisal and methods: In accordance with PRISMA guidelines, data were extracted on study characteristics, target mediators, sampling and assay techniques and the parameters associated with the PTF sampling and eluting protocol. A qualitative synthesis was conducted, and studies were critically appraised using a modified version of the Cochrane risk of bias tool. RESULTS: Study Characteristics: From 251 studies, 33 were eligible for inclusion. Sampling techniques included the use of paper points (n = 27), fine needle aspiration (n = 4) and filter strips (n = 2). Assay techniques included enzyme-linked immunosorbent assay (n = 18), quantitative polymerase chain reaction (n = 9), radioimmunoassay (n = 4), colorimetric assay (n = 2), immunofluorometric assay (n = 1) and cytometric bead array (n = 1). Forty-five different inflammatory mediators were targeted at the proteomic/metabolomic (n = 25) or transcriptomic level (n = 9). LIMITATIONS: Significant heterogeneity exists within the methodology, and only 5 studies disclosed unambiguous information about their PTF sampling and eluting protocols. CONCLUSIONS: Paper points and proteomic/metabolomic analysis are currently the preferred methods for studying and analysing PTF during root canal treatment. The most studied analytes were IL-1ß and TNF-α. IMPLICATIONS: Further research is required to develop an optimized PTF sampling and eluting protocol to overcome methodological heterogeneity, and future studies are advised to follow a standardized approach to reporting their methodology.

European Society of Endodontology position statement: Management of deep caries and the exposed pulp.

This position statement on the management of deep caries and the exposed pulp represents the consensus of an expert committee, convened by the European Society of Endodontology (ESE). Preserving the pulp in a healthy state with sustained vitality, preventing apical periodontitis and developing minimally invasive biologically based therapies are key themes within contemporary clinical endodontics. The aim of this statement was to summarize current best evidence on the diagnosis and classification of deep caries and caries-induced pulpal disease, as well as indicating appropriate clinical management strategies for avoiding and treating pulp exposure in permanent teeth with deep or extremely deep caries. In presenting these findings, areas of controversy, low-quality evidence and uncertainties are highlighted, prior to recommendations for each area of interest. A recently published review article provides more detailed information and was the basis for this position statement (Bjørndal et al. 2019, International Endodontic Journal, doi:10.1111/iej.13128). The intention of this position statement is to provide the practitioner with relevant clinical guidance in this rapidly developing area. An update will be provided within 5 years as further evidence emerges.

Growth factor release from dentine matrix by pulp-capping agents promotes pulp tissue repair-associated events.

AIM: To characterize growth factor release from dentine by pulp-capping agents and to determine the effects of liberated dentine extracellular matrix (dECM) components on pulp cells in the key wound healing processes of migration and cell growth. METHODOLOGY: Powdered human dentine was exposed to solutions of calcium hydroxide, white and grey mineral trioxide aggregate (MTA) (ProRoot, (Dentsply Tulsa, Tulsa, OK, USA) over 14 days. The solubilized dECM components were dialysed and lyophilized and characterized using multiplex quantitative ELISA. Following dECM component extraction dentine was analysed using Fourier-transformed infrared spectroscopy (FTIR). Primary rat dental pulp cells (RDPCs) were exposed to dECM components (0.1-100 µg mL-1 ) released by calcium hydroxide, white and grey MTA, and cell growth and chemotactic responses were assessed. Statistical differences between the experimental and control groups were determined using one-way anova. RESULTS: A broad range of growth factors, many not previously reported in dentine, were liberated by these pulp-capping agents, including SCF, M-CSF, GM-CSF, IGFBP-1, NGF and GDNF. White and grey MTA liberated more growth factors than calcium hydroxide. FTIR analysis of dentine exposed to pulp-capping agents showed partial depletion of amide bands I, II and III, with little alteration in phosphate peaks compared to untreated dentine. dECM components released by white and grey MTA induced significantly more cell growth at low-to-moderate concentrations (P â‰¦ 0.05) examined in this study and significantly enhanced cell chemotaxis at all concentrations compared with controls (P â‰¦ 0.05). CONCLUSIONS: White and grey MTA solubilize a broad range of bioactive molecules from dentine, which can induce proliferation and chemotaxis in pulp cells.

The impact of endodontic access on the biaxial flexure strength of dentine-bonded crown substrates - an in vitro study.

AIM: To investigate how preparation of a simulated access cavity into ceramic materials suitable for the manufacture of dentine-bonded crowns (DBCs) impacted on biaxial flexural strength (BFS) determined as a monolithic structure and in a more clinically representative resin-cemented form. METHODOLOGY: One hundred and twenty feldspathic and 120 leucite-reinforced ceramic disc-shaped specimens were divided into eight groups (n = 30). All groups received 'fit' surface treatments representative of pre-cementation modifications and of cementation prior to preparation of a representative endodontic access cavity through the sample. BFS was determined for both 'intact' and the 'annular' disc-shaped specimens which had received simulated endodontic access. Newly reported analytical solutions were used to calculate BFS of the 'annular' specimens. Statistical analysis included two-way anovas (α = 0.05) and Weibull analysis. Fractographic examination provided insight into the fracture mechanisms. RESULTS: A two-way anova identified a significant impact of material (P < 0.01) and of resin coating (P < 0.01) on the mean BFS of intact specimens. For the annular ceramic specimens, the substrate material significantly impacted on mean BFS (P < 0.01), but the effect of resin coating was dependent on the substrate type (P < 0.01). CONCLUSIONS: Endodontic access cavity preparation modified the critical defect population of the all-ceramic restorative materials investigated. The strength of a predominantly glassy ceramic following endodontic access can be maintained if adhesive cementation was used; however, the beneficial effects of adhesive cementation on ceramic reinforcement were lost on leucite-reinforced ceramics following access cavity preparation. Replacement restoration for these materials would be recommended clinically following endodontic access as opposed to repair of the access cavity using a direct restorative material.

Contemporary obturation of the root canal system.

This article will provide an overview of the rationale behind taking a contemporary approach to obturation of the root canal system. It will outline a broad range of obturation materials including those traditionally used and newer, advanced ones that are now available that have active physical and biological properties. Obturation techniques will be described in detail, including advice for those difficult cases such as managing obturation of a canal with a wide open apex. Assessment of obturation will also be discussed, considering those factors which affect outcome of root canal treatment.

Access cavity preparation.

Each stage of root canal treatment should be carried out to the highest possible standard. The access cavity is arguably the most important technical stage, as subsequent preparation of the root canal(s) can be severely comprised if this is not well executed. Inadequate access can lead to canals being left untreated, poorly disinfected, difficult to shape and obturate, and may ultimately lead to the failure of the treatment. This paper highlights common features in root canal anatomy and outlines basic principles for locating root canals and producing a good access cavity. It also explores each phase of the preparation in detail and offers suggestions of instruments that have been specifically designed to overcome potential difficulties in the process. Good access design and preparation will result in an operative environment which will facilitate cleaning, shaping and obturation of the root canal system in order to maximise success.

Dentin-pulp complex regeneration: from lab to clinic.

Dentistry is entering an exciting era in which many of the advances in biotechnology offer opportunities for exploitation in novel and more effective therapies. Pulp healing is complex and dependent on the extent of injury, among many other factors. Many of the molecular and cellular processes involved in these healing events recapitulate developmental processes. The regulation of odontoblast activity is clearly central to pulp healing, and an understanding of the mechanisms involved in these processes is necessary to enable laboratory studies to be translated to clinic application. Transcriptome analysis has identified changes in many odontoblast genes during the life-cycle of this cell and its responses to injurious challenge. The p38 MAPKinase pathway appears to be central to the transcriptional control of odontoblasts and may provide a key target for therapeutic intervention. The many recent advances in knowledge of pulpal stem cells and molecular signaling molecules within the tooth, now provide exciting opportunities for clinical translation to novel therapies. Such translation will require the partnership of researchers and skilled clinicians who can effectively apply advances in knowledge to appropriate clinical cases and develop novel therapies which can be realistically introduced into the clinic.

Dental regeneration and materials: a partnership.

Considerable focus on the biocompatibility of dental materials over the last three decades has provided a platform for a wealth of studies on the cellular and molecular responses of the cells of the pulp to injury, both from the disease process and from subsequent restorative intervention. These studies have been fundamental to understanding not only how we can achieve a biocompatible response during restoration of dental disease but also how we can exploit the pulpal cellular responses to achieve wound healing and tissue regeneration in the dentine-pulp complex. This article examines the responses of the pulp to injury and the events leading to tissue regeneration. As new biologically based regenerative therapies emerge for the dental tissues, it is important that these develop in partnership with more traditional approaches using dental materials.

Teaching posterior composite resin restorations in the United Kingdom and Ireland: consensus views of teachers.

Posterior composite resin restorations are an established feature of contemporary dental practice and all new dental graduates should be competent in providing such treatments for their patients. Surveys of educational curricula in this area in the United Kingdom and Ireland, as well as North America, have demonstrated variations both within and between dental schools. Such inconsistency does not help new dental school graduates, and may lead to confusion. At the British Association of Teachers of Conservative Dentistry Annual Conference held in Birmingham in September 2005, a session was devoted to the development of guidelines for dental schools on teaching posterior composite resin restorations to dental undergraduates. The theme of the conference concerned the teaching implications for changing from amalgam to composite. Two of the principal speakers at the meeting (Joost Roeters and Niek Opdam) were from the dental school at the University of Nijmegen in the Netherlands. This school was the first in Europe to discontinue the use of dental amalgam in its undergraduate curriculum over a decade ago. This paper reports the consensus views of those present on guidelines for teaching posterior composite resin restorations to dental undergraduate students.

An audit on periodontal assessment in general dental practice.

Thorough periodontal assessment is vital for diagnosis, treatment planning and monitoring the progression of periodontal disease. This is a report of a clinical audit that studied the periodontal assessment carried out at a large general dental practice in Shropshire, England. This audit was conducted over a 3-month period, analysing 700 patients for each audit cycle. A new protocol for periodontal assessment using the guidelines of the British Society of Periodontology was introduced. The results demonstrate a marked improvement in assessing the periodontal condition of patients in this general dental practice.