Apical root canal microbiome associated with primary and posttreatment apical periodontitis: A systematic review.

BACKGROUND: Microorganisms colonizing the apical root canal system are conceivably the ones directly involved with the causation and maintenance of apical periodontitis. OBJECTIVES: This article systematically reviews the reports on the microbiome occurring exclusively at the apical root canal of teeth with primary and posttreatment apical periodontitis. METHODS: The electronic databases PubMed, Embase, Web of Science, Science Direct, and Proquest were searched up to August 2023. Clinical studies using culture and molecular microbiology methods to identify the microbial taxa present exclusively in the apical root canal segment of infected teeth with apical periodontitis were included. Studies were critically assessed using the Joanna Briggs Institute Critical Prevalence Assessment Checklist. RESULTS: From 2277 articles initially detected, 52 were selected for full reading and 21 were eventually included in this review. Of these, molecular methods were used in 19 and culture in 2 studies. Ten studies evaluated primary infections, 8 evaluated posttreatment infections, and 3 included both. Cryopulverization of the apical root specimens was conducted in 11 studies. All studies evaluated the prevalence and diversity of bacteria, and only one also reported on fungi. Overall, the most frequent/abundant bacterial taxa found in the apical canal of primary infections were Pseudoramibacter alactolyticus, Olsenella uli, Fusobacterium species, Streptococcus species, Porphyromonas endodontalis, Prevotella species, Actinomyces species, Parvimonas micra, Treponema denticola, Synergistetes species, and an as-yet uncharacterized taxon. In posttreatment infections, the most prevalent/abundant bacterial taxa included species of Streptococcus, Enterococcus, Fusobacterium, Actinomyces, Pseudoramibacter, Pseudomonas, and Propionibacterium. At the phylum level, Firmicutes was the most represented. The average apical bacterial load ranged from 105 to 106 in primary infections and from 103 to 104 in posttreatment infections. DISCUSSION: Microbial diversity in the apical part of the root canal system was examined encompassing data from both primary and posttreatment infections. Heterogeneity amongst the studies, especially in sample collection and microbial identification methods, is an important limitation that prevented a meta-analysis. CONCLUSIONS: There is a pronounced bacterial diversity in the infected apical canal, with a high interindividual variability. Different microbiome compositions at the species/genus level are observed according to the infection type. REGISTRATION: PROSPERO CRD42021275886.

Intracanal removal and apical extrusion of filling material after retreatment using rotary or reciprocating instruments: A new approach using human cadavers.

AIM: This study compared intracanal removal of filling as well as the frequency and volume of extruded material after retreatment with either HyFlex or Reciproc instruments in mandibular teeth from cadavers. METHODOLOGY: The root canals of 14 pairs of contralateral single-rooted teeth in mandibles of cadavers were instrumented with Reciproc R40 and filled using lateral compaction. The mandibles were scanned in a micro-computed tomographic (micro-CT) device before and after retreatment procedures. The contralateral teeth were assigned to two groups (n = 14) according to the retreatment protocol using either HyFlex or Reciproc instrument systems. In the HyFlex group, the HyFlex Remover instrument was worked 3 mm short of the working length (WL), followed by HyFlex CM 40.04 and 50.04 at the WL. In the Reciproc group, the R50 instrument was worked up at the coronal two thirds, followed by two more cycles until the WL was reached. Pre- and post-operative micro-CT images were analysed for extrusion and intracanal removal of filling material. RESULTS: After retreatment, extrusion of filling material occurred in 11 (78%) and 14 (100%) teeth from HyFlex and Reciproc groups respectively (p > .05). A similar volume of extruded material was observed after retreatment with both systems (p > .05). A significant decrease in the intracanal filling volume was verified after retreatment with both tested systems (p < .05). However, residual filling material was found in all root canals, regardless of the system. The amount of filling material removed (HyFlex = 80.8%; Reciproc = 65.9%) and the operation time was similar between systems (p > .05). CONCLUSIONS: A high frequency of filling material extrusion was observed after retreatment with the two systems in a cadaver model, with no significant difference between them. Both protocols obtained similar efficacy in filling material removal procedures, although none completely cleaned the canals.

Impact of root canal preparation using two single-file systems on the intra-radicular microbiome of teeth with primary apical periodontitis.

OBJECTIVES: This study aimed to describe the effects of two single-file systems on the diversity of the endodontic microbiome of teeth with primary asymptomatic apical periodontitis. MATERIALS AND METHODS: The root canals from single-rooted teeth with apical periodontitis were prepared using either the Reciproc Blue (RB) or the XP-endo Shaper (XPS) instrument system. The latter was followed by a supplementary step with the XP-endo Finisher (XPF) instrument. For irrigation, 5.25% sodium hypochlorite was used. Root canal samples were taken at the baseline (S1), after preparation (S2), and after the supplementary step (S3). DNA was extracted and subjected to high-throughput sequencing using the MiSeq Illumina platform. RESULTS: Samples from 10 teeth from the RB and 7 from the XPS group were subjected to DNA sequencing. Initial samples differed significantly from post-preparation samples in bacterial diversity, with no significant difference when comparing the two instrument systems. The most dominant phyla in S2 were Bacteroidetes, Proteobacteria, Firmicutes, Fusobacteria, and Actinobacteria. The same phyla were found to dominate baseline samples and samples taken after using XPF, but with differences in the ranking of the most dominant ones. At the genus level, the most dominant genera identified after RB instrumentation were Bacteroidaceae [G-1], Fusobacterium, and Staphylococcus, while the most dominant genera after XPS instrumentation were Fusobacterium and Porphyromonas. These genera were also dominant in the initial samples. CONCLUSIONS: Both treatment protocols had measurable effects on the root canal microbial diversity, with no significant differences between them. Most of the dominant taxa involved in the primary infection and probably in the aetiology of apical periodontitis were eliminated or substantially reduced. CLINICAL RELEVANCE: The most dominant taxa that persisted after instrumentation were Fusobacterium, Porphyromonas, Staphylococcus, and Bacteroidaceae [G-1].

Shaping ability of BlueShaper, TruNatomy, DC Taper and HyFlex EDM in long oval canals: An ex vivo study.

AIM: This study evaluated and compared the shaping ability of four rotary instrument systems in long-oval root canals using microcomputed tomographic (micro-CT) evaluation. Currently, there is no data available on the canal shaping abilities of BlueShaper and DC Taper instruments. METHODOLOGY: Sixty-four single-rooted mandibular premolars were matched based on similar root canal morphologic features as determined by (micro-CT) and randomly assigned to 1 of 4 experimental groups (n = 16) according to the instrument system used: BlueShaper, TruNatomy, DC Taper and HyFlex EDM One File. Changes in the root canal surface and volume, remaining dentine thickness, and number of prepared areas were assessed. RESULTS: No significant differences were found amongst the four instrument systems for the parameters evaluated (p > .05). There was a significant reduction in the number of unprepared areas and the remaining dentine thickness after each increase in size of the instruments tested (p < .05). CONCLUSIONS: The four instrument systems perform similarly in long oval root canals. Although none could prepare all canal walls, larger preparations incorporated significantly more surfaces in the final shape.

Influence of brushing motions on the shaping of oval canals by rotary and reciprocating instruments.

OBJECTIVE: The effects of brushing on shaping with three different instruments were assessed in oval canals. DESIGN: Mandibular incisors were assigned to 6 groups (n = 12/group) according to the system, each one with or without brushing: Reciproc Blue, VDW.Rotate, and Race EVO. Micro-computed tomography was performed before and after preparation. RESULTS: Brushing strokes caused no increase in canal volume, surface area, and structure model index independently of the system (p > 0.05), except for RaCe EVO in the full canal surface area (p < 0.05). Brushing did not increase the prepared areas (p > 0.05), except for Reciproc in the apical canal (p < 0.05). Reciproc with no brushing exhibited less pericervical dentin than with brushing (p < 0.05), while RaCe EVO with brushing resulted in less remaining dentin (p < 0.05). CONCLUSIONS: The brushing motion had no effects on the overall shaping performance of the 3 instruments tested. An exception was the increase in prepared surface area in the apical canal segment when the Reciproc instrument was used with brushing strokes.

Present status and future directions: Microbiology of endodontic infections.

Apical periodontitis has a microbial aetiology and is one of the most common inflammatory diseases that affect humans. Fungi, archaea and viruses have been found in association with apical periodontitis, but bacteria are by far the most prevalent and dominant microorganisms in endodontic infections. Bacterial infection of the root canal system only occurs when the pulp is necrotic or was removed for previous treatment. In some specific cases, including acute and chronic abscesses, the bacterial infection may reach the periradicular tissues. Intracanal bacteria are usually observed as sessile multispecies communities (biofilms) attached to the dentinal root canal walls. Infection in the main root canal lumen can spread to other areas of the root canal system. Although more than 500 bacterial species have been detected in endodontic infections, a selected group of 20 to 30 species are most frequently detected and may be considered as the core microbiome. There is a high interindividual variability in the endodontic microbiome in terms of species composition and relative abundance. Obligate anaerobic species are more abundant in the intraradicular bacterial communities of teeth with primary apical periodontitis, while both anaerobes and facultatives dominate the communities in post-treatment apical periodontitis. Bacterial interactions play an essential role in determining the overall virulence of the community, which has been regarded as the unit of pathogenicity of apical periodontitis. This article reviews the microbiologic aspects of endodontic infections and provides perspectives for future research and directions in the field.

A critical analysis of research methods and experimental models to study the root canal microbiome.

Endodontic microbiology deals with the study of the microbial aetiology and pathogenesis of pulpal and periradicular inflammatory diseases. Research in endodontic microbiology started almost 130 years ago and since then has mostly focussed on establishing and confirming the infectious aetiology of apical periodontitis, identifying the microbial species associated with the different types of endodontic infections and determining the efficacy of treatment procedures in eradicating or controlling infection. Diverse analytical methods have been used over the years, each one with their own advantages and limitations. In this review, the main features and applications of the most used technologies are discussed, and advice is provided to improve study designs in order to properly address the scientific questions and avoid setbacks that can compromise the results. Finally, areas of future research are described.

Shaping ability of new reciprocating or rotary instruments with two cross-sectional designs: An ex vivo study.

AIM: This study compared the shaping ability of four new reciprocating and rotary nickel-titanium instruments, with triangular or S-shaped cross-section, in the mesial canals of mandibular molars using micro-computed tomographic (micro-CT) evaluation. METHODOLOGY: Twenty-four extracted mandibular molars with Vertucci's class IV configuration in the mesial root were selected for this study. The teeth were matched in fours according to anatomic similarities as revealed by micro-CT and then distributed into four groups of 12 mesial canals each according to the instrumentation technique: Reciproc Blue, R-motion, VDW.Rotate and RaCe EVO. The final apical size of instrumentation was 30/0.04 for three systems and 25/08 for the Reciproc Blue instrument. Micro-CT scans were taken before and after preparation to evaluate the canal volume, area and unprepared surface areas, as well as the centring ability, and the canal: root width ratio. RESULTS: Preparation with all systems significantly increased the volume and area of the canals (p < .05). There were no significant differences between groups regarding the amount of unprepared areas in both the apical and full canal lengths (p > .05). Variation in the centre of gravity showed no significant difference between groups either (p > .05). The canal: root width ratio at levels 0 and 4 mm apically to the coronal canal opening was significantly increased by all systems (p < .01), with no significant differences between groups (p > .05). The canal width never exceeded 40% of the root width. CONCLUSIONS: Reciprocating or rotary instruments, with a triangular or an S-shaped cross-section, performed equally in shaping Vertucci's class IV mesial canals of mandibular molars.

PRILE 2021 guidelines for reporting laboratory studies in Endodontology: explanation and elaboration.

Guidance to authors is needed to prevent their waste of talent, time and resources in writing manuscripts that will never be published in the highest-quality journals. Laboratory studies are probably the most common type of endodontic research projects because they make up the majority of manuscripts submitted for publication. Unfortunately, most of these manuscripts fail the peer-review process, primarily due to critical flaws in the reporting of the methods and results. Here, in order to guide authors, the Preferred Reporting Items for study Designs in Endodontology (PRIDE) team developed new reporting guidelines for laboratory-based studies: the Preferred Reporting Items for Laboratory studies in Endodontology (PRILE) 2021 guidelines. The PRILE 2021 guidelines were developed exclusively for the area of Endodontology by integrating and adapting the modified CONSORT checklist of items for reporting in vitro studies of dental materials and the Clinical and Laboratory Images in Publications (CLIP) principles. The process of developing the PRILE 2021 guidelines followed the recommendations of the Guidance for Developers of Health Research Reporting Guidelines. The aim of the current document is to provide authors with an explanation for each of the items in the PRILE 2021 checklist and flowchart with examples from the literature, and to provide advice from peer-reviewers and editors about how to solve each problem in manuscripts prior to their peer-review. The Preferred Reporting Items for study Designs in Endodontology (PRIDE) website (http://pride-endodonticguidelines.org/prile/) provides a link to the PRILE 2021 explanation and elaboration document as well as to the checklist and flowchart.

PRILE 2021 guidelines for reporting laboratory studies in Endodontology: A consensus-based development.

Reproducible, skilfully conducted and unbiased laboratory studies provide new knowledge, which can inform clinical research and eventually translate into better patient care. To help researchers improve the quality and reproducibility of their research prior to a publication peer-review, this paper describes the process that was followed during the development of the Preferred Reporting Items for Laboratory studies in Endodontology (PRILE) 2021 guidelines and which used a well-documented consensus-based methodology. A steering committee was created with eight individuals (PM, RO, OP, IR, JS, EP, JJ and SP), plus the project leaders (PD, VN). The steering committee prepared an initial checklist by combining and adapting items from the modified Consolidated Statement of Reporting Trials checklist for reporting in vitro studies of dental materials and the Clinical and Laboratory Images in Publications principles as well as adding several new items. The steering committee then formed a PRILE Delphi Group (PDG) and PRILE Online Meeting Group (POMG) to provide expert advice and feedback on the initial draft checklist and flowchart. The members of the PDG participated in an online Delphi process to achieve consensus on the items within the PRILE 2021 checklist and the accompanying flowchart for clarity and suitability. The PRILE checklist and flowchart developed by the online Delphi process were discussed further by the POMG. This online meeting was conducted on 12 February 2021 via the Zoom platform. Following this meeting, the steering committee developed a final version of the PRILE 2021 guidelines and flowchart, which was piloted by several authors when writing up a laboratory study for publication. Authors are encouraged to use the PRILE 2021 guidelines and flowchart to improve the clarity, completeness and quality of reports describing laboratory studies in Endodontology. The PRILE 2021 checklist and flowchart are freely available and downloadable from the Preferred Reporting Items for study Designs in Endodontology website (http://pride-endodonticguidelines.org/prile/).

Unprepared surface areas, accumulated hard tissue debris, and dentinal crack formation after preparation using reciprocating or rotary instruments: a study in human cadavers.

OBJECTIVES: This study using contralateral teeth from human cadavers assessed the amount of unprepared wall surface areas (USA), accumulated hard tissue debris (AHTD), and dentinal crack formation after root canal preparation using either reciprocating or rotary instruments. MATERIALS AND METHODS: Incisors with a single root canal from 11 human mandibles had their root canals prepared with Reciproc or Mtwo. Each contralateral tooth was treated with one of the test instruments in order to create matched pairs. Micro-CT scans were taken before and after preparation to instrument sizes 40 and 50 in both groups. USA and AHTD were evaluated in the full canal length or the apical portion only. Crack formation was also evaluated in the full root length. RESULTS: After size 40, no significant differences in USA and AHTD were observed in the full canal length (P > .05). Separate analysis of the apical 4-mm canal revealed no difference in AHTD, but significantly more USA with Mtwo than Reciproc (P < .05). After size 50, the amount of USA was similar between groups both in the full canal and in the apical canal (P > .05). More AHTD were observed in the full canal after using Mtwo size 50 (P < .05), but no difference occurred in the apical canal only (P > .05). Intragroup analyses showed a significant decrease of USA and a significant increase of AHTD when preparation size increased from 40 to 50 (P < .05). In teeth without any pre-existing detectable dentinal defect (n = 38), crack formation occurred in 4 teeth, 2 from each preparation system. CLINICAL RELEVANCE: Reciproc instrument size 40 resulted in more prepared areas in the apical canal than Mtwo instrument of the same size. However, the amount of AHTD did not differ between them at this file size. Canal enlargement to size 50 prepared more walls but created more debris. Dentinal defects were produced after preparation with both systems.

Distribution of sodium hypochlorite throughout the mesial root canal system of mandibular molars after adjunctive irrigant activation procedures: a micro-computed tomographic study.

OBJECTIVES: To evaluate and compare irrigant distribution throughout the root canal system of mesial roots of mandibular molars after application of distinct adjunctive irrigant activation procedures. MATERIALS AND METHODS: Fifteen extracted mandibular molars presenting Fan's type II, III, or IV isthmus configurations were selected. The canals were initially enlarged and cleaned. The same specimens were subjected to conventional positive pressure irrigation (PPI) followed by adjunctive irrigation approaches: passive ultrasonic irrigation (PUI), mechanical activation with the XP-endo Finisher instrument, and a control group in which the irrigant remained in the canal with no activation. Retention time for the irrigant was the same for all groups. Sodium hypochlorite mixed with a contrast medium (Omnipaque 300) was used as the irrigant. Before and immediately after the adjunctive irrigation, micro-computed tomographic (micro-CT) scans were taken to evaluate the volume of the entire root canal system that was filled by the irrigant. An exclusive analysis was also done for the isthmus area. RESULTS: Analysis of the entire root canal system showed that the XP-endo Finisher instrument promoted better distribution of the irrigant than PUI and the control group (p < 0.01). PUI was not significantly different from the control group (p > 0.05). A separate analysis of the isthmus area showed no significant difference between the adjunctive irrigation methods and the control (p > 0.05). CONCLUSIONS: The XP-endo Finisher instrument promoted better distribution of irrigant throughout the root canal system, especially in the apical canal segment, when compared to PUI. However, the tested approaches did not differ as to the ability to drive irrigants into the isthmus area. CLINICAL RELEVANCE: This study highlighted that the XP-endo Finisher instrument presented a better performance to distribute irrigant throughout the root canal system, especially in the apical canal segment compared to positive pressure irrigation and PUI.

Microbiology and treatment of acute apical abscesses.

Acute apical abscess is the most common form of dental abscess and is caused by infection of the root canal of the tooth. It is usually localized intraorally, but in some cases the apical abscess may spread and result in severe complications or even mortality. The reasons why dental root canal infections can become symptomatic and evolve to severe spreading and sometimes life-threatening abscesses remain elusive. Studies using culture and advanced molecular microbiology methods for microbial identification in apical abscesses have demonstrated a multispecies community conspicuously dominated by anaerobic bacteria. Species/phylotypes commonly found in these infections belong to the genera Fusobacterium, Parvimonas, Prevotella, Porphyromonas, Dialister, Streptococcus, and Treponema. Advances in DNA sequencing technologies and computational biology have substantially enhanced the knowledge of the microbiota associated with acute apical abscesses and shed some light on the etiopathogeny of this disease. Species richness and abundance and the resulting network of interactions among community members may affect the collective pathogenicity and contribute to the development of acute infections. Disease modifiers, including transient or permanent host-related factors, may also influence the development and severity of acute abscesses. This review focuses on the current evidence about the etiology and treatment of acute apical abscesses and how the process is influenced by host-related factors and proposes future directions in research, diagnosis, and therapeutic approaches to deal with this disease.

Dens invaginatus type II in an immature lateral incisor with apical periodontitis treated with regenerative and strategic antimicrobial procedures: a case report.

Dens invaginatus is a developmental dental anomaly that can predispose the tooth to pulp and periradicular disease. Management of this condition can be challenging because of anatomic and microbiologic issues. This case report describes the regenerative endodontic treatment (REP) using a strategic antimicrobial protocol for management of an immature maxillary lateral incisor with type-II dens invaginatus associated with apical periodontitis in a 13-year-old patient. The tooth presented with a complex anatomy and was associated with an active sinus tract. Because the true root canal was not negotiable in its coronal part due to the presence of the dens invaginatus, the closed end of the invagination (pseudocanal) was perforated to permit access to the apical segment of the true root canal for cleaning and disinfection. Both the invagination and the true canal were treated using an antimicrobial regimen based on chemomechanical preparation with sodium hypochlorite irrigation, supplementary disinfection with passive ultrasonic irrigation and interappointment calcium hydroxide medication. After two exchanges of calcium hydroxide medication, the sinus tract did not disappear, then the antimicrobial protocol was changed to include an antibiotic solution for irrigation and antibiotic paste for intracanal medication. After signs and symptoms disappeared, REP was performed by inducing blood clot formation within the root canal. The coronal canal segment including the invagination was filled with Biodentine. Follow-up including cone-beam computed tomography examination showed complete healing of the apical periodontitis lesion and mineralized tissue formation at the apical portion of the true root canal.

Factors influencing the outcome of nonsurgical root canal treatment and retreatment: A retrospective study.

This study assessed the influence of diverse variables on the outcome of nonsurgical root canal treatment/retreatment. In general, 304 teeth from 218 patients were treated/retreated and the outcome evaluated by the periapical index (PAI). Teeth with apical periodontitis lesions that have not completely healed were classified as success or failure based on lenient and rigid criteria, respectively. Findings were evaluated using a logistic regression analysis. The overall success rates were 74% and 82% using the PAI-rigid and lenient success criteria, respectively. Specifically for treatment, the success rates were 73% (rigid) and 82% (lenient), while for retreatment they were 78% (rigid) and 83% (lenient). The treatment outcome was negatively affected by overextension, presence of preoperative lesion, lesion size >10 mm, and higher number of treatment visits (with no intracanal medication). Regarding retreatment, the chance of success was greater for teeth with adequate coronal restorations.

Bacteriologic Conditions of the Apical Root Canal System of Teeth with and without Posttreatment Apical Periodontitis: A Correlative Multianalytical Approach.

INTRODUCTION: This study used a correlative multianalytical approach to investigate the bacteriologic conditions in the apical root canal system of treated teeth with or without apical periodontitis and their correlation with the technical quality of the previous root canal obturation and the presence and volume of apical periodontitis lesions. METHODS: Root apexes were obtained from recently extracted root canal-treated teeth with (n = 23) and without (n = 22) apical periodontitis lesions as demonstrated by cone-beam computed tomographic examination. The root apexes were sectioned and subjected to micro-computed tomographic (micro-CT) scanning. The specimens were cryopulverized, and DNA extracted from the powder was used as a template in real-time polymerase chain reaction assays to quantify total bacteria and members of the Streptococcus genus and Actinobacteria phylum. The bacteriologic findings were compared between the 2 groups and also evaluated for associations with cone-beam computed tomographic and micro-computed tomographic data. RESULTS: Bacteria were detected in all apical canal samples except 1. The mean counts of total bacteria, streptococci, and actinobacteria did not differ significantly between teeth with or without apical periodontitis (P > .05). Streptococcus levels were significantly lower by 80% in the apical canals of teeth with small lesions compared with those without lesions (P < .05). The limit of filling >2 mm short was significantly associated with more total bacterial counts compared with canals filled 0-2 mm short (P < .05). An adequate coronal restoration was significantly associated with lesser counts of Streptococcus (P < .05). CONCLUSIONS: Comparable bacterial loads were observed in the apical canal system of treated teeth with and without apical periodontitis, suggesting that factors other than only the total bacterial levels may also influence the development and progression of apical periodontitis. Bacteria were found in the apical canal in virtually all cases with a high prevalence of streptococci and actinobacteria. Streptococci counts were significantly higher in the apical canal of teeth with inadequate restorations and teeth with no lesions. Underfilled canals showed higher bacterial counts.

Correlation Between Proinflammatory Cytokine Expression and Clinical Data in Apical Granuloma.

INTRODUCTION: This study was intended to evaluate the expression of inflammatory cytokines commonly secreted by CD4+ T cells (IL-2, IL-5, IL-17, TGF-ß, TNF-α, and IFN-γ) in apical granulomas and correlate with the clinical conditions and time elapsed since root canal treatment. METHODS: Eighteen biopsy specimens obtained by periradicular surgery of teeth with post-treatment apical periodontitis and diagnosed as apical granuloma were available from the oral pathology laboratory. Silanized slides containing paraffin sections were used for immunohistochemical reactions. Images were analyzed by using an optical microscopy and each slide was subdivided into 5 fields at high magnification. RESULTS: IFN-γ and TGF-ß were the cytokines with the highest expression levels. There were statistically significant differences when comparing IL-2 and IFN-γ (P < .05), and IL-2 and TGF-ß (P < .05). Comparison between the detected cytokines and clinical data and time of treatment demonstrated significant correlation (P < .05) between lower expression of IL-2 and the presence of painful symptoms, absence of sinus tract, and treatments performed more than 4 years before. It was also possible to observe a significant correlation between lower expression of IL-5 and treatments performed less than 4 years before (P < .05). CONCLUSION: IFN-γ and TGF-ß were highly expressed in apical granulomas. However, only IL-2 and IL-5 levels were associated with clinical data and time since previous root canal treatment.

Effects of the irrigation needle design on root canal disinfection and cleaning.

INTRODUCTION: This ex vivo study evaluated the disinfecting and cleaning effects of root canal preparation using NaOCl irrigation with three different needle designs. METHODS: Mesial roots from extracted mandibular molars with Vertucci class II configuration were anatomically matched based on micro-computed tomography (micro-CT) analyses and distributed into three groups (n=18/group). The canals were contaminated with a mixed bacterial culture for 30 days and then subjected to preparation using 2.5% NaOCl irrigation with open-ended flat needle (3 mm short of the working length - WL), closed-ended side-vented irrigation needle (1 mm short of the WL) or a closed-ended plastic needle with two lateral outlets (TruNatomy) (1 mm short of the WL). Bacteriological samples were taken from the canals before (S1) and after preparation (S2). After another micro-CT scan, the roots were sectioned and samples were also taken from the apical canal segment (S3). Bacterial reduction was assessed by quantitative real-time polymerase chain reaction (qPCR). The amount of accumulated hard tissue debris was evaluated by micro-CT. RESULTS: A substantial reduction in bacterial counts was observed in all three groups when comparing S1 with S2 (99.95%, 98.93%, and 98.90% in the open-ended, closed-ended, and TruNatomy needle groups, respectively) (P < 0.01). No significant differences were observed between groups for bacterial reduction in the full and apical canal (P > 0.05). There were no intergroup differences in the amount of accumulated hard tissue debris either (P > 0.05). The open-ended needle group showed significantly more specimens with qPCR negative results for bacteria in S3 than TruNatomy (P < 0.05). CONCLUSION: When used up to an appropriate insertion depth, the three needle types result in similar antibacterial and cleaning performance, provided variables such as needle size and irrigant type, volume and flow rate are controlled. Exclusive analysis of the apical segment, including the isthmus, revealed that the open-ended needle yielded more cases negative for bacteria.

The Oral Microbiota in Health and Disease: An Overview of Molecular Findings.

Culture-independent nucleic acid technologies have been extensively applied to the analysis of oral bacterial communities associated with healthy and diseased conditions. These methods have confirmed and substantially expanded the findings from culture studies to reveal the oral microbial inhabitants and candidate pathogens associated with the major oral diseases. Over 1000 bacterial distinct species-level taxa have been identified in the oral cavity and studies using next-generation DNA sequencing approaches indicate that the breadth of bacterial diversity is even much larger. Nucleic acid technologies have also been helpful in profiling bacterial communities and identifying disease-related patterns. This chapter provides an overview of the diversity and taxonomy of oral bacteria associated with health and disease.

Microbial Community Profiling Using Terminal Restriction Fragment Length Polymorphism (T-RFLP) and Denaturing Gradient Gel Electrophoresis (DGGE).

In their natural environments, microorganisms usually live in organized communities. Profiling analysis of microbial communities has recently assumed special relevance as it allows a thorough understanding of the diversity of the microbiota, its behavior over time, and the establishment of patterns associated with health and disease. The application of molecular biology approaches holds the advantage of including culture-difficult and as-yet-uncultivated phylotypes in the profiles, providing a more comprehensive picture of the microbial community. This chapter focuses on two particular techniques, namely terminal restriction fragment length polymorphism (T-RFLP) and denaturing gradient gel electrophoresis (DGGE), both of which have been widely used in environmental studies and have been recently successfully used by the authors in the study of the oral microbial communities associated with conditions of health and disease.