Association between endodontic symptoms and root canal microbiota: a systematic review and meta-analysis of bacteroidetes, spirochaetes and fusobacteriales.

OBJECTIVES: This systematic review and meta-analysis aimed to assess the prevalence of Bacteroidetes, Spirochaetes, and Fusobacteriales in symptomatic versus asymptomatic apical periodontitis as a primary objective. The secondary objective was to evaluate the prevalence of these species independently in symptomatic and asymptomatic apical periodontitis. MATERIALS AND METHODS: An electronic search of the PubMed, Scopus, and Open-Grey databases was carried out from November 2022 to February 2023 and was later updated through July 2024..The risk of bias was assessed using the New Castle Ottawa scale. The quality of evidence was assessed using the Grading of Recommendations. Assessment, Development, and Evaluation. RESULTS: The initial search resulted in 968 records. Following the removal of duplicates and a review of titles and abstracts, 66 studies underwent full-texts analysis. Twenty studies were deemed eligible for inclusion. For the first outcome, a fixed-effects model was used. In a total of 7 studies with 193 participants, Spirochaetes were more prevalent in symptomatic apical periodontitis (p < .05) with a risk ratio of 1.91 [ 95% CI 1.25-2.92]. No significant difference was observed (p˃.05) in the prevalence of Bacteroidetes (12 studies with 451 participants) and Fusobacteriales (7 studies with 205 participants) in symptomatic vs asymptomatic apical periodontitis with a risk ratio of (.96), and (1.1), [95% CI, .78-1.19], and [95% CI, 0.84.1.44], respectively. The overall quality of evidence was low. CONCLUSIONS: The confirmation of the prevalence of Bacteroidetes and Fusobacteriales in symptomatic vs asymptomatic apical periodontitis remains uncertain. However, there is an association between Spirochetes and symptomatic apical periodontitis. Additional research is required to address the limitations of the current body of evidence. CLINICAL RELEVANCE: Identifying key pathogens in symptomatic apical periodontitis can help develop targeted interventions that address the underlying microbial causes.

Microbial diversity in primary endodontic infections: demographics and radiographic characteristics.

OBJECTIVE: To analyze if the microbiome community composition in primary endodontic infections is associated with clinical or radiographic factors. MATERIALS AND METHODS: Seventy-one patients with primary endodontic infections were evaluated for percussion tenderness, presence of a sinus tract, presence of caries, sex, probing depth > 4 mm, and age. Samples from the root canals were obtained and the microbiome was subsequently characterized by 16 S rRNA amplicon sequencing. For the radiographic analysis, a subset of 12 samples with a periapical index (PAI) ≤ 2 were compared with 19 samples with PAI of 5. The Shannon and Chao1 indices were used to measure alpha diversity. Differences in abundances of genera were evaluated using the Kruskal-Wallis test with Bonferroni's correction. Differences in community composition were evaluated using analysis of similarity (ANOSIM) with Bray-Curtis dissimilarity matrices. RESULTS: No significant differences in microbiome composition relative to clinical factors were found using ANOSIM. Teeth within the two categories of periapical index showed a similar number of species richness, and alpha diversity values P > 0.05. Community composition was significantly affected by the periapical index (ANOSIM P = 0.039, R = 0.10). Larger radiographic lesions demonstrated significant increase in Prevotellaceae, Olsenella, and the motile bacteria Oribacterium, Selenomonadaceae spp., and Treponema. CONCLUSION: Clinical factors associated with apical periodontitis have a limited impact on the root canal microbiome composition. Community composition appears to be affected in teeth with large apical lesions.

Efficacy of Glycyrrhizin as an Intracanal Medicament on Bacterial Load Reduction in Primary Infected Root Canals: A Randomized Clinical Trial.

AIM: This study was done to clinically investigate the efficacy of glycyrrhizin-based intracanal medication (ICM) compared to calcium hydroxide Ca(OH)2 and Ledermix on the bacterial counts inside the root canals of necrotic teeth with chronic periapical periodontitis. METHODOLOGY: Thirty-six patients having single-rooted and single-canaled necrotic teeth with chronic periapical periodontitis were enrolled in this research. Aseptic control measures were taken before clinical steps. Access cavity preparation was done. First bacteriological samples (S1) were collected immediately after access cavity preparation and before cleaning and shaping inside the root canals. Cleaning and shaping were performed on the root canals. Patients were randomly allocated into 3 groups according to the type of intracanal medicament used [Ca(OH)2, Ledermix, Glycyrrhizin]. Second bacteriological samples (S2) were collected after 1 week from placing the ICMs. S1 and S2 were transferred to the lab of microbiology for culturing on blood agar dishes in anaerobic conditions, and the bacteria on the plates were enumerated as colony-forming units (CFUs) by the manual counting method. The anti-bacterial efficacy of the ICM was estimated by the percentage reduction in the bacterial colonies from S1 to S2. RESULTS: All tested ICM documented a significant reduction in the CFUs from S1 to S2 (p < 0.05). Regarding S1, there was no significant difference between the three tested materials (p > 0.05). Regarding S2, Glycyrrhizin and Ledermix showed significantly lower CFUs than Ca(OH)2. There was no significant difference between Glycyrrhizin and Ledermix. CONCLUSION: Under the conditions of the present study, Glycyrrhizin and Ledermix showed comparable antimicrobial effects that were better than the commonly used Ca(OH)2 ICM. Glycyrrhizin may be a promising ICM. CLINICAL SIGNIFICANCE: Herbal medicine can be considered as an alternative antimicrobial material for root canal disinfection because it has many benefits, like low toxicity, absence of microbial resistance, and favorable antimicrobial efficacy. This study highlights the clinical efficacy of Glycyrrhizin as a promising ICM. How to cite this article: Eltantawi AR, Abdel-Razik GM, Elhawary YM, et al. Efficacy of Glycyrrhizin as an Intracanal Medicament on Bacterial Load Reduction in Primary Infected Root Canals: A Randomized Clinical Trial. J Contemp Dent Pract 2024;25(6):540-546.

Mustard Gel versus Chlorhexidine Gel as Root-Canal Medicament Against Enterococcus Faecalis: An in vitro Study.

OBJECTIVE: To evaluate the efficacy of mustard gel versus chlorhexidine (CHX) gel in treating the most resistant Enterococcus faecalis (E. faecalis) bacteria as intra-canal medications. STUDY DESIGN: Randomised controlled trial (in vitro study). Place and Duration of the Study: Department of Operative Dentistry, de'Montmorency College of Dentistry, in collaboration with Lahore General Hospital, between March and October 2021. METHODOLOGY: Forty single-rooted teeth, extracted and randomly allocated, were divided into two groups. Group 1 underwent treatment with CHX gel, while Group 2 was treated with Mustard gel. Subsequently, after disinfection, each tooth was horizontally divided into three equal parts. The middle section was standardised and inoculated with a controlled strain of E. faecalis. Intra-canal medicaments were administered for seven days. Bacterial colonies were quantified as colony forming units (CFU/mL) and subjected to analysis using Statistical Package for Social Sciences (SPSS) version 25. RESULTS: The median bacterial count observed was 1,550 (range 775 - 2,500) for Group 1, contrasting with 400 (range 200 - 775) for Group 2, indicating a significant difference in bacterial count between the groups (p <0.001). CONCLUSION: Mustard gel demonstrated significantly superior efficacy compared to chlorhexidine gel against E. faecalis in root canal procedures. KEY WORDS: Chlorhexidine, Enterococcus faecalis, Mustard, Root canal medicament, Antimicrobial drug-resistance, Root canal irrigants.

Effect of Active Oxygen Fluid (Blue®m) as a Root Canal Irrigant Against Enterococcus Faecalis.

PURPOSE: To evaluate the antimicrobial effect of a new active oxygen fluid (Blue®m) as a root canal irrigant against Enterococcus faecalis compared to sodium hypochlorite (NaOCl). MATERIAL AND METHODS: Forty-five extracted single-canaled human teeth were selected, received root canal preparation, autoclaved, and contaminated with Enterococcus faecalis. The specimens were randomly allocated into three groups: Group (A) served as the negative control, receiving irrigation with saline (n = 15); Group (B) was irrigated with 5.25% NaOCl (n = 15); and Group (C) was irrigated with 10 mL of Blue®m (n = 15). Microbial sampling from the root canals was performed before and after irrigation. The difference between the pre-irrigation and post-irrigation colony-forming units (CFU/mL) was calculated. The data was analysed using a one-way ANOVA followed by post-hoc Tukey tests. The significance level was set at 5%. RESULTS: Blue®m statistically significantly reduced the bacterial load compared to saline (p = 0.009), but NaOCl was most effective, outperforming both (p 0.0001). CONCLUSION: Irrigation with Blue®m demonstrated antibacterial efficacy against Enterococcus faecalis, but it was not as effective as NaOCl.

Comparative analysis of hydrophobicity and dentin adhesion ability in Candida albicans strains.

OBJECTIVE: Adhesion to dentin is a first step for a successful microbial root canal colonization. Cell hydrophobicity seems to have some influence in the Candida species adhesion to surfaces. To measure cell surface hydrophobicity and to investigate the adherence ability to human dentin among Candida albicans strains isolated from root canal and lingual dorsum via an in vitro study. METHODOLOGY: adhesion was quantified in function of dentin area covered by blastospores and/or hyphae presence detected by epifluorescence microscope. Cell surface hydrophobicity was estimated by assessing the percentage migration of cells from an aqueous phase to a hydrocarbon phase. Contact angles were measured by the sessile drop technique on the dentin surface using a contact angle measurements apparatus. We also examined the correlation between adhesion ability and hydrophobicity. RESULTS: although there was some intra-species variation in cell surface hydrophobicity, most isolates were characterized by moderate hydrophobicity. There was no significant difference in this parameter when the isolation niche was considered. Both root canal and lingual dorsum yeasts were able to adhere to dentin. No association was found between the strains' site of isolation and adhesion. Moreover, cell surface hydrophobicity and adhesion ability were not correlated. CONCLUSION: although hydrophobicity can influence Candida albicans virulence in many ways, this study suggests that this parameter by itself was not a good predictor of adhesion to dentin.

Optimized protocol for collecting root canal biofilms for in vitro studies.

Endodontic retreatment is often necessitated by several factors, including the persistence of microorganisms in the root canal system (RCS). Their complex organization in biofilms increases their pathogenic potential, necessitating new disinfection strategies. This study aimed to standardize a new in vitro protocol for collecting biofilm from the RCS. Thirty-four bovine incisors were used in the study, divided into two experimental groups with two collection steps each: (a) biofilm collection protocol and (b) absorbent paper points protocol. Twelve specimens from each group were selected for counting colony-forming units (CFUs), while eight specimens were prepared for scanning electron microscopy (SEM). Two additional specimens served as sterilization controls to ensure that experiments were free of contamination. The coronal region was removed and standardized at 15 mm. After preparation with ProTaper up to F5, the apical foramen was sealed with composite resin, and the roots were stabilized with acrylic resin in 1.5-mL Eppendorf tubes. The specimens were sterilized and inoculated with Enterococcus faecalis NTCT 775 every 24 h for 21 days. After this period, each group underwent biofilm collection protocols, and CFU and scanning electron microscopy (SEM) data were analyzed. The Shapiro-Wilk test was performed to assess the normality of log-transformed data, and the results indicated a normal distribution for all groups, allowing parametric testing. The Levene test was used to evaluate the equality of variances. The proposed biofilm collection method yielded significantly higher CFU counts compared with the absorbent paper points method, particularly when analyzed on a log10 scale. An independent samples t-test confirmed a statistically significant difference between the two methods (p < 0.0001). The proposed protocol achieved an efficiency rate of 95.85 % ± 1.15 %, whereas the absorbent paper points protocol yielded a lower efficiency of 5.46 % ± 1.37 %. Therefore, the biofilm collection protocol proposed in this study proved to be more effective for biofilm removal from the RCS.

Effectiveness of supplementary antimicrobial procedures in disinfecting lateral canals as evaluated by a novel ex vivo analytical approach.

This ex vivo study devised an analytical ex vivo method for infection/disinfection of simulated lateral canals located in the middle and apical segments of the root. The antibacterial effects of supplementary approaches were tested in this model. Extracted mandibular premolars had their main root canals enlarged and then two lateral canals (100 µm in diameter) were created in the root, one in the apical and the other in the middle portion. Micro-computed tomography was used for specimen selection and to confirm the quality of the simulated ramifications. The specimens were contaminated with a mixed bacterial culture from subgingival bacterial biofilm added to pure Enterococcus faecalis strain ATCC 29212 grown overnight, using special strategies to facilitate culture medium penetration within the lateral canals. The following procedures were tested for disinfection: NaOCl/passive ultrasonic irrigation (PUI), NaOCl/XP-endo Finisher, ozonated water/continuous ultrasonic irrigation (CUI), and NaOCl/conventional irrigation with 30-G needles (control). Bacteriological samples were taken from the main canal before (S1) and after (S2) each supplementary protocol, and also from each lateral canal after treatment (S3). DNA extracted from the samples was subjected to quantitative real-time polymerase chain reaction. All S1 main canal samples were positive for bacterial presence. Bacterial counts in the main root canal substantially decreased by 99.2% after PUI, 99.1% after ozone/CUI, 99% after XP-endo Finisher, and 96% in the control group (P < 0.01 for all groups). There were no significant differences between groups (P > 0.05). The same was observed when comparing the effects of the supplementary approaches in the apical and middle lateral canals (P > 0.05). Only a few lateral canals showed no detectable bacteria. The method proposed here proved effective for ex vivo infection/disinfection studies. All supplementary approaches induced a substantial bacterial reduction in the main canal, with no significant differences between them. However, in terms of lateral canal disinfection, none of the tested approaches showed significant effects when compared to the control group.

Strategies and mechanisms targeting Enterococcus faecalis biofilms associated with endodontic infections: a comprehensive review.

Enterococcus faecalis is one of the main microorganisms that infects root canals, ranking among the most prevalent microorganisms associated with endodontic treatment failure. Given its pervasive presence in persistent endodontic infections, the successful elimination of Enterococcus faecalis is crucial for effective endodontic treatment and retreatment. Furthermore, Enterococcus faecalis can form biofilms - defense structures that microbes use to fight environmental threats. These biofilms confer resistance against host immune system attacks and antibiotic interventions. Consequently, the presence of biofilms poses a significant challenge in the complete eradication of Enterococcus faecalis and its associated disease. In response, numerous scholars have discovered promising outcomes in addressing Enterococcus faecalis biofilms within root canals and undertaken endeavors to explore more efficacious approaches in combating these biofilms. This study provides a comprehensive review of strategies and mechanisms for the removal of Enterococcus faecalis biofilms.

Clinical Investigation of Bacteriome in Primary Endodontic Infections With Apical Periodontitis Using High-Throughput Sequencing Analysis.

INTRODUCTION: This study characterized the bacteriome in primary endodontic infection (PEI) with apical periodontitis (AP), identified core and rare bacteriome species and community diversity metrics, and analyzed the relationship between the bacteriome composition, diversity and features, and patient variables. METHODS: Twenty-seven patients with PEI and AP were sampled. The DNA was extracted and quantified using quantitative polymerase chain reaction. Raw V3-V4 amplicon sequencing data were processed with the DADA2 pipeline to generate amplicon sequence variants, and taxonomic assignment of the amplicon sequence variants up to the species level was done against the Human Oral Microbiome Database. Core bacteriome and differential abundance analyses were performed using ANCOM. Alpha diversity was determined using Chao1, Shannon, and Simpson indexes. LeFse analysis was used to identify abundant taxa. Sparse Estimation of Correlations among Microbiomes analysis estimated linear and nonlinear relationships among bacteria. RESULTS: Of 27, 24 root canal samples were analyzed, and 3 root canal sampling were filtered out with a low read count. The bacterial phyla with top mean relative abundance were Bacteroidetes, Firmicutes, Synergistetes, Fusobacteria, and Actinobacteria. A total of 113 genera and 215 species were identified. The samples were gathered into 3 clusters. LefSe analysis identified differences in abundant taxa between distinct age, gender, symptomatology, and lesion size groups. Sparse Estimation of Correlations among Microbiomes distance analysis indicated Slackia exigua as the node with the highest degree. CONCLUSIONS: The bacteriome in PEI with AP among the patients in this study was complex and displayed high microbial heterogeneity. Moreover, age, gender, symptomatology, and lesion size were associated with differences in bacteriome features in PEI with AP.

Comparative Evaluation of the Efficacy of ProTaper & Mtwo Rotary Retreatment System and Conventional Hand H-files for Removal of Gutta-percha from Root Canal - A Stereomicroscopic Study.

BACKGROUND: Root canal retreatment aims to eliminate or substantially reduce the microbial load from the root canal and removal of previous root filling materials is the primary goal of nonsurgical retreatment. AIM: To comparatively evaluate the effectiveness of ProTaper retreatment files, Mtwo retreatment files, and Hedstrom files along with Gates Glidden (GG) Drill by using a stereomicroscope for the detection of remaining gutta-percha after root canal retreatment using a stereomicroscope. MATERIALS AND METHODS: Forty-five test samples were divided into three experimental groups (15 samples each), such as Group A-ProTaper retreatment files; Group B-Mtwo retreatment files; Group C-Hedstrom files along with Gates Glidden Drill. After routine endodontic treatment in the test samples, gutta-percha was removed from the test samples as per the division of different retreatment file systems. Further, test samples were divided longitudinally and were evaluated under the stereomicroscope. Stereomicroscopic images were captured and analysed under Image Analyser Software-Image J. STATISTICAL ANALYSIS: One-way ANOVA test was used for comparison of gutta-percha removal between the groups and Tukey's post hoc test was used for pairwise comparisons. RESULTS: The least remaining filling material was seen in specimens retreated with the ProTaper universal system. The highest remaining filling material was seen in specimens retreated with Hedstrom file with Gates Glidden Drill. CONCLUSION: All experimental test techniques left gutta-percha remnants within the root canal. ProTaper universal rotary retreatment system proved to be an efficient method of removing gutta-percha and left relatively less volume in comparison with other test systems.

α-IRAK-4 Suppresses the Activation of RANK/RANKL Pathway on Macrophages Exposed to Endodontic Microorganisms.

Periapical lesions are common pathologies affecting the alveolar bone, often initiated by intraradicular lesions resulting from microbial exposure to dental pulp. These microorganisms trigger inflammatory and immune responses. When endodontic treatment fails to eliminate the infection, periapical lesions persist, leading to bone loss. The RANK/RANKL/OPG pathway plays a crucial role in both the formation and the destruction of the bone. In this study, the objective was to inhibit the RANK/RANKL pathway in vitro within exposed Thp-1 macrophages to endodontic microorganisms, specifically Enterococcus faecalis, which was isolated from root canals of 20 patients with endodontic secondary/persistent infection, symptomatic and asymptomatic, and utilizing an α-IRAK-4 inhibitor, we introduced endodontic microorganisms and/or lipoteichoic acid from Streptococcus spp. to cellular cultures in a culture plate, containing thp-1 cells and/or PBMC from patients with apical periodontitis. Subsequently, we assessed the percentages of RANK+, RANKL+, and OPG+ cells through flow cytometry and measured the levels of several inflammatory cytokines (IL-1ß, TNF-α, IL-6, IL-8, IL-10, and IL-12p70) in the cellular culture supernatant through a CBA kit and performed analysis by flow cytometry. A significant difference was observed in the percentages of RANK+RANKL+, OPG+ RANKL+ cells in thp-1 cells and PBMCs from patients with apical periodontitis. The findings revealed significant differences in the percentages of the evaluated cells, highlighting the novel role of the IRAK-4 inhibitor in addressing this oral pathology, apical periodontitis, where bone destruction is observed.

Comparative evaluation of selected concentrations of sodium hypochlorite on the outcome of endodontic therapy among Ghanaians.

BACKGROUND: Endodontic treatment is one of the main dental treatments to manage inflamed or infected root canal systems of teeth. The success of endodontic treatment principally depends on eradicating microorganisms in the root canal by chemo-mechanical debridement with irrigation solutions like sodium hypochlorite (NaOCl). NaOCl has been used in concentrations ranging from 0.5% to 5.25%. This study determined the antimicrobial effectiveness of selected concentrations (0.5%, 1.0%, 2.6%, and 5.2%) of NaOCl in endodontic treatment. METHODS: The study sites were the University of Ghana Dental School (UGDS) and Noguchi Memorial Institute for Medical Research (NMIMR). Sixty infected single-rooted single-canal teeth were used. Before (S1) and after (S2), root canal samples during the endodontic treatment with the selected concentrations of NaOCl were examined via anaerobic and aerobic cultures. The isolates were identified using Matrix Assisted Laser Desorption Ionization-Time Of Flight Mass Spectrometry (MALDI-TOF MS). RESULTS: All S1 samples were positive for cultivable bacteria. Fifty-three (53) different microbial species belonging to 20 different microbial genera were isolated. Streptococcus viridans was the most frequently isolated microbe. There were zero isolates in the root canals irrigated with 2.6% and 5.2% NaOCl. Two teeth had isolates in the groups irrigated with the lower concentrations (0.5% and 1.0%) of NaOCl. The persistent bacteria were one species each of Streptococcus mitis and Streptococcus oralis, respectively. CONCLUSION: Root canal treatments using chemo-mechanical preparation with the selected concentrations (0.5%, 1.0%, 2.6%, and 5.2%) of NaOCl were effective in significantly reducing the microbial load, and for the 5.2% and 2.6% concentrations, in eliminating all the microorganisms.

Molecular analyses indicate profuse bacterial diversity in primary and post- treatment endodontic infections within a cohort from the United Arab Emirates-A preliminary study.

OBJECTIVE: Endodontic microbiota appears to undergo evolutionary changes during disease progression from inflammation to necrosis and post-treatment. The aim of this study was to compare microbiome composition and diversity in primary and post-treatment endodontic infections from a cohort of patients from the UAE. DESIGN: Intracanal samples were collected from primarily infected (n = 10) and post-treatment infected (n = 10) root canals of human teeth using sterile paper points. Bacterial DNA was amplified from seven hypervariable regions (V2-V4 and V6-V9) of the 16S rRNA gene, then sequenced using next-generation sequencing technology. The data was analyzed using appropriate bioinformatic tools. RESULTS: Analyses of all the samples revealed eight major bacterial phyla, 112 genera and 260 species. Firmicutes was the most representative phylum in both groups and was significantly more abundant in the post-treatment (54.4%) than in primary (32.2%) infections (p>0.05). A total of 260 operational taxonomic units (OTUs) were identified, of which 126 (48.5%) were shared between the groups, while 83 (31.9%) and 51 (19.6%) disparate species were isolated from primary and post-treatment infections, respectively. A significant difference in beta, but not alpha diversity was noted using several different indices (p< 0.05). Differential abundance analysis indicated that, Prevotella maculosa, Streptococcus constellatus, Novosphigobium sediminicola and Anaerococcus octavius were more abundant in primary infections while Enterrococcus faecalis, Bifidobacterium dentium, Olsenella profusa and Actinomyces dentalis were more abundant in post-treatment infections (p <0.05). CONCLUSION: Significant differences in the microbiome composition and diversity in primary and post-treatment endodontic infections were noted in our UAE cohort. Such compositional differences of microbiota at various stages of infection could be due to both intrinsic and extrinsic factors impacting the root canal ecosystem during disease progression, as well as during their therapeutic management. Identification of the key microbiota in primarily and secondarily infected root canals can guide in the management of these infections.

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 sodium hypochlorite irrigation with 3 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 3 groups (n = 18/group). The canals were contaminated with a mixed bacterial culture for 30 days and then subjected to preparation using 2.5% sodium hypochlorite 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 2 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. The amount of accumulated hard tissue debris was evaluated by micro-CT. RESULTS: A substantial reduction in bacterial counts was observed in all 3 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 < .01). No significant differences were observed between groups for bacterial reduction in the full and apical canal (P > .05). There were no intergroup differences in the amount of accumulated hard tissue debris either (P > .05). The open-ended needle group showed significantly more specimens with quantitative real-time polymerase chain reaction negative results for bacteria in S3 than TruNatomy (P < .05). CONCLUSION: When used up to an appropriate insertion depth, the 3 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 Effectiveness of Three Different Irrigant Activation Methods in the Elimination of Enterococcus faecalis from Root Canals.

Background and Objective: Eliminating intracanal Enterococcus faecalis (E. faecalis) is challenging because of its ability to penetrate deep dentinal tubules and its high resistance to many chemicals. This study evaluated the effectiveness of conventional needle irrigation and three different irrigant activation methods in reducing E. faecalis. Methods: The root canals of extracted teeth were shaped, contaminated with E. faecalis, and incubated for three weeks. They were randomly allocated to four experimental groups of 15 teeth each according to the final irrigation method: group 1, conventional needle irrigation; group 2, passive ultrasonic (PU) irrigation; group 3, XP-endo Finisher (XPF); and group 4, laser-activated (LA) irrigation. Bacterial samples were taken and cultured before and after these final irrigation procedures. The colony-forming units were counted, and the bacterial reduction percentages of each group were calculated. The Kruskal-Wallis and Dunnet tests were used for statistical analysis. Results: All irrigant activation methods were significantly more effective than conventional needle irrigation. Although the LA group generated more negative samples than PU, there was no statistically significant difference between the LA and PU groups. LA was significantly more effective than the XPF, whereas PU and XPF were statistically similar. Conclusions: Within the limitations of this study, the final irrigation with LA and PU showed the best reductive effect on E. faecalis colonies. Considering that the LA group had more negative samples, it may be chosen as an alternative to enhance root canal disinfection, especially in difficult cases.

Revision of ex vivo endodontic biofilm model using computer aided design.

OBJECTIVE: Most endodontic diseases are bacterium-mediated inflammatory or necrotic process induced by contaminated dental pulp. Although great advances are being performed to obtain more efficient antibacterial strategies for persistent infections, most studies lack of representative models to test their antibacterial effects and their outcomes cannot be promptly translated to clinical practice. Therefore, this study aimed to refine an ex vivo endodontic biofilm model combining human tooth, computer guided design and 3D printing to obtain a more reproducible and predictable model. METHODS: Monoradicular teeth were cut using three different methods: hand-held (HCC), mechanical precision (MPC) and computer aid guided cutting (CGC). Then, blocks were reassembled. The different model preparations were assessed in terms of dimensional tolerance, surface analysis, liquid tightness and Enterococcus faecalis biofilm development for 21 days, which was studied by metabolic assays and confocal microscopy. Then, the proposed model was validated using different commercial disinfecting treatments. RESULTS: CGC exhibited significantly lower deviation and surface without defects compared to HHC and MPC, leading to superior liquid tightness. Similarly, mature biofilms with high metabolic activity and vitality were observed in all conditions, CGC showing the lowest variation. Regarding the model validation, all antibacterial treatments resulted in the complete eradication of bacteria in the standard 2D model, whereas commercial treatments exhibited varying levels of efficacy in the proposed ex vivo model, from moderately reduction of metabolic activity to complete elimination of biofilm. CONCLUSIONS: The novel guided approach represents a more reliable, standardized, and reproducible model for the evaluation of endodontic disinfecting therapies. CLINICAL SIGNIFICANCE: During antibacterial treatment development, challenging 3D models using teeth substrates to test antibacterial treatments novel guided approach represents a more reliable, standardized, and reproducible model for the evaluation of endodontic disinfecting therapies.

Antimicrobial Efficacy and Dentin Collagen Damage Caused by Calcium Hypochlorite and Sodium Hypochlorite.

This study aimed to evaluate the antimicrobial activity of calcium hypochlorite (Ca (OCl)2) and sodium hypochlorite (NaOCl) using confocal laser scanning microscopy (CLSM) and dentin organic matrix alteration by picrosirius staining and light microscopy (LM). Samples of human extracted teeth were infected with Enterococcus faecalis by centrifugation of the bacterial suspension and were treated with Ca(OCl)2 or NaOCl at 0.5%, 2.5%, and 6% for 15, 30, and 60 seconds. CLSM and viability staining were used to quantitatively analyze the proportions of dead/live bacteria in the canal lumen and border of the root canal. The data were analyzed by ANOVA and Fisher test. For LM analysis, one hundred bovine teeth were randomly divided into 10 test groups (n=10): G1- Without treatment; G2- 17% EDTA; G3- 6% NaOCl; G4- 6% NaOCl + EDTA; G5- 0.5% Ca(OCl)2; G6- 0.5% Ca(OCl)2 + EDTA; G7- 2.5% Ca(OCl)2; G8- 2.5% Ca(OCl)2 + EDTA; G9- 6% Ca(OCl)2; G10- 6% Ca(OCl)2 + EDTA. The samples were fragmented and stained with Picrosirius. Data were analyzed by Kruskal-Wallis and Dunn (P<0.05). There was a strong correlation between the results of the canal lumen and the border of the root canal (r=0.962). Both hypochlorites at a concentration of 0.5% showed less microbial reduction compared to 2.5% and 6% (P<0.05). There was less antimicrobial activity at 15 seconds compared to 30 and 60 seconds (P<0.05). Ca(OCl)2 and NaOCl showed similar results at the same concentrations (P>0.05). In conclusion, Ca(OCl)2 caused fewer alterations to the dentin organic matrix at concentrations of 0.5% and 2.5%. Ca(OCl)2 presents antimicrobial activity similar to NaOCl, and collagen damage is concentration-dependent.

Prevalence of species of yellow, purple and green microbial complexes in endo-perio lesions: a systematic review.

This review aimed to determine the prevalence of species of yellow, purple and green microbial complexes in root canals (RC) and periodontal pockets (PP) of teeth with endodontic-periodontal lesions. For this purpose, two reviewers searched the literature up to January 2022. Studies reporting the prevalence of species of the yellow, purple and green microbial complexes in teeth diagnosed with endodontic-periodontal lesions were included. The risk of bias of the included studies was assessed using the 14 criteria from the NIH Quality Assessment Tool. Of 1,611 references identified in the initial search, only four studies were eligible and included in the qualitative analysis. The profile and prevalence rates of bacterial species in RC and PP varied among the included studies: levels of Agregatibacter actinomycetemcomitans (12% RC, 58% PP), Capnocytophaga granulosa (10% RC, 35% PP), Capnocytophaga sputigena (15-70% RC, 0-30% PP), Streptococcus mitis (30% RC, 35% PP), Streptococcus sanguinis (30% RC, 35% PP), and Veillonella parvula (70% RC, 50% PP) were identified. The high methodological heterogeneity prevented grouping and quantitative analysis of data. The risk of bias was considered 'moderate' for all studies. The included studies identified the presence of seven bacterial species belonging to the yellow, purple, and green microbial complexes in RC and PP, but with different prevalence rates. Future clinical studies are encouraged to investigate the presence and role of these species in the occurrence and development of endodontic-periodontal lesions.

Effect of Disinfection Protocols on Bacterial Reduction in Mandibular Molars.

OBJECTIVE: Evaluate the bacterial reduction effect of disinfection protocols used in lower molars infected by Enterococcus faecalis. METHODS: Eighty extracted lower molars were prepared and inoculated with E. faecalis for 2 months. The teeth were then distributed into 2 control groups (n = 4) and 4 experimental groups (n = 18) in function of the disinfection protocol utilized: G1. WaveOne Gold (WOG), passive ultrasonic irrigation (PUI) and Ultracal; G2. WOG, PUI and calcium hydroxide (CH); G3. WOG, XP-Endo Finisher (XPF) and Ultracal; G4. WOG, XPF and CH. Bacteriological samples were collected previously (S1), after preparation (S2), final agitation (S3) and intracanal dressing (S4). Microbial growth was assessed according to culture turbidity and UV spectrophotometry. Statistical analyses used the Friedman test for paired samples and Kruskal-Wallis test for non-paired data (p<0.05). RESULTS: No protocol eliminated E. faecalis effectively. The S2, S3 and S4 samples were statistically different from the S1 samples in G1, G2, G3 and G4 (p<0.05). Statistical differences were observed in bacterial reduction between G1 and G2 and G1 and G3 after the intracanal medicament (S4) (p<0.05). CONCLUSION: The WOG + PUI + CH disinfection protocol showed higher percentages of bacterial reduction.