Is there any association between the presence of biomarkers and apical periodontitis? A systematic review.

This systematic review aimed to verify whether there is evidence of an association between apical periodontitis and the presence of systemic biomarkers. This study adhered to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses - PRISMA. For this, the acronym PECO was used; population (P) of adult humans exposed (E) to the presence of apical periodontitis, compared (C) to adult humans without apical periodontitis, and the outcome (O) of the presence of biomarkers was observed. The articles were searched in PubMed, Scopus, Web of Science, LILACS, Cochrane Library, OpenGray, and Google Scholar grey databases. Subsequently, studies were excluded based on title, abstract, and full article reading, following the eligibility criteria. The methodological quality of the selected studies was evaluated using the Newcastle-Ottawa qualifier. After exclusion, 656 studies were identified, resulting in 17 final articles that were divided into case-control, cross-sectional, and cohort studies. Eight studies were considered to have a low risk of bias, one had a medium risk of bias, and eight had a high risk of bias. In addition, 12 articles evaluated biomarkers in blood plasma, four evaluated them in saliva, and only one evaluated them in gingival crevicular fluid. The results of these studies indicated an association between apical periodontitis and the systemic presence of biomarkers. These markers are mainly related to inflammation, such as interleukins IL-1, IL-2, and IL-6, oxidative markers, such as nitric oxide and superoxide anions, and immunoglobulins IgG and IgM. Systematic review registration: https://www.crd.york.ac.uk/prospero/, identifier (CRD42023493959).

Nanoarchitectonics-Based Materials as a Promising Strategy in the Treatment of Endodontic Infections.

Endodontic infections arise from the interactive activities of microbial communities colonizing in the intricate root canal system. The present study aims to update the latest knowledge of nanomaterials, their antimicrobial mechanisms, and their applications in endodontics. A detailed literature review of the current knowledge of nanomaterials used in endodontic applications was performed using the PubMed database. Antimicrobial nanomaterials with a small size, large specific surface area, and high chemical activity are introduced to act as irrigants, photosensitizer delivery systems, and medicaments, or to modify sealers. The application of nanomaterials in the endodontic field could enhance antimicrobial efficiency, increase dentin tubule penetration, and improve treatment outcomes. This study supports the potential of nanomaterials as a promising strategy in treating endodontic infections.

Microbiota Association and Profiling of Gingival Sulci and Root Canals of Teeth with Primary or Secondary/Persistent Endodontic Infections.

INTRODUCTION: Microbiota associated with primary endodontic infection (PEI) and secondary/persistent endodontic infection (SPEI) must be characterized to elucidate pathogenesis in apical periodontitis and bacterial biomarkers identified for diagnostic and therapeutic applications. METHODS: This study analyzed the microbial community profiles of root canals and gingival sulci (sulcus-E) for teeth with PEI (n = 10) or SPEI (n = 10), using the Illumina MiSeq platform. Bacterial samples from gingival sulci (sulcus-C) of healthy contralateral teeth served as controls. RESULTS: There were 15 phyla, 177 genera, and 340 species identified. The number and diversity of bacteria in root canals did not differ significantly between PEI and SPEI. Proteobacteria, Firmicutes, Fusobacteria, Bacteroidetes, and Actinobacteria were the dominant phyla in both groups. At the genus level, Lancefieldella, Bifidobacterium, Stomatobaculum, and Schaalia were enriched in root canals with SPEI. Of significance, Lancefieldella was observed in both root canals and sulcus-E of teeth with SPEI. At the species level, Neisseria macacae, Streptococcus gordonii, Bifidobacterium dentium, Stomatobaculum longum, and Schaalia odontolytica were increased significantly in root canals with SPEI compared to PEI. Oribacterium species, Streptococcus salivarius, Lancefieldella parvula, Prevotella denticola, and Oribacterium asaccharolyticum were more abundant in sulcus-E of teeth with SPEI compared to PEI. CONCLUSIONS: There were distinctive and differing predominant bacterial species associated with the root canals and gingival sulci between teeth with PEI and SPEI. Specific bacteria identified in sulcus-E and root canals of teeth with SPEI could serve as noninvasive diagnostic biomarkers for detecting SPEI.

Antibiotic Over-Prescription by Dentists in the Treatment of Apical Periodontitis: A Systematic Review and Meta-Analysis.

After pulp infection and necrosis, the passage of microbial antigens into the periapical space causes apical periodontitis (AP). Most of the clinical forms of AP can be managed without prescribing antibiotics, only with root canal treatment and abscess drainage or, where appropriate, tooth extraction. However, the scientific literature provides evidence of inappropriate antibiotic prescriptions by dentists in the management of apical disease. OBJECTIVES: The aim of this systematic review and meta-analysis was to analyze the global pattern of antibiotic prescription in the treatment of apical disease. METHODS: PRISMA Guidelines were followed to carry out this systematic review. The research question was as follows: What is the pattern of antibiotic prescription by dentists in the treatment of the different clinical forms of apical periodontitis? A systematic search was conducted on MEDLINE/PubMed, Wiley Online Database, Web of Science and Scopus. All studies reporting data about the pattern of antibiotic prescription by dentists in the treatment of apical disease were included. The meta-analyses were calculated using the Open Meta Analyst version 10.10 software. Random-effects meta-analyses were performed. The risk of bias was assessed using the Newcastle-Ottawa Scale. The certainty of evidence was assessed using GRADE. RESULTS: The search strategy identified 96 articles and thirty-nine cross-sectional studies fulfilled the inclusion criteria. The overall percentage of antibiotic prescriptions by dentists in cases of symptomatic AP was 25.8%, and 31.5% in cases of asymptomatic AP with sinus tract present. The percentage of dentists prescribing antibiotics in cases of acute apical abscess with no/mild symptoms was 47.7%, whereas, in cases of acute apical abscess with moderate/severe symptoms, 88.8% of dentists would prescribe antibiotics. Endodontists prescribe antibiotics at a lower rate than general practitioners. The total risk of bias was considered moderate, and the final rating for the certainty of the evidence was low. CONCLUSIONS: Dentists worldwide are over-prescribing antibiotics in the management of apical disease. It is necessary to improve antibiotic prescribing habits in the treatment of endodontic infections, as well as educational initiatives to encourage the rational and appropriate prescription of antibiotics in periapical diseases.

Unveiling Therapeutic Potential: Targeting Fusobacterium nucleatum's Lipopolysaccharide Biosynthesis for Endodontic Infections-An In Silico Screening Study.

Complex microbial communities have been reported to be involved in endodontic infections. The microorganisms invade the dental pulp leading to pulpitis and initiating pulp inflammation. Fusobacterium nucleatum is a dominant bacterium implicated in both primary and secondary endodontic infections. Drugs targeting the molecular machinery of F. nucleatum will minimize pulp infection. LpxA and LpxD are early acyltransferases involved in the formation of lipid A, a major component of bacterial membranes. The identification of leads which exhibit preference towards successive enzymes in a single pathway can also prevent the development of bacterial resistance. A stringent screening strategy utilizing physicochemical and pharmacokinetic parameters along with a virtual screening approach identified two compounds, Lomefloxacin and Enoxacin, with good binding affinity towards the early acyltransferases LpxA and LpxD. Lomefloxacin and Enoxacin, members of the fluoroquinolone antibiotic class, exhibit wide-ranging activity against diverse bacterial strains. Nevertheless, their effectiveness in the context of endodontic treatment requires further investigation. This study explored the potential of Lomefloxacin and Enoxacin to manage endodontic infections via computational analysis. Moreover, the compounds identified herein serve as a foundation for devising novel combinatorial libraries with enhanced efficacy for endodontic therapeutic strategies.

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].

Effect of supplementary sodium hypochlorite agitation techniques on an ex vivo oral multispecies biofilm during passive disinfection of simulated immature roots.

AIM: To compare the effect of different sodium hypochlorite (NaOCl) agitation techniques on an ex vivo oral multispecies biofilm during passive disinfection of simulated immature roots. METHODOLOGY: Extracted human teeth were prepared to simulate immature roots. They were infected with a dental plaque-derived multispecies biofilm and cultured for 14 days. The roots were randomly designated into four groups: (1) negative control (PBS), (2) 1.5% NaOCl (CNI), (3) CNI + Ultrasonic activation (UA), (4) CNI + EasyClean agitation (ECA), (5) CNI + XP-endo finisher agitation (XPF), and (6) positive control (6% NaOCl). Biofilm samples were collected from the root canals and used to determine the number of viable cells (colony-forming units), scanning electron microscopy, and 16S rRNA gene sequencing. The mean colony-forming units per mL (CFU/mL) were analysed using One-way anova. 16S rRNA sequencing data were analysed for alpha (observed OTUs, Shannon index, and Chao1) and beta diversity (Bray-Curtis dissimilarities). The LEfSe analysis was used to determine the effect of treatment procedures on the abundance of root canal microbiota. The significance was set at .05. RESULTS: PBS and CNI samples had significantly higher CFU/mL counts than UA, ECA, XPF, and 6% NaOCl samples (p < .05). The pre-treatment, PBS, and CNI groups had significantly greater alpha diversity than the UA, ECA, XPF, and 6% NaOCl groups (p < .05). NaOCl agitation groups and the 6% NaOCl group achieved a more pronounced reduction in bacteria from the genera Fusobacterium, Actinomyces, Porphyromonas, and Capnocytophaga. CONCLUSIONS: The effectiveness of passive disinfection protocols was enhanced by NaOCl agitation techniques, suggesting that this supplementary method can improve the outcome of revitalization procedures.

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.

Photoinactivation of Enterococcus faecalis Biofilm: In Vitro Antimicrobial Effect of Photoexcited Rutin-Gallium(III) Complex via Visible Blue Light.

INTRODUCTION: Endodontic infection is a common problem that can result in tooth loss if not effectively treated. This study focused on investigating the use of rutin-gallium (Ga)(III) complex-mediated antimicrobial photodynamic therapy (aPDT) for the photoinactivation of Enterococcus faecalis biofilm. METHODS: The minimum biofilm eradication concentration of the rutin-Ga(III) complex and the minimum biofilm eradication dose of light-emitting diode against E. faecalis were evaluated. The antimicrobial effect of rutin-Ga(III) complex-mediated aPDT against E. faecalis was assessed. Additionally, the expression of genes associated with E. faecalis virulence, such as ace, gelE, and esp, as well as the production of reactive oxygen species within the cells were evaluated. RESULTS: The minimum biofilm eradication concentration of the rutin-Ga(III) complex was determined to be 25 µmol/L, whereas the minimum biofilm eradication dose of light-emitting diode irradiation was defined as 5 minutes with an energy density of 300-420 J/cm2. Rutin-Ga(III) complex-mediated aPDT demonstrated a significant dose-dependent reduction in the growth of E. faecalis biofilms. Moreover, aPDT led to increased intracellular reactive oxygen species generation in treated E. faecalis cells. Furthermore, the messenger RNA levels of ace, gelE, and esp genes were significantly down-regulated in E. faecalis treated with rutin-Ga(III) complex-mediated aPDT (P < .05). CONCLUSIONS: Rutin-Ga(III) complex-mediated aPDT effectively reduces E. faecalis biofilm growth by disrupting biofilm structure and down-regulating virulence genes. These findings highlight the potential of aPDT with the rutin-Ga(III) complex as an adjuvant therapeutic approach against E. faecalis biofilms.

Bacterial diversity in primary infected root canals of a Chinese cohort: analysis of 16 S rDNA sequencing.

PURPOSE: To characterize the bacterial community in the primarily infected root canals. METHODS: A total of 13 samples were collected from the primarily infected root canals. 16 S rDNA sequencing was performed to define bacterial community. Taxonomic annotation, bacterial hierarchical structures, community richness and diversity, and inter-subject variability of the bacterial community in the root canal samples were analyzed. Gender, age, and duration of the toothache-specific bacterial community associated with the patient groups were analyzed. RESULTS: A total of 359 Species were annotated and identified in the whole study cohort. The Alpha diversity analysis showed that the species diversity and detection rate of the 13 samples were high, which reflected the authenticity of sequencing results. The Beta diversity analysis was used to compare the degree of difference between different root canal samples. The 13 samples were divided into two groups according to the results, group A was samples I1-I12, and group B was samples I13. The bacterial species of group A samples were analyzed with the clinical characteristics of patients, and it was found that gender, and duration specific differences in bacterial species, and there was no significant difference in species types among different ages of patients. CONCLUSION: There were a wide diversity and inter-subject variability in the bacterial community in the primary infected root canals. While Porphyromonas gingivalis was the most abundant species, Fusobacterium nucleatum was the most variable species in the bacterial community of the root canal. The bacterial community at different taxonomic levels varied from sample to sample, despite consistent disease diagnoses. There was gender, duration-specific differences in the bacterial species in the primary infected root canals.

Asymptomatic Apical Periodontitis Lesions and Their Association With Systemic Inflammatory Burden: A Preliminary Prospective Clinical Study.

Background: Apical periodontitis (AP) is an inflammatory disorder of the periapical tissues caused by the persistence of a microbial infection within the root canal system of the affected tooth. Clinically, it is symptomatic or asymptomatic depending on several factors such as the type of microorganisms, bacterial load, immunological reaction, and local tissue mediators. Chronic or asymptomatic infections may initiate and modulate intravascular accumulation of inflammatory cells resulting in endothelial dysfunction which subsequently represents a possible systemic inflammatory burden. Aim: The present study aimed to evaluate the relationship between asymptomatic AP and systemic inflammatory burden by assessing the levels of chronic inflammatory cells. Methodology: A total of 25 patients diagnosed with asymptomatic AP who showed a negative response to the pulp sensitivity test with no history of any systemic diseases were included in this preliminary prospective observational study. Blood samples were collected at each phase of the study, and a complete hemogram was carried out. All hematological parameters were recorded before and after root canal therapy and they were analyzed for statistical significance at p <.05 using IBM SPSS Statistics for Windows, Version 21 (Released 2012; IBM Corp., Armonk, New York, United States). Results: Evaluation of the mean total leukocyte count (TLC), lymphocyte, and eosinophil cell count showed a significant reduction in the number of cells before and after root canal therapy treatment respectively (p<.05). One-way analysis of variance also revealed statistical significance at p < .05 with a weak positive correlation between the TLC, lymphocyte, and eosinophil count before and after treatment. Conclusion: The present study showed that systemically healthy individuals with asymptomatic AP had increased inflammatory burden in the circulation, and thus, it is essential to identify and quantify the risk associated. It was evident that complete healing of the asymptomatic AP lesions results in an overall reduction of systemic inflammatory cells ultimately reducing the burden and risk of associated systemic inflammatory diseases.

Bifidobacterium animalis subsp. lactis HN019 has antimicrobial activity against endodontic pathogens in vitro.

The aim of the present study was to evaluate, in vitro, the antimicrobial activity of the probiotic Bifidobacterium animalis subsp. lactis HN019, through the well technique, against 10 microorganisms can be found involved in endodontic infections. The antimicrobial activity of the probiotic was performed on Streptococcus mutans, Streptococcus sobrinus, Lacticaseibacillus casei, Enterococcus faecalis, Staphylococcus aureus, Candida albicans, Porphyromonas gingivalis, Porphyromonas endodontalis, Fusobacterium nucleatum and Prevotella intermedia. For the control group, it was used non-pathogenic bacteria Escherichia coli, Saccharomyces cerevisiae, and Kocuria rizhopilla. After 48 to 72 h of incubation of the petri dishes containing the culture medium, the microorganism strains, and the probiotic, the plates were examined to assess the uniformity of microbial growth, presence of contaminants, and the halo of inhibition. After visual inspection, the reading of the halo of inhibition was performed with the aid of a digital caliper using a reflected light source to illuminate the inverted plate on a black, opaque background after removing the cap. Thus, 3 values were obtained from each bacterial inoculum, which were added and divided by three to obtain the average of the values. The results of the in vitro study demonstrated that the probiotic B. animalis subsp. lactis HN019 promoted the inhibition of all strains of the pathogens evaluated, with the exception of Candida albicans, demonstrating antimicrobial activity on these microorganisms.

Unraveling the Endodontic Archaeome: A Systematic Review with Meta-Analysis.

INTRODUCTION: The controversial issue of whether the Archaea domain plays a role in endodontic infections is the focus of this systematic review with meta-analysis. The aim is to emphasize the significance of minority microbial domains in oral dysbiosis by evaluating the prevalence of archaea in root canals and its association with clinical parameters such as symptomatology and type of endodontic infection. METHODS: The search strategy involved researching 6 databases and the gray literature. Publications were accepted in any year or language that identified archaea in samples from endodontic canals. A 2-step selection process narrowed the final choice to 16 articles. The methodological quality of the studies was evaluated using tools from the Joanna Briggs Institute, and the certainty of evidence was assessed using the Grading of Recommendations, Assessment, Development, and Evaluation (GRADE) approach. RESULTS: The results showed that archaea were present in 20% (95% [confidence interval] CI = 8%-32%) of individuals with endodontic samples analyzed. The samples were about twice as likely to be archaeal-positive if collected from individuals with primary vs. persistent/secondary infection (odds ratio = 2.33; 95% CI = 1.31-4.14; I2 = 0%), or individuals with self-reported vs. symptom-free infections (odds ratio = 2.67; 95% CI = 1.47-4.85; I2 = 0%). Methanogenic archaea were reported in 66% of the included studies. Representative members of phyla Thaumarchaeota and Crenarchaeota were also identified. CONCLUSIONS: Archaea are present in about one-fifth of the infected root canals. Recognized biases in experimental approaches for researching archaea must be addressed to understand the prevalence and roles of archaea in endodontic infections, and to determine whether the decontamination process should include the elimination or neutralization of archaea from root canals (International Prospective Register of Systematic Reviews protocol = CRD42021264308).

Real-time optical detection of endodontic infection using bacterial autofluorescence.

OBJECTIVES: For successful root canal treatment (RCT), it is essential to objectively assess the presence and activity of bacteria in the root canal system. However, current methods rely on subjective observations of root canal exudates. This study aimed to confirm whether real-time optical detection using bacterial autofluorescence can evaluate endodontic infection status by assessing the red fluorescence (RF) detected from root canal exudates. METHODS: During RCT, endodontic paper points were used to collect root canal exudates scored using conventional organoleptic tests to assess the severity of root canal infections. RF on the paper points was assessed using quantitative light-induced fluorescence (QLF) technology. RF intensity and area from the paper points were quantified, and their correlations with infection severity were assessed using their organoleptic scores. The oral microbiome composition of RF samples was compared with non-red fluorescent (non-RF) samples. RESULTS: The RF detection rate was nil and >98% in the non-infectious and severe groups. The RF intensity and area significantly increased with infection severity (p<0.001) and showed strong correlations with organoleptic scores (r=0.72, 0.82, respectively). The diagnostic accuracy for detecting root canal infection using RF intensity was good to excellent (AUC = 0.81-0.95) and increased with infection severity. The microbial diversity of the RF samples was significantly lower than that of the non-RF samples. Gram-negative anaerobic bacteria such as Prevotella and Porphyromonas were more predominant in RF samples. CONCLUSIONS: Optical detection using bacterial autofluorescence can objectively evaluate endodontic infection status in real-time by assessing the RF of endodontic root canal exudates. CLINICAL SIGNIFICANCE: This real-time optical technology can be utilised to detect endodontic bacterial infection without conventional incubation, allowing clinicians to determine the endpoint of chemomechanical debridement and increase the positive outcomes of RCTs.

Bacterial quantity and interleukin-17 expression in necrotic teeth with apical periodontitis from type II diabetic patients.

Diabetes mellitus (DM) increases the susceptibility to infection. A plausible association between apical periodontitis (AP) and DM has been reported, but the underlying mechanism is not yet elucidated. AIM: To investigate the bacterial quantity and the expression of interleukin-17 (IL-17) in necrotic teeth with AP in type 2 DM (T2DM), pre-diabetic and non-diabetic control patients. METHODOLOGY: In all, 65 patients with necrotic pulp and AP [periapical index (PAI) scores ≥3] were included. The age, gender, medical history and medications list, including metformin and statin intake, were recorded. Glycated haemoglobin (HbA1c) was analysed, and the patients were divided into three groups: T2DM (n = 20), pre-diabetics (n = 23) and non-diabetic (n = 22). Bacterial samples (S1) were collected by file and paper points. Bacterial DNA was isolated and quantified using 16S ribosomal RNA gene-targeted quantitative real-time polymerase chain reaction (qPCR). For IL-17 expression, (S2) samples were collected from the periapical tissue fluid using paper points passing through the apical foramen. The IL-17 total RNA was extracted, and reverse transcription (RT-qPCR) analysis was performed. Comparisons between the three study groups were conducted using one-way anova and Kruskal-Wallis test to explore the relationship between bacterial cell counts and IL-17 expression in each group. RESULTS: The distributions of PAI scores were equivalent among the groups, p = .289. T2DM patients had higher bacterial counts and IL-17 expression than other groups, but these differences were not statistically significant, p = .613 and p = .281, respectively. T2DM patients taking statin appear to have lower bacterial cell count than those who do not take statin, approaching the significance level, p = .056. CONCLUSION: T2DM patients had a non-significant higher bacterial quantity and IL-17 expression compared to pre-diabetic and healthy controls. Although these findings indicate a weak association, it may impact the clinical outcome of endodontic diseases in diabetic patients.

Gram-negative quorum sensing signalling enhances biofilm formation and virulence traits in gram-positive pathogen Enterococcus faecalis.

Acyl-homoserine lactones (AHLs) are typical quorum-sensing molecules of gram-negative bacteria. Recent evidence suggests that AHLs may also affect gram-positives, although knowledge of these interactions remains scarce. Here, we assessed the effect of AHLs on biofilm formation and transcriptional regulations in the gram-positive Enterococcus faecalis. Five E. faecalis strains were investigated herein. Crystal violet was employed to quantify the biomass formed, and confocal microscopy in combination with SYTO9/PI allowed the visualisation of biofilms' structure. The differential expression of 10 genes involved in quorum-sensing, biofilm formation and stress responses was evaluated using reverse-transcription-qPCR. The AHL exposure significantly increased biofilm production in strain ATCC 29212 and two isolates from infected dental roots, UmID4 and UmID5. In strains ATCC 29212 and UmID7, AHLs up-regulated the quorum-sensing genes (fsrC, cylA), the adhesins ace, efaA and asa1, together with the glycosyltransferase epaQ. In strain UmID7, AHL exposure additionally up-regulated two membrane-stress response genes (σV, groEL) associated with increased stress-tolerance and virulence. Altogether, our results demonstrate that AHLs promote biofilm formation and up-regulate a transcriptional network involved in virulence and stress tolerance in several E. faecalis strains. These data provide yet-unreported insights into E. faecalis biofilm responses to AHLs, a family of molecules long-considered the monopole of gram-negative signalling.

Correlation between bacterial type/bacterial quantity and bone loss detected by cone beam computed tomography (CBCT) in primary endodontic infections / Correlación entre el tipo y cantidad de bacterias y la pérdida ósea detectada por Tomografía Computarizada de Haz Cónico (CBCT) en infecciones endodóncicas primarias

Abstract Periapical lesions (PL) of endodontic origin are one of the most common pathological conditions that affect peri-radicular tissues. The main objective of this study was to evaluate the amount and species of microorganisms isolated from necrotic pulps, establish a correlation between these and the size of periapical lesions, and how the amount and species of microorganisms decreased with non-surgical root canal treatment. Twenty-seven patients with a clinical diagnosis of dental pulp necrosis and chronic periapical lesions were selected; a Cone Beam Computed Tomography (CBCT) and microbial samples of the root canal system were taken previous to a disinfection protocol, a post-instrumentation/ disinfection protocol, and a post-medication placement. Samples were processed for colony-forming unit (CFU) counting, Gram staining technique, and bacterial identification by the API-20 Strep/API-20A system. The API system identified 21 species of microorganisms in the pre-instrumentation samples, 11 species in the post-instrumentation samples, and 11 in the post-medication samples. There was a correlation coefficient of 0.598% between the initial size of the lesion and the number of bacteria, with a coefficient of determination up to 35.7%, a correlation coefficient of 0.486% and a determination coefficient of 23.6% between the size of the periapical lesion and the number of CFUs. This study contributes to the knowledge of the amount and species of microorganisms isolated and identified from necrotic pulps, establishes a correlation between the amount and species of microorganisms and the size of the periapical lesions, and shows how the decrease of microorganisms contributes to the healing of PL, corroborating the importance of an adequate disinfection protocol.

Resumen Las lesiones periapicales (LP) de origen endodóncico son la condición patológica más común que afectan los tejidos perirradiculares. El objetivo principal de este estudio es evaluar la cantidad y especie de bacterias aisladas de pulpas necróticas, correlacionar la cantidad y especies bacterianas con el tamaño de la lesión, y cómo disminuyen la cantidad y especies de microorganismos con el tratamiento de conductos. A 27 pacientes con diagnóstico de necrosis pulpar y lesión periapical crónica detectada con CBCT se les tomaron muestras microbianas del sistema de conductos antes y después del protocolo de desinfección y de la medicación intraconducto. Las muestras se procesaron para el recuento de unidades de formación de colonias (UFC), tinción de Gram e identificación mediante el sistema API-20 Strep/API-20A. Se identificaron 21 especies en las muestras pre-instrumentación, 11 en las muestras post-instrumentación y 11 en las muestras post-medicación; se observó un coeficiente de correlación del 0,598% entre el tamaño inicial de la lesión y la cantidad de bacterias, con un coeficiente de determinación hasta el 35,7%, un coeficiente de correlación del 0,486% y un coeficiente de determinación del 23,6% entre el tamaño de la lesión periapical y el número de UFCs. Este estudio contribuye al conocimiento sobre la cantidad y especies de microorganismos aislados e identificados a partir de pulpas necróticas, establece una correlación entre la cantidad y especies de microorganismos y el tamaño de las lesiones periapicales y exhibe cómo la disminución de microorganismos contribuye a la curación de LP, corroborando la importancia de un adecuado protocolo de desinfección.

Amplicon-based next-generation sequencing for comparative analysis of root canal microbiome of teeth with primary and persistent/secondary endodontic infections.

OBJECTIVES: To compare the root canal microbiome profiles of primary and persistent/secondary infections using high-throughput sequencing with the help of a reliable bioinformatics algorithm. MATERIALS AND METHODS: Root canal samples of 10 teeth in the primary endodontic infection (PEI) group and 10 teeth in the persistent/secondary endodontic infection (SEI) group were included resulting in a total of 20 samples. After DNA extraction from the samples, sequencing was performed on the Illumina MiSeq platform. Pair-end Illumina reads were imported to QIIME 2; amplicon sequence variants (ASVs) generated by DADA2 were mapped to GreenGenes database. Weighted UniFrac distances were calculated and principal coordinates analysis (PCoA) was used to compare beta diversity patterns. The multiple response permutation procedure (MRPP), the analysis of similarities (ANOSIM), and permutational multivariate analysis of variance (adonis) were conducted for testing group differences. Linear discriminant analysis effect size (LEfSe) analysis was utilized to identify differentially abundant taxa between the groups. The linear discriminant analysis (LDA) score threshold was set to 4.0. RESULTS: Within the Gram-negative facultative anaerobic Gammaproteobacteria class outgroup, two orders (Pasteurellales, Vibrionales) and two families (Pasteurellaceae, Vibrionaceae) were significantly more abundant in the PEI group, whereas Gram-positive bacteria, Actinomycetales order, and Gram-positive anaerobic taxa, one genus (Olsenella) and one species (Olsenella uli), were identified as significantly more abundant in the SEI group. CONCLUSIONS: A few taxa were differentially abundant within either the PEI or SEI group. CLINICAL RELEVANCE: Reliable bioinformatic tools are needed to define microbial profiles of endodontic infections. Based on a limited number of samples, no distinct variation was determined between the bacterial diversity of initial and recurrent endodontic infections.

Antimicrobial effect of grape seed extract as a potential intracanal medicament combined with Nd:YAG laser.

This study evaluated the antimicrobial efficacy of grape seed extract medicament combined with Nd:YAG laser, against Enterococcus faecalis, Staphylococcus aureus and Candida albicans biofilms. Root canals infected with 4-week-old biofilms were divided into five groups (n = 11): calcium hydroxide, 6.5% GSE, Nd:YAG laser (1064 nm, 1.5 w, 15 Hz and 100 mj) and 6.5% GSE followed by Nd:YAG laser and normal saline (control). Dentin chips were collected using Gates-Glidden and cultured to obtain colony-forming units. Statistical analysis was performed using Kruskal-Wallis and Mann-Whitney U test. GSE showed higher antibacterial activity against all species investigated compared to Ca(OH)2 . However, the lowest microbial count was obtained using a combination of GSE and Nd:YAG laser (p < 0.001). No significant difference in the susceptibility of tested pathogens to each of the protocols was observed (p > 0.05). Application of Nd:YAG laser following GSE medicament is efficient against endodontic biofilms; also, GSE can be considered as an alternative to Ca(OH)2 dressing.