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).

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.

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).

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.

Infecções odontogênicas - patogênese e repercussões sistêmicas: revisão de literatura / Odontogenic infections pathogenesis and systemic repercussions

A região de cabeça e pescoço pode ser acometida por doenças infecciosas de origem não odontogênica ou odontogênica - sendo essa geralmente de causa endodôntica. A saúde bucal faz parte do sistema corporal de um indivíduo, desse modo, a negligência com seus cuidados gera um impacto sistêmico. Sendo assim, o objetivo do presente trabalho é analisar o desenvolvimento das infecções odontogênicas desde a necrose pulpar às possíveis repercussões sistêmicas, como: Angina de Ludwing, Mediastinite Necrosante, Fasceíte Necrosante, Endocardite Bacteriana, Sinusite do Seio Maxilar e Osteomielite. É de suma importância que o profissional cirurgião dentista conheça a patogênese dessas infecções, para que este seja capaz de elaborar um diagnóstico precoce e preciso, bem como saiba utilizar os protocolos de prevenção e condutas, a fim de evitar graves complicações sistêmicas. A partir dessa elucidação, será possível evitar o adoecimento do corpo gerado por uma doença da cavidade oral.

The head and neck region can be affected by infectious diseases of non-odontogenic or odontogenic origin - the latter usually having an endodontic cause. Oral health is part of an individual's body system, thus, neglecting its care generates a systemic impact. Therefore, the aim of this study is to analyze the development of odontogenic infections from pulpal necrosis to possible systemic repercussions, such as: Ludwing's Angina, Necrotizing Mediastinitis, Necrotizing Fasciitis, Bacterial Endocarditis, Maxillary Sinusitis and Osteomyelitis. It is extremely important that the dental surgeon knows the pathogenesis of these infections, so that he is able to make an early and accurate diagnosis, as well as know how to use prevention protocols and conducts, in order to avoid serious systemic complications. From this elucidation, it will be possible to avoid the illness of the body generated by a disease of the oral cavity.

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.

Regenerative Endodontic Therapy for Management of an Immature Permanent Tooth with Recurrent Post-treatment Apical Periodontitis: A Case Report.

This case report describes the treatment outcome and further retreatment of an immature permanent maxillary right central incisor with necrotic pulp and chronic apical abscess using regenerative endodontic therapy (RET). The patient had a history of traumatic injury. The initial periapical radiographic and cone-beam computed tomographic (CBCT) examinations revealed tooth #8 had incomplete root formation, thin dentinal walls, and pulp necrosis associated with a large apical periodontitis lesion. RET was conducted in two visits and included a disinfection protocol with 5.25% NaOCl irrigation and medication with a double antibiotic paste (metronidazole and ciprofloxacin). At the second visit, a blood clot was induced, and the cervical third was sealed with a mineral trioxide aggregate plug and the coronal portion with light-cure composite. The tooth was asymptomatic at the 12-, 24-, and 36-month follow-ups, and radiographs showed continued root development with healed periradicular tissues. However, the 4-year radiographic follow-up revealed a recurrent apical periodontitis lesion. A second attempt of RET was conducted in one visit using 1% NaOCl irrigation and stimulation of a blood clot. A double seal with silicate-based cement and composite was placed. At the 24-month follow-up, the tooth remained asymptomatic, and both radiographic and CBCT examinations showed apical closure and complete repair of the periradicular tissues. When a tooth develops recurrent apical periodontitis, a second attempt of RET is a feasible option to control infection, helping to promote tooth retention associated with healthy periradicular conditions.

Curcumin reduces inflammation in rat apical periodontitis.

AIM: The aim of the study was to evaluate the effect of systemic curcumin administration on the severity of apical periodontitis (AP). METHODOLOGY: Forty male Wistar rats weighing 250-280 g each, age 2.5 months, were distributed into four groups (n = 10): control untreated rats (C), control rats treated with curcumin (CUR), rats with pulp exposure-induced apical periodontitis (AP) and rats with pulp exposure-induced apical periodontitis treated with curcumin (AP-CUR). Curcumin treatment was administered orally once daily for 15 days before pulp exposure and continued for 30 days after pulp exposure. The rats were sacrificed at 30 days, and the jaws were collected and reconstructed in a programme specific for micro-CT. The jaws were processed for analysis of the inflammatory process using haematoxylin and eosin staining and immunohistochemical assays for interleukin tumour necrosis factor alpha (TNF-α), interleukin (Il)-6 and Il-1ß. Tartrate-resistant acid phosphatase (TRAP) and osteocalcin (OCN) staining were used to analyse the resorptive process on the bone surface of periapical area. Kruskal-Wallis with Dunn's test was performed for nonparametric data and anova with Tukey's test for parametric data, p < .05. RESULTS: Micro-CT revealed no statistically significant differences in bone resorption between the AP and AP-CUR groups (p > .05). The levels of inflammatory cell infiltration and immunoreactivity for the proinflammatory cytokines TNF-α, Il-6 and Il-1ß were significantly higher in the periapical lesions of the AP group than in the AP-CUR group (p < .05). The number of TRAP-positive multinucleated cells was higher in the AP group than in the AP-CUR group (p < .05). In OCN-positive cells, no differences were observed between the AP and AP-CUR groups (p > .05). CONCLUSIONS: Oral supplementation with curcumin had a significant effect on the AP severity in rats, suggesting an anti-inflammatory effect of curcumin on AP development.

In vitro physicochemical characterization of five root canal sealers and their influence on an ex vivo oral multi-species biofilm community.

AIM: To evaluate the physicochemical properties of five root canal sealers and assess their effect on an ex vivo dental plaque-derived polymicrobial community. METHODOLOGY: Dental plaque-derived microbial communities were exposed to the sealers (AH Plus [AHP], GuttaFlow Bioseal [GFB], Endoseal MTA [ESM], Bio-C sealer [BCS] and BioRoot RCS [BRR]) for 3, 6 and 18 h. The sealers' effect on the biofilm biomass and metabolic activity was quantified using crystal violet (CV) staining and MTT assay, respectively. Biofilm community composition and morphology were assessed by denaturing gradient gel electrophoresis (DGGE), 16S rRNA sequencing and scanning electron microscopy. The ISO6876:2012 specifications were followed to determine the setting time, radiopacity, flowability and solubility. Obturated acrylic teeth were used to assess the sealers' effect on pH. Surface chemical characterization was performed using SEM with coupled energy-dispersive spectroscopy. Data normality was assessed using the Shapiro-Wilk test. One-way anova and Tukey's tests were used to analyze data from setting time, radiopacity, flowability and solubility. Two-way anova and Dunnett's tests were used for the data analysis from CV, MTT and pH. 16S rRNA sequencing data were analyzed for alpha (Shannon index and Chao analysis) and beta diversity (Bray-Curtis dissimilarities). Differences in community composition were evaluated by analysis of similarity (p < .05). RESULTS: The sealers significantly influenced microbial community composition and morphology. All sealers complied with ISO6876:2012 requirements for setting time, radiopacity and flowability. Although only AHP effectively reduced the biofilm biomass, all sealers, except BRR, reduced biofilm metabolic activity. CONCLUSION: Despite adequate physical properties, none of the sealers tested prevented biofilm growth. Significant changes in community composition were observed. If observed in vivo, these changes could affect intracanal microbial survival, pathogenicity and treatment outcomes.

Nature and Prevalence of Bacterial Taxa Persisting after Root Canal Chemomechanical Preparation in Permanent Teeth: A Systematic Review and Meta-analysis.

INTRODUCTION: Culture-independent molecular studies have shown a broad spectrum of bacterial taxa that persist after chemomechanical procedures (CMP). Therefore, this study systematically reviewed these reports to explore the prevalence of bacteria in post-instrumentation samples of root canals from permanent teeth, especially of as-yet-uncultivated/difficult-to-culture bacteria. METHODS: Electronic databases were searched from 2007 to January 2021. Clinical studies using culture-independent molecular methods to identify species-level taxa before and after CMP were included. Studies were critically appraised using the Joanna Briggs Institute Prevalence Critical Appraisal Checklist and the funnel plot analysis. The meta-analysis was performed on the prevalence of as-yet-uncultivated/difficult-to-culture bacterial taxa using RStudio. RESULTS: A total of 3781 titles were screened, but only 20 studies were included. The most frequent species in post-instrumentation samples were Streptococcus spp., Leptotrichia buccalis, Fusobacterium nucleatum, and Capnocytophaga ochracea. The detection frequency of some species increased after CMP, including mainly Firmicutes members such as streptococci, Enterococcus faecium, Selenomonas noxia, and Solobacterium moorei. The prevalence (confidence interval) of difficult-to-culture species was as follows: Dialister invisus, 17% (7%-29%); Solobacterium moorei, 14% (8%-23%); Bacteroidaceae [G-1] bacterium HMT 272, 13% (5%-23%); and Filifactor alocis, 11% (3%-23%). CONCLUSIONS: The prevalence of as-yet-uncultivated/difficult-to-culture bacterial taxa in post-instrumentation samples was low. The persistent species belonged mainly to the phylum Firmicutes, and streptococci were the major members. Future larger clinical studies on the composition of the whole bacterial community that persist after CMP are still necessary for a better understanding of bacterial interactions and their clinical significance in the treatment outcome.

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.

The effect of photodynamic therapy on postoperative pain in teeth with primary endodontic infection.

INTRODUCTION: The objective of this study was to investigate the action of photodynamic therapy on pain control after endodontic treatment in asymptomatic teeth with a primary infection, within a single visit. METHODS: Sixty (60) single-rooted teeth with pulp necrosis and periapical lesions were selected and randomly divided into two (2) groups (n = 30), according to the protocol; a control group (CG) and a group using photodynamic therapy (aPDT). The canals were instrumented with Reciproc files # 25 up to 40 along the entire length of the canal, using 2% chlorhexidine gel as the auxiliary chemical substance, followed by irrigation with sterile saline. aPDT consited of 0.005% methylene blue as photosensitizer, using AsGaAl diode laser, 660 nm wavelength, 100 mW of power and 9 J of energy, using optical fibers with 365 µm in diameter. The canals were filled with Endomethasone N cement. RESULTS: Pain intensity was assessed at 8, 12, 24, 48, 72 h and 1 week after endodontic treatment using a visual analogue scale. The level of pain was classified as none (0), mild (1-3), moderate (4-7) or severe (8-10). The data were at a significance level of 5%. There was a statistically significant difference (p<0.05) in the periods of 8, 12, 24, 48 and 72 h between the control group and the aPDT group. After 1 week, there was no statistically significant difference. CONCLUSIONS: It is concluded that photodynamic therapy had a significant effect on decreasing post-endodontic treatment pain in teeth with necrotic pulp and asymptomatic periapical lesions.

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.

Clinical and radiographic outcome of the root canal treatment of infected teeth with associated sinus tract: A retrospective study.

This retrospective study evaluated the outcome of the endodontic treatment/retreatment of teeth with intraoral sinus tracts. Seventy-two teeth from 70 individuals were included. One operator treated all teeth over about 20 years. Time for the sinus tract to heal was recorded. Treatment outcome was evaluated after at least 1 year and classified as healed, healing or diseased. The overall healed rate of periradicular lesions was 73.6% (74.1% for treatment and 72.2% for retreatment). In 6 (8.3%) teeth, lesions were healing. Thirteen teeth (18.1%) exhibited persistent disease. In 68/72 (94.4%) cases, the sinus tract healed in less than 1 month. No preoperative/intraoperative variable affected the outcome (P > 0.05). Missing restorations had a negative impact (P < 0.05). Healing of both the sinus tract quickly after treatment and the periradicular lesion later were usually achieved. Closure of the sinus tract in the first month was a good predictor of lesion healing.

Immunological profile of periapical endodontic infection in patients undergoing haematopoietic transplantation.

OBJECTIVES: To evaluate the mRNA expression levels of cytokines interferon-γ, tumour necrosis factor-α, interleukin IL-1ß, IL-10, and the chemokine CCL2/MCP-1, CCL4, and CXCR4 in the periapical interstitial fluid from root canal infections before and after bacterial load reduction in patients undergoing haematopoietic stem cell transplantation (HSCT). MATERIALS AND METHODS: The case group was composed of 10 patients undergoing HSCT, and our control group included 10 healthy patients. Clinical samples were taken from teeth with pulp necrosis. Three paper points were placed in the RCS and maintained for 2 min for microbial evaluation before cleaning and shaping procedures. After cleaning and drying the canal, three paper points were introduced into the root canal, passing passively through the root apex (2 mm) into the periapical tissues for 1 min. Samples were collected immediately after root canal cleaning and 7 days later (restrained root canal bacterial load) to characterize gene expression using real-time PCR. RESULTS: The results showed significantly reduction in the microbial load on day 7. An increased expression level of TNF-α and IFN-γ on day 7 in control and case groups was observed (p < 0.05). The mRNA levels of IL-1ß and IL-10 in the pre-HSCT group increased in the samples from day 7 (p < 0.05). The chemokine CCL-2/MCP-1 was not detected in pre-HSCT group. Chemokine receptor CXCR4 levels increased in samples obtained from the day 7 in the control group (p < 0.05). CONCLUSIONS: Individuals undergoing HSTC presented similar cytokine and chemokine mRNA expression compared with healthy individuals. However, it was observed the total absence of mRNA MCP-1/CCL2 expression in those individuals undergoing HSCT. CLINICAL RELEVANCE: Patients undergoing HSCT are at higher risk of infection. No study has analysed the periapical immune responses to root canal infections in HSCT individuals.

Analysis of Active Bacteria Persisting after Chemomechanical Procedures: An RNA- and DNA-based Molecular Study.

INTRODUCTION: Because active cells present higher abundance of ribosomal RNA (rRNA) than rDNA (rRNA genes), data obtained with rDNA-based quantitative polymerase chain reaction (qPCR) and rRNA-based qPCR (RT-qPCR) were correlated to search for active bacteria after chemomechanical procedures (CMP). In addition, the ability of both assays to detect bacteria in endodontic samples was evaluated. METHODS: Samples were taken from 40 teeth with primary endodontic infections before (S1) and after CMP (S2). DNA and cDNA (synthetized from RNA) were used as templates for qPCR using universal primers for bacteria and species-specific primers for Bacteroidaceae sp. HOT-272, Cutibacterium acnes, Selenomonas spp., and Enterococcus faecalis. RESULTS: After CMP, there was a drastic reduction in the number of total bacteria, Selenomonas spp., and E. faecalis, whereas no significant difference was observed for the levels of Bacteroidaceae sp. HOT-272 and C. acnes. The concentration of rRNA copies in S2 samples was significantly higher than the corresponding levels of rDNA for assays targeting total bacteria, Bacteroidaceae sp. HOT-272, and C. acnes (P < .05), indicating persistence of active bacteria. The rDNA-based qPCR presented low sensitivity and high specificity when compared with RT-qPCR. For most assays, samples positive for rDNA were also positive for rRNA (positive predictive value = 100%). CONCLUSIONS: CMP was effective in reducing levels but not the metabolic activity of total bacteria. Bacteroidaceae sp. HOT-272 and C. acnes were active members of the persistent community. Although less sensitive than RT-qPCR, most rDNA-based qPCR assays had a low risk of providing false-positive results in postinstrumentation samples.

Assessment of the antibacterial, antivirulence, and action mechanism of Copaifera pubiflora oleoresin and isolated compounds against oral bacteria.

The microorganisms that constitute the oral microbiome can cause oral diseases, including dental caries and endodontic infections. The use of natural products could help to overcome bacterial resistance to the antimicrobials that are currently employed in clinical therapy. This study assessed the antimicrobial activity of the Copaifera pubiflora oleoresin and of the compounds isolated from this resin against oral bacteria. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) assays provided values ranging from 6.25 to > 400 µg/mL for the C. pubiflora oleoresin and its isolated compounds. The fractional inhibitory concentration index (FICI) assay showed that the oleoresin and chlorhexidine did not act synergistically. All the tested bacterial strains formed biofilms. MICB50 determination revealed inhibitory action: values varied from 3.12-25 µg/mL for the oleoresin, and from 0.78 to 25 µg/mL for the ent-hardwickiic acid. Concerning biofilm eradication, the C. pubiflora oleoresin and hardwickiic acid eradicated 99.9 % of some bacterial biofilms. Acid resistance determination showed that S. mutans was resistant to acid in the presence of the oleoresin and ent-hardwickiic acid at pH 4.0, 4.5, and 5.0 at all the tested concentrations. Analysis of DNA/RNA and protein release by the cell membrane demonstrated that the oleoresin and hardwiickic acid damaged the bacterial membrane irreversibly, which affected membrane integrity. Therefore, the C. pubiflora oleoresin and ent-hardwickiic acid have potential antibacterial effect and can be used as new therapeutic alternatives to treat oral diseases such as dental caries and endodontic infections.

Next-Generation Sequencing to Assess Potentially Active Bacteria in Endodontic Infections.

INTRODUCTION: Because active bacteria present a higher abundance of ribosomal RNA (rRNA) than DNA (rRNA gene), the rRNA/DNA ratio of next-generation sequencing (NGS) data was measured to search for active bacteria in endodontic infections. METHODS: Paired complementary DNA and DNA samples from 5 root canals of teeth with apical periodontitis were subjected to polymerase chain reaction with bar-coded primers amplifying the 16S rRNA gene hypervariable regions V4-V5. High-throughput sequencing was performed using MiSeq (Illumina, San Deigo, CA), and data were analyzed using Quantitative Insights Into Microbial Ecology and Human Oral Microbiome Database. Statistical analysis was performed for relative abundance of bacteria in the DNA- and rRNA-based NGS data using the Mann-Whitney test, whereas differences in the diversity and richness indexes were assessed using a nonparametric 2-sample t test (P < .05). For bacterial taxa detected in both approaches, the rRNA/DNA ratios were calculated by dividing the average abundance of individual species in the respective analysis. RESULTS: Although no significant difference was found in the indexes of bacterial richness and diversity, the relative abundance of bacterial members varied in both analyses. Comparing rRNA with DNA data, there was a significant decrease in the relative abundance of Firmicutes (P < .05). The bacterial taxa Bacteroidales [G-2] bacterium HMT 274, Porphyromonas endodontalis, Tannerella forsythia, Alloprevotella tannerae, Prevotella intermedia, Pseudoramibacter alactolyticus, Olsenella sp. HMT 809, Olsenella sp. HMT 939, Olsenella uli, and Fusobacterium nucleatum subsp. animalis were both dominant (DNA ≥ 1%) and active (rRNA/DNA ≥ 1). CONCLUSIONS: The integrated DNA- and rRNA-based NGS strategy was particularly important to disclose the activity of as-yet-uncultivated or difficult-to-culture bacteria in endodontic infections.

Prevalence of Candida albicans in primary endodontic infections associated with a higher frequency of apical periodontitis in type two diabetes mellitus patients.

To identify the prevalence of C. albicans in primary endodontic infections of type two diabetes mellitus (T2DM) patients and compare their clinical and radiographical characteristics with a non-diabetic control group, establishing the possible relationship between primary endodontic infection, T2DM, and C. albicans, since diabetes mellitus (DM), influences the development, course, and response to the treatment of apical periodontitis, but the presence of Candida albicans (C. albicans) has not been considered before. A total of 120 patients were selected and divided into two groups: 60 T2DM diagnosed patients and 60 non-diabetic controls. A clinical examination and radiographic analysis were performed to establish a periapical index score (PAI). Root canal samples were taken. Deoxyribonucleic acid (DNA) was extracted, and specific primers were used to identify C. albicans by polymerase chain reaction (PCR). A twofold increase in the prevalence of C. albicans in T2DM patients was observed in contrast to control patients (p = 0.0251). Sixty-five percent of T2DM patients with positive C. albicans scored a ≥ 3 PAI, while only 27% of the patients without C. albicans had a ≥ 3 PAI score (p = 0.0065). Long-term DM patients presented C. albicans more frequently (p < 0.0001). In this study, long-term T2DM patients carried C. albicans in their root canals more frequently when having a primary endodontic infection. Furthermore, this C. albicans presence seems to be related to a higher frequency of apical periodontitis.