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.

Role of oral microbiota in irreversible pulpitis - Current strategies and future perspectives.

Irreversible pulpitis is an inflammation of the tooth pulp caused by an opportunity-driven invasion of the pulp space by oral microbiota typically prevalent in the oral cavity. Microbial organisms are extensively recognised to be the fundamental cause of endodontic infections and treatment failures. Previously, bacterial species responsible for these infections were largely recognised using conventional microbial culture techniques, lending credence to the widely held belief that anaerobic Gram-negative bacteria frequently enter the pulp space and trigger endodontic infections. The advent of novel technologies grants the advantage of detecting and studying microbial populations via an amalgamation of the modern "Omics" techniques and meticulous bioinformatics analysis, additionally detecting the metatranscriptome, metaproteome and metabolome along with the metagenome. Amongst these analytical strategies, metagenomic analyses are essentially pragmatic for investigating the oral microbiome. Metagenomics favor not only assessment of microbial composition in diseased conditions, but also contributes to detection of novel, potentially pathogenic species inclusive of non-viable bacteria. The present review describes current knowledge of root canal microbiome, including its composition and functional attributes, the novel strategies available for detection of microbiome as well as challenges associated and provides some crucial pointers for areas of future research.

Expression of Toll-like receptor 2, Dectin-1, and Osteopontin in murine model of pulpitis.

OBJECTIVES: This in vivo animal study aimed to develop a murine model of pulpitis induced by pulp exposure with or without application of zymosan in Naval Medical Research Institute (NMRI) mice and observe expressions of Toll-like receptor (TLR)-2, TLR-4, Dectin-1, Osteopontin (OPN), tumor necrosis factor alpha (TNF-α), interleukin (IL)-6, and IL-1ß. MATERIAL AND METHODS: A total of 168 NMRI mice were divided into two groups, i.e., group A (n = 84) (pulpitis induced by pulp exposure only) and group B (n = 84) (pulpitis induced by pulp exposure and zymosan application). Right maxillary molar pulps were exposed with » round bur, and animals were sacrificed at 0, 6, 9, 12, 24, 48, and 72 h. The exposed teeth were obtained for real-time polymerase chain reaction (qRT-PCR) analysis and histological and immunohistochemistry (IHC) analysis. RESULTS: Histological evaluation revealed a time-dependent steady increase in inflammation. Similar time-dependent increase in the expression of inflammatory cytokines was noted. Group A exhibited an increase in TLR-4, Dectin-1, and OPN at 6 h, while TLR-2 was expressed at 24 h. Group B expressed TLR-2, Dectin-1, and OPN at 9, 48, and 72 h, respectively (p ≤ 0.05). Expression of OPN and TNF-α exhibited a similar pattern in both groups. IHC also detected expression of TLR-2, Dectin-1, TLR4, and CD68 in some cells at 6 and 9 h. CONCLUSIONS: NMRI mice provided for a stable pulp inflammation model. Zymosan may be used to develop pulp inflammation model and study inflammatory response towards fungal antigens. Dental pulp expressed Dectin-1 receptor. OPN and TNF-α exhibited a similar expression pattern. CLINICAL RELEVANCE: Innate immunity of dental pulp is capable of detecting fungal pathogens.

Stimulation phosphatidylinositol 3-kinase/protein kinase B signaling by Porphyromonas gingivalis lipopolysacch aride mediates interleukin-6 and interleukin-8 mRNA/protein expression in pulpal inflammation.

BACKGROUND/PURPOSE: The signaling mechanisms for Porphyromonas gingivalis lipopolysaccharide (PgLPS)-induced inflammation in human dental pulp cells are not fully clarified. This in vitro study aimed to evaluate the involvement of phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) pathway in PgLPS-induced pulpal inflammation. METHODS: Human dental pulp cells (HDPCs) were challenged with PgLPS with or without pretreatment and coincubation with a PI3K/Akt inhibitor (LY294002). The gene or protein levels of PI3K, Akt, interleukin (IL)-6, IL-8, alkaline phosphatase (ALP), osteocalcin and osteonectin were analyzed by reverse transcription polymerase chain reaction (PCR), real-time PCR, western blotting, and immunofluorescent staining. In addition, an enzyme-linked immunosorbent assay was used to analyze IL-6 and IL-8 levels in culture medium. RESULTS: In response to 5 µg/ml PgLPS, IL-6, IL-8, and PI3K, but not Akt mRNA expression of HDPCs, was upregulated. IL-6, IL-8, PI3K, and p-Akt protein levels were stimulated by 10-50 µg/ml of PgLPS in HDPCs. PgLPS also induced IL-6 and IL-8 secretion at concentrations higher than 5 µg/ml. Pretreatment and co-incubation by LY294002 attenuated PgLPS-induced IL-6 and IL-8 mRNA expression in HDPCs. The mRNA expression of ALP, but not osteocalcin and osteonectin, was inhibited by higher concentrations of PgLPS in HDPCs. CONCLUSION: P. gingivalis contributes to pulpal inflammation in HDPCs by dysregulating PI3K/Akt signaling pathway to stimulate IL-6 and IL-8 mRNA/protein expression and secretion. These results are useful for understanding the pulpal inflammation and possible biomarkers of inflamed pulp diagnosis and treatment.

The Microbiota Profile Analysis of Combined Periodontal-Endodontic Lesions Using 16S rRNA Next-Generation Sequencing.

OBJECTIVE: The primary aim of this investigation was to analyze the microbiome in patients with combined periodontal-endodontic lesions. METHOD: Patients with loose and/or painful teeth referred for treatment from March 2020 to December 2020 in the First People's Hospital of Jinzhong were recruited. Samples were collected from teeth diagnosed as chronic periodontics (PE), ulcerative pulpitis (PU), and retrograde pulpitis (RE). Genomic DNA was extracted. The quantitative polymerase chain reaction, targeting the 16S ribosomal RNA (rRNA), was adopted for the quantification of bacteria. Then, the V3-V4 hypervariable regions of the 16S rRNA gene were amplified and subjected to next-generation sequencing. The statistical analysis was performed by R software (V3.5.1). RESULTS: A total of 57 qualified samples were collected from 48 patients and analyzed (7 PE, 21 PU, and 19 RE). By linear discriminant analysis effect size, Kingella and Barnesiella were significantly increased in the periodontal pocket of retrograde pulpitis (RE-PE), compared with PE. The relative abundance of Clostridiales Incertae Sedis XI, Fusobacteriaceae, Fusobacterium, Parvimonas, Micrococcaceae, and Rothia was significantly increased in the pulp of retrograde pulpitis (RE-PU) than PU and RE-PE. Prevotella, Leptotrichia, Porphyromonas, Streptococcus, and Fusobacterium are consistently at a high abundance, across PU, RE-PE, and RE-PU. CONCLUSION: The current study highlighted the evidence that a specific microbial community is associated with the occurrence of retrograde pulpitis. The microenvironment of the root canal and pulp chamber will select microbiota. This study offered insights into the pathogenesis of retrograde pulpitis.

Diagnosing odontogenic sinusitis of endodontic origin: A multidisciplinary literature review.

PURPOSE: Endodontic disease is one of the most common causes of bacterial odontogenic sinusitis (ODS). Diagnosing ODS of endodontic origin involves otolaryngologists confirming sinusitis, and dental specialists confirming endodontic sources. The purpose of this study was to conduct a multidisciplinary literature review to highlight clinical and microbiological features of ODS, and the most optimal diagnostic modalities to confirm endodontic disease. METHODS: An extensive review of both medical and dental literature was performed by rhinologists, endodontists, and an infectious disease specialist. Frequencies of various clinical and microbiological features from ODS studies were collected, and averages were calculated. Different endodontic testing and imaging modalities were also evaluated on their abilities to confirm endodontic disease. RESULTS: ODS patients most often present with unilateral sinonasal symptoms for over 3 months, purulence on nasal endoscopy, and overt dental pathology on computed tomography (CT). Subjective foul smell, and maxillary sinus cultures demonstrating anaerobes and α-streptococci (viridans group) may be more specific to ODS. For endodontic evaluations, cold pulp testing and cone-beam CT imaging are most optimal for confirming pulpal and periapical disease. CONCLUSION: Diagnosing ODS requires collaboration between otolaryngologists and dental specialists. Clinicians should suspect ODS when patients present with unilateral sinonasal symptoms, especially foul smell. Patients will generally have purulent drainage on nasal endoscopy, and both sinus opacification and overt dental pathology on CT. However, some patients will have subtle or absent dental pathology on CT. For suspected endodontic disease, endodontists should be consulted for at least cold pulp testing, and ideally cone-beam CT.

Presence of bacterial DNA in thrombotic material of patients with myocardial infarction.

Infectious agents have been suggested to be involved in etiopathogenesis of Acute Coronary Syndrome (ACS). However, the relationship between bacterial infection and acute myocardial infarction (AMI) has not yet been completely clarified. The objective of this study is to detect bacterial DNA in thrombotic material of patients with ACS with ST-segment elevation (STEMI) treated with Primary Percutaneous Coronary Intervention (PPCI). We studied 109 consecutive patients with STEMI, who underwent thrombus aspiration and arterial peripheral blood sampling. Testing for bacterial DNA was performed by probe-based real-time Polymerase Chain Reaction (PCR). 12 probes and primers were used for the detection of Aggregatibacter actinomycetemcomitans, Chlamydia pneumoniae, viridans group streptococci, Porphyromonas gingivalis, Fusobacterium nucleatum, Tannarella forsythia, Treponema denticola, Helycobacter pylori, Mycoplasma pneumoniae, Staphylococus aureus,  Prevotella intermedia and Streptococcus mutans. Thus, DNA of four species of bacteria was detected in 10 of the 109 patients studied. The most frequent species was viridans group streptococci (6 patients, 5.5%), followed by Staphylococus aureus (2 patients, 1.8%). Moreover, a patient had DNA of Porphyromonas gingivalis (0.9%); and another patient had DNA of Prevotella intermedia (0.9%). Bacterial DNA was not detected in peripheral blood of any of our patients. In conclusion, DNA of four species of endodontic and periodontal bacteria was detected in thrombotic material of 10 STEMI patients. Bacterial DNA was not detected in the peripheral blood of patients with bacterial DNA in their thrombotic material. Bacteria could be latently present in plaques and might play a role in plaque instability and thrombus formation leading to ACS.

Identification of microorganisms in irreversible pulpitis and primary endodontic infections with respect to clinical and radiographic findings.

OBJECTIVES: The aim of this study was to evaluate the composition of microbiota of irreversible pulpitis and primary endodontic infections with respect to clinical and radiographic findings by performing cultures and 16s rDNA sequencing in Iranian patients. MATERIAL AND METHODS: In this prospective cross-sectional study, samples were collected from 41 root canals for 4 main groups of patients. Bacterial identification was performed by the polymerase chain reaction (PCR) and 16s rDNA sequencing of aerobic and anaerobic cultivable colonies taken from patients' culture plates. Additionally, the presence of 13 bacterial species and 3 nonbacterial species was also explored using PCR and species-specific primers. RESULTS: Sixteen microbial species, 1 fungus (Candida albicans), and 1 virus (Herpes simplex virus) were discovered and isolated. Species with the highest prevalence were Dialister invisus (68.3%), Porphyromonas gingivalis (58.8%), Streptococcus salivarius (58.5%), and Treponema denticola (56.1%). Lysinibacillus fusiformis (19.1%) was detected in the root canals for the first time. Candida albicans was seen in 11 cases (26.8%). Herpes simplex virus (HSV) was seen in 4 patients (9.8%). CONCLUSIONS: Our results suggest that Gram-negative anaerobic oral bacteria are the majority of the microbes in primary endodontic infections. Various combinations of bacterial species were related to different clinical and radiographic conditions. Lysinibacillus fusiformis was detected for the first time in primary endodontic infections. CLINICAL RELEVANCE: The results of this investigation might help clinicians choose to identify suspected endodontic pathogens in the etiology of each form of pulpal and periradicular diseases to determine the best therapeutic measures.

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.

Role(s) of cytokines in pulpitis: Latest evidence and therapeutic approaches.

Pulpitis is known as a typical inflammation of dental pulp tissue, and microorganisms of the oral microbiome are involved in this opportunistic infection. Studies indicated that several factors related to host response have a crucial role in pulpitis. Among these factors, inflammatory mediators of the immune system such as cytokines and chemokines contribute to pulpal defense mechanisms. A wide range of cytokines have been observed in dental pulp and these small molecules are able to trigger inflammation and participate in immune cell trafficking, cell proliferation, inflammation, and tissue damage in pulp space. Therefore, the aim of this review was to describe the role of cytokines in the pathogenesis of pulpitis.

The Susceptibility to Calcium Hydroxide Modulated by the Essential walR Gene Reveals the Role for Enterococcus faecalis Biofilm Aggregation.

INTRODUCTION: Enterococcus faecalis is considered a predominant pathogen for persistent periapical infections and in addition is reportedly resistant to calcium hydroxide medication. The WalRK 2-component system of E. faecalis is essential for environmental adaptation, survival, and virulence. The goal of this study was to investigate the potential roles of walR in the regulation of biofilm aggregation, alkaline stress, and susceptibility to calcium hydroxide (CH) medication. METHODS: Antisense walR RNA (aswalR) overexpression strains were constructed. Exopolysaccharide (EPS) production and bacterial viability of E. faecalis biofilms were evaluated by confocal laser scanning microscopy. Quantitative real-time polymerase chain reaction was used to investigate the expressions of virulent factor genes. The proportion of viable bacteria and EPS production in dentin were assessed after CH medication. RESULTS: We showed that walR interference by aswalR RNA leads to a reduction in the dextran-dependent aggregation in E. faecalis biofilm. The overexpression of aswalR reduced the transcripts of the virulence genes and alkaline stress tolerance ability. Furthermore, the down-regulation of walR sensitized E. faecalis in infected canals to CH medication associated with inhibiting EPS synthesis. CONCLUSIONS: The data suggest a role for the walR regulator in the susceptibility to CH associated with dispelling the EPS matrix, which could be explored as a potential supplementary therapy for the management of root canal infection.

Microbiome of Deep Dentinal Caries from Reversible Pulpitis to Irreversible Pulpitis.

INTRODUCTION: This study examined the identity of the microbiome of deep dentinal caries and its correlation with the inflammation status of caries-induced pulpitis. METHODS: Seventy-five cases were diagnosed based on the American Association of Endodontics's diagnostic criteria and divided into 4 groups: normal pulp with deep caries (NP; n = 13), reversible pulpitis with only cold-evoked pain (CRP; n = 17), reversible pulpitis with both cold/heat-evoked pain (CHRP; n = 24), and symptomatic irreversible pulpitis (SIP; n = 21). Samples were sequenced by 16S rDNA. Alpha and beta diversity were determined. Linear discriminant analysis effect size (LEfSe) analysis was used to detect intergroup differences, and receiver operating characteristic (ROC) curves were generated to assess the role of the caries microbiome in caries-induced pulpitis. RESULTS: The 16S rDNA sequencing yielded 9100 operational taxonomic units. Lactobacillus had the highest relative abundance at the genus level among the 4 groups. There were significant differences in the distribution of the microbiome among the groups. In an alpha diversity analysis, species richness differed between the CRP group and the other groups. In a beta diversity analysis, the distribution of microorganisms in the SIP group was significantly different from those in the other 3 groups. LEfSe analysis indicated substantial differences in the microbiome among the groups, and the areas under the ROC curves (AUC) were all high (AUC: 0.734-0.952). CONCLUSIONS: Characterization of the caries microbiome has the potential to become an auxiliary method for the diagnosis of pulpitis. This finding may prompt new research on diagnostic strategies for caries-induced pulpitis.

Kaurenoic acid and its sodium salt derivative: antibacterial activity against Porphyromonas gingivalis and their mechanism of action.

AIM: To evaluate the antibacterial activity of 12 kaurane-type diterpenes against a panel of bacteria that cause endodontic infection. METHODS & MATERIALS: We conducted tests against bacteria in the planktonic or in the sessile mode, cytotoxic assays for the most promising compounds against human normal lung fibroblast cells, and Porphyromonas gingivalis (ATCC 33277) proteomic analysis. RESULTS & CONCLUSION: Kaurenoic acid and its salt exhibited satisfactory antibacterial action against the evaluated bacteria. Proteomic analysis suggested that these compounds might interfere in bacterial metabolism and virulence factor expression. Kaurane-type diterpenes are an important class of natural products and should be considered in the search for new irrigating solutions to treat endodontic infections.

Comparative Molecular Analysis of Gram-Negative Bacteria in Primary Teeth with Irreversible Pulpitis or Periapical Pathology.

Purpose: The purpose of this study was to compare the Gram-negative pathogens identified in the root canals of primary teeth with irreversible inflammatory pulpitis and in teeth showing apical periodontitis. Methods: Samples were collected from 123 root canals of primary teeth from three- to seven-year-old patients. Root canals were assigned to either group one (irreversible inflammatory pulpitis; n equals 63) or group two (pulp necrosis and apical periodontitis; n equals 60). Total number of cells of selected Gram-negative microorganisms was determined by the checkerboard DNA-DNA hybridization technique. Demographic data were compared using either chi-squared or t tests. Total numbers of microorganisms were compared using the Mann-Whitney test (α equals 0.05). Results: There were no significant intergroup differences in gender, age, and tooth group distribution (P>0.05). Among the 123 samples, 17 were discarded due to salivary contamination. The total numbers of Prevotella nigrescens, Treponema denticola, Fusobacterium nucleatum polymorphum, Fusobacterium nucleatum spp nucleatum, Aggregatibacter actinomycetemcomitans serotype a, Aggregatibacter actinomycetemcomitans serotype b, Porphyromonas gingivalis, Prevotella intermedia, and Prevotella melaninogenica were higher in teeth with apical periodontitis compared to those with irreversible inflammatory pulpitis (P<0.05). Conclusion: Higher numbers of Gram-negative bacteria were found in teeth with apical periodontitis compared to teeth with irreversible in- flammatory pulpitis.

Roles of Oral Infections in the Pathomechanism of Atherosclerosis.

Oral infections occur frequently in humans and often lead to chronic inflammations affecting the teeth (i.e., caries), the gingival tissues surrounding the teeth (i.e., gingivitis and endodontic lesions), and the tooth-supporting structures (i.e., periodontitis). At least four basic pathogenic mechanisms have been proposed that involve oral inflammations in the pathogenesis of atherosclerosis: (1) low level bacteremia by which oral bacteria enter the blood stream and invade the arterial wall; (2) systemic inflammation induced by inflammatory mediators released from the sites of the oral inflammation into the blood stream; (3) autoimmunity to host proteins caused by the host immune response to specific components of oral pathogens; (4) pro-atherogenic effects resulting from specific bacterial toxins that are produced by oral pathogenic bacteria. In this narrative review, we summarize published experimental evidence related to these four mechanisms and discuss their impact on the pathogenesis of atherosclerosis.

Frecuencia de Candida en conductos radiculares de dientes con infección endodóntica primaria y persistente / Frequency of Candida in root canals of teeth with primary and persistent endodontic infections

Antecedentes: La identificación microbiológica en infecciones endodónticas se ha enfocado principalmente a la caracterización bacteriana sin dar relevancia a las levaduras que, por sus factores de virulencia, pueden afectar el resultado del tratamiento clínico realizado. Objetivos: Determinar la frecuencia de Candida en condiciones anaerobias en conductos radiculares con infecciones endodónticas primarias y persistentes, y evaluar un método de muestreo microbiológico por lavado y aspiración en comparación con el método tradicional por absorción con puntas de papel. Métodos: Se tomaron 50 muestras microbiológicas de dientes con infección endodóntica primaria y persistente provenientes de 47 pacientes que requirieron tratamiento endodóntico. Se emplearon dos métodos de toma de muestra microbiológica: un método por aspiración y un método por absorción con puntas de papel, ambos con dos tipos de caldo de cultivo (M1-M4). Las muestras fueron cultivadas en condiciones de anaerobiosis hasta lograr una turbidez de 0,5 en la escala de McFarland, y se resembraron en placas de agar dextrosa Sabouraud y agar sangre enriquecido para anaerobios. Se realizó una observación macroscópica y microscópica de las colonias formadas. Las pruebas de producción de tubo germinal, crecimiento en CHROMagar e identificación bioquímica se realizaron a los aislamientos levaduriformes obtenidos. Resultados: De los 50 dientes evaluados, 18 de ellos (36%) mostraron infección por levaduras. En los casos de infección primaria se encontraron levaduras en 15 de 36 dientes (41,6%) y en casos de infección persistente en 3 de 14 (21,4%). El método por lavado y aspiración con caldo de dextrosa Sabouraud recuperó mayor diversidad de especies. Conclusiones: La frecuencia de levaduras fue más alta en los dientes con infección primaria en comparación con los dientes con infección persistente. La especie de levadura predominante fue Candida albicans. El método de toma de muestra por lavado y aspiración fue más eficiente en la recuperación de aislamientos de Candida que el método tradicional por absorción con puntas de papel

Background: Microbiological identification in endodontic infections has focused mainly on bacteria without giving much attention to yeasts, which, due to their virulence factors, can affect the outcomes of root canal treatment. Aims: To determine the frequency of Candida in anaerobic conditions in root canals with primary and persistent endodontic infection, as well as to evaluate a microbiological sampling method using aspiration compared to the traditional absorption method with paper points. Methods: Fifty microbiological samples were obtained from teeth of 47 patients requiring endodontic treatments, due to either primary or persistent infections. Two microbiological sampling methods were used: an aspiration method, and the traditional paper point absorption method. In each of these methods, two types of medium were used (M1-M4). Samples were cultured under anaerobic conditions until reaching 0.5 McFarland turbidity, and then inoculated on Sabouraud dextrose, as well as on anaerobic enriched blood agar plates. Macroscopic and microscopic observations of the colonies were performed. The germ-tube test, growth on CHROMagar, and biochemical identification were performed on the isolated yeasts. Results: Fungal infection was found in 18 (36%) samples out of the 50 teeth evaluated. In the 18 samples positive for fungal infection, 15 out of 36 (41.6%) teeth were taken from a primary infection, and 3 out of 14 (21.4%) from a persistent infection. The aspiration method using Sabouraud dextrose medium recovered a greater diversity of species. Conclusions: Yeasts frequency was higher in teeth with primary infections compared to teeth with persistent infections. The predominant yeast species was Candida albicans. The aspirating sampling method was more efficient in the recovery of Candida isolates than the traditional absorption method

Molecular and clinical analyses of Helicobacter pylori colonization in inflamed dental pulp.

BACKGROUND: Recently, dental pulp has been considered a possible source of infection of Helicobacter pylori (H. pylori) in children. We previously developed a novel PCR system for H. pylori detection with high specificity and sensitivity using primer sets constructed based on the complete genome information for 48 H. pylori strains. This PCR system showed high sensitivity with a detection limit of 1-10 cells when serial dilutions of H. pylori genomic DNA were used as templates. However, the detection limit was lower (102-103 cells) when H. pylori bacterial DNA was detected from inflamed pulp specimens. Thus, we further refined the system using a nested PCR method, which was much more sensitive than the previous single PCR method. In addition, we examined the distribution and virulence of H. pylori in inflamed pulp tissue. METHODS: Nested PCR system was constructed using primer sets designed from the complete genome information of 48 H. pylori strains. The detection limit of the nested PCR system was 1-10 cells using both H. pylori genomic DNA and bacterial DNA isolated from inflamed pulp specimens. Next, distribution of H. pylori was examined using 131 inflamed pulp specimens with the nested PCR system. In addition, association between the detection of H. pylori and clinical information regarding endodontic-infected teeth were investigated. Furthermore, adhesion property of H. pylori strains to human dental fibroblast cells was examined. RESULTS: H. pylori was present in 38.9% of inflamed pulp specimens using the nested PCR system. H. pylori was shown to be predominantly detected in primary teeth rather than permanent teeth. In addition, samplings of the inflamed pulp were performed twice from the same teeth at 1- or 2-week intervals, which revealed that H. pylori was detected in most specimens in both samplings. Furthermore, H. pylori strains showed adhesion property to human dental fibroblast cells. CONCLUSION: Our results suggest that H. pylori colonizes inflamed pulp in approximately 40% of all cases through adhesion to human dental fibroblast cells.

[Frequency of Candida in root canals of teeth with primary and persistent endodontic infections]. / Frecuencia de Candida en conductos radiculares de dientes con infección endodóntica primaria y persistente.

BACKGROUND: Microbiological identification in endodontic infections has focused mainly on bacteria without giving much attention to yeasts, which, due to their virulence factors, can affect the outcomes of root canal treatment. AIMS: To determine the frequency of Candida in anaerobic conditions in root canals with primary and persistent endodontic infection, as well as to evaluate a microbiological sampling method using aspiration compared to the traditional absorption method with paper points. METHODS: Fifty microbiological samples were obtained from teeth of 47 patients requiring endodontic treatments, due to either primary or persistent infections. Two microbiological sampling methods were used: an aspiration method, and the traditional paper point absorption method. In each of these methods, two types of medium were used (M1-M4). Samples were cultured under anaerobic conditions until reaching 0.5 McFarland turbidity, and then inoculated on Sabouraud dextrose, as well as on anaerobic enriched blood agar plates. Macroscopic and microscopic observations of the colonies were performed. The germ-tube test, growth on CHROMagar, and biochemical identification were performed on the isolated yeasts. RESULTS: Fungal infection was found in 18 (36%) samples out of the 50 teeth evaluated. In the 18 samples positive for fungal infection, 15 out of 36 (41.6%) teeth were taken from a primary infection, and 3 out of 14 (21.4%) from a persistent infection. The aspiration method using Sabouraud dextrose medium recovered a greater diversity of species. CONCLUSIONS: Yeasts frequency was higher in teeth with primary infections compared to teeth with persistent infections. The predominant yeast species was Candida albicans. The aspirating sampling method was more efficient in the recovery of Candida isolates than the traditional absorption method.

Enterococcus faecalis Demonstrates Pathogenicity through Increased Attachment in an Ex Vivo Polymicrobial Pulpal Infection.

This study investigated the host response to a polymicrobial pulpal infection consisting of Streptococcus anginosus and Enterococcus faecalis, bacteria commonly implicated in dental abscesses and endodontic failure, using a validated ex vivo rat tooth model. Tooth slices were inoculated with planktonic cultures of S. anginosus or E. faecalis alone or in coculture at S. anginosus/E. faecalis ratios of 50:50 and 90:10. Attachment was semiquantified by measuring the area covered by fluorescently labeled bacteria. Host response was established by viable histological cell counts, and inflammatory response was measured using reverse transcription-quantitative PCR (RT-qPCR) and immunohistochemistry. A significant reduction in cell viability was observed for single and polymicrobial infections, with no significant differences between infection types (∼2,000 cells/mm2 for infected pulps compared to ∼4,000 cells/mm2 for uninfected pulps). E. faecalis demonstrated significantly higher levels of attachment (6.5%) than S. anginosus alone (2.3%) and mixed-species infections (3.4% for 50:50 and 2.3% for 90:10), with a remarkable affinity for the pulpal vasculature. Infections with E. faecalis demonstrated the greatest increase in tumor necrosis factor alpha (TNF-α) (47.1-fold for E. faecalis, 14.6-fold for S. anginosus, 60.1-fold for 50:50, and 25.0-fold for 90:10) and interleukin 1ß (IL-1ß) expression (54.8-fold for E. faecalis, 8.8-fold for S. anginosus, 54.5-fold for 50:50, and 39.9-fold for 90:10) compared to uninfected samples. Immunohistochemistry confirmed this, with the majority of inflammation localized to the pulpal vasculature and odontoblast regions. Interestingly, E. faecalis supernatant and heat-killed E. faecalis treatments were unable to induce the same inflammatory response, suggesting E. faecalis pathogenicity in pulpitis is linked to its greater ability to attach to the pulpal vasculature.

Rotary versus reciprocating shaping files in bacterial elimination from long oval canals.

A comparison of the abilities of rotary versus reciprocating files to eliminate viable Enterococcus faecalis populations from the long oval root canals of extracted human teeth. Fifty teeth were contaminated and randomly distributed into two groups (n = 25 each): BT-RaCe group and WaveOne group. Two microbial samples were obtained from each tooth before (S1) and after (S2) instrumentation. The CFUs from the S1 and S2 measurements were calculated and compared between the groups. Both groups showed significantly fewer CFUs in the S2 samples (P < 0.001). In the S2 intragroup comparison, BT-RaCe resulted in significantly fewer CFUs than WaveOne (P = 0.010). In the direct comparison between the rotary multiple file shaping system and the reciprocating single-file system, the multiple file system was more efficient at reducing the microbiological load of viable E. faecalis from long oval root canals.