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

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

A Multimethod Assessment of a New Customized Heat-Treated Nickel-Titanium Rotary File System.

This study aimed to compare three endodontic rotary systems. The new Genius Proflex (25/0.04), Vortex Blue (25/0.04), and TruNatomy (26/0.04v) instruments (n = 41 per group) were analyzed regarding design, metallurgy, and mechanical performance, while shaping ability (untouched canal walls, volume of removed dentin and hard tissue debris) was tested in 36 anatomically matched root canals of mandibular molars. The results were compared using one-way ANOVA, post hoc Tukey, and Kruskal−Wallis tests, with a significance level set at 5%. All instruments showed symmetrical cross-sections, with asymmetrical blades, no radial lands, no major defects, and almost equiatomic nickel−titanium ratios. Differences were noted in the number of blades, helical angles, cross-sectional design, and tip geometry. The Genius Proflex and the TruNatomy instruments had the highest and lowest R-phase start and finish temperatures, as well as the highest and lowest time and cycles to fracture (p < 0.05), respectively. The TruNatomy had the highest flexibility (p < 0.05), while no differences were observed between the Genius Proflex and the Vortex Blue (p > 0.05). No differences among tested systems were observed regarding the maximum torque, angle of rotation prior to fracture, and shaping ability (p > 0.05). The instruments showed similarities and differences in their design, metallurgy, and mechanical properties. However, their shaping ability was similar, without any clinically significant errors. Understanding these characteristics may help clinicians to make decisions regarding which instrument to choose for a particular clinical situation.

Cytotoxicity, antibacterial and physicochemical properties of a new epoxy resin-based endodontic sealer containing calcium hydroxide.

BACKGROUND: This study evaluated the cytotoxicity, antibacterial and physicochemical properties of a new epoxy resin-based endodontic sealer containing calcium-hydroxide Sealer Plus. AH Plus was used as a reference for comparison. MATERIAL AND METHODS: Cytotoxicity evaluation was performed according to ISO-10993-5 specifications using MTT assay to check the 3T3 cells viability at 1- to 4-week periods. Antibacterial activity was evaluated using a direct contact test against Enterococcus faecalis. Radiopacity, solubility and flow evaluations were performed according to ISO-6876/2012 specifications. Setting time was assessed following the ANSI/ADA-standard-n.57. The pH level was measured at 3,24,48,72, and 168 hours. Data were statistically analyzed using t-test. The significance level adopted was P<0.05. RESULTS: AH Plus and Sealer Plus showed cytotoxic effects after 24 hours and 1 week of manipulation and become noncytotoxic after 2 weeks. No differences were observed in the cytotoxicity of both tested sealers (P>0.05). Direct contact results revealed that both freshly prepared sealers has antibacterial effects against Enterococcus faecalis. However, after 7 days both sealers had lost much of their antibacterial effects. Although AH Plus presented higher radiopacity and flow than Sealer Plus (P<0.05), both sealers showed minimum required values. No differences were observed in the solubility of both sealers (P>0.05). AH Plus showed a higher setting time when compared to Sealer Plus (P<0.05). AH Plus and Sealer Plus showed a neutral pH during all tested periods (P<0.05). CONCLUSIONS: It can be concluded that Sealer Plus showed suitable properties to be used as an endodontic sealer, comparable with those obtained by AH Plus. Key words:Cytotoxicity, root canal filling materials, root canal obturation.

The Apical Root Canal System of Teeth with Posttreatment Apical Periodontitis: Correlating Microbiologic, Tomographic, and Histopathologic Findings.

INTRODUCTION: This study evaluated the microbiological conditions of the apical root canal system of teeth with posttreatment apical periodontitis and correlated them with observations from cone-beam computed tomographic (CBCT) imaging, micro-computed tomographic (micro-CT) imaging, and histopathology. METHODS: Root apices were obtained from 36 root canal-treated teeth subjected to periradicular surgery. CBCT examination was available before surgery. The apical root specimens were scanned in a micro-CT device and then cryopulverized. The powder was subjected to DNA extraction for real-time polymerase chain reaction quantification of total bacteria, Streptococcus species, members of the phylum Actinobacteria, and Enterococcus faecalis. Microbiological findings were evaluated for associations with CBCT, micro-CT, and histopathologic data. An association between lesion size and the proportion of unfilled apical canal system volume was also assessed. RESULTS: All cryopulverized specimens were positive for total bacteria. Actinobacteria and streptococci occurred in 35 and 33 specimens, respectively, and were usually dominant in the community. Actinobacteria counts were 2.23 times higher in granulomas than in cysts. Streptococci were significantly more present in small lesion cases. E. faecalis was detected in only 7 samples, always as a dominant community member. The association of total bacteria, streptococci, and Actinobacteria counts with the unfilled canal volume was significant in the univariate analyses but not confirmed in the adjusted analyses. Large lesions were significantly associated with a higher volume of unfilled apical canals. CONCLUSIONS: Bacterial infection occurred in all root apices, with high prevalence and dominance of Actinobacteria and streptococci. The volume of the unfilled apical canal system was significantly associated with the lesion size and possibly with bacterial counts. Findings illustrate the need to thoroughly disinfect and fill the apical root canal of infected teeth during endodontic therapy.

Molecular Analysis of the Antibacterial Effects of Photodynamic Therapy in Endodontic Surgery: A Case Series.

In this case series, bacterial reduction promoted by antimicrobial photodynamic therapy (PDT) used during endodontic surgery was evaluated. Cases were also followed up, and the surgical outcome was reported. The study consisted of 19 teeth with posttreatment apical periodontitis that were consecutively treated by endodontic surgery. After apicoectomy, the root end was treated with PDT using methylene blue as the photosensitizer. Bacteriologic samples were taken from both the cut root surface and the root-end cavity before and after PDT. Samples were analyzed for the total bacterial and Streptococcus group counts using quantitative real-time polymerase chain reaction. EndoSequence BC-RRM Putty (Brasseler, Savannah, GA) was used as the root-end filling. Patients were followed up, and the surgical outcome was evaluated. The reduction in bacterial counts after the PDT approach was analyzed using the nonparametric Wilcoxon signed rank test. PDT significantly reduced the total bacterial and streptococcal counts in both root-end cavities and resected root surfaces (P < .05). The success rate for 15 teeth that were available for recall after 12 to 21 months was 93% under a loose evaluation criterion and 73% under a rigid one. Used during endodontic surgery, PDT significantly enhanced disinfection of the cut surface area and root-end cavity. Cases treated with PDT showed a high healing rate.

Human Exoproteome in Acute Apical Abscesses.

INTRODUCTION: An acute apical abscess is a severe response of the host to massive invasion of the periapical tissues by bacteria from infected root canals. Although many studies have investigated the microbiota involved in the process, information on the host factors released during abscess formation is scarce. The purpose of this study was to describe the human exoproteome in samples from acute apical abscesses. METHODS: Fourteen pus samples were obtained by aspiration from patients with an acute apical abscess. Samples were subjected to protein digestion, and the tryptic peptides were analyzed using a mass spectrometer and ion trap instrument. The human proteins identified in this analysis were classified into different functional categories. RESULTS: A total of 303 proteins were identified. Most of these proteins were involved in cellular and metabolic processes. Immune system proteins were also very frequent and included immunoglobulins, S100 proteins, complement proteins, and heat shock proteins. Polymorphonuclear neutrophil proteins were also commonly detected, including myeloperoxidases, defensins, elastases, and gelatinases. Iron-sequestering proteins including transferrin and lactoferrin/lactotransferrin were found in many samples. CONCLUSIONS: The human exoproteome included a wide variety of proteins related to cellular processes, metabolism, and immune response. Proteins involved in different mechanisms against infection, tissue damage, and protection against tissue damage were identified. Knowledge of the presence and function of these proteins using proteomics provides an insight into the complex host-pathogen relationship, the host antimicrobial strategies to fight infections, and the disease pathogenesis.

Microbiome in the Apical Root Canal System of Teeth with Post-Treatment Apical Periodontitis.

INTRODUCTION: Bacteria present in the apical root canal system are directly involved with the pathogenesis of post-treatment apical periodontitis. This study used a next-generation sequencing approach to identify the bacterial taxa occurring in cryopulverized apical root samples from root canal-treated teeth with post-treatment disease. METHODS: Apical root specimens obtained during periradicular surgery of ten adequately treated teeth with persistent apical periodontitis were cryogenically ground. DNA was extracted from the powder and the microbiome was characterized on the basis of the V4 hypervariable region of the 16S rRNA gene by using paired-end sequencing on Illumina MiSeq device. RESULTS: All samples were positive for the presence of bacterial DNA. Bacterial taxa were mapped to 11 phyla and 103 genera composed by 538 distinct operational taxonomic units (OTUs) at 3% of dissimilarity. Over 85% of the sequences belonged to 4 phyla: Proteobacteria, Firmicutes, Fusobacteria and Actinobacteria. In general, these 4 phyla accounted for approximately 80% of the distinct OTUs found in the apical root samples. Proteobacteria was the most abundant phylum in 6/10 samples. Fourteen genera had representatives identified in all cases. Overall, the genera Fusobacterium and Pseudomonas were the most dominant. Enterococcus was found in 4 cases, always in relatively low abundance. CONCLUSIONS: This study showed a highly complex bacterial community in the apical root canal system of adequately treated teeth with persistent apical periodontitis. This suggests that this disease is characterized by multispecies bacterial communities and has a heterogeneous etiology, because the community composition largely varied from case to case.

Host-Bacterial Interactions in Post-treatment Apical Periodontitis: A Metaproteome Analysis.

INTRODUCTION: This study evaluated the bacterial and human metaproteome of root apexes and the matched inflammatory lesions from teeth with post-treatment apical periodontitis. METHODS: Root apexes and matched inflammatory lesions from root canal-treated teeth with apical periodontitis were obtained during periradicular surgery. All root canal fillings were rated as adequate on the basis of radiographs and cone-beam computed tomography. The specimens were cryopulverized and subjected to metaproteomic analysis for human and bacterial proteins by using a mass spectrometry platform that is based on nanoflow liquid chromatography coupled with linear ion trap quadrupole Velos Orbitrap. RESULTS: The metaproteome analyses revealed the presence of viable and metabolically active human and bacterial cells in both apexes and lesions. Several bacterial proteins of interest for pathogenicity and therapeutics were identified in both apexes and lesions, including proteins related to antibiotic resistance, proteolytic function, stress response, adhesion, and virulence. Many human proteins related to immune defense mechanisms were also detected in both root apex and lesion specimens, including immunoglobulins, complement system, and proteins linked to T-cell and B-cell activation, neutrophil microbicidal processes, antigen recognition/presentation, bone resorption, and protection against tissue damage. CONCLUSIONS: Occurrence of host defense factors from the innate and adaptive immune responses and bacterial virulence, survival, and resistance proteins in matched root apexes/periradicular inflammatory lesions indicates a complex and dynamic host-pathogen interaction in teeth with post-treatment apical periodontitis.

Infection Control in Retreatment Cases: In Vivo Antibacterial Effects of 2 Instrumentation Systems.

INTRODUCTION: This in vivo study compared the antibacterial effects of 2 instrumentation systems in root canal-treated teeth with apical periodontitis. METHODS: Forty-eight teeth with a single root and a single canal showing post-treatment apical periodontitis were selected for this study. For retreatment, teeth were randomly divided into 2 groups according to the instrumentation system used: Self-Adjusting File (SAF; ReDent-Nova, Ra'anana, Israel) and Twisted File Adaptive (TFA; SybronEndo, Orange, CA). In both groups, 2.5% sodium hypochlorite was the irrigant. Bacteriological samples were taken before (S1) and after chemomechanical preparation (S2). In the TFA group, passive ultrasonic irrigation (PUI) was performed after instrumentation, and samples were also taken after this supplementary step (S2b). DNA was extracted from the clinical samples and subjected to quantitative real-time polymerase chain reaction to evaluate the levels of total bacteria, streptococci, and Enterococcus faecalis. Statistical analyses from quantitative real-time polymerase chain reaction data were performed within groups using the Wilcoxon matched pairs test and between groups using the Mann-Whitney U test and the Fisher exact test with the significance level set at P < .05. RESULTS: Bacteria were detected in S1 samples from 43 teeth, which were then included in the antibacterial experiment. Both SAF and TFA instrumentation protocols showed a highly significant intracanal bacterial reduction (P < .001). Intergroup quantitative comparisons disclosed no significant differences between TFA with or without PUI and SAF (P > .05). PUI did not result in significant improvement in disinfection (P > .05). CONCLUSIONS: Both instrumentation systems/treatment protocols were highly effective in significantly reducing the intracanal bacterial counts. No significant difference was observed between the 2 systems in disinfecting the canals of teeth with post-treatment apical periodontitis.

Total and Specific Bacterial Levels in the Apical Root Canal System of Teeth with Post-treatment Apical Periodontitis.

INTRODUCTION: Most studies of the microbiota in root canal-treated teeth focused only on the main canal, not distinguishing regions nor incorporating the intricate anatomy in the analysis. Moreover, most of them provided only prevalence data. This study was designed to evaluate the total bacterial counts and the presence, levels, and relative abundance of candidate endodontic pathogens exclusively in the apical root canal system associated with post-treatment apical periodontitis. METHODS: Apical root specimens obtained during periradicular surgery of 27 adequately treated teeth with persistent apical periodontitis were cryogenically ground. DNA was extracted from the powder, and real-time polymerase chain reaction was used to quantify the total bacteria and 7 bacterial taxa. RESULTS: Samples from 21 teeth were positive for bacteria. Streptococcus species were the most prevalent (76%) followed by members of the Actinobacteria phylum (52%) and Pseudoramibacter alactolyticus (19%). The mean total bacterial load in the apical root segments was 5.7 × 10(4) cell equivalents per root apex (or 2.1 × 10(4)/100 mg root powder). Streptococci comprised from 0.02%-99.9% of the total bacterial counts, Actinobacteria from 0.02%-84.7%, and P. alactolyticus from 67.9%-99%. Although Enterococcus faecalis was found in only 3 (14%) cases, it was dominant in 2. CONCLUSIONS: Streptococcus species, members of the Actinobacteria phylum, and P. alactolyticus were the most prevalent taxa in the apical canal system and dominated the bacterial populations in many cases of post-treatment apical periodontitis.

Estudo epidemiológico das fraturas coronárias em pacientes atendidos em um projeto de trauma dental em um período de 6 anos / Epidemiological study of coronal fractures in patients treated in a dental trauma project in a 6 years periods

O presente estudo teve como objetivo realizar um levantamento epidemiológico sobre os casos de fratura coronária em pacientes que foram atendidos no Projeto Trauma Dental da Escola de Odontologia da Unigranrio, no período de 2005 a 2011. O estudo foi baseado nos dados coletados dos prontuários e as informações obtidas estão relacionadas ao gênero, idade, etiologia da fratura, local do pronto atendimento, tempo decorrido até o pronto atendimento, dentes acometidos, situação clínica e a necessidade de tratamento endodôntico. Foi observada uma maior incidência em pacientes do gênero masculino, com a idade mais afetada entre 6 e 10 anos. Os resultados mostram a distribuição etiológica, clínica e demográfica das fraturas coronárias em um projeto de trauma dentário na cidade de Duque de Caxias.

The aim of the present study was to conduct an epidemiological survey on coronal fracture cases in patients treated at the Dental Trauma Project at Unigranrio School of Dentistry, from 2005 to 2011. This study was based on data collected from records and information related to gender, age, fracture cause, prompt service, elapsed time from the accident, affected teeth, clinical condition of the teeth and the need of endodontic treatment. It was observed a higher incidence in male patients, with the most affected age between 6 to 10 years. The results show the etiological, clinical and demographic distribution of coronal fractures in a project of dental trauma in Duque de Caxias city.