Radiographic outcome of endodontic treatment and retreatment of teeth with apical periodontitis using two different root canal irrigants. A prospective cohort study.

AIM: The aim of this prospective cohort study was to compare the radiographic outcome of endodontic treatment and retreatment of teeth with apical periodontitis using either 1% sodium hypochlorite (NaOCl) or 2% chlorhexidine digluconate (CHX) for root canal irrigation. MATERIALS AND METHODS: In the years from 2013 to 2015 standard irrigation varied by semester between NaOCl and CHX at the Department of Endodontics at the Faculty of Dentistry. During that time, 912 teeth received treatment for apical periodontitis in 744 patients, of whom 532 responded to the request for a 1-year follow-up. Only one tooth per person (the most distally located) were included; 285 teeth treated with NaOCl and 247 with CHX. One hundred cases were then randomly sampled from each irrigation group and analysed for outcome by periapical index (PAI) scoring using criteria for success, uncertain and failure. Clinical and other radiographic parameters were scored or recorded and analysed for associations with radiographic outcome using chi-square, ANOVA and regression analyses. RESULTS: Success rates (PAI score 1 or 2 at control) were nearly identical for the two irrigation liquids. The use of irrigating solution also did not significantly influence the outcome in chi-square analyses of subgroups of teeth or regression analyses with other variables included. Ordinal regression analysis established that preoperative lesion size or preoperative PAI score were significantly associated with outcome, and teeth in the mandible had significantly better outcomes than in the maxilla. CONCLUSIONS: No significant differences in the radiographic outcome using either 1% NaOCl or 2% CHX as irrigants were found. The outcome was better for teeth with small lesions or lower PAI scores at completion of treatment and for mandibular teeth.

Endodontic sealers after exposure to chlorhexidine digluconate: An assessment of physicochemical properties.

OBJECTIVES: Final root canal irrigation should ideally maintain the physicochemical stability of root canal sealers. We seek to assess the effect of contact with 2% chlorhexidine digluconate (CHX) on the physicochemical properties of AH Plus, BioRoot™ RCS, and Pulp Canal Sealer (PCS). METHODS: Mixed sealers were placed in cylindrical teflon molds and allowed to set for 1.5x the manufacturers' setting time. Half of the specimens had their free surface in contact with CHX for the first minute of their setting period. Solubility, radiopacity, surface roughness, microhardness and wettability of the sealers were assessed up to 28 days after setting. Elemental analysis of sealer surfaces and their leachates together with pH measurements were also performed. Appropriate parametric and non-parametric analysis with post hoc tests were performed (p < 0.05). RESULTS: Exposure to CHX had no effect on solubility and radiopacity of all sealers. CHX altered the surface roughness of PCS and BioRoot RCS (p < 0.05). Contact with CHX reduced the microhardness of AH Plus and PCS (p < 0.05). AH Plus was more hydrophilic after CHX contact, whereas PCS became more hydrophobic (p < 0.05). AH Plus and PCS surfaces appeared to adsorb CHX as exhibited by chlorine peaks after contact with CHX. Sealer leachates' alkalinity was not affected. CHX increased elution of silicon and zirconium for BioRoot and zinc for PCS leachates. SIGNIFICANCE: In our study, CHX affected sealers' physicochemical properties to various extents. Further studies are needed to confirm the obtained results by investigating various final irrigation strategies and correlating to biological properties.

Antibacterial Activity of Root Repair Cements in Contact with Dentin-An Ex Vivo Study.

This study assessed the antibacterial characteristics of the dentin/material interface and dentin surfaces exposed to experimental hydraulic calcium silicate cement (HCSC) with or without bioactive glass (BG) replacement (20% or 40%) or mixed with a silver nanoparticle (SNP) solution (1 or 2 mg/mL), and Biodentine, TotalFill BC RRM putty and Intermediate Restorative Material (IRM). Human root dentin segments with test materials were assessed at 1 or 28 days. In one series, the specimens were split to expose the dentin and material surfaces. A 24 h direct contact test was conducted against three-day established Enterococcus faecalis and Pseudomonas aeruginosa monospecies biofilms. In another series, the dentin/material interface of intact specimens was exposed to biofilm membranes for 3 days and the antibacterial activity was assessed via confocal microscopy. The interface was additionally characterised. All one-day material and dentin surfaces were antibacterial. Dentin surfaces exposed to HCSC with 40% BG-replacement, Biodentine and IRM had decreased antibacterial properties compared to those of the other cements. The HCSC mixed with a 2 mg/mL SNP solution had the highest antimicrobial effect in the confocal assay. The interfacial characteristics of HCSCs were similar. The test materials conferred antibacterial activity onto the adjacent dentin. The BG reduced the antibacterial effect of dentin exposed to HCSC; a 2 mg/mL SNP solution increased the antibacterial potential for longer interaction periods (three-day exposure).

Are comorbidities associated with the cytokine/chemokine profile of persistent apical periodontitis?

OBJECTIVES: This study aimed to identify disease-related markers in persistent apical periodontitis (PAP) biopsies and examine whether these were associated with comorbidities like rheumatoid arthritis (RA) and cardiovascular diseases (CVD). MATERIALS AND METHOD: The levels of the cytokines/chemokines GM-CSF, IFN-γ, IL-2, IL-6, IL-9, IL-10, IL-13, IL-15, IL-17E/IL-25, IL-21, IL-23, IL-27, IL-28A/IFN -λ2, IL-33, MIP-3α/CCL20, and TNF-α were determined in lesions from patients with PAP (n = 20) and compared to healthy bone samples (n = 20). RESULTS: We identified eleven cytokines to be differently expressed, and among them, IL-2, IL-6, IL-17E, IL-21, and IL-27 appeared to drive the discrepancy between the disease and healthy groups. The levels of T follicular helper (Tfh) cell promoting cytokines (IL-21, IL-6, IL-27) were enhanced while T helper (Th) 1 cell promoting cytokine (IL-2), Th2 cell promoting cytokine (IL-13), and Th17 cell promoting cytokine (IL-17E) were reduced in the PAP group. The data also indicate that Tfh cell differentiation (IL-21), along with Th1 (GM-CSF, IFNγ), Th2 (IL-13), and Th17 (GM-CSF) cell differentiation, might be increased in the subpopulation of patients suffering from RA, whereas no differences were found in patients with CVD. CONCLUSIONS: Levels of cytokines/chemokines in PAP were identified, and cluster analyzes indicated that these markers may be associated with the differentiation of different T cell populations. Patients with PAP and RA comorbidities showed elevated levels of markers reinforcing this association. CLINICAL RELEVANCE: Molecular analyses of PAP may result in identification of prognostic markers.

Effect of exposure conditions on chemical properties of materials for surgical endodontic procedures.

This study investigated the role of aging and changes in environmental conditions on selected properties of a prototype radiopacified calcium silicate-based cement (TZ-base) with or without incorporation of silver nanoparticles or bioactive glass, and two commercial materials, Biodentine and intermediate restorative material. Materials were immersed in ultrapure water or fetal bovine serum for 28 days and were characterized with scanning electron microscopy and energy dispersive x-ray analysis. Immersion media were either replaced weekly or not replenished at all and were assessed for alkalinity and calcium release after 1, 7, 14, 21, and 28 days; antibacterial effect against 2-day monospecies biofilms; and cytotoxicity by the 3-(4,5 dimethylthiazolyl-2-yl)-2,5-diphenyl tetrazolium bromide assay after 1, 7, or 28 days. Alkalinity, calcium release, antibacterial activity, and cell cytotoxicity increased over time when the medium was not changed but decreased with medium replenishment. Immersion in fetal bovine serum resulted in lower alkalinity, less bactericidal properties, and lower cytotoxicity of prototype cements and Biodentine than did water immersion. Biodentine and 20% bioactive glass-containing cement had overall lower alkalinity, calcium release, and antibacterial activity than TZ-base, and Biodentine was less cytotoxic than TZ-base. In conclusion, exposure conditions and cement modifications significantly affected materials' leaching properties. Exposure conditions warrant consideration when evaluating cements' clinical properties.

Periapical and endodontic status among 65-year-old Oslo-citizens.

AIM: This cross-sectional study aimed to investigate the prevalence of apical periodontitis (AP) and root-filled teeth in a 65-year-old population in Oslo, Norway, and to investigate associations of pathosis and endodontic treatment with selected individual risk indicators and technical quality of root fillings. MATERIAL AND METHODS: A random sample of 450 65-year-olds in Oslo answered a questionnaire and underwent a clinical and radiological examination (52% men and 48% women). Periapical radiographs were taken of all root-filled teeth and of teeth with apical radiolucency, and periapical status was evaluated using the Periapical Index. Apex-to-filling distance and homogeneity were assessed for all root fillings. Analyses on individual level and tooth level were performed. The outcome variables were 'non-root-filled tooth with AP' ('untreated AP'), 'root-filled tooth', and 'root-filled tooth with AP'. The explanatory variables were gender, education, dental attendance pattern, smoking, remaining teeth (n), tooth group, and root filling quality. Chi-square test and logistic regression analyses were used to assess the associations between outcome variables and explanatory variables. The level of significance was set to p < 0.05. RESULTS: The mean number of remaining teeth was 26 (SD: 4). AP was present in 45% of the individuals. Sixteen percent of the individuals had untreated AP and 38% had at least one root-filled tooth with AP. Sixty-six percent of the individuals had one or more root-filled teeth. Untreated AP was significantly associated with a decreasing number of remaining teeth and smoking. All the outcome variables were significantly more prevalent in molars compared with premolars and anterior teeth. Thirty-five percent of the root-filled teeth had AP, and AP was more prevalent in teeth with too short apex-to-filling distance (53%) or unsatisfactory homogeneity (46%). CONCLUSIONS: The remaining number of teeth was high, and AP and root-filled teeth were prevalent in the present young-elderly population. A notable amount of untreated AP was observed, especially in smokers. The findings in the present study indicate a substantial need for dental care associated with endodontic conditions in the future elderly.

Surface characteristics and bacterial adhesion of endodontic cements.

OBJECTIVES: To investigate the effect of inclusion of silver nano-particles (SNP) or bioactive glass (BG) on the surface characteristics and bacterial adhesion of prototype tricalcium silicate (TCS)-based cements alongside two commercial cements, under different aging periods and exposure conditions. MATERIALS AND METHODS: A basic formulation of radio-opacified TCS without (TZ-base) and with additions of SNP (0.5, 1, or 2 mg/ml) or BG (10 or 20%) was used. Biodentine and intermediate restorative material (IRM) served as reference materials. Material disks were immersed in ultrapure water or fetal bovine serum (FBS) for 1, 7, or 28 days. Surface roughness (n = 3), microhardness (n = 9), and wettability (n = 6) were analyzed by standard procedures. Adhesion of Enterococcus faecalis was assessed by fluorescence microscopy (n = 5). Data from these assays were evaluated for normality and comparisons among groups were conducted with statistical procedures (p < 0.05 for significance). RESULTS: The surface morphology of SNP- and BG-containing cements had higher roughness values than TZ-base after 28 days (p < 0.05). No differences in microhardness were observed among prototype cements (p > 0.05). Biodentine presented smooth surface characteristics and the highest hardness values (p < 0.05). The FBS-immersion resulted in surface reactions in prototype materials and Biodentine, depicted with scanning electron microscopy. All 1- and 7-day prototype cements showed negligible bacterial adhesion, while in Biodentine and IRM, noticeable E. faecalis adherence was observed from day 1 (p < 0.05). CONCLUSIONS: Incorporation of SNP or BG did not improve the antibacterial effect of the experimental cement; all 28-day aged materials failed to inhibit bacterial adherence. The measured physical parameters did not appear to be related to the degree of bacterial adhesion. Exposure of TCS-based cements in FBS resulted in surface reactions, which did not affect bacterial adhesion. CLINICAL RELEVANCE: Changes in the surface characteristics of prototype TCS-based cements by inclusion of SNP and BG or exposure to different environments did not affect bacterial adhesion. All experimental materials showed inferior physical properties and higher antibacterial effect than Biodentine.

Effect of chlorhexidine digluconate on antimicrobial activity, cell viability and physicochemical properties of three endodontic sealers.

OBJECTIVE: Assess the biological and physicochemical properties of AH Plus, BioRoot RCS and Pulp Canal Sealer (PCS) leachates with and without chlorhexidine (CHX). METHODS: The sealers were studied in no contact and 1-minute contact with CHX. For biological properties (antibacterial activity and cytotoxicity), leachates were formed in saline of freshly mixed, 1-, 7- and 28 days set sealers. The antibacterial properties of sealer leachates were investigated for planktonic and biofilm growth of E. faecalis, S. mutans, S.epidermidis and S.aureus. The 3-(4,5 dimethylthiazolyl-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay was used to evaluate murine fibroblast cell viability after exposure to the leachates. The physical properties (water uptake, sorption, solubility, porosity, surface characteristics) of sealers and the pH of the immersion liquid (saline or distilled water) were also assessed over a 28-days period. RESULTS: CHX improved the antibacterial properties of the sealer leachates and reduced cell viability for all sealer leachates, except for freshly mixed PCS. BioRoot RCS leachates presented the highest antibacterial properties and cell viability with and without CHX contact. PCS was the material most affected by CHX in terms of physical properties, whereas for AH Plus, solubility was increased. CHX did not affect the physical properties of BioRoot RCS, except for solubility that was decreased. CHX contact did not change sealers' alkalinity in distilled water whereas it increased it for AH Plus and BioRoot RCS in saline. SIGNIFICANCE: CHX improved the antibacterial efficacy of sealer leachates and either compromised or did not affect cell viability. CHX affected to various extent sealers' physicochemical properties.

The dentine-sealer interface: Modulation of antimicrobial effects by irrigation.

AIM: Assess whether sodium hypochlorite (NaOCl) or chlorhexidine (CHX) and two irrigation protocols may alter the antibacterial properties of dentine and three endodontic sealers using a novel ex vivo tooth model. METHODOLOGY: Prior to antibacterial testing, the tooth model was validated by means of scanning electron microscopy (SEM) to evaluate the separation between dentine and sealer surfaces. Root blocks prepared from extracted human roots were pre-treated with 17% EDTA + 0.9% saline and subsequently treated with 1% NaOCl (G1), 2% CHX (G2) or no irrigant (G3). Two irrigation protocols were further investigated, "1% NaOCl + 17% EDTA" (P1) and "1% NaOCl + 17% EDTA + 2% CHX" (P2). Following irrigation, the root blocks were either filled with AH Plus, BioRoot RCS and Pulp Canal Sealer (PCS), or left empty. All groups were incubated for 1, 7 and 28 days. Direct contact tests for planktonic E. faecalis and 48 h E. faecalis biofilms were performed at the level of dentine and sealer surfaces. Statistical analysis was performed on the bacterial survival between irrigants (G1, G2 and G3) and between irrigation protocols (P1 and P2); p < .05. RESULTS: The model was considered reproducible as SEM examination of dentine samples indicated consistent separation between dentine and sealer surfaces. Irrigation with CHX (G2) and irrigation protocol P2 enhanced the antibacterial properties of dentine without sealer application as well as dentine in contact with all three sealers tested, especially against planktonic E. faecalis. G2 and P2 also improved the antibacterial effect of AH Plus surfaces for all three incubation times. No irrigation groups (G1, G2) or irrigation protocols (P1, P2) altered the antibacterial properties of BioRoot RCS surfaces against planktonic bacteria or biofilms. Only BioRoot RCS surfaces eliminated the planktonic E. faecalis in all irrigation groups (G1, G2, G3) and protocols (P1, P2) investigated whilst PCS surfaces eliminate E. faecalis in biofilms in all groups up to 7 days. CONCLUSIONS: The tooth model was reproducible. CHX improved the antibacterial activity upon both sealer and dentine surfaces. Amongst sealers, BioRoot RCS was less affected by NaOCl and CHX, and exhibited high antibacterial properties regardless the irrigation applied.

Effects of an individualized training course on technical quality and periapical status of teeth treated endodontically by dentists in the Public Dental Service in Norway: An observational intervention study.

AIM: To investigate the effect of a continuing education course on technical quality and treatment outcome for root filled permanent teeth in Møre and Romsdal County, Norway. METHODOLOGY: Fifty-two dentists employed in the Public Dental Service in Møre and Romsdal county, Norway, completed a two-day continuing education course in root canal treatment. Periapical radiographs of root filled teeth treated before and after the course, with at least one-year follow-up were identified and scored for technical quality and periapical status. Technical root filling quality was assessed by density and length and the treatment outcome by PAI scores. Treatment information was achieved from the county's electronic dental record system. Information regarding the dental practitioners' background and treatment procedure routines was collected by a questionnaire. Descriptive statistics analyses and mixed-effect logistic regression analyses were performed to evaluate the effect of the course. RESULTS: Radiographs were available for 224 teeth root filled before and for 221 teeth after the course. The proportion of teeth with adequate root filling quality was significantly lower after the course (p = .006), associated mainly with short root fillings (p < .001). No significant differences were observed in treatment outcome. There were, however, large differences in treatment outcome amongst subgroups of dentists. Further, there was evidence of effect modification by the continuing education course on periapical outcome by patient's age (pinteraction  = .0023) suggesting that teeth in patients ≤18 years healed relatively better post-course compared to patients >18 years. CONCLUSIONS: A two-day continuing education course in root canal treatment attended by Public Dental Service dentists in Norway did not improve the technical quality of root fillings or periapical status associated with root filled teeth.

Effects of an individualised training course in endodontics on the knowledge and insights of dentists in Public Dental Service in Norway.

OBJECTIVE: To evaluate the knowledge and insights of general dental practitioners regarding endodontic diagnosis and treatment principles before and after attending a 2-days continuing education course. MATERIALS AND METHODS: Sixty-seven dentists employed in the Public Dental Service in Møre and Romsdal county, Norway, were invited to a continuing education course in endodontics. Before and after the course, they answered a questionnaire weighing the importance of factors influencing the prognosis of endodontic treatment. The same questionnaire was answered by specialists in endodontics (n = 56; Spec Group) and a reference group consisting of general dental practitioners from both private and public practice (n = 21; Ref Group). The Test Group answered both before and after the course, while the Spec and Ref Groups completed the questionnaire once. The responses were compared using the Wilcoxon Sign test and Mann-Whitey U test. RESULTS: Of the 67 dentists in the Test Group, 49 (73%) completed the course and answered both questionnaires. Before attending the course, the Test Group differed significantly from the Spec Group in 18 out of 27 prognostic factors (p ≤ .05). After the course, there was only moderate improvement in the Test Group responses. On only three factors, they agreed significantly more with the specialists than before attending the course (p ≤ .05). After the Test Group participants had attended the course, their responses were comparable to the responses of the Ref Group. CONCLUSIONS: A two-days continuing education course only marginally improved dentists' level of knowledge and insights regarding the influence of prognostic factors in endodontics.

Antimicrobial and physicochemical characterization of endodontic sealers after exposure to chlorhexidine digluconate.

OBJECTIVES: Assess the antibacterial, physical and chemical properties of AH Plus, BioRoot RCS and Pulp Canal Sealer (PCS) in contact with 2% chlorhexidine digluconate (CHX) used as final irrigant prior to root canal obturation. METHODS: The antimicrobial properties were investigated by direct contact tests for planktonic and biofilm growth of E. faecalis, S. mutans, S.epidermidis and S.aureus in vitro. The setting time, wettability, microhardness and surface roughness were also assessed. The sealers were studied in no contact, 1-minute (short-term) and continuous contact (long-term) with CHX. Chemical characterization of sealers was performed by scanning electron microscopy, X-ray diffraction analysis and Fourier-transform infrared spectroscopy after CHX or saline used as the last irrigant in an ex vivo tooth model and in endo training blocks. RESULTS: CHX increased the antibacterial activity of all the sealers investigated against planktonic bacteria and biofilms with PCS exerting the highest antimicrobial activity with and without the presence of CHX. The setting of AH Plus and BioRoot RCS was retarded, while for PCS accelerated in the presence of CHX. AH Plus and PCS were more hydrophilic after contact with CHX, whilst BioRoot RCS was hydrophobic in a time-dependent manner. The microhardness of sealers was compromised and the surface roughness increased after CHX exposure for AH Plus and BioRoot RCS, and decreased for PCS. CHX did not affect the sealers' chemistry, but PCS that exhibited two extra phases. SIGNIFICANCE: CHX improved the antibacterial efficacy of endodontic sealers but further evidence is needed to confirm its suitability as a final irrigant prior to root canal obturation.

Maintenance of the aseptic working field during endodontic treatment.

Objective: The aim of this clinical quality study was to determine whether the aseptic working field is maintained during the endodontic procedure. Materials and methods: Bacterial samples were collected from the rubber dam of 27 patients during endodontic treatment performed by postgraduate students at the Department of Endodontics, University of Oslo. A bacterial sample was first obtained immediately after disinfection of the working field (A), and the second sample was collected just before obturation or dressing with calcium hydroxide cement (B). Aerobic cultivation technique and PCR were used for detection of bacterial growth and species. Results: All samples were negative on culturing except in one case, which showed positive results with cultivation in both sample A and B. Specie detected with cultivation technique were Streptococcus mitis. With PCR technique, 6 samples in 5 patients (11%), showed positive results. Species detected with PCR technique were Bacteroidales spp. Propionibacterium spp., Bacteroidetes spp., Prevotella nigrescens, Haemophilus parainfluenzae, Neisseria elongata, Alloprevotella tannerae, Capnocytophaga granulosa, Cardiobacterium hominis, Fusobacterium nucleatum and Streptococcus mitis. Conclusion: The present study showed that an aseptic working field was maintained throughout the endodontic procedure in 81% (22/27) of the cases after disinfection of the rubber dam.

Antibacterial Activity of Endodontic Sealers against Planktonic Bacteria and Bacteria in Biofilms.

INTRODUCTION: The aim of this study was to investigate the antibacterial activity of 4 endodontic sealers against bacteria planktonic grown or in biofilms commonly detected from persistent and secondary endodontic infections. METHODS: The antibacterial activity of the sealers AH Plus, TotalFill BC sealer, RoekoSeal, and Guttaflow 2 was investigated for planktonic grown and 24-hour-old biofilms of Enterococcus faecalis, Staphylococcus epidermidis, Staphylococcus aureus, and Streptococcus mutans. RESULTS: AH Plus had high antibacterial activity toward all species investigated, both planktonic and in biofilms. However, the antibacterial activity was lost after 24 hours. TotalFill BC sealer showed marked antibacterial effect on planktonic bacteria up to 7 days after setting. TotalFill BC sealer had lower antibacterial activity against biofilms of S. aureus and E. faecalis compared with AH Plus when direct contact between the sealer and biofilm was investigated and for all species investigated when a membrane was used to separate the biofilm and sealer. Guttaflow 2 and RoekoSeal had no antibacterial activity against planktonic bacteria or bacteria in biofilms. CONCLUSIONS: Bacteria in biofilms showed higher susceptibility for AH Plus compared with TotalFill BC sealer during the first 24 hours after setting. Investigating the antibacterial activity of endodontic sealers and materials against bacteria in biofilms is highly important to evaluate the materials' ability to eradicate bacteria from the infected root canal.

Anatomic Comparison of Contralateral Premolars.

INTRODUCTION: The comparative anatomy of contralateral premolars has not been previously studied. The purpose of this micro-computed tomography investigation was to qualitatively and quantitatively assess and compare the morphology of contralateral premolars in terms of length, canal width, dentinal thicknesses, accessory canals, root canal configurations, isthmi, C-shapes, root canal orifices, and apical foramina. The null hypothesis (H0) is that contralateral premolars are more morphologically similar than randomly assigned pairs for simple morphometric measurements (lengths, canal widths, and dentinal thickness). METHODS: Forty-one intact premolar pairs (n = 82) extracted from 28 patients were scanned with micro-computed tomography and reconstructed. Quantitative comparative assessment of measurements was performed by pairwise statistical analysis of contralateral and random pairs with Student t test and one-sample t test. All measured parameters (lengths, widths, and thicknesses) were normalized by Z score so that they could all be compared on a common scale. A correlation study was also performed. Canal configurations and isthmi were classified according to preexisting classification schemes. The number and location of accessory canals and apical foramina were registered and compared. RESULTS/CONCLUSIONS: Contralateral premolar pairs demonstrated a high degree of similarity in terms of the linear measurements (lengths, widths, and thicknesses). The apical portion (foramina, C-shapes, and accessory canals) did not demonstrate bilateral symmetry. The comprehensive statistical analysis of the normalized parameters by Z score showed no statistically significant differences between the contralateral premolar pairs. The null hypothesis (H0) was accepted.

Human cytomegalovirus and Epstein-Barr virus in apical and marginal periodontitis: a role in pathology?

Periodontitis is presumably caused by bacterial infection, but it has been shown recently that affected tissue often contains human cytomegalovirus (HCMV) and Epstein-Barr virus (EBV). The present study was initiated to evaluate the role of these viruses in the pathogenesis of periodontitis. HCMV and EBV were quantified in 40 apical and 25 marginal periodontitis samples using real time PCR. In situ hybridization or immunohistochemistry was carried out on apical samples to detect viral presence within cells. A possible association with relevant bacteria was examined. Of the apical periodontitis samples, 50% contained EBV, while none contained HCMV. Of the marginal periodontitis samples, 40% were positive for EBV and 12% for HCMV. With one exception, however, the amount of virus was close to the detection limits. EBV was only detected in 1 out of 15 healthy periodontium samples. Immunohistochemistry and in situ hybridization were all negative. Significant associations were found between periodontal EBV and the presence of Aggregatibacter actinomycetemcomitans and Porphyromonas gingivalis. Although there was an obvious association of the virus with clinical samples, it seems unlikely that these viruses play a major role in the pathogenesis of periodontitis of the average patient. Their presence may reflect that the clinical samples contain more blood or saliva compared to controls, or an accumulation of lymphoid cells harboring virus in the inflamed tissue.

Patient with severe periodontitis and subgingival Epstein-Barr virus treated with antiviral therapy.

It has been suggested that the presence of Epstein-Barr virus (EBV) can increase the severity of marginal periodontitis. We administered antiviral treatment (Valtrex for a period of 10 days) to a patient with recurrent periodontal disease and a high EBV load subgingivally. The antiviral treatment decreased the presence of EBV to the detection limit and the periodontal condition improved dramatically. One year after treatment, the periodontal condition was still stable and the virus barely detectable. The case suggests that virus screening and subsequent antiviral therapy may be useful as an adjunct to conventional periodontal therapy.

Microbiota of periapical lesions refractory to endodontic therapy.

The periapical microbiota of 36 teeth with refractory apical periodontitis was investigated. None of the teeth had responded to conventional endodontic or long-term (> 6 months), calcium-hydroxide treatment. Eight patients had received antibiotics systemically. After anaerobic culture, a total of 148 microbial strains were detected among 67 microbial species. One of the 36 lesions was culture-negative. Approximately half (51.0%) of the bacterial strains were anaerobic. Gram-positive species constituted 79.5% of the flora. Facultative organisms, such as Staphylococcus, Enterococcus, Enterobacter, Pseudomonas, Stenotrophomonas, Sphingomonas, Bacillus, or Candida species were recovered from 27 of the lesions (75%). Sulfur granules were found in 9 lesions (25%). In these granules Actinomyces israelii, A. viscosus, A. naeslundii, and A. meyeri were identified. Other bacterial species, both gram-positive and gram-negative, were detected in the granules as well. Two sulfur granules did not contain Actinomyces. Scanning electron microscopy demonstrated rod- and spirochete-like cells in the granules, and transmission electron microscopy revealed organisms with copious amounts of extracellular material. Outer membrane vesicles were also seen. Some of the granules were calcified. This study demonstrated a wide variety of microorganisms, particularly gram-positive ones, in the periapical lesions of teeth with refractory apical periodontitis.