Rogério de Castilho Jacinto

Microbial Analysis of Canals of Root-filled Teeth with Periapical Lesions Using Polymerase Chain Reaction

The objective of the present study was to investigate the presence of nine bacterial species in root-filled teeth associated with periapical lesions using a polymerase chain reaction analysis and to correlate these species with clinical features of the cases. DNA was extracted from 45 canal samples of root-filled teeth with periapical lesions. A PCR assay using species-specific primers of 16S rDNA and the downstream intergenic spacer region was used for microbial detection. Enterococcus faecalis was the most prevalent species, detected in 77.8% of the study teeth, followed by Peptostreptococcus micros, detected in 51.1%. Porphyromonas gingivalis, Porphyromonas endodontalis, Prevotella intermedia, and Prevotella nigrescens were detected in 35.6%, 22.2%, 11.1%, and 11.1% of the sampled teeth, respectively. Moreover, PCR detected Filifactor alocis in 26.7%, Treponema denticola in 24.4%, and Tannerella forsythia in 4.4% of the samples. T. denticola and P. micros were statistically associated with tenderness to percussion (p < 0.05). P. nigrescens was associated with the presence of spontaneous pain and abscess (p < 0.05). P. endodontalis and P. nigrescens were associated with purulent exudates (p < 0.05). Synergistic relationship was also observed between some species. The results of this study indicated that E. faecalis was the most frequently identified test species by PCR in teeth with failing endodontic treatment.

Evaluation of Cytotoxicity and Physicochemical Properties of Calcium Silicate-based Endodontic Sealer MTA Fillapex

INTRODUCTION The aim of the study was to evaluate the cytotoxicity, radiopacity, pH, and flow of a calcium silicate-based and an epoxy resin-based endodontic sealer, MTA Fillapex (Angelus, Londrina, PR, Brazil) and AH Plus (Dentsply, Konstanz, Germany), respectively. METHODS Cytotoxicity, radiopacity, and flow evaluation were performed following ISO requirements. The pH level was measured at periods of 3, 24, 72, and 168 hours. Cytotoxicity was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay to check the Balb/c 3T3 cells viability at 1- to 4-week periods. Data were statistically analyzed by analysis of variance and the Tukey test with a significance level of 5%. RESULTS In all tested periods, MTA Fillapex was more cytotoxic than AH Plus (P < .05). Although AH Plus presented higher radiopacity than MTA Fillapex (P < .05), both sealers showed minimum required values. MTA Fillapex presented alkaline pH in all experimental times, whereas AH Plus cement showed a slightly neutral pH and a flow significantly lower than that of MTA Fillapex (P < .05). CONCLUSIONS Although MTA Fillapex was more cytotoxic than AH Plus, it showed suitable physicochemical properties for an endodontic sealer.

In Vitro Evaluation of the Antimicrobial Activity of Five Root Canal Sealers

The aim of the present study was to analyze the antimicrobial properties of five endodontic sealers: Endo Fill, Endomethasone, Endomethasone N, Sealer 26 and AH-Plus, against the following microorganisms: Candida albicans, Staphylococcus aureus, Enterococcus faecalis, Streptococcus sanguis and Actinomyces naeslundii. The sealers were tested immediately, 24 h, 48 h and 7 days after manipulation.The direct contact method through the observation of the microbial growth in liquid medium and the agar diffusion test were used to evaluate the antimicrobial properties of the sealers. The results, in both methodologies used, showed that immediately after manipulation, Endo-Fill and Endomethasone demonstrated the highest antimicrobial activity, with no statistically significant difference between them. Sealer 26 demonstrated the lowest antimicrobial activity. At all other times after manipulation, there were no statistically significant differences among all the sealers tested. In conclusion, none of the sealers totally inhibited the growth of the microorganisms. Furthermore, the antimicrobial activity of each sealer decreased with time and was dependent upon the microbial susceptibility to them.

Frequency, Microbial Interactions, and Antimicrobial Susceptibility of Fusobacterium nucleatum and Fusobacterium necrophorum Isolated from Primary Endodontic Infections

This study assessed the prevalence and microbial interactions of Fusobacterium nucleatum and Fusobacterium necrophorum in primary endodontic infections from a Brazilian population and their antimicrobial susceptibility to some antibiotics by the E-test. One hundred ten samples from infected teeth with periapical pathologies were analyzed by culture methods. Five hundred eighty individual strains were isolated; 81.4% were strict anaerobes. F. nucleatum was found in 38 root canals and was associated with Porphyromonas gingivalis, Prevotella spp., and Eubacterium spp. F. necrophorum was found in 20 root canals and was associated with Peptostreptococcus prevotii. The simultaneous presence of F. nucleatum and F. necrophorum was not related to endodontic symptoms (p > 0.05). They were 100% susceptible to amoxicillin, amoxicillin/clavulanate, and cephaclor. Fusobacterium spp. is frequently isolated from primary-infected root canals of teeth with periapical pathologies. Amoxicillin is a useful antibiotic against F. nucleatum and F. necrophorum in endodontic infections and has been prescribed as the first choice in Brazil.

Persistent Extraradicular Infection in Root-filled Asymptomatic Human Tooth: Scanning Electron Microscopic Analysis and Microbial Investigation after Apical Microsurgery

INTRODUCTION Procedural accidents have a negative effect on healing and might contribute to the persistence of infections in inaccessible apical areas, requiring surgical intervention. This report describes a case of persistent apical periodontitis of a lower left first molar associated with the sinus tract and a periapical lesion that required nonsurgical endodontic retreatment and apical surgery for resolution. METHODS The tooth had received endodontic treatment 3 years ago and had to be retreated using the crown-down technique with chemical auxiliary substance (2% chlorhexidine gel), foramen patency, and enlargement and was filled in a single appointment. The occlusal access cavity was immediately restored with composite resin. After 1 month, it could be observed that the sinus tract persisted and, radiographically, the lesion remained unaltered. Therefore, endodontic microsurgery was indicated. Apical microsurgery was performed under magnification with the use of a dental operating microscope including apicectomy, root end with ultrasound, and sealing with mineral trioxide aggregate. A microbiological sample was collected from the apical lesion. The resected distal root apex was observed by scanning electron microscopy. RESULTS The following species were detected: Actinomyces naeslundii and Actinomyces meyeri, Propionibacterium propionicum, Clostridium botullinum, Parvimonas micra, and Bacteroides ureolyticus; scanning electron microscopic analysis revealed bacterial biofilm surrounding the apical foramen and external radicular surface. Gutta-percha overfilling at the apex because of a zip caused during initial endodontic treatment could be observed. A 6-month follow-up showed apparent radiographic periapical healing, which progressed after 24 months. CONCLUSION Gram-positive anaerobic bacteria and extraradicular biofilm seem to participate in the maintenance of persistent periapical pathology, and endodontic retreatment followed by periapical microsurgery proved to be a successful alternative in the resolution of persistent extraradicular infections.

Clustering Behavior in Microbial Communities from Acute Endodontic Infections

INTRODUCTION Acute endodontic infections harbor heterogeneous microbial communities in both the root canal (RC) system and apical tissues. Data comparing the microbial structure and diversity in endodontic infections in related ecosystems, such as RC with necrotic pulp and acute apical abscess (AAA), are scarce in the literature. The aim of this study was to examine the presence of selected endodontic pathogens in paired samples from necrotic RC and AAA using polymerase chain reaction (PCR) followed by the construction of cluster profiles. METHODS Paired samples of RC and AAA exudates were collected from 20 subjects and analyzed by PCR for the presence of selected strict and facultative anaerobic strains. The frequency of species was compared between the RC and the AAA samples. A stringent neighboring clustering algorithm was applied to investigate the existence of similar high-order groups of samples. A dendrogram was constructed to show the arrangement of the sample groups produced by the hierarchical clustering. RESULTS All samples harbored bacterial DNA. Porphyromonas endodontalis, Prevotella nigrescens, Filifactor alocis, and Tannerela forsythia were frequently detected in both RC and AAA samples. The selected anaerobic species were distributed in diverse small bacteria consortia. The samples of RC and AAA that presented at least one of the targeted microorganisms were grouped in small clusters. CONCLUSIONS Anaerobic species were frequently detected in acute endodontic infections and heterogeneous microbial communities with low clustering behavior were observed in paired samples of RC and AAA.

Cast metal vs. glass fibre posts: A randomized controlled trial with up to 3 years of follow up

OBJECTIVE This randomized controlled trial compared the survival of glass fibre and cast metal dental posts used to restore endodontically treated teeth with no remaining coronal wall. METHODS Fifty-four participants (45 women) and 72 teeth were evaluated during a follow-up period of up to 3 years. Teeth were randomly allocated to the glass-fibre and cast-metal post groups. All teeth were restored with single metal-ceramic crowns. Survival probabilities were analyzed using Kaplan-Meier statistics (p≤0.05). RESULTS The 3-year recall rate was 92.3% and the survival rates of glass fibre and cast metal posts were similar (97.1% and 91.9%, respectively; p=0.682). Four failures were observed: two glass fibre posts in a premolar and anterior tooth debonded, one glass fibre post in a premolar debonded in association with root fracture, and one root fracture occurred in a molar with a cast metal post. CONCLUSIONS Glass fibre and cast metal posts showed similar clinical performance in teeth with no remaining coronal wall after 3 years. CLINICAL SIGNIFICANCE Posts are used to restore most endodontically treated teeth with no remaining coronal wall. This randomized controlled trial, one of few to compare glass fibre and cast metal posts in such teeth, showed that post type did not significantly influence the survival of restorations. These results can help dentists respond to the important question of how best to rehabilitate endodontically treated teeth with no remaining coronal wall.

Analysis of the antimicrobial susceptibility of anaerobic bacteria isolated from endodontic infections in Brazil during a period of nine years.

INTRODUCTION The purpose of this study was to analyze the susceptibility of some anaerobic species isolated from a Brazilian population at different periods of time by determining a pattern of development of resistance to frequently prescribed antibiotics in endodontics. METHODS Root canal samples were collected from infected teeth at different periods of time (2000-2002, 2003-2005, and 2007-2008) and microbiologically identified with conventional culture techniques. The susceptibility of Prevotella intermedia/nigrescens, P. oralis, Fusobacterium nucleatum, and P. micra isolated strains was determined by the minimum inhibitory concentration (MIC) of amoxicillin, amoxicillin + clavulanate, benzylpenicillin, clindamycin, erythromycin, and metronidazole by using the E-test method. RESULTS Amoxicillin and amoxicillin + clavulanate were effective against the majority of species at the different periods of study. Overall, there were low statistical differences regarding the microbial susceptibility between the experimental periods. However, an increase in the anaerobic resistance to penicillin G and clindamycin was observed. Resistance to erythromycin was observed in all species, and there were statistically significant differences between 2000-2002 and 2003-2005 periods for F. nucleatum (P < .05) and between 2003-2005 and 2007-2008 periods for P. intermedia/nigrescens and P. oralis (P < .05). CONCLUSIONS The antimicrobial resistance of anaerobes isolated from primary endodontic infections showed an increase throughout a period of time regarding a specific Brazilian population.

Influence of 2% chlorhexidine gel on calcium hydroxide ionic dissociation and its ability of reducing endotoxin.

OBJECTIVE The aim of this study was to evaluate the influence of 2% chlorhexidine gel (CHX) on pH, calcium release, and Ca(OH)(2) capability of reducing endotoxin. STUDY DESIGN Calcium release was verified by atomic-absorbance spectrophotometry, and pH was measured with a pHmeter. For endotoxin quantification, extracted human teeth previously contaminated with standard endotoxin were filled with: group I, Ca(OH)(2) + saline solution; group II, Ca(OH)(2) + CHX; and group III, CHX for 14 days. The remaining endotoxin was quantified by using chromogenic quantitative test. Statistical analysis was performed with analysis of variance and Tukey post hoc test (P < .05). RESULTS Group II released more calcium than group I (P < .05) after 15 days. Groups I and II showed alkaline pH in all periods, and group I showed higher pH values than group II (P < .05) after 30 days. Groups II and III showed significantly more decreased endotoxin than group I. CONCLUSIONS CHX did not interfere with the chemical properties of Ca(OH)(2), in fact even improving its properties of reducing the endotoxin content in root canals in vitro.

Treponema species detected in infected root canals and acute apical abscess exudates.

INTRODUCTION Different microbial communities have been associated with acute endodontic infections. The majority of the microorganisms are as yet uncultivable or difficult to grow under current laboratory conditions. Treponema species are strict anaerobic bacteria that are involved in several oral diseases. The aim of this study was to detect the presence of Treponema species in infected root canals (RCs) and exudates related to acute apical abscesses (AAAs) as well as to determine positive association between targeted species and clinical signs/symptoms. METHODS Paired samples of infected RCs and AAAs were collected from 20 subjects. Nested polymerase chain reaction assay with species-specific primers for 16S rDNA and downstream intergenic spacer region was used for microbial detection. The frequency of species and statistical associations between species and signs/symptoms of endodontic origin as well as their simultaneous detection in both milieus were investigated. RESULTS The most frequently detected species were T. socranskii (RC, 17/20; AAA, 15/20), T. denticola (RC, 8/20; AAA, 11/20); T. medium (RC, 6/20; AAA, 9/20); and T. amylovorum (RC, 5/20; AAA, 9/20). Positive correlation was found for simultaneous presence of T. denticola in both RCs and AAAs (p = 0.01). Positive association was observed between T. medium and T. vincentii (p = .037). No positive statistical association was observed between the targeted species and signs/symptoms. CONCLUSIONS The high incidence of Treponema species in RC and AAA samples from the same tooth indicated that they are important pathogens in acute endodontic infections.