Pulp revascularization for immature replanted teeth: a case report.

Immature avulsed teeth are not usually treated with pulp revascularization because of the possibility of complications. However, this therapy has shown success in the treatment of immature teeth with periapical lesions. This report describes the case of an immature replanted tooth that was successfully treated by pulp revascularization. An 8-year-old boy suffered avulsion on his maxillary left lateral incisor. The tooth showed incomplete root development and was replanted after 30 minutes. After diagnosis, revascularization therapy was performed by irrigating the root canal and applying a calcium hydroxide paste and 2% chlorhexidine gel for 21 days. In the second session, the intracanal dressing was removed and a blood clot was stimulated up to the cervical third of the root canal. Mineral trioxide aggregate was placed as a cervical barrier at the entrance of the root canal and the crown was restored. During the follow-up period, periapical repair, apical closure and calcification in the apical 4 mm of the root canal was observed. An avulsed immature tooth replanted after a brief extra-alveolar period and maintained in a viable storage medium may be treated with revascularization.

Thickness of dentine in mesial roots of mandibular molars with different lengths.

AIM: To measure the minimum thickness of the distal (furcal) root dentine associated with the buccal and lingual canals of the mesial roots of mandibular first molars with different lengths. METHODOLOGY: The mesial roots of 285 mandibular first molars were allocated into three groups according to their length: group I - long (24.14 mm +/- 0.85), group II - medium (22.10 mm +/- 0.65) and group III - short (19.97 mm +/- 0.75). The minimum thickness of the distal (furcal) root dentine associated with the buccal and lingual canals of the mesial roots 2 mm below the furcation was measured. The distance between the buccal and lingual canals, and the depth of concavity in the distal surface of the mesial roots were also measured. anova and Tukey-Kramer were used to test for significant differences among the groups. RESULTS: The minimum thickness of the distal wall of the mesiobuccal canal was significantly different (P < 0.05) between group I (long) and III (short), with long teeth having the smallest mean values. No significant difference was found in the thickness of the distal wall of the mesiolingual canal among the groups studied (P > 0.05). The shortest distance between the mesiobuccal and the mesiolingual canals was observed in group III (P < 0.05). The distal (furcal) concavity was deeper in group I (P < 0.05) when compared with the other groups. CONCLUSION: There was a significant difference in the minimum thickness of the distal (furcal) root wall of the mesiobuccal canal of mandibular first molars 2 mm below the furcation between group I (long) and group III (short) teeth. The thinnest walls were found in the longest teeth. The deepest concavities in the distal (furcal) walls of the mesial roots were found in the longest roots.

Degradation of trans-polyisoprene after root filling with thermoplasticized techniques.

AIM: To evaluate ex vivo degradation of gutta-percha following six thermoplastic obturation techniques. METHODOLOGY: Ninety human-extracted mandibular premolars were selected and divided randomly into nine groups for filling. Group 1: thermomechanical compaction for 3 s with Konne gutta-percha points (Konne Ind. e Com. de Mat. Odontol., Belo Horizonte, MG, Brazil); Group 2: thermomechanical compaction for 3 s with Dentsply TP gutta-percha points (Dentsply Indústria e Comércio Ltda, Petrópolis, R.J. Brazil); Group 3: thermomechanical compaction for 10 s with Konne; Group 4: thermomechanical compaction for 10 s with Dentsply TP; Group 5: warm vertical condensation using System B (EIE/Analytic, Richmond, WA, USA) with Konne; Group 6: warm vertical condensation using System B with Dentsply TP; Group 7: vertical condensation with Konne; Group 8: vertical condensation with Dentsply TP; Group 9: Microseal cone (Analytic Endodontics, Glendora, CA, USA). A further four groups were assessed without using teeth, Group 10: Microseal microflow (Analytic Endodontics); Group 11: Obtura (Obtura Corporation, Penton, MO, USA); Group 12: Obtura flow (Obtura Corporation); Group 13: Thermafil (Dentsply Maillefer, Tulsa, OK, USA). The filling material was removed from the root canal and trans-1,4-polyisoprene isolated by solubilization of the root filling remnants in chloroform followed by filtration and centrifugation. By gel permeation chromatography and infrared spectroscopy, the occurrence and degree of degradation were assessed. The results were analysed statistically using the Kruskal-Wallis test. With differential scanning calorimetry, the thermal behaviour of the gutta-percha was determined. RESULTS: A significant decrease in polymer molar mass and the production of carboxyl and hydroxyl groups in the polymer were observed with thermomechanical compaction used for 10 s and vertical condensation filling techniques (P = 0.0001 and P = 0.0005, respectively). Other techniques caused no polymer degradation. CONCLUSION: Polyisoprene degrades with high temperature. Thermomechanical compaction for 10 s and vertical condensation were associated with the greatest degradative process.

Ex vivo antimicrobial activity of several bleaching agents used during the walking bleach technique.

AIM: To investigate ex vivo the antimicrobial activity of a paste of sodium perborate associated with various vehicles comparing it with 37% carbamide peroxide and 35% hydrogen peroxide. METHODOLOGY: The antimicrobial activity of these agents was evaluated against three microorganisms: Enterococcus faecalis, Streptococcus mutans and Candida albicans. One millilitre of each tested substance was placed on the bottom of wells of 24-well cell culture plates. Six wells were used for each time period and group. Two millilitres of the microbial suspension was ultrasonically mixed for 10 s with the bleaching pastes and placed in contact with them for 10, 30, 45 s; 1, 3, 5, 10, 20, 30 min; and 1 and 2 h. After each period of time, 1 mL from each well was transferred to tubes containing 2 mL of freshly prepared brain heart infusion agar + neutralizers. Agar plates were inoculated in appropriate gaseous conditions. Data were analysed statistically by the Kruskal-Wallis test with the level of significance set at P < 0.05. RESULTS: In all groups containing chlorhexidine (groups 3, 5 and 7), the antimicrobial activity of the bleaching paste was significantly increased when compared with groups with other kinds of vehicle (groups 1, 2, 4, 6 and 8). For all tested groups, the most resistant microorganism was E. faecalis. CONCLUSIONS: Chlorhexidine when used as a vehicle for sodium perborate enhanced its antimicrobial activity.

Effect of root canal procedures on endotoxins and endodontic pathogens.

BACKGROUND/AIMS: The purpose of this study was to determine the amount of endotoxin (lipopolysaccharide) and cultivable bacteria in human necrotic root canals before (S1) and after chemo-mechanical preparation using chlorhexidine (CHX) gel as auxiliary chemical substance (S2), and after 7 days of intracanal dressing (S3) in order to evaluate the anti-endotoxin and antimicrobial effects of endodontic procedures. METHOD: Twenty-four teeth were selected for the present study. Chemo-mechanical preparation was performed using 2% CHX gel and three different intracanal medicaments [CaOH2 paste; 2% CHX gel; and CaOH2 + 2% CHX gel]. A quantitative chromogenic Limulus amoebocyte lysate assay was used to measure the amount of endotoxin. Aerobic and anaerobic techniques were used to isolate and identify bacteria, and to determine the bacterial reduction by counting colony-forming units (CFU). RESULTS: Endotoxins and bacteria were present in 100% of the initial samples, with endotoxin concentration ranging from 62.93 to 214.56 UE/ml and CFU ranging from 4 x 10(5) to 2.6 x 10(6). After chemo-mechanical preparation a mean endotoxin reduction of 44.4% was found. Eight (33.3%) root canals were still positive by culture analysis with a mean reduction of bacteria (CFU) of 99.96%. After 7 days of intracanal dressing, endotoxin concentration decreased by only 1.4% compared with S2, and residual bacteria were recovered by culture analysis in 13 cases (54.1%). No significant difference was found among different intracanal medicaments. CONCLUSION: Relatively high values of endotoxin were still present in the root canal after chemo-mechanical preparation although the majority of bacteria were eliminated. No improvement was achieved by 7 days of intracanal dressing.

Filling of artificial lateral canals and microleakage and flow of five endodontic sealers.

AIM: To evaluate the flow characteristics of AH Plus, Epiphany Root Canal Sealant, Endométhasone, Pulp Canal Sealer (EWT) and Sealapex and their ability to fill artificial lateral canals and prevent microleakage. METHODOLOGY: Flow of the sealers was analysed using the American Dental Association (ADA) 57 and the International Standards Organization (ISO) 6,876 specifications. Two lateral canals were produced in the middle and apical third of 64 roots using 0.1 mm cylindrical drills. Lateral condensation of gutta-percha or Resilon and one or other of the sealers were used to fill the root canals. Buccal-lingual digital radiographs were exposed. After the sealer had set, the roots were immersed in Indian ink and cleared in methyl salicylate. The extent of filling and dye penetration were measured on the buccal and lingual root surfaces under 30 x magnification with a stereoscope. RESULTS: AH Plus, Epiphany and Pulp Canal Sealer (EWT) complied with ADA 57 and ISO 6,876 specifications. Sealapex complied with the ADA 57 specification but not with ISO 6,876. Endométhasone did not comply with either specification. Filling of lateral canals was similar for the five sealers tested. Dye leakage demonstrated that AH Plus, Epiphany and Sealapex permitted less leakage than Pulp Canal Sealer (EWT) (P < 0.05). CONCLUSIONS: All the sealers flowed into the 0.1 mm artificial lateral canals. AH Plus, Epiphany and Sealapex allowed less linear leakage than Pulp Canal Sealer (EWT). The flow of Endométhasone did not comply with either ADA 57 or ISO 6,876 specifications and Sealapex did not comply with ISO 6,876.

Degradation of trans-polyisoprene over time following the analysis of root fillings removed during conventional retreatment.

AIM: To evaluate in vivo degradation of root filling materials over time. METHODOLOGY: Thirty-six root filled teeth with or without periapical lesions were selected. Teeth with poor coronal restoration were not included. The teeth had been root filled 3-30 years previous and were scheduled for conventional retreatment. The association of root canal treatment, age, periapical lesion and root filling degradation was investigated. The filling material was removed from the root canal using files and no solvent. Trans-1,4-polyisoprene was isolated through solubilization of root filling remnants in chloroform followed by filtration and centrifugation. Gel permeation chromatography (GPC) and infrared spectroscopy (FT-IR) were utilized to study the occurrence and degree of degradation. The GPC and FT-IR data were collected for each sample and analysed statistically using the Kruskal-Wallis test. RESULTS: Degradation of trans-1,4-polyisoprene was a slow process. The process was identified as an oxidation reaction through the production of carboxyl and hydroxyl groups. Compared with the control group, significant molar mass decrease was noted after 15 years (P = 0.0146) in teeth with no periapical lesions. However, in teeth associated with periapical lesions the number of years for significant degradation was reduced to 5 (P = 0.0009). CONCLUSION: Polyisoprene degrades inside root canals as an oxidative process. The presence of periapical lesions was associated with a more rapid onset of degradation.

In vitro antimicrobial activity of sodium hypochlorite and chlorhexidine against selected single-species biofilms.

AIM: To investigate the antimicrobial activity of 2.5% and 5.25% sodium hypochlorite and 2.0% chlorhexidine gel and liquid as endodontic-irrigating substances against selected single-species biofilms. METHODS: Single-species biofilms of Enterococcus faecalis, Staphylococcus aureus, Candida albicans, Prevotella intermedia, Porphyromonas gingivalis, Porphyromonas endodontalis and Fusobacterium nucleatum were generated on a cellulose nitrate membrane placed on agar medium. The biofilms were then immersed in the endodontic-irrigating substances for 30 s and also for 5, 10, 15, 30 and 60 min, with and without mechanical agitation. Sterile saline was used as control. After each time period, the membrane filters were then transferred to tubes containing 2 mL of fresh broth medium plus neutralizers (in order to prevent the residual action of the tested substances). The micro-organisms were suspended using a vortex, and the inoculum was serially diluted 10-fold. Aliquots of the dilutions were plated on 5% sheep blood agar medium, and incubated under adequate gaseous conditions. Colony-forming units were calculated. The samples were compared using the Friedman and Tukey test, when necessary, at a significance level of P < 0.05. RESULTS: Mechanical agitation promoted the effectiveness of the antimicrobial agents, resulting in less time to eliminate the same micro-organisms, except for S. aureus with 2.5% NaOCl. Antimicrobial agents in liquid presentation, especially 5.25% NaOCl and 2% chlorhexidine, killed the tested micro-organisms more rapidly. Saline did not inhibit the growth of any of the tested micro-organisms, with or without agitation, being statistically different (P < 0.05) from NaOCl and chlorhexidine. P. intermedia, P. gingivalis, P. endodontalis and F. nucleatum were eliminated in 30 s by all antimicrobial agents, with our without agitation, in contrast with the facultative and aerobe strains. CONCLUSIONS: Mechanical agitation improved the antimicrobial properties of the chemical substances tested using a biofilm model, favouring the agents in liquid presentation, especially 5.25% NaOCl and 2% chlorhexidine.

Assessment of different gutta-percha brands during the filling of simulated lateral canals.

AIM: To compare the ability of five different commercially available gutta-percha points to fill simulated lateral canals when subjected to warm vertical compaction. METHODOLOGY: Fifty clear plastic teeth with a lateral canal in each third of the root were used. All teeth were filled using warm vertical compaction. Backfilling was completed with a sealer and the same gutta-percha point used during the apical condensation. After this, they were horizontally sectioned using a diamond disc adapted to a low-speed saw. The resulting sections were embedded in epoxy resin. The extent of gutta-percha and sealer filling were measured in each lateral canal using an IMAGE-PRO 4.0 software system. The voids in each canal were measured using the same system. Data were ranked and analysed using the Kruskal-Wallis statistical test. RESULTS: The mean percentage of the three lateral canals filled with gutta-percha and sealer were respectively: Konne (68.23% and 24.50%), Analytic (67.90% and 25.28%), Obtura (63.80% and 29.60%), Tanari (49.42% and 45.86%) and Dentsply (44.60% and 47.05%). There was significantly (P < 0.05) more gutta-percha in the lateral canal filled with Analytic, Obtura and Konne points than with Tanari and Dentsply points. CONCLUSIONS: The brand of gutta-percha cone had an influence on the length of filling within lateral canals. This may be a reflection of the chemical formulation of the gutta-percha points.

Efficacy of various concentrations of NaOCl and instrumentation techniques in reducing Enterococcus faecalis within root canals and dentinal tubules.

AIM: To evaluate the efficacy of 0.5%, 2.5% and 5.25% sodium hypochlorite (NaOCl) as intracanal irrigants associated with hand and rotary instrumentation techniques against Enterococcus faecalis within root canals and dentinal tubules. METHODOLOGY: A total of 180 extracted human premolar teeth were infected for 21 days with E. faecalis. The specimens were divided into 12 groups, as follows: group 1: 5.25% NaOCl + Hybrid technique (Valdrighi et al. 1998); group 2: 5.25% NaOCl + nickel-titanium (NiTi) rotary technique 4 mm shorter than the apex (by FOP-UNICAMP); group 3: 5, 25% NaOCl + NiTi rotary technique (Hero 642); group 4: 2.5% NaOCl +Hybrid technique; group 5: 2.5% NaOCl + NiTi rotary technique 4 mm shorter than the apex; group 6: 2.5% NaOCl + NiTi rotary technique (Hero 642); group 7: 0.5% NaOCl + Hybrid technique; group 8: 0.5% NaOCl + NiTi rotary technique 4 mm shorter than the apex; group 9: 0.5% NaOCl + NiTi rotary technique (Hero 642); group 10: sterile saline solution + Hybrid technique; group 11: sterile saline solution + NiTi rotary technique 4 mm shorter than the apex; group 12: sterile saline solution + NiTi rotary technique (Hero 642). Canals were sampled before and after preparation. After serial dilution, samples were plated onto brain heart infusion (BHI) agar, and the colony forming units (CFU) that were grown were counted. The teeth were sectioned into three thirds and dentine chips were removed from the canals with conical burs. The samples obtained with each bur were immediately collected into test tubes containing BHI broth, and were incubated at 37 degrees C and plated onto BHI agar. The CFU were counted and analysed. RESULTS: At all depths and thirds of the root canals and for all techniques used, 5.25% NaOCl was shown to be the most effective irrigant solution tested when dentinal tubules were analysed, followed by 2.5% NaOCl. No differences among concentrations in cleaning the canals were found. CONCLUSIONS: Especially at higher concentrations, NaOCl, was able to disinfect the dentinal tubules, independent of the canal preparation technique used.

Incidence and antimicrobial susceptibility of Porphyromonas gingivalis isolated from mixed endodontic infections.

AIM: To investigate the prevalence of Porphyromonas gingivalis in root canals of infected teeth with periapical abscesses and to investigate the antimicrobial susceptibility of this species to some frequently prescribed antibiotics. METHODOLOGY: Samples were obtained from 70 root canals of abscessed teeth. Microbial sampling, isolation and bacterial identification were accomplished using appropriate culture methods for anaerobic species. The antimicrobial susceptibility of the 20 strains of P. gingivalis isolated was determined by using the E-test. The antimicrobial agents tested were amoxicillin, amoxicillin + clavulanate, azythromycin, benzylpenicillin, cephaclor, clindamycin, erythromycin, metronidazole and tetracycline. RESULTS: A total of 352 individual strains, belonging to 69 different species, were isolated. Eighty three percent of the strains were strict anaerobes and 47.5% of the isolated bacteria were Gram-negative. Porphyromonas gingivalis was found in 20 root canals and was most frequently found in symptomatic cases. Statistically, the presence of P. gingivalis was related to purulent exudates and pain on palpation (both P < 0.05). All P. gingivalis strains were sensitive to amoxicillin, amoxicillin + clavulanate, cephaclor, clindamycin, benzylpenicyllin, metronidazole and tetracycline. The lowest range of minimum inhibitory concentration (MIC) (0.026-0.125 microg mL(-1)) was observed against amoxicillin + clavulanate and clindamycin. The lowest MIC 90 was observed against clindamycin (0.064 microg mL(-1)). One strain was resistant to erythromycin and eight strains were resistant to azythromycin. CONCLUSION: Porphyromonas gingivalis pathogen is isolated with frequency from root canals of infected teeth with periapical abscesses. Amoxicillin, as well as amoxicillin-clavulanic acid and benzylpenicillin were effective against P. gingivalis.

Recovery of Enterococcus faecalis after single- or multiple-visit root canal treatments carried out in infected teeth ex vivo.

AIM: To assess the presence of Enterococcus faecalis after root canal treatment in single or multiple visits in an ex vivo model. METHODOLOGY: Forty-five premolar teeth were infected ex vivo with E. faecalis for 60 days. The canals were then prepared using a crowndown technique with System GT and Gates-Glidden burs and irrigated with 2% chlorhexidine gel. The specimens were divided into five groups (G1, G2, G3, G4 and G5) according to the time elapsed between chemical-mechanical preparation and root canal filling, the irrigant solution used and the use or nonuse of a calcium hydroxide intra-canal medicament. The teeth were then root-filled and incubated for 60 days at 37 degrees C. Dentine chips were removed from the canal walls with sequential sterile round burs at low speed. The samples obtained with each bur were immediately collected in separate test tubes containing Brain-Heart Infusion broth. These samples were placed onto agar plates and colony forming units were counted after 24 h at 37 degrees C. Data were ranked and analysed using the Kruskal-Wallis statistical test. RESULTS: Enterococcus faecalis was recovered from 20% (three of 15 specimens) of G1 (chlorhexidine irrigation and immediate root filling in a single visit), 25% (four of 15 specimens) of G2 (chlorhexidine irrigation and filling after 14 days use of a calcium hydroxide dressing in multiple visits), 40% (two of five specimens) of G3 (chlorhexidine irrigation and filling after 7 days), 60% (three of five specimens) of G4 (saline irrigation and filling after 7 days) and from 100% (five of five specimens) of G5 (saline irrigation and immediate filling without sealer). CONCLUSIONS: Neither single- nor multiple-visit root canal treatment ex vivo, eliminated E. faecalis completely from dentinal tubules. Up to 60 days after root filling, E. faecalis remained viable inside dentinal tubules. When no sealer was used, E. faecalis presented a higher growth rate.

Porphyromonas gingivalis, Porphyromonas endodontalis, Prevotella intermedia and Prevotella nigrescens in endodontic lesions detected by culture and by PCR.

he aim of this study was to investigate the presence of four black-pigmented bacteria, Porphyromonas gingivalis, Porphyromonas endodontalis, Prevotella intermedia and Prevotella nigrescens, in endodontic infections by culture and polymerase chain reaction (PCR) analyses. Microbial samples were obtained from 50 teeth with untreated necrotic pulps (primary infection) and from 50 teeth with failing endodontic treatment (secondary infection). Microbiological strict anaerobic techniques were used for serial dilution, plating, incubation, and identification. For PCR detection, the samples were analyzed using species-specific primers of 16S rDNA and the downstream intergenic spacer region. Culture and PCR detected the test species in 13/100 and 50/100 of the study teeth, respectively. The organisms were cultured from 11/50 (22%) of primarily infected root canal samples and from 2/50 (4%) of secondary root canal samples. PCR detection identified the target species in 32/50 (64%) and 18/50 (36%) of primary and secondary infections, respectively. P. gingivalis was rarely isolated by culture methods (1%), but was the most frequently identified test species by PCR (38%). Similarly, P. endodontalis was not recovered by culture from any tooth studied, but was detected by PCR in 25% of the sampled teeth. PCR-based identification also showed higher detection rates of P. intermedia (33%) and P. nigrescens (22%) than culture (13%). In conclusion, P. gingivalis, P. endodontalis, P. intermedia, and P. nigrescens were identified more frequently in teeth with necrotic pulp than in teeth with failing endodontic treatment. Also, a higher frequency of black-pigmented species was detected by PCR than by culture.

Antimicrobial susceptibility of Enterococcus faecalis isolated from canals of root filled teeth with periapical lesions.

AIM: To test, in vitro, the susceptibility to different antibiotics of Enterococcus faecalis isolates from canals of root filled teeth with periapical lesions. METHODOLOGY: Twenty-one E. faecalis isolates, from canals of root filled teeth with persisting periapical lesions, were tested for their antibiotic susceptibilities. The following antibiotics were used: benzylpenicillin, amoxicillin, amoxicillin-clavulanic acid, erythromycin, azithromycin, vancomycin, chloramphenicol, tetracycline, doxycycline, ciprofloxacin and moxifloxacin. Minimal inhibitory concentrations (MICs) for the antimicrobial agents were determined using the E-test System (AB BIODISK, Solna, Sweden), and the E. faecalis strains classified as susceptible or resistant according to the guidelines of National Committee for Clinical Laboratory Standards (NCCLS). The strains were also tested for beta-lactamase production with nitrocefin (Oxoid, Basingstoke, UK). RESULTS: All strains were susceptible to penicillins in vitro, however, the MICs of amoxicillin and amoxicillin-clavulanic acid (MIC(90) = 0.75 microg mL(-1)) were lower than for benzylpenicillin (MIC(90) = 3.0 microg mL(-1)). All strains studied were also susceptible to vancomycin and moxifloxacin, whilst 95.2% were susceptible to chloramphenicol. Amongst the isolates, 85.7% were susceptible to tetracycline and doxycycline and 80.9% to ciprofloxacin. The MIC of erythromycin ranged from 0.38 to >256 microg mL(-1); only 28.5% of the strains were susceptible (MIC < or = 0.5 microg mL(-1)). Limited susceptibility was also observed with azithromycin which was active against only 14.2% of isolates. No strains produced beta-lactamase. CONCLUSION: Enterococcus faecalis isolates were completely susceptible, in vitro, to amoxicillin, amoxicillin-clavulanic acid, vancomycin and moxifloxacin. Most isolates were susceptible to chloramphenicol, tetracycline, doxycycline or ciprofloxacin. Erythromycin and azithromycin were least effective.

Resolution of persistent periapical infection by endodontic surgery.

AIM: To examine the surfaces of a root tip removed during surgical endodontic treatment for the presence of microorganisms. SUMMARY: The present clinical case illustrates an endodontic retreatment of a maxillary premolar tooth with a fistula and periapical reaction. The case was under treatment for 1 year, during which an intracanal medicament was replaced several times. As the lesion did not decrease and exudate was persistent through the fistula and root canal, root end resection with root end filling was performed. Microbiological samples were collected from the fistula, where Propionibacterium acnes, a species associated with endodontic failures, was detected by appropriate anaerobic technique. The resected root apex was observed by scanning electron microscopy (SEM), which revealed cocci and fungal forms surrounding one of the foramina. After 12 months, the periapical lesion had reduced.

Microbiological examination of infected dental root canals.

OBJECTIVES: The aim of this study was to investigate the root canal microbiota of primary and secondary root-infected canals and the association of constituent species with specific endodontic signs and symptoms. METHODS: Microbial samples were taken from 60 root canals, 41 with necrotic pulp tissues (primary infection) and 19 with failed endodontic treatment (secondary infection). Strict anaerobic techniques were used for serial dilution, plating, incubation and identification. RESULTS: A total of 224 cultivable isolates were recovered belonging to 56 different bacterial species. Individual root canals yielded a maximum of 10 bacterial species. Of the bacterial isolates, 70% were either strict anaerobes or microphilic. The anaerobes most frequently isolated were: Peptostreptococcus micros (35%), Fusobacterium necrophorum (23.3%), Fusobacterium nucleatum (11.7%), Prevotella intermedia/nigrescens (16.7%), Porphyromonas gingivalis (6.7%) and Porphyromonas endodontalis (5%). The root canal microflora of untreated teeth with apical periodontitis was found to be mixed, comprising gram-negative and gram-positive and mostly anaerobic microorganisms and usually containing more than 3 species per canal. On the other hand, facultative anaerobic and gram-positive bacteria predominated in canals with failed endodontic treatment, which harbored 1-2 species per canal. Suggested relationships were found between anaerobes, especially gram-negatives, and the presence or history of pain, tenderness to percussion and swelling (P<0.05). In particular, associations were found between: a) pain (n=29) and P. micros (P<0.01), P. intermedia/nigrescens and Eubacterium spp. (both P<0.05); b) history of pain (n=31) and P. micros (P<0.01) Porphyromonas and Fusobacterium spp. (P<0.05); c) tenderness to percussion (n=29) and Porphyromonas spp. (P<0.01), Peptostreptococcus and Fusobacterium spp. (P<0.001); d) swelling (n=20) and Peptostreptococcus spp. (P<0.01), Porphyromonas and Enterococcus spp. (P<0.05); e) wet canals (n=33) and Porphyromonas and Fusobacterium spp. (P<0.05); f) purulent exudate (n=20) and Porphyromonas, Peptostreptococcus and Fusobacterium spp. (P<0.05); previous endodontic treatment and Enterococcus faecalis, Streptococcus spp., P. micros, F. necrophorum (P<0.05). CONCLUSIONS: Our findings indicate potential complex interactions of species resulting in characteristic clinical pictures which cannot be achieved by individual species alone. They also indicate that the microbiota of primary infected canals with apical periodontitis differs in number and in species from the secondary infected canals by using the culture technique.

Evaluation of time required for recontamination of coronally sealed canals medicated with calcium hydroxide and chlorhexidine.

AIM: To determine in vitro the time required for recontamination of coronally sealed canals medicated with either calcium hydroxide (CaOH2), 2% chlorhexidine gel (CG) or with a combination of both. METHODOLOGY: Eighty intact, caries-free, premolar teeth with straight roots and mature apices were selected for the study. After biomechanical preparation of 75 teeth, they were randomly divided into nine groups according to the intracanal medicament and the coronal seal with 'Intermediate Restorative Material' (IRM) as follows: (i) 10 teeth medicated with CG, coronally unsealed; (ii) 10 teeth medicated with CaOH2, coronally unsealed; (iii) 10 teeth medicated with CaOH2 + CG, coronally unsealed; (iv) 10 teeth medicated with CG + coronal seal; (v) 10 teeth medicated with CaOH2 + coronal seal; (vi) 10 teeth medicated with CG + CaOH2 + coronal seal; (vii) 10 teeth without intracanal medicament and coronally sealed; (viii) 5 teeth without intracanal medicament and coronally unsealed, used as the positive control group (PC); (ix) 5 teeth with intact crowns used as the negative control group (NC). Glass flasks were filled with Brain Heart Infusion broth (BHI), so that only the root apex was in contact with the broth, while the crown was immersed in human saliva + BHI (3:1). The flasks were then incubated at 37 degrees C in an atmosphere of 10% CO2, and microbial growth was checked daily. RESULTS: All specimens of the PC showed contamination within 1 day of incubation, while the NC showed no evidence of broth turbidity. Recontamination was detected after an average time of 3.7 days in the unsealed canals medicated with CG, 1.8 days in the group medicated with CaOH2 and 2.6 days in the group medicated with CaOH2 + CG. When the crowns were sealed with IRM, recontamination was detected within 13.5 days in the canals medicated with CG, after 17.2 days in the group medicated with CaOH2 and after 11.9 days in the group medicated with CG + CaOH2. The group with no medication, but sealed with IRM, showed recontamination after 8.7 days. There were statistically significant differences between the teeth with or without coronal seal (P<0.05). CONCLUSION: The coronal seal delayed but did not prevent leakage of microorganisms. There was no difference between the various medicaments.

Investigation of the marginal adaptation of root-end filling materials in root-end cavities prepared with ultrasonic tips.

AIM: To compare the surface topography of root apices following ultrasonic root-end preparation, and again after root-end fillings submitted to three different finishing techniques. METHODOLOGY: Eighty-one root-end cavities prepared ultrasonically in human canines, were divided at random into three test groups of 27 each. The cavities were filled with Super-EBA, IRM, or ProRoot-MTA and finished by ball burnishing. Eighteen roots from each group received a final finish with either a 30-fluted tungsten carbide finishing bur, or a Zekrya carbide 28 mm bur after storage in water at 37 degrees C for 24 h. The root-end surface topographies were reproduced by means of polyvinylsiloxane impressions and epoxy resin replicas. Scanning electron micrography (SEM) images of each replica were taken prior to and after root-end filling. An image analysis system was used to compare the alteration of the marginal chipping areas and to calculate the gaps located in the dentine/root-end filling interface. RESULTS: When a bur was used to finish the set materials, a significant (P < 0.05) area of marginal chipping was eliminated. The finishing technique did not significantly (P > 0.05) affect the incidence of gaps in groups root-end filled with MTA or IRM. Super-EBA and IRM retrofillings finished with a ball burnisher or a Zekrya bur displayed a significantly (P < 0.05) larger calculated gap area than roots filled with MTA. CONCLUSION: Under this in vitro study, the marginal adaptation of MTA was good with or without finishing procedures. Applying a finishing bur over the condensed and set IRM and Super-EBA created better marginal adaptation.

Effectiveness of 2% chlorhexidine gel and calcium hydroxide against Enterococcus faecalis in bovine root dentine in vitro.

AIM: To evaluate the effectiveness of 2% chlorhexidine gluconate gel and calcium hydroxide (Ca(OH)2) as intracanal medicaments against Enterococcus faecalis. METHODOLOGY: One hundred and eighty dentine tubes prepared from intact freshly extracted bovine maxillary central incisors were infected in vitro for 7 days with E. faecalis. The specimens were divided into four groups, according to the intracanal medicament used, as follows: Group 1: 2% chlorhexidine gluconate gel; Group 2: calcium hydroxide in a viscous vehicle (polyethyleneglycol 400); Group 3: 2% chlorhexidine gluconate gel + calcium hydroxide and Group 4: Brain Heart Infusion (BHI) broth (control group). The medicaments were placed into the canal lumen and left there for experimental times of 1, 2, 7, 15 and 30 days. After each period, irrigation with sterile saline to remove the medicament was performed and the canals were dried with sterile paper points. Dentine chips were removed from the canals with sequential sterile round burs at low speed. The samples obtained with each bur were immediately collected in separate test tubes containing BHI broth. The tubes were incubated at 37 degrees C and daily observed for microbial growth, visualized by the medium turbidity. RESULTS: Chlorhexidine gel alone completely inhibited the growth of E. faecalis after 1, 2, 7 and 15 days. Calcium hydroxide allowed microbial growth at all experimental times. The combination of chlorhexidine and Ca(OH)2 was effective after 1 and 2 days demonstrating 100% antibacterial action; however, its antibacterial activity reduced between 7 and 15 days. CONCLUSION: Under the conditions of this study, it can be concluded that 2% chlorhexidine gel alone was more effective against E. faecalis than calcium hydroxide (P < 0.05). However, its antibacterial activity depended on how long it remained inside the root canal.

A preliminary in vitro study of the incidence and position of the root canal isthmus in maxillary and mandibular first molars.

AIM: To investigate in vitro the incidence and position of the root canal isthmus in extracted mesiobuccal roots of maxillary and mesial roots of mandibular first molars. METHODOLOGY: Fifty maxillary and 50 mandibular molars were included in the study. The mesiobuccal roots of maxillary molars and the mesial roots of mandibular molars were sectioned from their crowns in the furcation region and embedded in clear resin. Transverse serial 1-mm-thick sections from the apical 6 mm were prepared. The apical side of each section was stained with India ink and observed through a light microscope. The sample images were saved to disk using a digital camera and the root canals in terms of the number present and the incidence and classification of isthmuses. RESULTS: In the mesiobuccal root of the maxillary first molars, 70% had one canal, whereas 29.5% had two canals. In the mesial root of mandibular molars, 41% had one canal, whereas 59% had two canals. In some sections, more than two canals were found close to the apical foramen. The isthmus incidence was greatest 3-5 mm from the apex. In teeth having two canals, a complete or partial isthmus was frequently observed in the sections between 3 and 4 mm from the apex. Of the isthmuses present, 22% were complete and 37% partial in mandibular molars and 17.3% were complete and 11.7% partial in maxillary molars. CONCLUSIONS: The incidence of isthmus in the mesiobuccal root of the maxillary first molars and in the mesial root of the mandibular first molars was high, particularly in sections 3-5 mm from the apex. Cleaning the isthmus is a major challenge during root canal treatment.