Primary Root Canal Infections Research Articles

In vitro susceptibility of e.faecalis and C.albicans isolates from apical periodontitis to common antimicrobial agents, antibiotics and antifungal medicaments

The aim of this study was to evaluate in vitro antimicrobial activity of 4 antibiotic agents (for E.faecalis) and 4 antifungal agents (for C.albicans) by agar dilution method. Additionally, modified strip diffusion method was used for detection of in vitro antimicrobial activities of 5% NaOCl, 2.5% NaOCl, 17% EDTA and 2% CHX and agar diffusion method for detection of in vitro susceptibilities of three intracanal medicaments for 18 E.faecalis and 18 C.albicans isolates from primary and secondary root canal infection. Isolates were recovered from 231 endodontic samples of patients, with the need of root canal treatment and retreatment. All tested E.faecalis isolates showed resistance to antibiotics. For irrigation solutions, 2% CHX was more effective in eliminating E.faecalis but 5% NaOCl showed larger inhibition zone than 2.5% NaOCl, 17% EDTA and 2% CHX. For intracanal medication, Ca(OH)2-CHX worked efficiently in killing E.faecalis isolates compared to Ca(OH)2-Steril saline solution, Ca(OH)2-Glycerin. For C.albicans, 18 isolates were susceptible to amphotericin B, nistatin, fluconazole but showed resistance to ketoconazole. 5% NaOCl was more effective in eliminating and produced larger inhibition zone compared to 2.5% NaOCl, 17% EDTA and 2% CHX. Ca(OH)2-Glycerin intracanal medication was better in eliminating C.albicans isolates and produced larger inhibition zone compared to other Ca(OH)2 medicaments. Key words:E.faecalis, C.albicans, antimicrobial, antibiotic, antifungal.

Analysis of Symptomatic and Asymptomatic Primary Root Canal Infections in Adult Norwegian Patients

Introduction This molecular study analyzed the microbiota of primary root canal infections from adult Norwegian patients. Methods Samples were taken from the necrotic root canals of teeth with symptomatic (n = 13) or asymptomatic (n = 21) apical periodontitis and chronic apical abscesses (n = 9). DNA was extracted from samples, and bacterial identifications were performed by a closed-ended reverse-capture checkerboard approach targeting 50 candidate endodontic pathogens. Results Bacterial DNA was detected in all cases. In teeth with asymptomatic apical periodontitis, the most frequent taxa were Dialister invisus (71%), Fusobacterium nucleatum (62%), and Porphyromonas endodontalis (62%). In chronic apical abscesses, the most prevalent taxa were P. endodontalis (100%), D. invisus (89%), Parvimonas micra (78%), and Solobacterium moorei (78%). In teeth with symptomatic apical periodontitis, the most prevalent taxa were D. invisus, P. endodontalis, S. moorei, Propionibacterium acnes, and Streptococcus species (all in 69%). None of the targeted taxa were significantly associated with either sinus tract or pain ( P > .05), except for Selenomonas sputigena, which was more frequently found in painful cases ( P = .04). No taxa were found in significantly higher levels in any conditions ( P > .05). Cluster analyses revealed bacterial groupings that differed between cases with and without pain. Conclusions Although basically the same species were highly prevalent in the different conditions examined and none of the most prevalent taxa were positively associated with symptoms, results revealed that species formed different partnerships and associations in samples from teeth with or without pain. Therefore, it is possible that more virulent multispecies communities can form as a result of overall bacterial combinations and give rise to acute inflammation.

Clinical Investigation of the Efficacy of Chemomechanical Preparation with Rotary Nickel-Titanium Files for Removal of Endotoxin from Primarily Infected Root Canals

Introduction This clinical study was conducted to investigate the ability of chemomechanical preparation with 2.5% NaOCl + 17% ethylenediaminetetraacetic acid (EDTA) and rotary nickel-titanium (NiTi) system in removing endotoxin from primary root canal infection with apical periodontitis. Methods Twenty-one root canals with necrotic pulps were selected. Samples were collected before (s1) and after chemomechanical preparation (s2). The limulus amebocyte lysate (LAL) assay was used to quantify endotoxins. Results The LAL assay indicated that endotoxins were present in 100% of the root canals investigated (19/19) before (s1) and after chemomechanical preparation (s2). Analyses of the quantitative data revealed that the endotoxin content was significantly reduced at s2 (98.06%) compared with that at s1 ( P < .05). Conclusions Our findings indicate that chemomechanical preparation with 2.5% NaOCl + 17% EDTA and rotary NiTi files was effective in reducing endotoxin load in the root canal infection from primarily infected teeth with apical periodontitis.

Antigenic Activity of Bacterial Endodontic Contents from Primary Root Canal Infection with Periapical Lesions against Macrophage in the Release of Interleukin-1β and Tumor Necrosis Factor α

Introduction Periradicular tissue chronic stimulation by endotoxin may cause apical periodontitis. This study investigated the microbial profile and the levels of endotoxin found in primary root canal infection with apical periodontitis, determined their antigenicity against macrophages through the levels of interleukin (IL)-1β and tumor necrosis factor α (TNF-α), and evaluated their relationship with clinical and radiographic findings. Methods Samples were taken from 21 root canals with primary endodontic infection and apical periodontitis with paper points. PCR technique (16S rDNA) was used for the detection of the target bacteria. Limulus Amebocyte Lysate (LAL) was used to measure endotoxin. The amounts of IL-1ß/ TNF-alpha in macrophages supernatants were measured by enzyme-linked immunosorbent assay – Duoset-kit (ELISA). Results Prevotella nigrescens (13/21), Porphyromonas endodontalis (6/21), and Treponema socranskii (6/21) were the most frequently detected gram-negative bacterial species. The presence of the sinus tract (2/21) was related to the detection of Filifactor alocis (p < 0.05), whereas a tooth with a radiolucent area ≥2 mm was related to the detection of Treponema denticola. A correlation was found between the number of gram-negative bacteria and the levels of IL-1β/TNF-α (p < 0.05). Increased levels of endotoxin were followed by TNF-α release (p < 0.05). Higher levels of IL-1β (p < 0.05) and endotoxin contents were related to the larger size of the radiolucent area. Conclusion The antigenicity of the endodontic contents is not only related to the amount of endotoxin found in the root canal but also to the number of different species of gram-negative bacteria involved in the infection. Moreover, a larger size (≥2 mm) of the radiolucent area was related to IL-1β and endotoxin.

Susceptibility of Endodontic Pathogens to Antibiotics in Patients with Symptomatic Apical Periodontitis

Introduction The aim of this study was to evaluate susceptibility of predominant endodontic pathogens isolated from teeth with symptomatic apical periodontitis to most commonly prescribed antibiotics. Methods Among 58 patients with symptomatic apical periodontitis, 47 and 11 cases were caused by primary and secondary root canal infection, respectively. The microbial samples were taken either from the root canals (35 cases) or by aspiration from apical abscesses (23 cases). Culture methods were used to identify the microorganisms present in the samples. Antibiotic susceptibilities of all isolates were evaluated by using the E-test method. Results Microorganisms were isolated from 49 of the 58 samples studied and included facultative and obligate anaerobes. Streptococci and obligate anaerobes were the predominant microorganisms in cases of primary infection. Enterococcus faecalis dominated in cases of secondary infection. All tested microorganisms were highly sensitive to penicillin G, amoxicillin, and ampicillin. Susceptibilities to clindamycin and erythromycin were 73.8% and 54.7%, respectively. About 40% of the isolates were resistant to tetracycline. More than 50% of all anaerobes were resistant to metronidazole. All E. faecalis isolates were resistant to clindamycin. Conclusions Based on the study results, penicillin and amoxicillin are suitable antibiotics for treatment of endodontic infection when conventional root canal treatment alone is insufficient. Clindamycin could be advised for penicillin-allergic patients with primary endodontic infections.

Disinfection of the root canal system during root canal re‐treatment

In this narrative review, the differences between primary root canal treatments and re‐treatments are explored in view of optimal disinfection of the root canal system. A critical appraisal of the literature raises doubt as to whether the microbiota found in re‐treatment cases per se is more resistant to antiseptics than the counterpart found in primary infections. In reality, primary, refractory, and persisting endodontic infections are all biofilm‐related; their microbial composition is dictated by local ecological factors rather than treatment history. Furthermore, their resistance to antimicrobials is most likely similar. The true difficulty in disinfecting root canal systems during re‐treatment cases is to achieve access to the infected areas. Iatrogenic alterations in canal anatomy and the presence of root filling material hamper the diffusion of disinfectants to these target areas. Consequently, cleaning the canal systems of foreign material and creating a canal shape that can properly be disinfected should be the initial aims. Ways of achieving these goals are discussed. Subsequently, the disinfection regimen can be similar to that in primary root canal infections. However, time is a crucial factor in re‐treatments, and thus a multiple‐visit approach is preferable in more complex cases in order to ensure more complete disinfection has been achieved.

Microbiological evaluation of primary endodontic infections in teeth with and without sinus tract

To examine the microbiological status of primary endodontic infections in teeth with and without a sinus tract. Samples were collected by means of a size 15 H-type file and two sterile paper points from 30 cases of primary endodontic infections with (n = 15) or without (n = 15) a sinus tract. The presence of 40 bacterial species was determined by the checkerboard DNA-DNA hybridization method. The species found at the highest levels and prevalence were Fusobacterium nucleatum sp. vincentii, Porphyromonas gingivalis, Veillonella parvula, Enterococcus faecalis, Campylobacter gracilis and Neisseria mucosa. Total bacterial counts were similar between teeth with (44 x 10(5)) and without (50 x 10(5)) a sinus tract (t-test: P > 0.05). E. faecalis, Streptococcus anginosus, Capnocytophaga sputigena and Capnocytophaga gingivalis had significantly higher counts in the absence of sinus tract (Mann-Whitney test, P < 0.05). Higher levels of P. gingivalis and Fusobacterium nucleatum sp. nucleatum were observed in cases with a sinus tract. Leptotrichia buccalis (OR = 1.83; CI 95%) and Porphyromonas endodontalis (OR = 2.15; CI 95%) were associated with an increased chance of subjects having a sinus tract. Primary endodontic infections were associated with a large variety of bacterial species. Specific differences between the composition of the microbiota of primary root canal infections were observed in cases with or without a sinus tract.

Molecular evaluation of residual endodontic microorganisms after instrumentation, irrigation and medication with either calcium hydroxide or Septomixine

The correct choice of antimicrobial agents as inter-appointment medicaments is as important as the instrumentation and irrigation to remove pathogens from infected root canals. Calcium hydroxide [Ca(OH)2] and framycetin sulfate (Septomixine) are common endodontic medicaments. Therefore, we evaluated the efficacy of either calcium hydroxide or Septomixine in eliminating residual intra-canal bacteria, particularly Actinomyces spp., during inter-appointment interval in endodontic therapy using molecular methods. A total of 31 single-rooted teeth with primary root canal infections were studied immediately after opening the canals and subsequently after instrumentation, irrigation with sterile saline and 1-week medication with either Ca(OH)2 (n = 25) or Septomixine (n = 6). Whole bacterial genomic DNA was isolated directly from samples and PCR with universal primers performed to detect total intra-canal bacteria. The variable regions of 16S rDNA of bacteria were amplified and labeled with digoxigenin for further hybridization to detect Actinomyces spp. A total of seven oligonucleotide probes specific for A. bovis, A. gerencseriae, A. israelii, A. meyeri, catalase-negative A. naeslundii (genospecies 1 and 2), catalase-positive A. naeslundii genospecies 2 and A. odontolyticus were used to detect Actinomyces spp. in 22 of 31 medicated root canals [Ca(OH)2: n = 17; Septomixine: n = 5]. The PCR results showed that 25 of 31 examined canals were positively detected with residual microorganisms after instrumentation, irrigation with sterile saline and 1-week medication with either Ca(OH)2 (n = 20) or Septomixine (n = 5). Thus, only six canals [Ca(OH)2: n = 5, Septomixine: n = 1] were aseptic after treatment. Hybridization results showed higher detection frequency of both A. odontolyticus and A. gerencseriae after treatment. Significant correlation was found between exposed pulp before treatment and positive detection of Actinomyces spp., particularly A. odontolyticus on the second visit (P < 0.05). The conventional, 1-week medication of either Ca(OH)2 or Septomixine in endodontic therapy may not effectively inhibit residual bacterial growth in all root canals during inter-appointment intervals. Further investigations using, for instance quantitative real-time PCR analyses, are required to substantiate the present findings.

Gram‐positive organisms in endodontic infections

Culture‐based studies in endodontics have more or less overlooked the significance of Gram‐positive facultative bacteria in recent decades. By contrast, Gram‐negative anaerobes have been extensively studied because of their frequent recovery in primary root canal infections and their association with acute manifestations of apical periodontitis. More recent years have seen a renewed interest in Gram‐positive facultatives as these organisms are common in samples from root‐filled teeth affected by apical periodontitis. Structural components of the robust bacterial cell wall of Gram positives protect them from noxious environmental factors. Additionally, the majority of these organisms express fast‐adaptive properties when exposed to extreme conditions, thus making them potentially interesting as causal elements in post‐treatment endodontic disease. This review relates to different aspects of Gram‐positive bacteria and their adaptive responses when being exposed to stressful conditions such as endodontic treatment procedures.

Fungi in endodontic infections

Fungi are chemoorganotroph eukaryotic microorganisms that can take part in endodontic infections and thereby may participate in the etiology of periradicular diseases. They possess virulence attributes—including adaptability to a variety of environmental conditions, adhesion to a variety of surfaces, the production of hydrolytic enzymes, morphologic transition, biofilm formation, and evasion and immunomodulation of the host defense—that may play a role in the pathogenesis of periradicular diseases. Fungi have occasionally been found in primary root canal infections, but they seem to occur more often in the root canals of obturated teeth in which treatment has failed. Candida albicans is by far the fungal species most commonly isolated from infected root canals, and this species has been considered a dentinophilic microorganism because of its invasive affinity to dentin. C albicans has also been discovered to be resistant to some intracanal medicaments, such as calcium hydroxide. Its ability to invade dentinal tubules and resistance to commonly used intracanal medicaments may help to explain why C albicans has been associated with cases of persistent root canal infections. Some medicaments, such as chlorhexidine digluconate, calcium hydroxide combinations (with camphorated paramonochlorophenol or chlorhexidine), and EDTA, have the potential to be used as effective intracanal medications for patients in whom fungal infection is suspected.

Fungi in endodontic infections

Fungi are chemoorganotroph eukaryotic microorganisms that can take part in endodontic infections and thereby may participate in the etiology of periradicular diseases. They possess virulence attributes—including adaptability to a variety of environmental conditions, adhesion to a variety of surfaces, the production of hydrolytic enzymes, morphologic transition, biofilm formation, and evasion and immunomodulation of the host defense—that may play a role in the pathogenesis of periradicular diseases. Fungi have occasionally been found in primary root canal infections, but they seem to occur more often in the root canals of obturated teeth in which treatment has failed. Candida albicans is by far the fungal species most commonly isolated from infected root canals, and this species has been considered a dentinophilic microorganism because of its invasive affinity to dentin. C albicans has also been discovered to be resistant to some intracanal medicaments, such as calcium hydroxide. Its ability to invade dentinal tubules and resistance to commonly used intracanal medicaments may help to explain why C albicans has been associated with cases of persistent root canal infections. Some medicaments, such as chlorhexidine digluconate, calcium hydroxide combinations (with camphorated paramonochlorophenol or chlorhexidine), and EDTA, have the potential to be used as effective intracanal medications for patients in whom fungal infection is suspected.

Direct detection of Actinomyces spp. from infected root canals in a Chinese population: a study using PCR-based, oligonucleotide-DNA hybridization technique.

The poor sensitivity of phenotypic identification techniques has hampered the taxonomic differentiation of Actinomyces. Hence we developed a sensitive and specific, PCR-based oligonucleotide-DNA hybridization technique to detect Actinomyces spp. and, used this method to detect these organisms in samples directly obtained from infected root canals. A total of 32 samples from 28 Chinese patients, with primary root canal infections, aseptically exposed at the first patient visit, were studied. Whole bacterial genomic DNA was isolated directly from paper point samples. The variable regions of 16S ribosomal DNA of bacteria were amplified and labeled with digoxigenin for further hybridization and detection. A total of seven oligonucleotide probes specific for A. bovis, A. gerencseriae, A. israelii, A. meyeri, catalase-negative A. naeslundii (genospecies 1 and 2), catalase-positive A. naeslundii genospecies 2 and A. odontolyticus were used. 16 of the 32 teeth were infected with one or more Actinomyces species. The prevalence rates of the examined species were: A. odontolyticus 31.3%, A. meyeri 9.4%, A. naeslundii 9.4%, A. israelii 6.3% and A. gerencseriae 3.1%; no A. bovis was detected in any of the canals. Furthermore, A. odontolyticus was isolated more frequently from root canals with caries or a history of caries (Fisher's exact test: P=0.0496; Odds ratio=9.00, 95% confidence interval: 0.97-83.63), and A. naeslundii was significantly associated with traumatized teeth (Fisher's exact test: P=0.0121; Odds ratio=57.00, 95% confidence interval: 2.10-1546.90). However, no significant correlation was found between Actinomyces spp. and clinical symptoms and signs, such as pain, swelling, percussion to tenderness, sinus and periapical radiolucency. Actinomyces spp. may be important pathogens of root canal infections. A. naeslundii in particular may be related with traumatized teeth. A. odontolyticus appears to be involved in infections related to caries, exposure of dentinal tubules during cavity preparation and/or leaking restoration, but further clarification with large samples is necessary.

PCR-based identification of Treponema maltophilum, T. amylovorum, T. medium, and T. lecithinolyticum in primary root canal infections

Objective: Molecular genetic methods have significantly contributed to the knowledge about the microbiota associated with infected root canals. Albeit spirochetes have been commonly observed in primary root canal infections, only recently they have been identified. The purpose of the present study was to investigate the occurrence of four treponemes— Treponema maltophilum, Treponema lecithinolyticum, Treponema amylovorum, and Treponema medium—in cases of primary endodontic infections associated with different forms of periradicular diseases through a 16S rDNA-based nested PCR assay. Design: Samples were taken from thirty-one infected root canals associated with either asymptomatic or symptomatic apical periodontitis. DNA extracted from the samples was initially amplified using universal 16S rDNA primers, followed by a second round of amplification using the first PCR products to detect a specific fragment of the 16S rDNA of each target Treponema species. Results: All cases were positive for the universal bacterial primers, indicating that samples contained bacterial DNA. Of the four target species, T. maltophilum was the most prevalent, being detected in 39% of the cases (33% of the asymptomatic cases and 50% of the symptomatic cases). T. lecithinolyticum was the next more prevalent among the species tested, being found in 26% of the samples (33% of asymptomatic cases and 10% of the symptomatic cases). T. amylovorum was found in 7% of the cases (5% of the asymptomatic cases and 10% of the symptomatic cases), while T. medium was in 13% of the cases (14% of the asymptomatic cases and 10% of the symptomatic cases). None of the species tested was significantly associated with clinical symptoms. Conclusions: This was possibly the hitherto first study to report the occurrence of T. lecithinolyticum, T. amylovorum, and T. medium in infections of endodontic origin. Overall, findings suggested that these oral treponemes, particularly T. maltophilum and T. lecithinolyticum, can be involved in the pathogenesis of periradicular diseases.

Campylobacter gracilis and Campylobacter rectus in primary endodontic infections.

A species-specific nested polymerase chain reaction (PCR) assay was used to investigate the occurrence of Campylobacter gracilis and C. rectus in primary root canal infections. Samples were collected from 57 single-rooted teeth with carious lesions, necrotic pulps and radiographic evidence of periradicular disease. Twenty-eight cases were diagnosed as chronic asymptomatic periradicular lesions, 12 cases as acute apical periodontitis, and 17 cases as acute periradicular abscess. DNA was extracted from the samples and initially amplified using universal 16S rDNA primers. A second round of amplification using the first PCR products was performed to specifically detect C. gracilis or C. rectus in the samples. Campylobacter gracilis and C. rectus were, respectively, detected in 21.4 (6 of 28) and 30% (6 of 20) of the root canals associated with chronic asymptomatic periradicular lesions. Campylobacter gracilis was found in 16.7% (2 of 12) of the cases diagnosed as acute apical periodontitis, whilst C. rectus was found in 33.3% (two of six cases). In the abscessed cases, C. gracilis and C. rectus were detected in 23.5 (4 of 17) and 11.8% (2 of 17) of the cases, respectively. No association of these species with clinical symptoms was observed (P > 0.01) In general, species-specific nPCR allowed the detection of C. gracilis in 21.1% (12 of 57) and C. rectus in 23.3% (10 of 43)of the samples taken from primary endodontic infections. Findings confirmed the assertion that both C. gracilis and C. rectus participate in infections of endodontic origin and suggest a pathogenetic role with regard to periradicular diseases.

Peptostreptococcus micros in Primary Endodontic Infections as Detected by 16S rDNA-based Polymerase Chain Reaction

A 16S rDNA-based polymerase chain reaction (PCR) method was used to detect Peptostreptococcus micros in primary root canal infections. Samples were collected from 50 teeth having carious lesions, necrotic pulps, and different forms of periradicular diseases. DNA extracted from the samples was amplified using the PCR assay, which yielded a specific fragment of P. micros 16S rDNA. P. micros was detected in 6 of 22 root canals associated with asymptomatic chronic periradicular lesions (27.3%), 2 of 8 teeth with acute apical periodontitis (25%), and 6 of 20 cases of acute periradicular abscess (30%). In general, P. micros was found in 14 of 50 cases (28%). There was no correlation between the presence of P. micros and the occurrence of symptoms. Findings suggested that P. micros can be involved in the pathogenesis of different forms of periradicular lesions.

Oral treponemes in primary root canal infections as detected by nested PCR.

To investigate the prevalences of four Treponema species in primary root canal infections using a nested PCR assay. Samples were obtained from 32 infected root canals. Twenty-two cases showed chronic asymptomatic periradicular lesions and 10 symptomatic cases were diagnosed as acute apical periodontitis. DNA extracted from the samples was initially amplified using universal 16S rDNA primers. A second round of amplification used the first PCR products to detect a specific fragment of the 16S rDNA of each Treponema denticola, T. socranskii, T. vincentii and T. pectinovorum. Bacteria were present in all cases sampled. T. denticola was detected in 77.3% of the asymptomatic cases, T. socranskii in 40.9%, T. vincentii in 18.2% and T. pectinovorum in 13.6%. In the cases diagnosed as acute apical periodontitis, T. denticola was detected in 80%, whilst T. socranskii and T. vincentii were detected in 40% and 10% of cases, respectively. No symptomatic case yielded T. pectinovorum. In general, nested PCR detected T. denticola in 78.1% of the cases, T. socranskii in 40.6%, T. vincentii in 15.6% and T. pectinovorum in 9.4%. At least one of the four Treponema species was found in 84.4% of the cases examined. The species T. denticola was detected in a large number of the cases examined: the prevalence of T. socranskii was also relatively high. The species T. vincentii and T. pectinovorum were also found, but in a smaller number of cases. Based on these data, the recognized pathogenicity of these microorganisms and their involvement with other oral diseases, they should be included in the restricted set of putative endodontic pathogens.

Dialister pneumosintes can be a suspected endodontic pathogen.

Dialister pneumosintes is a nonmotile, nonfermentative, non-spore-forming, obligately anaerobic, gram-negative bacillus that has been associated with some infections in the human body. A species-specific nested polymerase chain reaction assay was used to investigate the occurrence of D pneumosintes in primary root canal infections. Samples were collected from 32 teeth with carious lesions, necrotic pulps, and radiographic evidence of periradicular bone destruction. Twenty-two teeth were asymptomatic, and 10 cases were diagnosed as acute apical periodontitis. DNA extracted from the samples was initially amplified with universal 16S ribosomal DNA primers. A second round of amplification used the first polymerase chain reaction products to specifically detect D pneumosintes. This bacterial species was detected in 17 of 22 asymptomatic cases (77.3%) and in 4 of 10 root canals associated with acute apical periodontitis (40%). No relationship was found between the presence of this bacterial species and the occurrence of symptoms. In general, D pneumosintes was detected in 21 of 32 root canal samples (65.6%). This study is the first to report a high prevalence of D pneumosintes in root canal infections of humans. Because of this high prevalence and the apparent pathogenicity of the microorganism, we suggest the inclusion of D pneumosintes in the selected group of putative endodontic pathogens.

Actinomyces Species, Streptococci, and Enterococcus faecalis in Primary Root Canal Infections

The purpose of this study was to evaluate the prevalence of Actinomyces species, streptococci, and Enterococcus faecalis in primary root canal infections by using a molecular genetic method. Samples were obtained from 53 infected teeth, of which 27 cases were diagnosed as acute periradicular abscesses. DNA was extracted to evaluate the occurrence of 13 bacterial species by using whole genomic DNA probes and checkerboard DNA-DNA hybridization. Polymerase chain reaction using an ubiquitous bacterial primer was undertaken to check the presence of bacterial DNA in clinical samples. All root canal samples contained bacteria as demonstrated by polymerase chain reaction. The checkerboard DNA-DNA hybridization assay allowed the detection of streptococci in 22.6% of the samples, Actinomyces species in 9.4%, and E. faecalis in 7.5%. The most prevalent species were members of the Streptococcus anginosus group. With regard to the asymptomatic lesions, the most prevalent species were S. intermedius (11.5% of the cases), E. faecalis (11.5%), and S. anginosus (7.7%). S. constellatus was the most prevalent species in pus samples (25.9% of the cases). The other most prevalent species in abscessed teeth were A. gerencseriae (14.8%), S. gordonii (11.1%), S. intermedius (11.1%), A. israelii (7.4%), S. anginosus (7.4%), and S. sanguis (7.4%). S. constellatus was the only species positively associated with acute periradicular abscess (p < 0.01).

Comparison of the Effectiveness of Bacterial Culture, 16S rDNA Directed Polymerase Chain Reaction, and Checkerboard DNA-DNA Hybridization for Detection of Fusobacterium nucleatum in Endodontic Infections

Fusobacterium nucleatum is a Gram-negative, non-spore-forming, nonmotile, obligatory anaerobic rod that is normally isolated from the oral cavity. Epidemiological studies have shown that this species is one of the most prevalent in primary root canal infections. The purpose of this study was to compare the effectiveness of bacteriological culture, 16S rDNA directed polymerase chain reaction and checkerboard DNA-DNA hybridization for detection of F. nucleatum strains in infected teeth associated with periradicular lesions. Thirteen single-root teeth from adult patients, all having carious lesions, necrotic pulps, and radiographic evidence of periradicular bone loss were included in this study. Combining all methods, the results indicated that F. nucleatum was present in approximately 31% (4 of 13) of the specimens. Incidence of F. nucleatum in root canal infections, as evaluated in this study by polymerase chain reaction, culture, and DNA-DNA hybridization, was 15.4%, 15.4%, and 10.0%, respectively. Our data demonstrated that no method used herein could be considered superior for detecting F. nucleatum directly from clinical samples. However, the small number of samples examined and the low prevalence that was observed should be considered.

Patterns of microbial colonization in primary root canal infections

The purpose of this study was to investigate the patterns of microbial infection of root canals in untreated teeth associated with chronic periradicular lesions by use of scanning electron microscopy. Fifteen extracted teeth with extensive carious lesions, radiolucent lesions of varying sizes, and attached periradicular lesions after extraction were selected for study. After fixation, lesions were removed and the teeth were split into 2 halves. The teeth were then dehydrated, sputter-coated with gold, and then examined for the patterns of microbial colonization of the root canal system by using a scanning electron microscope. All examined root canals were infected, and bacterial cells were seen in practically all areas of the root canal system. The pattern of colonization was not uniform between specimens and even within the same specimen. Most of the root canal walls of all specimens were heavily colonized by a root canal microbiota consisting of cocci and/or rods, often forming mixed communities. Spirilla were occasionally observed as single cells or as small clusters between other bacterial forms. Bacteria were often observed penetrating the dentinal tubules. Although a shallow penetration was the most common finding, bacterial cells could be observed reaching approximately 300 microm in some specimens. Yeastlike cells were observed in 1 specimen together with bacteria. The root canals of teeth associated with periradicular lesions were heavily infected by bacteria and occasionally by fungi. The pattern of colonization of the root canal microbiota often showed the characteristic of a climax community, which may require special considerations regarding its elimination and prevention of clinical problems. In addition, the observed propagation of the infection to the entire root canal system in teeth associated with periradicular lesions suggests that proper therapeutic measures may be necessary to eliminate the root canal infection completely.