Effect of maleic acid and ethylenediaminetetraacetic acid on the shear bond strength of RealSeal SE sealer to root canal dentin.

Aim of this study was to evaluate the effect of 7% maleic acid and 17% ethylenediaminetetraacetic acid (EDTA) on the shear bond strength of RealSeal SE sealer to root canal dentin. Twenty incisors were split into coronal, middle and apical third and were treated in the following manner: Group 1: 5 ml of saline (1 minute). Group 2: 2.5% sodium hypochlorite (5 ml/min) followed by 79% maleic acid (5 ml/min). Group 3: 2.5% sodium hypochlorite (5 ml/min) followed by 17% EDTA (5 ml/min). Sealer was the placed on the root dentin and samples were subjected to bond strength measurement. There was no significant difference in bond strength between maleic acid and 17% EDTA in coronal & middle third. However, in apical third, 7% maleic acid showed higher bond strength. Least bond strength was observed with saline. Bond strength was maximum in apical third for both 7% maleic acid and 17% EDTA.

Tooth pulp inflammation increases brain-derived neurotrophic factor expression in rodent trigeminal ganglion neurons.

Nociceptive pathways with first-order neurons located in the trigeminal ganglion (TG) provide sensory innervation to the head, and are responsible for a number of common chronic pain conditions, including migraines, temporomandibular disorders and trigeminal neuralgias. Many of those conditions are associated with inflammation. Yet, the mechanisms of chronic inflammatory pain remain poorly understood. Our previous studies show that the neurotrophin brain-derived neurotrophic factor (BDNF) is expressed by adult rat TG neurons, and released from cultured newborn rat TG neurons by electrical stimulation and calcitonin gene-related peptide (CGRP), a well-established mediator of trigeminal inflammatory pain. These data suggest that BDNF plays a role in activity-dependent plasticity at first-order trigeminal synapses, including functional changes that take place in trigeminal nociceptive pathways during chronic inflammation. The present study was designed to determine the effects of peripheral inflammation, using tooth pulp inflammation as a model, on regulation of BDNF expression in TG neurons of juvenile rats and mice. Cavities were prepared in right-side maxillary first and second molars of 4-week-old animals, and left open to oral microflora. BDNF expression in right TG was compared with contralateral TG of the same animal, and with right TG of sham-operated controls, 7 and 28 days after cavity preparation. Our ELISA data indicate that exposing the tooth pulp for 28 days, with confirmed inflammation, leads to a significant upregulation of BDNF in the TG ipsilateral to the affected teeth. Double-immunohistochemistry with antibodies against BDNF combined with one of nociceptor markers, CGRP or transient receptor potential vanilloid type 1 (TRPV1), revealed that BDNF is significantly upregulated in TRPV1-immunoreactive (IR) neurons in both rats and mice, and CGRP-IR neurons in mice, but not rats. Overall, the inflammation-induced upregulation of BDNF is stronger in mice compared to rats. Thus, mouse TG provides a suitable model to study molecular mechanisms of inflammation-dependent regulation of BDNF expression in vivo.

Bacterial community profiles of endodontic abscesses from Brazilian and USA subjects as compared by denaturing gradient gel electrophoresis analysis.

This study compared the bacterial community profiles of the microbiota associated with acute apical abscesses from Brazilian and USA patients using denaturing gradient gel electrophoresis (DGGE). DNA was extracted from purulent exudate aspirates and part of the 16S rRNA gene was amplified by polymerase chain reaction and separated by DGGE. The resulting banding patterns, which were representative of the bacterial community structures in samples from the two locations, were then compared. Distinct DGGE banding patterns were observed from different samples. Ninety-nine bands with distinct positions in the gels were detected, of which 27 were found only in the USA samples and 13 were exclusive to Brazilian samples. Four of the 59 shared bands showed very discrepant findings with regard to prevalence in the two locations. Cluster analysis of DGGE banding profiles showed a great variability in the bacterial populations associated with teeth with abscesses regardless of the geographical location. Two big clusters, one for each location, were observed. Other clusters contained a mixture of samples from the two locations. The results of the present study demonstrated a great variability in the bacterial community profiles among samples. This indicates that the bacterial communities of abscesses are unique for each individual in terms of diversity. The composition of the microbiota in some samples showed a geography-related pattern. Furthermore, several bands were exclusive for each location and others were shared by the two locations and showed great differences in prevalence.

Searching for Archaea in infections of endodontic origin.

Archaea is a highly diverse group of prokaryotes, whose members have been traditionally recognized as extremophiles. Recently, some of these microorganisms have also been found to thrive in nonextreme environments, including the human body. Methanogenic archaea have been detected in samples from subgingival plaque associated with periodontal disease and a pathogenetic role is suspected. The purpose of this study was to survey samples taken from different types of endodontic infections for the presence of archaea. Samples were taken from untreated and treated root canals associated with asymptomatic chronic periradicular lesions as well as from cases diagnosed as acute periradicular abscesses. Overall, 96 samples were obtained. DNA from samples was extracted by using two different protocols and used as template for polymerase chain reaction amplification using oligonucleotide universal primers for the domains Archaea or Bacteria. Samples were also checked for the presence of spirochetes by making use of a group-specific primer. While bacteria were present in all samples, no case yielded archaeal DNA. Spirochetes occurred in a high number of cases. Our findings suggested that members of the Archaea domain are not members of the microbiota present in different types of endodontic infections and thereby may not be implicated in the etiology of apical periodontitis.

Geographical differences in bacteria detected in endodontic infections using polymerase chain reaction.

The polymerase chain reaction (PCR) is an innovative nucleic acid-based assay that has the highest sensitivity of any microbiological technique for the detection of bacteria. The purpose of this study was to use PCR to detect the presence of specific species of bacteria in samples collected from two geographical locations. Microbial samples from abscesses of endodontic origin were collected from patients in Portland, Oregon, and Rio de Janeiro, Brazil. PCRs with species-specific oligonucleotide primers for the 16S ribosomal RNA gene were used for detection of the bacteria after DNA extraction from each clinical sample. Statistical analysis revealed that there was a significant difference in detection of the bacteria between the two geographical locations for Prevotella intermedia, Prevotella nigrescens, Prevotella tannerae, Fusobacterium nucleatum, and Porphyromonas gingivalis, but not for Porphyromonas endodontalis, Fusobacterium necrophorum, and Enterococcus faecalis. These results suggest that differences in bacteria detected or cultured in studies can be associated with geographical location.

Radicular temperatures associated with thermoplasticized gutta-percha.

Thermoplasticized gutta-percha has been used to obturate root canals. The continuous wave of condensation technique uses the System B Heat Source with the choice of different-sized pluggers. The purpose of this study was to measure the temperatures within the root canal and on the root surface at different radicular levels while using the System B Heat Source. Fine, Fine-Medium, and Medium pluggers were evaluated at temperature settings of 200 degrees C, 250 degrees C, and 300 degrees C. The Obtura II gutta-percha delivery system following the manufacturer's instructions and ultrasonically thermoplasticized gutta-percha were used for comparative purposes. The highest mean temperature change on the internal root surface was 74.19 degrees C with the system B at the 6 mm level (6 mm coronal to working length) when the Fine-Medium plugger was set at 300 degrees C. The lowest mean temperature change on the internal root surface was 2.09 degrees C at the 0 mm level (at working length) when the F plugger was set at 200 degrees C. With the Obtura II, the lowest mean internal temperature change was 5.22 degrees C at the 0 mm level, whereas the highest mean internal temperature change was 26.63 degrees C at the 6 mm level. With ultrasonic lateral compaction the lowest mean internal temperature change was 5.01 degrees C at the 0 mm level, whereas the highest mean internal temperature change was 28.95 degrees C at the 6 mm level. At no time did the System B, the Obtura II, or ultrasonic delivery of warm gutta-percha exceed an increase of 10 degrees C at any thermocouple level on the external root surface.

A clinical comparison of pain perception to the Wand and a traditional syringe.

OBJECTIVES: This study compared the pain response of a group of 40 volunteers to the Wand with the response to syringe injections. STUDY DESIGN: Of 240 total injections given, 120 were with the Wand system, and 120 were with the traditional aspirating syringe. Three injections were evaluated: injections to the middle superior alveolar (MSA) of the maxillary right first premolar and the maxillary left first premolar; palatal injections of the maxillary right first premolar and the maxillary left first premolar; and inferior alveolar nerve injection (IAN) of both the right and the left sides. Each volunteer received 6 injections, 3 on one side with the Wand and 3 on the opposite side with the syringe (control). All injections were given by the primary investigator without the use of a topical anesthetic. Pain perception levels were recorded with a 4-point visual analog scale: no pain, mild pain, moderate pain, and severe pain. The results were evaluated statistically by using an unpaired t test. RESULTS: The results showed injections with the syringe were statistically more painful than injections with the Wand in 4 of 18 evaluations: MSA of the maxillary left first premolar, female volunteers responses to MSA of the maxillary left first premolar, IAN left side injections, and male responses to IAN left injections (P =.01, P =.05, P =.05 and P =.05, respectively). CONCLUSIONS: The Wand generally seemed to provide less painful injections; however, the mean ratings of pain were mostly mild pain for both injections. Therefore, the clinical significance of the results should be interpreted with caution. The operator technique and tactile skill in syringe injections and site of injection (right or left) could be important factors that were not evaluated in this study.

Isolation and identification of Prevotella tannerae from endodontic infections.

Black-pigmented bacteria are often isolated from endodontic infections. Five strains of black-pigmented bacteria isolated from endodontic infections could not be identified in our laboratory. The purpose of this study was to sequence the 16S rRNA gene of the five unknown isolates and identify the organisms. The 16S rRNA genes from the unknown organisms were cloned, sequenced, and determined to be Prevotella tannerae. In addition, samples from endodontic infections were surveyed for the presence of the organism. When 118 samples from endodontic infections were examined using polymerase chain reaction with specific primers for P. tannerae, 60% of the samples were positive for the presence of the organism. This suggests that P. tannerae is commonly present in endodontic infections and could be a potential pathogen.

Occurrence of Candida albicans in infections of endodontic origin.

Microorganisms are recognized as the etiological agent for the majority of pulpal and periradicular disease. Although bacteria have been the most studied, fungi have also been associated with infected root canals. The purpose of this study was to evaluate the contents of infected root canals and aspirates of cellulitis/abscesses of endodontic origin for the presence of Candida albicans using the polymerase chain reaction (PCR). PCR primers specific for the 18S ribosomal RNA gene of C. albicans were used to survey 24 samples taken from infected root canals and 19 aspirates from periradicular infections of endodontic origins. The presence of C. albicans was detected in 5 of 24 (21%) samples taken from root canals, but none was detected in the periradicular aspirates. The results indicate that PCR is an extremely sensitive molecular method that may be used to identify C. albicans directly in samples from infections of endodontic origin.

A comparative study of smear layer removal using different salts of EDTA.

Three solutions of EDTA--a 15% concentration of the alkaline salt, a 15% concentration of the acid salt, and a 25% concentration of the alkaline salt--were evaluated for smear layer removal in root canal systems. All solutions were adjusted to pH 7.1 using either NaOH or HCl. When the EDTA solutions were alternately used for root canal irrigation with 5.25% NaOCl, they completely removed the smear layer in the middle and coronal thirds of canal preparations, but were less effective in the apical third. None of the EDTA solutions by themselves were effective at completely removing the smear layer at any level. The alkaline tetrasodium salt, pH adjusted with HCl, is more cost effective and performed equally as well as the more commonly used disodium salt.

Association of black-pigmented bacteria with endodontic infections.

Black-pigmented bacteria (BPB) have been associated with endodontic infections. The purpose of this study was to evaluate further the presence of BPB with the clinical signs and symptoms associated with endodontic infections. Microbial samples were collected from the root canals of 40 intact teeth with necrotic pulps and apical periodontitis. Conventional laboratory methods were used for identification of the strains of BPB isolated in pure culture. In addition, the polymerase chain reaction and specific primers for 16S r-RNA genes were used to differentiate Prevotella nigrescens from Prevotella intermedia. Twenty-two (55%) samples were positive for the growth of BPB. Of those, 11 of 22 (50%) were identified as P. nigrescens, 8 of 22 (36%) were P. intermedia, 2 of 22 (9%) were Porphyromonas gingivalis, and 1 of 22 (5%) was Prevotella melaninogenica. Sixteen of the 22 root canals positive for the growth of BPB were associated with purulent drainage either from the root canal or an associated sinus tract. Statistical analysis did not show a significant relationship for the presence of BPB with clinical signs and symptoms.

An analysis of canal centering using mechanical instrumentation techniques.

The purpose of this study was to compare canal transportation in moderately curved canals using mechanical instrumentation systems. Mesial roots of mandibular first or second molars were mounted in resin using a modified Bramante muffle system and divided into four groups. The roots were cross-sectioned 2 mm from the working length and at the height of root curvature. Tracings of the canal were made from preinstrumentation slides of the cross-sections. The canals were prepared using ProFile Series 29 rotary instruments, Quantec 2000 rotary instruments, Flex-R files in the Endo Gripper contra-angle handpiece, and Shaping Hedstrom files in the M4 contra-angle handpiece. Tracings of the prepared canals were made onto the originals from postinstrumentation slides. A canal centering ratio was calculated along the line of maximum transportation. Quantec 2000 rotary instruments yielded significantly greater transportation at the apical level when compared with the Profile Series 29 system. There were no other significant differences in transportation at either level. There were no differences in the direction of canal transportation between instrument systems, and the direction of canal transportation was not related to the direction of canal curvature. Canal preparation time was shortest with Profile Series 29 system followed by Flex-R files in the Endo Gripper, Quantec 2000, and Shaping Hedstrom files in the M4.

Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and polymerase chain reaction for differentiation of Prevotella intermedia and Prevotella nigrescens.

Isolates previously thought to be Prevotella intermedia have been shown to be a closely related species now known as Prevotella nigrescens. The purpose of this study was to determine the efficacy of sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and polymerase chain reaction (PCR) to differentiate endodontic isolates of P. nigrescens from P. intermedia. Fifty-six strains of black-pigmented bacteria isolated from endodontic infections and conventionally identified as P. intermedia were used in this study. Using SDS-PAGE, novel polypeptide bands were used to differentiate P. nigrescens from P. intermedia. PCR was accomplished with specific primers for the 16S ribosomal RNA gene of both strains. Of 56 endodontic isolates, 41 (73%) strains were identified by SDS-PAGE as P. nigrescens and 15 (27%) strains as P. intermedia. Of the 41 strains of P. nigrescens identified by SDS-PAGE, PCR identified 37 strains as P. nigrescens. Restriction endonuclease digestion of amplified 16S ribosomal RNA genes indicated that the remaining four strains originally identified by SDS-PAGE as P. nigrescens were actually strains of Prevotella distinct from P. nigrescens and P. intermedia. Of 15 strains of P. intermedia identified by SDS-PAGE, PCR identified 14 strains as P. intermedia; but, one strain was identified as P. nigrescens. The results indicated that PCR was a more precise method than SDS-PAGE to differentiate P. intermedia from P. nigrescens. This study confirms that P. nigrescens is more commonly isolated in pure culture from endodontic infections than P. intermedia.

Effect of calcium hydroxide and four irrigation regimens on instrumented and uninstrumented canal wall topography.

The topography of instrumented and uninstrumented canal walls exposed to calcium hydroxide and four different irrigation regimens was observed by scanning electron microscopy. After chemomechanical debridement, one tooth in each matched pair was medicated with calcium hydroxide. One week later, the teeth were irrigated and split longitudinally for evaluation. When no calcium hydroxide was used, predentin and pulpal debris covered the dentinal tubules of the uninstrumented surfaces in specimens irrigated with water or EDTA, but was absent on uninstrumented surfaces in specimens irrigated with NaOCl or NaOCl and EDTA. A typical smear layer was absent in instrumented specimens irrigated with NaOCl and EDTA, but covered the dentinal tubules of the instrumented surfaces of the EDTA irrigated specimens (partially) and the water or NaOCl irrigated specimens (completely). Calcospherites or their remnants were seen on the uninstrumented canal walls of specimens irrigated with NaOCl or NaOCl and EDTA, respectively. Calcium hydroxide use did not alter the surface topography in specimens irrigated with water, EDTA, or NaOCl, but seemed to erode the intertubular dentin in specimens irrigated with NaOCl and EDTA. All irrigants seemed to effectively remove most of the calcium hydroxide.

Survey for collagenase gene prtC in Porphyromonas gingivalis and Porphyromonas endodontalis isolated from endodontic infections.

Collagenase is a potential virulence factor shown to be expressed by Porphyromonas gingivalis associated with periodontal disease. The purpose of this study was to use the polymerase chain reaction (PCR) to detect the presence of the collagenase gene (prtC) in 21 strains of Porphyromonas species isolated from endodontic infections. Type strains for P. gingivalis (ATCC 33277), P. endodontalis (ATCC 35406), Prevotella intermedia (ATCC 25611), and Prevotella nigrescens (ATCC 33563) were used as controls. When PCR primers specific for the 16S ribosomal RNA gene of P. gingivalis or P. endodontalis were used, 16 of the strains were identified as P. gingivalis, and five strains were identified as P. endodontalis. The presence of the prtC gene for collagenase was detected using PCR. Amplicons were analyzed by agarose gel electrophoresis, with an 815 bp amplicon representing the presence of the collagenase gene. Type strain ATCC 33277 and all 16 clinical isolates of P. gingivalis produced the collagenase gene amplicon. Neither type strain ATCC 35406 nor the five strains from clinical isolates of P. endodontalis produced the collagenase gene amplicon. These results indicate that P. gingivalis from endodontic infections possesses the prtC gene. P. endodontalis does not seem to exhibit prtC. The virulence of P. gingivalis may be related to its production of collagenase.

Evaluation of diamond-coated ultrasonic instruments for root-end preparation.

Ultrasonic instrumentation has been associated with cracking of the dentin in the area of the root-end preparation. The purpose of this study was to evaluate root-end preparations for cracking and to describe cavosurface morphology after the use of diamond-coated instruments. Forty teeth were inspected for intradentin cracks, incomplete canal cracks, and complete canal cracks before and after preparation with a stainless steel CT-5 ultrasonic instrument and again after root-end preparation with an S12D/90 degrees diamond-coated instrument. Six teeth had polyvinylsiloxane impressions taken of the root ends after preparation with the CT-5 and again after preparation with the diamond-coated instrument. Replicas were made, split, sputter-coated, and inspected using the scanning electron microscope. This study indicates that use of the diamond-coated instrument for root-end preparation does not result in significant root-end cracking and that it can remove cracks created by a prior instrument's use. The use of the diamond-coated instrument resulted in a heavily abraded, debris-covered cavosurface that may affect the apical seal.

Longitudinal study of the microleakage of two root-end filling materials using a fluid conductive system.

A comparison of the microleakage of mineral trioxide aggregate (MTA) and a high copper admix amalgam (Valiant PhD) in root-end preparations was made using a fluid conductive device. Thirty-three bilaterally matched pairs of extracted, single-rooted teeth were prepared and obturated using lateral compaction of gutta-percha. The root-ends were resected and 3-mm-deep class I cavity preparations were made. The root-end preparations were filled with either amalgam or MTA. Guttapercha coronal to the root-end fillings was removed leaving only the amalgam or MTA present as a barrier to fluid movement. In the fluid-conductive device, the root canals were filled with a phosphate-buffered saline solution at a pressure of 10 psi. The flow of fluid was measured and compared at 1, 2, 3, 4, 8, 12, 16, 20, and 24 wk. The results showed amalgam to have significantly higher microleakage after 4 wk and higher variability compared with the MTA group.

Black-pigmented bacteria in coronal and apical segments of infected root canals.

Black-pigmented bacteria (BPB) have been associated with infections of endodontic origin. The purpose of this study was to culture and identify BPB from the apical and coronal segments of infected root canals to understand better their ecological relationships. Teeth with a periapical radiolucency were extracted and immediately placed in reduced transport fluid for transport to an anaerobic chamber. Of 18 sampled roots, 12 were positive for the growth of BPB. Eight of the 12 roots with BPB had a carious exposure of the pulp chamber. Seven roots had Prevotella nigrescens in both the apical and the coronal segments. Six of these seven teeth had carious exposures of the pulp chamber. Of the 12 roots infected with BPB, six roots had two different species of BPB, with P. nigrescens always being one of the species. P. nigrescens was the most often isolated BPB from both the coronal and apical segments of infected root canals.