Biological tests of a temporary crown and bridge material.
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The biological compatibility of a silicophosphate cement (Fluoro-Thin) and a zinc phosphate cement (de Trey's Improved) has been assessed in in vitro (cell culture) and in vivo (monkey teeth) tests. In the in vitro tests both materials were toxic when freshly prepared. In experiments with prolonged cell-material contact time, with set specimens of the materials, the zinc phosphate cement appeared to be non-toxic, whereas the silicophosphate cement was clearly toxic. The in vivo experiments confirmed that a possible pulp reaction cauded by zinc phosphate cement is of a mild nature. The silicophosphate cement, however, cauded a moderate or severe reaction in the pulp after 8 days of observation, and there was chronic inflammation in most teeth after 36 or 72 days. It was concluded, therefore, that Fluoro-Thin should not be used as a luting agent or for restorative purposes in direct contact with vital dentine.
Twenty-nine strains from the Actinomyces species were tested for a range of surface properties. Results show considerable heterogeneity both between different species and within some of the species, especially Actinomyces naeslundii. Two commonly used A. naeslundii strains, T14V and ATCC 12104, fell within the low (salivary aggregation and collagen binding by T14V), moderate (surface charge and haemagglutination) or high range of values (hydrophobicity, saliva-coated hydroxyapatite adhesion, polystyrene binding by T14V, fibrinogen binding by T14V and collagen binding by A. naeslundii ATCC 12104). Both strains adhered well to saliva-coated hydroxyapatite; T14V bound the highest amount of fibrinogen, ATCC 12104 had the highest number of cells bound to collagen and T14V was not bound at all. The heterogeneity of these characteristics highlights the need to include a range of strains of Actinomyces in studies on their pathogenicity. Statistical correlations were found between a number of properties, for example saliva-coated hydroxyapatite adhesion and hydrophobicity, and between haemagglutination and hydrophobicity.
BACKGROUND/AIMS: Enterococci have been implicated in persistent root canal infections but their role in the infection process remains unclear. This study investigated the virulence, phenotype and genotype of 33 endodontic enterococcal isolates. METHODS: Phenotypic tests were conducted for antibiotic resistance, clumping response to pheromone, and production of gelatinase, hemolysin and bacteriocin. Genotype analysis involved polymerase chain reaction amplification of virulence determinants encoding aggregation substances asa and asa373, cytolysin activator cylA, gelatinase gelE, gelatinase-negative phenotype ef1841/fsrC, adherence factors esp and ace, and endocarditis antigen efaA. Physical DNA characterization involved pulsed-field gel electrophoresis of genomic DNA, and plasmid analysis. RESULTS: Potential virulence traits expressed included production of gelatinase by Enterococcus faecalis (n=23), and response to pheromones in E. faecalis culture filtrate (n=16). Fourteen strains produced bacteriocin. Five strains were resistant to tetracycline and one to gentamicin, whereas all were susceptible to ampicillin, benzylpenicillin, chloramphenicol, erythromycin, fusidic acid, kanamycin, rifampin, streptomycin and vancomycin. Polymerase chain reaction products encoding efaA, ace, and asa were detected in all isolates; esp was detected in 20 isolates, cylA in six isolates, but asa373 was never detected. The gelatinase gene (gelE) was detected in all isolates of E. faecalis (n=31) but not in Enterococcus faecium (n=2); a 23.9 kb deletion sequence corresponding to the gelatinase-negative phenotype was detected in six of the eight E. faecalis isolates that did not produce gelatinase. Pulsed-field gel electrophoresis and plasmid analyses revealed genetic polymorphism with clonal types evident. Plasmid DNA was detected in 25 strains, with up to four plasmids per strain and a similar (5.1 kb) plasmid occurring in 16 isolates. CONCLUSIONS: Phenotypic and genotypic evidence of potential virulence factors were identified in endodontic Enterococcus spp., specifically production of gelatinase and response to pheromones.
AIMS: To investigate the occurrence of Olsenella uli and Olsenella profusa in samples taken from infections of endodontic origin using a devised 16S RNA gene-based nested PCR protocol. METHODS AND RESULTS: DNA extracted from clinical samples was initially amplified using universal 16S rRNA gene primers followed by a second round of amplification using the first PCR products to detect a specific fragment of either O. uli or O. profusa 16S rRNA gene. Olsenella uli was detected in 27% of samples from teeth with asymptomatic periradicular lesions and in 20% of pus samples taken from symptomatic (abscess) lesions. Olsenella profusa was detected in 7% of asymptomatic teeth and in 13% of symptomatic teeth. A new Olsenella phylotype was also detected. CONCLUSIONS: Olsenella species can take part in the microbiota associated with infections of the root canal of human teeth and a role in the pathogenesis of periradicular diseases is suspected. SIGNIFICANCE AND IMPACT OF THE STUDY: The present study demonstrates that molecular genetic methods keep expanding the list of candidate endodontic pathogens to include Olsenella species.
Necrotic dental root canal infections are polymicrobial infections dominated by anaerobic bacteria. The number of different species in one canal is usually low, approx. 4-7 species. The species isolated most frequently belong to the genera Prevotella, Porphyromonas, Fusobacterium, Peptostreptococcus, Eubacterium and Streptococcus. The frequency of isolation of black-pigmented Gram-negative anaerobes in endodontic infections varies from 25% to > 50%. Pr. intermedia is the most commonly found pigmented species, followed by Pr. denticola and two Porphyromonas species, P. gingivalis and P. endodontalis. Several studies have shown that P. gingivalis and P. endodontalis are closely related to the presence of acute symptoms in endodontic infections, whereas other black-pigmented Gram-negative anaerobes are not. However, several other species may also be involved in acute infections. Moreover, Porphyromonas species have occasionally been isolated from cases with no symptoms. Although Porphyromonas spp. are clearly related to symptoms at the beginning of therapy, they are not important for the prognosis of the treatment.
Citric acid conditioning of dentine has been proposed as an efficacious treatment for promoting re-attachment after periodontal surgery. This study investigated the pulpal reactions to periodontal surgery combined with citric acid (pH 1) demineralization of exposed dentine for 3 min. 9 cats were used as the experimental animals, with each cat providing 1 negative and 1 positive control and 2 experimental canine teeth. Evaluation was over 3 observation periods, with 3 cats in each period being sacrificed at 4, 21 and 83 days. Positive control teeth, which underwent periodontal surgery combining the removal of approximately 3 mm2 of labial bone and vigorous root planning of the exposed root surfaces, displayed mild to moderate pulpal reactions during the short- and intermediate-term periods and only mild to no reactions in the long-term period. The addition of citric acid demineralization, however, resulted in 5 experimental teeth (28%) becoming abscessed or totally necrotic in the intermediate- and long-term periods. At 21 days, the frequency of adverse pulpal responses to surgery and citric acid conditioning was significantly (p less than 0.05) greater than to surgery alone. Although 4 experimental teeth were relatively uninflamed at the long-term period of observation, all showed irritational dentine formation. Penetration of exposed dentinal tubules by plaque bacteria was observed only in the citric acid treated teeth.
A series of cases demonstrating the destruction of periapical periodontal structures, without pulpal involvement has been presented. Treatment using both surgery and antibiotics resulted in extensive healing without any concommitant endodontic therapy. The results suggest that lesions affecting the apical periodontium are either periodontal or pulpal in origin. Careful diagnosis allows the maintenance of pulp vitality in cases where apical destruction has a source other than an infected pulp. The commonly held belief that lateral and accessory canals are a significant source of pulpal contamination from deep periodontal pockets has been questioned.
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The healing capacity of mechanically exposed and bacterially contaminated dental pulps was assessed in monkeys after capping with 2 commercial Ca(OH)2 containing compounds. One hundred eighty teeth in 7 monkeys were employed, 45 as untreated controls and 135 as treated exposures. Class V buccal cavity preparations resulting in pulpal exposure were prepared, left open to the oral cavity for 0, 1, 24 h or 7 days and employed as controls, or debrided, capped, restored with amalgam and left undisturbed for 5 weeks as treated exposures. Zero and 1 h untreated exposures presented damage from the mechanical trauma only, whereas 24 h and 7 day pulp wounds exhibited pronounced infiltrations of polymorphonuclear and mononuclear leukocytes. In addition, the 7 day exposures demonstrated several teeth with partial and total necrosis. Treated 0, 1 and 24 h exposures demonstrated wound healing, minimal pulp tissue inflammation, reorganization of soft tissue and formation of new hard tissue at the exposure site in 86 of 99 teeth. Treated 7 day exposures healed less frequently, showing signs of dentin bridging in 15 of 27 teeth. This study indicated that mechanically exposed and orally contaminated dental pulps in monkeys have a high capacity to resolve inflammation and initiate healing with new dentin formation at the exposure site when treated as described.
The pulpal response to Protemp (ESPE Gmbh) was compared to zinc oxide-eugenol, Super Syntrex (de Trey) and Protemp plus zinc oxide-eugenol liner in 144 vervet monkey (Cercopithecus pygerythus) incisor teeth, according to ADA Specification No. 41. Cellular displacement into dentinal tubules, superficial and deep inflammation were present only in the 3-day specimens in all materials. Mean scores for the test material were lower than both negative and positive controls. In the 30-day specimens the prevalence of reparative dentine under the test material was similar to the negative control, while at 90 days it was less than the negative control, although the severity in affected specimens was more than the positive control. Use of a zinc oxide-eugenol liner reduced the severity of reparative dentine formation.
Four adult Rhesus monkeys provided 120 teeth for buccal Class V cavities. Twenty-nine were non-exposed controls and 91 were exposed for 3 intervals. All 120 teeth were capped with a hard set Ca(OH)2 medicament, restored with amalgam, 57 evaluated after 1 year and 63 after 2 years. Of the 91 exposed pulps, 45 showed complete healing, 25 showed pulpal inflammation varying from acute to chronic, 12 showed severe pulpal breakdown and abscess formation and 9 were necrotic. No difference was observed in the healing response between the 3 exposure times. New hard tissue formed at, or subjacent to, the medicament in 77 of 91 exposed pulps with a tunnel defect frequently present, running from the medicament interface to the pulp. This study demonstrates that recurring pulp inflammation observed after 1 & 2 year direct pulp capping, is associated with bacterial contamination.
The pulpal response to an experimental adhesion promoter (PTP) used in conjunction with a composite resin material, Biogloss, is described, in lined and unlined cavities. Dycal was the lining and negative control material. Standardised Class V cavities were prepared in ferret canine teeth and the pulpal responses studied after 1, 2 and 4 weeks and 6 months post-operatively. Pre-treatment of cavities with PTP causes a reduction in pulpal inflammation at 7 and 14 days compared with Biogloss. There was no obvious difference at the longer time intervals. Pre-treatment with PTP delays the ingress of microorganisms at the cavity wall/material interface but does not prevent it in the long term in this situation. The effect of PTP is not altered if the fluid is allowed to dry naturally rather than by blow drying.
A conventional low copper amalgam was placed in cavities with and without smear layer removal and the amalgam restorations were covered with a resin overlay. The restored teeth were retrieved at 2, 28 and 56 days postoperatively in order to test for leakage and then pulpal reactions. More inflammation was seen in the teeth from which the smear layer had been removed. Abscesses were present in 6 teeth at 28 days but only one was associated with demonstrable bacteria. This study does not support smear layer removal before insertion of low copper amalgam restorations.
The aim of the experiment was to study the immune reaction of cells in both healthy and inflamed human dental pulps. For this it was necessary to identify, in particular, populations of lymphocytes in the pulp, and also other cells involved in the immune reaction. By using monoclonal antibodies which specifically recognize subpopulations of the T lymphocytes, we were able: 1) to directly determine the phenotype of the cells of the pulp, 2) to examine the interactions of the different cell populations. The study was based on 30 permanent human dental pulps and the method was that of indirect immunofluorescence on frozen sections. The monoclonal antibodies used were the antibodies OKT and anti-Leu of types anti-Leu1, anti Leu3a, OKT4, OKT8 and OK1a. The results obtained were evaluated statistically. Two important points stand out from this study: 1) in the healthy samples there is preferential localization of lymphocytes at the periphery of the pulp and 2) there is a predominance of the OKT8+ cytotoxic suppressor subpopulation in both the healthy and the inflamed pulps.
The fine structure of tissue changes during the first 14 days following pulp exposure and capping with a hard setting Ca(OH)2 cement has been studied. The early changes included hemorrhage and moderate inflammation which were largely resolved during the first week. During the second week differentiation of cells occurred at the wound surface. These cells had the characteristic features of odontoblasts and formed a predentin-like collagen matrix. The capping material was closely adapted to cellular structures at the wound surface or to the predentin-like matrix at all observation periods. Dentin fragments displaced into the pulp tissue during cavity preparation, acted as sites for pulp stone formation.
In order to study the osteogenic action of hydroxylapatite (HA) on the dental pulp, a pulp capping experiment was designed using the rat upper molar. Under general anesthesia, molar teeth in 14 male Sprague-Dawley rats were pulp capped with Osteogen (HA) or with Dycal as a control material. After pulp capping, the maxillary molars cavities were restored with amalgam and a pedodontic steel crown was adjusted and sealed over the molar teeth on either side of the maxilla. After 7 days, the areas of necrosis and acute inflammation were more evident in the pulps treated with Dycal than with Osteogen. Hard tissue formation began to appear around dentinal chips in the pulp and extended from the cavity walls into the pulp regardless of the material that was used. Furthermore, this calcified material was scattered throughout the pulp when Osteogen was used, but was not observed in the Dycal treated pulps. The hard tissue formation was thought to be due to the putative fibroblasts and odontoblasts found in the pulp. After 28 days dense dentinal tissue was observed bridging the exposure site when Dycal was used. The dentinal tissues formed with Osteogen was always of a globular type, and showed an irregular distribution. Since Osteogen tends to cause areas of dystrophic calcification in the pulp, its use is not be recommended for pulp capping purposes in humans, because these areas of calcification would make future endodontic treatment difficult.
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