R. Nissan

The Comparative Sealing Ability of Hydroxyapatite Cement, Mineral Trioxide Aggregate, and Super Ethoxybenzoic Acid as Root-End Filling Materials

A comparison was made of the ability of hydroxyapatite cement, mineral trioxide aggregate, and super ethoxybenzoic acid to prevent the leakage of bacteria from root canals, when used as root-end filling materials. The materials were tested in a double-chamber device in which a root segment connects the upper (delivery) chamber and the lower (receiving) chamber. The root segment was prepared by having the root canal instrumented to a #45 file, and a 3-mm-deep, root-end preparation placed at the apical foramen. The canal of each root segment was filled with gutta-percha, and the root-end preparation was filled with one of three test materials, mixed according to the manufacturers directions. Negative controls were constructed with sticky wax sealing the apical foramen. A titered suspension of radioactively (3H-thymidine)-labeled bacteria (Enterococcus fecalis) was placed into the delivery chamber, and sterile saline was placed into the receiving chamber such that the apical third of each root section was immersed. At various time points, samples were taken from the receiving chamber, and measured for 3H activity. The results indicated that (a) all the test materials leaked significantly compared with the negative controls; and (b) there was no significant difference found between the leakage rates of the three materials tested.

Ability of bacterial endotoxin to diffuse through human dentin

An in vitro system was developed to determine whether bacterial endotoxin is capable of diffusing through dentin without the use of filtration pressure. Cavities were prepared in five third molar teeth in order to produce a split chamber device consisting of occlusal and pulpal chambers with 0.5 mm of intervening dentin. Endotoxin was introduced into the occlusal chamber and the effluent in the pulpal chamber was sampled every 30 min for 5 h and at 24 h using the limulus lysate assay. In four specimens the initial appearance of endotoxin in the effluent ranged from 15 min to 4 1/2 h. In two specimens the concentration of endotoxin in the effluent leveled off in 4 1/2 and 5 h, respectively, whereas in another two the concentration continued to increase throughout the experiment. In one specimen no endotoxin was detected. The results indicate that endotoxin is capable of passing through 0.5 mm of dentin.

Dual wavelength spectrophotometry as a diagnostic test of the pulp chamber contents

The purpose of this in vitro study was to determine the feasibility of using dual wavelength spectrophotometry to identify teeth with pulp chambers that are either empty, filled with fixed pulp tissue, or filled with oxygenated blood. In phase I of the experiment, a human third molar was prepared so that its pulp space could be filled with oxygenated blood and later emptied. In phase II, the lower jaw of a beagle dog was removed and placed in formalin, thereby fixing the pulps of the teeth. The pulp of the right canine was removed via an apical approach, and attachments were placed in a similar position to those on the human tooth, to allow filling and emptying of the pulp space. Cavit was placed over the exposed fixed pulp in the left canine. Ten readings, which were separated by light source and detector removal and replacement, were taken of the right canine pulp space when it was empty or filled with oxygenated blood, or the left canine pulp space when it was filled with fixed tissue. Distinct and reproducible changes were measured for pulp spaces filled with air, tissue, or oxygenated blood. In phase III, simulated pulp testing on a dog tooth model was performed. Blood was introduced into the root canal space, the chamber was rinsed with water and replaced with air, according to a predetermined code. Spectrophotometer readings were recorded. The identification of pulpal contents was correctly determined in all 20 of the predetermined conditions. The findings indicate that continuous wave spectrophotometry may become a useful pulp testing method.

Antibacterial effect of genetically-engineered bacteriophage ϕEf11/ϕFL1C(Δ36)PnisA on dentin infected with antibiotic-resistant Enterococcus faecalis

OBJECTIVE Enterococcus faecalis is a gram-positive facultative anaerobic bacterium, which is present in 30-89% of teeth with postendodontic treatment failures. E. faecalis is capable of penetrating dentinal tubules and surviving as a monoculture after conventional endodontic therapy, indicating that it is resistant to commonly used endodontic disinfection protocols. Different E. faecalis strains have shown resistance to several antibiotics, and have been associated with both dental pathology and systemic infections. The aim of this study was to evaluate the efficacy of a genetically engineered bacteriophage to disinfect dentin infected with antibiotic resistant strains of E. faecalis. METHODS Extracted human dentin root segments were cemented into sealable two-chamber devices, fabricated from syringe needle caps to form in vitro infected-dentin models. The models were inoculated with an overnight suspension of either E. faecalis V583 (vancomycin resistant strain) or E. faecalis JH2-2 (fusidic acid and rifampin resistant, vancomycin sensitive strain). After 7days of incubation at 37°C, a suspension of a genetically engineered phage, ϕEf11/ϕFL1C(Δ36)PnisA, was added to the root canal of each infected dentin segment, and the incubation was continued for an additional 72-h. Dentin was harvested from the walls of each root canal and assayed for the residual titer of E. faecalis cells. RESULTS The recovered E. faecalis titer was reduced by 18% for the JH2-2 infected models, and by 99% for the V583 infected models. CONCLUSION Treatment: of E. faecalis-infected dentin with bacteriophage ϕEf11/ϕFL1C(Δ36)PnisA consistently resulted in a decrease in the residual bacterial population of both vancomycin-sensitive and resistant strains.

Whole Genomic DNA Probe for Detection of Porphyromonas endodontalis

The purpose of the present study was to develop a DNA probe for Porphyromonas endodontalis. Pure cultures of P. endodontalis were grown in TYP medium, in an anaerobic chamber. DNA was extracted from the P. endodontalis and labeled using the Genius System by Boehringer Mannheim. The labeled P. endodontalis DNA was used in dot-blot hybridization reactions with homologous (P. endodontalis) and unrelated bacterial samples. To determine specificity, strains of 40 other oral bacterial species (e.g. Porphyromonas gingivalis, Porphyromonas asaccharolytica, and Prevotella intermedia) were spotted and reacted with the P. endodontalis DNA probe. None of the panel of 40 oral bacteria hybridized with the P. endodontalis probe, whereas the blot of the homologous organism showed a strong positive reaction. To determine the sensitivity of the probe, dilutions of a P. endodontalis suspension of known concentration were blotted onto a nylon membrane and reacted with the probe. The results of our investigation indicate that the DNA probe that we have prepared specifically detects only P. endodontalis and can detect at least 3 x 10(4) cells.