Impact of Streptococcus mutans on the generation of fluorescence from artificially induced enamel and dentin carious lesions in vitro.

The purpose of this study was to examine whether Streptococcus mutans is implicated in the generation of fluorescence detected in carious lesions. Enamel surfaces and dentin cavities of extracted human teeth were subjected to artificial caries generation by exposing them either to a culture medium containing S. mutans or to a lactic acid buffer for 2 weeks. Fluorescence from the lesions was detected with confocal laser scanning microscopy or fluorescence microscopy at various excitation wavelengths, and maximum fluorescence radiance was computed using imageanalyzing software. Culture media of S. mutans were also examined for fluorescence generation. The results demonstrated that S. mutans-induced enamel and dentin lesions exhibited increased fluorescence in the red and green spectral regions, with the signal stronger in the red region. In the blue region, however, fluorescence signals in the corresponding area were below the background level. Significantly weaker or virtually no fluorescence was detected in lactic acid-demineralized lesions at all excitation wavelengths. Neither bacterial cells nor culture media generated any fluorescence. These results indicate that, although the presence of S. mutans may be a prerequisite for the emission of fluorescence from carious lesions, some interaction of S. mutans with exposed tooth matrix elements may also be required for the generation or unmasking of fluorophores.

Odontoblast responses to GaAlAs laser irradiation in rat molars: an experimental study using heat-shock protein-25 immunohistochemistry.

Pulpal responses to gallium-aluminum-arsenide (GaAlAs) laser irradiation applied to the tooth remains to be elucidated. This study aimed to evaluate the effect of the GaAlAs laser on odontoblasts using immunohistochemistry for heat-shock protein (HSP)-25, which labels mature and newly differentiated odontoblasts. The mesial surface of the upper right first molar of 8-wk-old Wistar rats was lased at an output power of 0.5-1.5 W for 180 s. The animals were perfusion-fixed at intervals of 6 h to 30 d after irradiation. At 6 h to 7 d, the intensity of HSP-25-immunoreactivity was found to be disturbed in the coronal odontoblast-layer in an energy-dependent manner. At 30 d, tertiary dentin with/without bone-like tissue was formed abundantly in the dental pulp. Statistical analysis revealed that the area occupied by the new hard tissues was significantly wider in 1.5 W-lased specimens than in 0.5 W-lased specimens. An intense HSP-25 immunoreactivity was seen in the odontoblasts underlying the tertiary dentin, whereas immunoreactivity was weak around the bone-like tissue. It was concluded that the GaAlAs laser may induce the formation of tertiary dentin by influencing the secretory activity of odontoblasts. However, higher energies may cause irreversible changes to the pulp, often leading to the formation of an intrapulpal bone-like tissue.

Cariology in Japanese dental schools.

AIM: The aim of this study was to evaluate the teaching of cariology in Japanese dental schools. DESIGN: Postal questionnaires were sent to all Japanese dental schools. PARTICIPANTS: Twenty-five of the 29 Japanese dental schools (response rate: 86%). RESULTS: The results were in many areas similar to those reported from North America. The non-surgical approach to the management of primary caries predominated, but no schools used bacteriological tests in caries diagnosis and only one school recommended antibacterial treatment. Four of the 25 responding schools advocated the treatment of primary enamel lesions by surgical intervention while three awaited operative intervention until the lesions had reached the middle third of the dentine. Recommendation of fluoride treatment was less common than in North America. The use of electronic devices to diagnose occlusal caries lesions was more common than in North America, but 'explorer catch' was the most frequently used method to diagnose primary and secondary caries in dental schools in both regions. CONCLUSION: Research in cariology over the years has led to altered views on how and when to treat caries lesions. The interpretation of research findings and the transfer of knowledge by teachers in dental schools have an impact on the practice of operative dentistry, but the diversity in the teaching of cariology indicates that the interpretation and implementation of modern principles in the treatment of caries differ quite markedly in Japanese dental schools.

EPMA observation between dentin and resin interfaces. Part 1. Comparison of wet and dry technique after short-term stored in water.

The study was to evaluate the marginal leakage of wet or dry dentin condition after restoration on short-term. In the study we used Photobond adhesive system and Single Bond adhesive system. The specimens were prepared from premolars. A class V cavity was prepared at the CEJ with a high-speed hand piece and #010 round diamond point. The cavity's walls and floors were etched with phosphoric acids of the test materials by wet and dry bonding techniques. The specimens were cross-sectioned longitudinally through the center of the cavities with a low speed diamond micro-cutter and polished with carbide paper (#600-1200) after storage in distilled water for 1 day, and silver nitrate staining. Specimens were viewed with EPMA for elemental distribution of calcium, nitrogen and silver on the resin-dentin interface. The uptake of silver particles was less in samples treated with the wet-bonding technique when compared with dry-bonding technique. This in vitro study showed that bonding technique is important in establishing a seal along the restoration margins to control marginal leakage.

Caries diagnosis using a laser fluorescence system--observation of autofluorescence of dental caries--.

The purpose of this study was to evaluate the usefulness of autofluorescence of carious lesions on caries diagnosis. The observation of the micromorophology of caries lesions was conducted using a confocal laser scanning microscope, a fluorescence microscope and a WDX type electron probe X-ray microanalyzer. Observation of autofluorescence under Cy5 and UV fields showed clearly specific images of autofluorescence in the carious lesion. However, observation of autofluorescence under FITC field showed images of autofluorescence with unclear boundaries in the carious lesion. EPMA images showed decreases in Ca and P in the carious areas. As a result of the observation of autofluorescence and the EPMA images in the carious lesion, a correlation was noted between autofluorescence under the Cy5 field as the laser fluorescence apparatus for caries diagnosis and demineralized areas. The usefulness of autofluorescence of carious lesion on caries diagnosis was suggested.

An immunohistochemical study on hard tissue formation in a subcutaneously transplanted rat molar.

While dental pulp undergoes calcification following tooth replantation or transplantation, we actually know little about these mechanisms. We therefore conducted histological and immunohistochemical evaluations of mineralized tissue that formed in the pulp of rat maxillary molar transplanted into abdominal subcutaneous tissue. One, 2, 3, and 4 weeks post-transplantation, the teeth were investigated immunohistochemically using antibodies to osteocalcin (OCN), osteopontin (OPN), bone sialoprotein (BSP), dentin sialoprotein (DSP), and tissue non-specific alkaline phosphatase (TNAP). In the 1st week after transplantation, cell-rich hard tissue was formed at the root apex. At 2 weeks, formations of hard tissue, with few cells in the root canals and bone-like tissue in the coronal pulp chamber, were noted. After 3 and 4 weeks, the amounts of these hard tissues were increased. The immunolocalization of OCN, OPN, and BSP was seen strongly in coronal and apical hard tissues, but weakly in the root hard tissue. Conversely, DSP localized in the root hard tissue, but not in other newly formed hard tissues. At 1 week, TNAP localized along the periphery of the apical hard tissue and the lower surfaces of root predentin. These results demonstrate that the newly formed hard tissues in the pulp cavity of subcutaneously transplanted molars could be classified into three types, suggesting that these might be formed by type-specific cells.

A study of cavity preparation by Er:YAG laser. Effects on the marginal leakage of composite resin restoration.

The purpose of this study was to evaluate marginal leakage of composite resin restoration from cavities prepared by Er:YAG laser. The observation of the dentin surface after the application of laser irradiation was performed by LSM, the cutting surface showed a rough surface similar to scales, and exposed dentinal tubules were observed without striations or a smeared layer formation that were observed when using a rotary cutting device. Leakage tests revealed no significant differences in the marginal seal for both enamel and dentin between cavities prepared by Er:YAG laser irradiation and when using an air-turbine. In this study, the usefulness of cavity preparation by Er:YAG laser irradiation in composite resin restoration was suggested.

Odontoblast processes in human dentin revealed by fluorescence labeling and transmission electron microscopy.

In the present undertaking, the distribution of odontoblast processes in human dentin was determined through the DiI carbocyanine dye fluorescent staining of the cell membrane, while F-actin was identified by rhodamine-phalloidin. Confocal laser scanning microscopy revealed intense labeling for both agents in inner dentin, while transmission electron microscopy (TEM) identified dentinal tubules including odontoblast processes in this area, each process being surrounded by a cell membrane and containing an abundance of filamentous structures. Electron-dense "lamina limitans" lined the dentinal tubules. Individual cell processes became narrower toward the middle area, and their overall numbers decreased as well under TEM. Labeling for F-actin was absent in both middle and outer dentin, while faint labeling for DiI was visible along the dentinal tubules as far as the dentino-enamel junction (DEJ), where it was also recognized within the tubules themselves. Under TEM, the dentinal tubules lined with electron-dense structures were, in fact, empty in the middle and outer dentin. Immediately below the DEJ, however, the tubules manifested dense concentrations of fine granular material. Our study, therefore, appears to suggest that odontoblast processes do not extend beyond the inner dentin of fully erupted human premolars.

Immunohistochemical study on pulpal response in rat molars after cavity preparation by Er:YAG laser.

While Er:YAG laser systems are in extensive use for caries removal and cavity preparation, the effects of such treatment on pulp tissue remain unclear. This study evaluates these systems using immunohistochemical methods and compares the results with information gained from treatment using conventional burs. Cervical cavities were prepared in the upper first molars of rats, using either an Er:YAG laser or a conventional tungsten-carbide bur. At intervals of 5 min, 6 h, 12 h, 1 d, 3 d and 7 d after cavity preparation, the teeth were processed for immunohistochemical analyses of tissue non-specific alkaline phosphatase, OX6-positive major histocompatibility complex class II antigen-expressing cells and PGP 9.5-immunoreactive nerve fibers. DNA fragmentation was detected by the terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) method. Tissue non-specific alkaline phosphatase was observed mainly in the subodontoblastic layer under the cavity lesion, from 5 min, in both groups. The immunoreactivity was more pronounced in the laser group, but by 7 d no significant differences were recognizable. At 12 h, TUNEL-positive cells were detected around the odontoblastic layer in both groups. From 3 d to 7 d, a limited number of positive cells were still visible in the group that underwent standard treatment. Clear similarities in the distribution patterns of OX6-immunopositive cells and PGP 9.5-immunoreactive nerve fibers were also noted. From 12 h to 1 d, OX6-positive cells accumulated along the pulp-dentin border, extending their processes into the dentinal tubules. Numerous bead-like PGP 9.5-immunoreactive nerve fibers were observed under the odontoblastic layer at 7 d. These results demonstrated that there was no appreciable difference in the manner in which pulp tissue responded to treatment with either Er:YAG laser or a conventional drill. This would seem to indicate the usefulness of the Er:YAG laser system in the removal of caries and cavity preparation.

A study of cavity preparation by Er:YAG laser--observation of hard tooth structures by laser scanning microscope and examination of the time necessary to remove caries.

The purpose of this study was to observe and measure the morphological changes that occur in the hard tissue after the application of Er:YAG laser. Another objective was to evaluate and compare the duration of application of both the laser apparatus and a conventional cutting device. In this study, sound and newly extracted carious tissues were used. The morphological changes in hard tooth structures produced by Er:YAG laser irradiation were examined by using a laser scanning microscope. Results showed that appropriate laser irradiation was 100 mJ/pulse for dentin, and 200 mJ/pulse for enamel. Also, the laser scanning microscope images were less damaged than the SEM images due to pretreatment of the specimens. The time taken to remove carious enamel by laser irradiation was slightly longer than the compared rotary cutting device; however, no differences between the two methods were observed in case of carious dentin removal.

A comparative study of fluoride-releasing adhesive resin materials.

One of the most important and exciting properties of recently introduced dental restorative materials is their ability to release fluoride ions, as this has several advantageous effects on tooth structures. They have been extensively used as fluoride-releasing filling and luting materials. Recently, fluoride-releasing adhesive resins and fluoride-releasing adhesive resin cement have been developed and introduced for clinical use. The purpose of this study was to evaluate the fluoride release from these adhesive resins and the fluoride uptake by both enamel and dentin, as well as the acid-resistance of these tooth structures. Based on our results, we conclude that fluoride-releasing adhesive resins and luting cements are useful for the prevention of initial or secondary caries, especially along the margins of restorations.

Effect of fluoride mouth rinse on fluoride releasing and recharging from aesthetic dental materials.

This study evaluated fluoride-release and recharging with the fluoride mouth rinsing technique on fluoridated materials. Three fluoride containing materials and one non-fluoride containing composite resin were used for this study. Samples for each material consisted of 15 discs, 9 mm diameter with a thickness of 1 mm. Initial fluoride release was assessed over a 60-day period. After that, 15 discs for each material were divided into 3 groups: distilled water group, 450-ppm and 900-ppm mouth rinsing groups. Fluoride release increased in combination with fluoride mouth rinse, and fluoride was higher in the 900-ppm group than the 450-ppm group. Moreover, S-PRG or F-PRG fillers materials released fluoride in higher than fluoroaluminosilicate glass fillers materials. In addition fluoride release from control samples was not observed. Therefore, only fluoride release material takes up fluoride. The findings of the present investigation suggest that the rate of fluoride release was different for each material, because they contained different function fillers. The results showed the importance of the fluoride mouth rinsing technique for fluoride-releasing restorative materials for the prevention of secondary caries.