External Dentin Stimulation Induces ATP Release in Human Teeth.

ATP is involved in neurosensory processing, including nociceptive transduction. Thus, ATP signaling may participate in dentin hypersensitivity and dental pain. In this study, we investigated whether pannexins, which can form mechanosensitive ATP-permeable channels, are present in human dental pulp. We also assessed the existence and functional activity of ecto-ATPase for extracellular ATP degradation. We further tested if ATP is released from dental pulp upon dentin mechanical or thermal stimulation that induces dentin hypersensitivity and dental pain and if pannexin or pannexin/gap junction channel blockers reduce stimulation-dependent ATP release. Using immunofluorescence staining, we demonstrated immunoreactivity of pannexin 1 and 2 in odontoblasts and their processes extending into the dentin tubules. Using enzymatic histochemistry staining, we also demonstrated functional ecto-ATPase activity within the odontoblast layer, subodontoblast layer, dental pulp nerve bundles, and blood vessels. Using an ATP bioluminescence assay, we found that mechanical or cold stimulation to the exposed dentin induced ATP release in an in vitro human tooth perfusion model. We further demonstrated that blocking pannexin/gap junction channels with probenecid or carbenoxolone significantly reduced external dentin stimulation-induced ATP release. Our results provide evidence for the existence of functional machinery required for ATP release and degradation in human dental pulp and that pannexin channels are involved in external dentin stimulation-induced ATP release. These findings support a plausible role for ATP signaling in dentin hypersensitivity and dental pain.

Expression of ecto-ATPase NTPDase2 in human dental pulp.

Dental pulpal nerve fibers express ionotropic adenosine triphosphate (ATP) receptors, suggesting that ATP signaling participates in the process of dental nociception. In this study, we investigated if the principal enzymes responsible for extracellular ATP hydrolysis, namely, nucleoside triphosphate diphosphohydrolases (NTPDases), are present in human dental pulp. Immunohistochemical and immunofluorescence experiments showed that NTPDase2 was predominantly expressed in pulpal nerve bundles, Raschkow's nerve plexus, and in the odontoblast layer. NTPDase2 was expressed in pulpal Schwann cells, with processes accompanying the nerve fibers and projecting into the odontoblast layer. Odontoblasts expressed the gap junction protein, connexin43, which can form transmembrane hemichannels for ATP release. NTPDase2 was localized close to connexin43 within the odontoblast layer. These findings provide evidence for the existence of an apparatus for ATP release and degradation in human dental pulp, consistent with the involvement of ATP signaling in the process of dentin sensitivity and dental pain.

Effect of CO2 laser on pulpal temperature and surface morphology: an in vitro study.

OBJECTIVES: The objective of this study was to evaluate the potential effects on underlying dental hard tissues of a high pulse rate carbon dioxide (CO2) laser that was designed for soft tissue surgery. METHODS: Eighteen extracted human teeth were sectioned longitudinally, cleaned, and varnished, leaving nine exposed windows on each: six on the coronal surface (enamel) and three on the root surface (cementum, dentin). The CO2 irradiation conditions used were: wave length 10.6 microm; 1.2-2.6J/cm(2) fluence per pulse; repetition rate 120-1000Hz; 100-200ms pulse duration; and cumulative fluences ranging from 14 to 2200J/cm(2). Each window was irradiated with a 0.3mm beam diameter at one of nine power settings for 0.1, 0.5, or 1.0s. The pulp chamber temperature was measured with a microthermocouple. The irradiated teeth were evaluated by Polarized Light Microscopy (PLM) and Scanning Electron Microscopy (SEM). RESULTS: The pulp chamber temperature rise ranged from 0.5 to 19 degrees C depending on the location of the window and distance to pulp chamber. SEM revealed crystal fusion in both enamel and dentin at all cumulative fluences. At cumulative fluences of 40J/cm(2), 200 pulses/second and higher, measurable tissue loss was observed with PLM both in dentin and enamel. CONCLUSIONS: These results indicate there are threshold conditions above which pulsed CO2 laser light used for soft tissue surgery may cause detrimental changes to underlying oral hard tissue and to the pulp.

Effect of smokeless tobacco use in humans on mucosal immune factors.

To assess the effects of smokeless tobacco on the secretory immune system and dental caries, we examined users of smokeless tobacco and non-tobacco users. There were no significant differences in the prevalence of DMFS between users and non-users. There was significantly more salivary IgA, IgA2 and J-chain in users. Levels of salivary lysozyme and lactoferrin were significantly lower in users than controls. Because there was no difference in levels of secretory component in relation to the increased IgA levels of smokeless tobacco users, this suggests an effect of smokeless tobacco on secretory epithelial cells responsible for synthesis of secretory component, lysozyme and lactoferrin, and for the packaging of secretory component on IgA. There were only slight differences in salivary or serum antibody levels to Streptococcus mutans. These findings indicate that although smokeless tobacco has a significant influence on the synthesis of secretory IgA, the numbers of DMFS were similar between smokeless tobacco users and controls.

Function of anti-Streptococcus mutans antibodies: inhibition of virulence factors and enzyme neutralization.

The levels of parotid salivary IgA and serum IgG antibodies from dental caries-resistant (CR) and caries-susceptible (CS) individuals to Streptococcus mutans antigens were determined. In general, the levels of salivary IgA and serum IgG antibodies to S. mutans antigens were significantly higher in CR subjects than in CS individuals. There were significantly higher levels of IgA2, but not IgA1, salivary antibodies to S. mutans whole cells in CR subjects than in CS individuals. These results led us to investigate the functional effects parotid saliva and sera containing these antibodies had on several factors associated with S. mutans virulence. Parotid saliva and sera from CR subjects significantly inhibited S. mutans growth, adherence, acid production, glucosyltransferase and glucose-phosphotransferase activities to a greater extent than saliva and sera from CS individuals. The data suggest that neutralization of S. mutans enzymes and inhibition of S. mutans virulence factors by saliva and serum may be responsible for the lower numbers of carious lesions in CR subjects.

Osseo-integrated implant treatment of a patient with rapidly progressive periodontitis. A case report.

The case report presented describes placement of osseointegrated fixtures in a patient with a probable history of rapidly progressive periodontitis. A 12-year history is presented. The post-operative sequelae of placing fixtures in a fully edentulous maxilla and partially edentulous mandible were the worst seen by the Implant Team at Emory University. The hypothesis presented is that patients with aggressive forms of periodontal disease should be subjected to appropriate mechanical and antimicrobial therapy to produce a healthy oral flora before any implants are placed.

Immunodominant antigens of Streptococcus mutans in dental caries-resistant subjects.

Previously, we reported that dental caries-resistant subjects, who have significantly fewer Streptococcus mutans in whole saliva than caries-susceptible patients, have significantly higher levels of naturally occurring binding and neutralizing parotid salivary immunoglobulin A and serum immunoglobulin G antibodies to native S. mutans antigens than caries-susceptible patients. Recent animal studies indicated that the immunogenicity of swallowed S. mutans may be altered by either saliva-coating or stomach acid-denaturation. These results suggest a difference not only in the quantity of antibody to S. mutans, but also in the antigenic epitopes that caries-resistant subjects synthesize antibody to as compared with caries-susceptible patients. In the present report, sodium dodecyl sulfate-polyacrylamide gel electrophoresis/immunoblotting studies indicate that caries-resistant subjects produce salivary immunoglobulin A and serum immunoglobulin G antibodies to several different (molecular weight: 94, 80, 40, and 35 kilodaltons) as well as several similar (molecular weight: 67, 55, and 30 kilodaltons) S. mutans epitopes as compared to caries-susceptible patients. This provides additional confirmation for our previous binding and functional antibody studies, indicating that caries-resistant subjects synthesize antibodies of different specificities than caries-susceptible patients. This study supports the concept of immune regulation of dental caries by naturally occurring antibodies induced by swallowing S. mutans antigens in saliva.

Effects of smokeless tobacco on the ability of secretory component to bind to the IgA/J chain complex.

Previously, we reported that smokeless tobacco users have significantly higher levels of immunoglobulin A and J chain in whole saliva than non-tobacco users. Because there was no difference in levels of secretory component between the two groups, the proportion of secretory component/immunoglobulin A was significantly lower in users than non-users. There was no significant difference in antibody function. In the present study, we examined immunoglobulin A from whole saliva of users and non-users to determine the effect of smokeless tobacco on the ability of secretory component to bind to immunoglobulin A containing J chain. Whole saliva was passed over an affinity chromatography filter unit coupled with anti-alpha heavy chain-specific antibody followed by passage over a molecular sieve high-performance liquid chromatography column. Peaks were collected and examined for immunoglobulin A, J chain and secretory component by enzyme-linked immunosorbent assay. Saliva from users had three significantly larger peaks (3-4 fold) at 280 nm than non-users, confirming the presence of a higher concentration of immunoglobulin A in users. The secretory component/J chain and secretory component/immunoglobulin A ratios for the largest peak were significantly less in users. This indicates that smokeless tobacco has an effect on the ability of secretory component to bind to immunoglobulin A without a loss in antibody function. This may occur either prior to immunoglobulin A/J chain binding to secretory component receptors on secretory epithelial cells or internal to the epithelial cells. These studies provide further evidence for the role of secretory component in mucosal immunity.

Dental management of patients with hereditary angioedema: report of case.

Dental management and treatment of a patient with hereditary angioedema can be accomplished safely and satisfactorily in an outpatient setting by a dentist in collaboration with a physician. However, as acute attacks can occur as a complication of treatment, we recommend that dental treatment be provided in a hospital environment with adequate medical emergency resources.