Quantitative detection of periodontal pathogens using real‐time polymerase chain reaction with TaqMan probes

Quantitative analysis, with identification of periodontopathic bacteria, is important for the diagnosis, therapeutic evaluation and risk assessment of periodontal disease. We developed a highly sensitive and specific method using real‐time polymerase chain reaction (PCR) to detect and quantify six periodontal bacteria: Porphyromonas gingivalis, Tannerella forsythia, Actinobacillus actinomycetemcomitans, Treponema denticola, Prevotella intermedia, and Prevotella nigrescens. Species‐specific TaqMan probe/primer sets were designed according to 16S ribosomal RNA gene sequences. Plaque and tongue debris specimens were collected from 10 patients with advanced periodontitis and 10 periodontal healthy individuals and analyzed. All species, except for P. nigrescens, were detected in samples from diseased sites in significantly greater numbers than in those from healthy sites, whereas greater numbers of P. nigrescens were found in the controls. These results suggest that the present real‐time PCR method with the designed probe/primer sets enabled sensitive detection of the six periodontal bacteria, and may also assist future microbial studies of periodontal diseases.     

Quantitative detection of periodontal pathogens using real-time polymerase chain reaction with TaqMan probes

Quantitative analysis, with identification of periodontopathic bacteria, is important for the diagnosis, therapeutic evaluation and risk assessment of periodontal disease. We developed a highly sensitive and specific method using real-time polymerase chain reaction (PCR) to detect and quantify six periodontal bacteria: Porphyromonas gingivalis, Tannerella forsythia, Actinobacillus actinomycetemcomitans, Treponema denticola, Prevotella intermedia, and Prevotella nigrescens. Species-specific TaqMan probe/primer sets were designed according to 16S ribosomal RNA gene sequences. Plaque and tongue debris specimens were collected from 10 patients with advanced periodontitis and 10 periodontal healthy individuals and analyzed. All species, except for P. nigrescens, were detected in samples from diseased sites in significantly greater numbers than in those from healthy sites, whereas greater numbers of P. nigrescens were found in the controls. These results suggest that the present real-time PCR method with the designed probe/primer sets enabled sensitive detection of the six periodontal bacteria, and may also assist future microbial studies of periodontal diseases.     

Impact of Different Elastomer Formulations on Moisture Permeation through Stoppers Used for Lyophilized Products Stored under Humid Conditions

The purpose of this study was to evaluate the effect of moisture permeability of different elastomer formulation stoppers, which had different moisture absorption abilities, on the increase of moisture content inside lyophilized vials during long-term storage under humid conditions. Two different elastomer formulation stoppers (high-moisture and low-moisture uptake stoppers) were compared. The increased amount of moisture content inside lyophilized vials fitted with high-moisture stoppers was higher than those fitted with low-moisture stoppers during the early stage of storage. However, this trend was reversed during the later stage of storage. Our data show that the moisture increase inside the lyophilized vials at the early stage was caused by moisture transfer from the stoppers, whereas the later moisture increase was caused by external moisture permeation through the stoppers. Results indicate that the difference in the moisture uptake profile inside the lyophilized vials at each period of storage was caused by the moisture absorption ability and moisture permeation ability of the two elastomer formulation stoppers. In terms of long-term storage stability under humid conditions, our data indicate that external moisture permeating through the stopper into the lyophilized vial during the late stage was the more important factor. In addition, the increase in moisture content at the early stage was controlled by stopper drying time. Furthermore, stopper drying time did not have an effect on moisture permeation at the late stage. Moisture permeation during the storage period appears to be dependent on the different elastomer formulations of the stoppers. The moisture permeation of different elastomer stoppers was an important factor in terms of the increased moisture content inside the lyophilized vials during the late stage of long-term storage under humid conditions. For lyophilized products stored at room temperature, the moisture permeation ability of the stopper is one of the most important factors for long-term storage stability.     

Development of Web-based intervention system for periodontal health: a pilot study in the workplace

BACKGROUND: It has recently been accepted that periodontal disease is a risk factor for not only tooth loss but also systemic diseases. An effective system of public intervention for periodontal health to enable continuous intervention of dental professionals has been sought. We developed a Web-based intervention system regarding periodontal health and evaluated the effects of the system in the workplace. METHODS: The system was capable of storage and display of personalized oral health records including video images pertaining to toothbrush manipulation within their own oral cavities based on instructions by dental professionals. The system enabled clients to view movement of their own skill, and repeatedly. Thirteen workers of a company were randomized to either an experimental or control group. The control group received face-to-face toothbrushing instruction at the company and follow-up via telephone. The experimental group received follow-up through our system in addition to those components employed in the control group. MAIN RESULTS: The workers in the experimental group benefited in terms of improvements of not only plaque removal but also periodontal health over 3 months. CONCLUSIONS: These results indicate that implementation of an Web-based approach for periodontal health affords the possibility of remote instruction and produces additional public benefit.     

Characterization of binding of Streptococcus oralis glyceraldehyde-3-phosphate dehydrogenase to Porphyromonas gingivalis major fimbriae

Binding of Streptococcus oralis glyceraldehyde-3-phosphate dehydrogenase (GAPDH) to Porphyromonas gingivalis fimbriae was characterized via a biomolecular interaction analysis system. The interaction was specific, and the association constant value was 4.34 x 10(7) M(-1), suggesting that S. oralis GAPDH functions as a dominant receptor for P. gingivalis and contributes to P. gingivalis colonization.     

P29 The effects of essential oils on periodontopathic bacteria and oral halitosis

Oral halitosis has been associated with periodontal disease. Hydrogen sulphide, methyl mercaptan and dimethyl sulphide are the major components of halitosis originating in the oral cavity. These compounds are mainly produced by periodontopathic bacteria in the oral cavity. In the absence of the bacteria, the odoriferous compounds are not generated. We have demonstrated the antibacterial effects of essential oils on the periodontopathic bacteria such as Porphyromonas gingivalis, Actinobacillus actinomycetemcomitans and Fusobacterium nucleatum. As essential oils, manuka oil, tea tree oil, eucalyptus oil, lavandula oil, and romarinus oil were used, and their minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were determined. The essential oils inhibited the growth of the bacteria tested, and manuka oil did so most effectively. MBC values showed that lavandula oil acted bacteriostatically, and the remaining oils bactericidally. Bacterial strains tested were killed completely by exposure for 30 s to 0.2% manuka oil, tea tree oil or eucalyptus oil. This study showed that among the essential oils tested, tea tree oil and manuka oil, particularly the latter, had strong antibacterial activity against periodontal disease associated bacteria. In the present study, we investigated the clinical effects of essential oils in the treatment of oral halitosis. When the degree of halitosis before and after the treatment was evaluated, the essential oils demonstrated efficacy in reducing the level of the three major components of halitosis. From the data of the present study, we consider that essential oils can be used in oral health management. This study was supported by grant from Kanpou Science Foundation.