Influence of artificial saliva contamination on adhesion in class V restorations.

This study aimed to evaluate the effects of artificial saliva contamination on a glass ionomer cement (GIC), a resin-modified GIC (RMGIC), and a composite resin (CR) that was used with two different etching adhesive systems. Three surface conditions were created on bovine teeth using artificial saliva: control (group I), mild saliva contamination (group II), and severe saliva contamination (group III). The microtensile bond strength (µTBS) of CR with dentin was significantly lower in group III than in group I. However, the µTBS of GIC and RMGIC with both enamel and dentin showed no significant intergroup differences. Moreover, CR exhibited significantly greater microleakage on cementum in group III than in group I, whereas both GIC and RMGIC showed no significant differences for both enamel and cementum. Thus, GIC and RMGIC may be suitable for preventing secondary caries after class V restorative treatments when contamination by saliva cannot be avoided.

Gender differences in adolescents' perceptions toward dentists using the Japanese version of the dental beliefs survey: a cross-sectional survey.

BACKGROUND: While adult women show greater dental anxiety than adult men, few studies have examined gender differences in adolescent perceptions of dentists. Therefore, this cross-sectional study aimed to evaluate the gender differences in adolescents' perceptions toward dentists by using the Japanese version of the Dental Beliefs Survey (DBS) and the factor structure of the DBS. METHODS: We conducted surveys at schools, and 957 Japanese adolescents (403 girls and 554 boys, aged between 13 and 15 years) participated in this study. To assess their confidence in dentists, participants were asked to complete the self-reported, 15-item Japanese version of the DBS. We performed a Welch's t-test and a one-way analysis of variance to assess differences in DBS scores by gender and age. Factor analysis (principal components, varimax rotation) was used to assess the scale's factor structure. RESULTS: A significant gender difference was observed in the DBS scores (P = 0.018), suggesting that boys exhibit greater negative perceptions toward the behavior of dentists than girls. However, there was no significant difference found among ages. The factor analysis yielded two results: Factor 1, "trust" (seven items); and Factor 2, "lack of control" (five items). Notably, the factor structure differed according to gender. As such, by including only factors with eigenvalues above 1.0, the DBS for girls comprised "trust" (seven items) and "communication" (three items), while that for boys comprised "lack of control" (six items) and "belittlement" (six items). CONCLUSIONS: This study identified two factors of differing strengths pertaining to the confidence of Japanese adolescents in dentists. Gender differences in perceptions toward dentists were observed. Accounting for these differences may improve the effectiveness of strategies to lower dental anxiety and foster positive dental beliefs in young patients.

Reliability, validity, and sex differences in a quantitative gag reflex measurement method.

A reliable assessment method is required to manage the gag reflex. We tested the inter- and intra-examiner reliability and validity of a quantitative measurement method and evaluated the differences between sexes. This study included 21 healthy adults (10 women, 11 men; mean age, 27.1 ± 9.9 years). An examiner inserted a standard saliva ejector slowly down the participant's throat to determine the maximum tolerance of the gag reflex; the insertion depth was used as an index of gag reflex. The reflex was measured by one examiner during two sessions (S1 and S3) and by a second examiner during one session (S2) under the same conditions. The galvanic skin response (GSR) and heart rate (HR) were recorded during each session to test the method validity. Data were analysed using the intraclass correlation coefficient (ICC), paired t test, and two-way repeated-measures analysis of variance. The ICC of the gag reflex measurements was 0.91 between S1 and S2, and 0.93 between S1 and S3. The GSR and HR increased significantly after the maximum tolerance of the reflex in S1, S2, and S3 (GSR: P < 0.001, P = 0.004, P = 0.001, respectively; HR: all, P < 0.001). Differences in the measurements according to sex and session were not significant. Our method exhibited excellent inter- and intra-examiner reliability and was unaffected by the participants' sex. Measurement revealed significant autonomic reactivity, which is common for this reflex. Consequently, this method may be used in clinical and laboratory-based applications.

Contribution of Streptococcus gordonii Hsa Adhesin to Biofilm Formation.

Adhesion of oral mitis group streptococci, such as Streptococcus gordonii, to acquired pellicle on the tooth surface is the first step in oral biofilm formation. S. gordonii strain DL1 possesses an Hsa adhesin, which recognizes the terminal sialic acid of host sialoglycoconjugates. The aim of the present study was to investigate the role of the Hsa adhesin in biofilm formation. The biofilm-forming ability of a S. gordonii hsa mutant on microtiter plates pre-coated with saliva, fetuin, or mucin was significantly lower than that of wild-type strain DL1. In contrast, no significant difference in biofilm-forming ability was observed in plates pre-coated with bovine serum albumin, which does not contain sialic acid. The biofilm-forming ability of strain DL1 in saliva-coated microtiter plates was also significantly reduced when the plate was pre-treated with neuraminidase. The sialic acid-dependent biofilm-forming ability of different wild-type S. gordonii strains varied. However, Southern and western blot analyses showed that all the tested wild-type strains possessed and expressed hsa homologs, respectively. These results indicate that the binding of Hsa adhesin to sialoglycoconjugates is associated with biofilm formation of S. gordonii DL1, and imply variation in the contribution of Hsa and its homologs to S. gordonii biofilm formation.

Effects of surface reaction-type pre-reacted glass ionomer on oral biofilm formation of Streptococcus gordonii.

Streptococcus gordonii, a bacterium involved in the initial colonization of tooth surfaces, contributes to dental biofilm formation and is an important cause of infective endocarditis. This study aimed to investigate the influence of surface reaction-type pre-reacted glass ionomer (S-PRG) filler on oral bacterial growth and aggregation of S. gordonii. The effect of various concentrations of S-PRG eluate on the growth and the biofilm formation of S. gordonii and other oral microorganisms (Streptococcus mutans, Streptococcus oralis, Lactobacillus acidophilus, and Candida albicans) was assessed. In addition, the effect of S-PRG eluate on coaggregation of S. gordonii with both S. oralis and Fusobacterium nucleatum was assessed. The effect of S-PRG eluate treatment on autoaggregation of S. gordonii was also evaluated. Our results indicate that S-PRG eluate treatment reduced both for the growth and for biofilm of all organisms in a dose-dependent manner. Coaggregation of S. gordonii with both S. oralis and F. nucleatum was inhibited by S-PRG eluate, whereas autoaggregation of S. gordonii increased at certain concentrations of S-PRG eluate. These results indicate that the S-PRG filler possesses antimicrobial activity that is mediated by inhibiting growth and biofilm of oral microorganisms, and by suppressing coaggregation of S. gordonii. In addition, these findings indicate that coaggregation of S. gordonii with other bacteria is inhibited by increased autoaggregation of S. gordonii.

Prevalence and association of self-reported anxiety, pain, and oral parafunctional habits with temporomandibular disorders in Japanese children and adolescents: a cross-sectional survey.

BACKGROUND: Associations between temporomandibular disorder (TMD) and psychological variables, pain conditions, and daily activities have been reported more commonly in middle-aged individuals than in children. However, to determine factor-specific preventive programs for TMD, it is important to evaluate the associations between multiple factors and TMD symptoms during childhood. The aim of this study was to assess the relationship between TMD symptoms and other orofacial pain conditions, daily activities, and trait anxiety in a population-based cross-sectional survey of Japanese children and adolescents. METHODS: A total of 1,415 subjects (11-15 years old) self-reported their TMD symptoms, headache, neck pain, and toothache, and completed questionnaire scales that assessed 15 daily activities. Trait anxiety was assessed using the State Trait Anxiety Inventory for Children-Trait (STAIC-T) scale. Subjects were dichotomized into a TMD group or control group, based on whether they reported at least 1 TMD symptom: the TMD group (≥1 TMD symptom, n = 182) and the control group (no TMD symptoms, n = 1,233). Data were analyzed using the chi-square test and multivariate logistic regression analysis. RESULTS: The prevalence rates for headache and neck pain were significantly higher in the TMD group than in the control group (44.0% vs. 24.7% and 54.4% vs. 30.0%, respectively; both P < 0.001). The odds ratios for TMD symptoms in subjects with neck pain and frequent diurnal clenching were 2.08 (P < 0.001) and 3.69 (P = 0.011), respectively. Moreover, high STAIC-T scores were weakly associated with TMD symptoms. CONCLUSIONS: In this young Japanese population, TMD symptoms were associated with other orofacial pain conditions, particularly neck pain, although they were only weakly associated with trait anxiety. Diurnal clenching was strongly associated with TMD symptoms. Health professionals should carefully consider these factors when developing appropriate management strategies for TMD in children and adolescents.

Comparison of self-reported pain intensity, sleeping difficulty, and treatment outcomes of patients with myofascial temporomandibular disorders by age group: a prospective outcome study.

BACKGROUND: Subjective symptoms of temporomandibular disorders (TMDs) have rarely been studied by age group. We aimed to compare self-reported pain intensity, sleeping difficulty, and treatment outcomes of patients with myofascial TMDs among three age groups. METHODS: The study population included 179 consecutive patients (151 women and 28 men) who underwent comprehensive clinical examinations at a university-based orofacial pain center. They were classified into myofascial pain subgroups based on the Research Diagnostic Criteria for Temporomandibular Disorders. They were stratified by age group: M1, under 20 years; M2, 20-39 years; and M3, 40 years and older. The patients scored their pretreatment symptoms (first visit) and post-treatment symptoms (last visit) on a form composed of three items that assessed pain intensity and one item that assessed sleeping difficulty. Their treatment options (i.e., pharmacotherapy, physical therapy, and orthopedic appliances) and duration were recorded. All variables were compared between sexes in each group and between the age groups by using the Kruskal-Wallis test, the Mann-Whitney U test, the chi-square test, and analysis of variance (p < 0.05). RESULTS: No significant sex differences were found in any age group. Only sleeping difficulty was significantly different before treatment (p = 0.009). No significant differences were observed in the treatment options or treatment duration. After treatment, the intensity of jaw/face pain and headache and sleeping difficulty was significantly reduced in groups M2 and M3, but only the intensity of jaw/face pain was significantly decreased in group M1. The changes in the scores of pain intensity and sleeping difficulty were not different between the groups. CONCLUSIONS: Pain intensity does not differ by age group, but older patients with myofascial TMDs had greater sleeping difficulties. However, there were no differences between the age groups in the treatment outcomes. Clinicians should carefully consider the age-related characteristics of patients with myofascial TMDs when developing appropriate management strategies.

Effect of artificial saliva contamination on adhesion of dental restorative materials.

The purpose of this study was to evaluate the effects of artificial saliva contamination on three restorative materials, namely, a glass ionomer cement (GIC), a resin-modified GIC (RMGIC), and a composite resin (CR), for which two different etching adhesive systems were used. Thus, three surface conditions were created on bovine teeth using artificial saliva: control, mild saliva contamination, and severe saliva contamination. The dentin bond strength for CR was significantly lower after artificial saliva contamination. There were, however, no significant differences among the three surface conditions in terms of the dentin and enamel bond strengths of GIC and RMGIC. Moreover, CR exhibited significantly greater microleakage after artificial saliva contamination, whereas no significant differences were found in GIC and RMGIC. The results showed that artificial saliva contamination did not affect the shear bond strengths of GIC and RMGIC or their degrees of microleakage.

Evaluation of the caries-preventive effect of three orthodontic band cements in terms of fluoride release, retentiveness, and microleakage.

This in vitro study was undertaken to evaluate the caries-preventive effect of three orthodontic band cements (a dual-curing resinmodified glass ionomer cement [RMGIC] and two light-curing polyacid-modified composite resin [compomer] cements) in terms of fluoride release, retentiveness, and microleakage after thermocycling. The RMGIC (Ortholy Band Paste [GC Ortholy, Inc., Tokyo, Japan]) showed a significantly higher amount of cumulative fluoride release over 180 days (p<0.001) and significantly greater tensile bond strength (p<0.001) than the compomer cements (Transbond Plus [3M Unitek, Monrovia, CA, USA] and Ultra Band-Lok [Reliance Orthodontic Products, Inc., Itasca, IL, USA]). Its bond strength was unaffected by thermocycling (2,000 cycles), indicating good retentiveness, whereas that of the compomer cements significantly decreased after thermocycling. Moreover, it had lower dyepenetration scores, indicative of less microleakage. These findings suggest that the RMGIC may have a better caries-preventive effect than the compomer cements and is suitable for long-term orthodontic banding.

Caries-preventive effect of fissure sealant containing surface reaction-type pre-reacted glass ionomer filler and bonded by self-etching primer.

OBJECTIVE: We aimed to evaluate the caries-preventive effect of a fissure sealant containing surface reaction-type pre-reacted glass ionomer (S-PRG) filler and bonded by self-etching primer versus those of 2 conventional resin-based sealants bonded by acid etching in terms of its impact on enamel demineralization and remineralization, enamel bond strength, and integrity of debonded enamel surfaces. MATERIALS AND METHOD: Demineralization, remineralization, and bond strength on untreated enamel and enamel subsurface lesions of bovine incisors were assessed among the sealants by polarizing microscopy and microradiography; debonded enamel surfaces were examined by scanning electron microscopy. RESULTS: The conventional resin-based sealants bonded by acid etching caused surface defects on the enamel subsurface lesions and significantly increased the lesion depth (p = 0.014), indicative of enamel demineralization. However the S-PRG filler-containing sealant bonded by self-etching primer maintained the enamel surface integrity and inhibited enamel demineralization. No difference in bond strength on both untreated enamel and enamel subsurface lesions was noted among the sealants. CONCLUSIONS: An S-PRG filler-containing fissure sealant bonded by self-etching primer can prevent enamel demineralization, microleakage, and gaps without the tags created by acid etching regardless of the enamel condition. Such sealants are suitable for protecting the pits and fissures of immature permanent teeth.

Comparison of subjective symptoms of temporomandibular disorders in young patients by age and gender.

The authors assessed the subjective symptoms of temporomandibular disorders (TMDs) in 167 young patients using self-reported forms, with five ratings for pain intensity and six ratings for difficulty in activities of daily living (ADL), to compare TMD symptoms according to gender and three age groups: group 1: 6- to 12-year-olds (juvenile); group 2: 13- to 15-year-olds (early adolescent); group 3: 16- to 18-year-olds (late adolescent). No significant gender differences were found in the symptoms among the groups, except for headache and neck pain in group 3. Pain intensity and tightness in the jaw/face, headache, and neck pain, as well as the ADL-related difficulty in prolonged jaw opening, eating soft/hard foods, and sleeping significantly differed among the groups (p < 0.01, Kruskal-Wallis test). Therefore, late adolescent patients with TMDs have higher pain intensity in the orofacial region and greater difficulty in ADL than do early adolescent and juvenile patients with TMDs.

Contribution of phosphoglucosamine mutase to determination of bacterial cell morphology in Streptococcus gordonii.

Phosphoglucosamine mutase (GlmM; EC 5.4.2.10) catalyzes the interconversion of glucosamine-6-phosphate to glucosamine-1-phosphate, an essential step in the biosynthetic pathway leading to the formation of the peptidoglycan precursor uridine 5'-diphospho-N-acetylglucosamine. We have recently identified the gene (glmM) encoding the enzyme of Streptococcus gordonii, an early colonizer on the human tooth and an important cause of infective endocarditis, and indicated that the glmM mutation in S. gordonii appears to influence bacterial cell growth, morphology, and sensitivity to penicillins. Moreover, the glmM mutation results in increased sensitivity to polymorphonuclear leukocyte (PMN)-dependent killing. In the present study, we observed similarities in the utilization of sugar between the wild-type strain and the glmM mutant of S. gordonii when cultivated with medium containing 0.2% glucose, fructose, lactose, or sucrose. Morphological analyses clearly indicated that the glmM mutation causes marked elongation of the streptococcal chains, enlargement of bacterial cells, increased distortion of the bacterial cell surface, and defects in cell separation. These results suggest that mutations in glmM appear to influence bacterial cell growth and morphology, independent of the carbon source.

Evaluation of the ion-releasing and recharging abilities of a resin-based fissure sealant containing S-PRG filler.

We evaluated the concentration of fluoride release at the initial stage and after recharging the fluoride from a resin-based sealant containing surface reaction-type pre-reacted glass-ionomer (S-PRG) filler and conventional resin-based sealants. The concentration of other ions released from the sealants in the surrounding distilled water was also examined. At the initial stage, fluoride was released from all sealants. For the sealant containing S-PRG filler, a significantly higher amount of Sr and B ions was detected relative to those from other sealants. After sealant samples were soaked in a sodium fluoride solution, a significant amount of fluoride and other ions was released from the sealant containing S-PRG filler. These findings suggest that sealants containing S-PRG filler enhance the enamel demineralization-inhibiting effect and the remineralization-promoting effect for long-term dental care.

Inter- and intraobserver reliability of the hand-held monitor for measuring salivary alpha-amylase level.

BACKGROUND: Alpha-amylase levels can be used to quantify the psychological stress. A hand-held salivary alpha-amylase (sAA) monitor was recently developed for measuring sAA levels. This study investigated the inter- and intraobserver reliability of this novel instrument. METHODS: Saliva was collected from 32 healthy volunteers. To determine the interobserver reliability, three observers measured the sAA levels in 10 samples. To determine the intraobserver reliability, one observer performed consecutive sAA measurements in 32 samples. The sAA levels in these 32 samples were also measured by a conventional laboratory enzymatic analysis, and these measurements were compared with those obtained using the hand-held monitor. RESULTS: Intraclass correlation coefficients indicated excellent inter- and intraobserver reliability (0.99 and 0.92, respectively). The results from the monitor and the lab assay showed a significant positive correlation (r = 0.96, p < 0.001). CONCLUSIONS: These results suggest that the hand-held monitor is highly reliable for measuring the sAA level.

Combined effect of paste containing casein phosphopeptide-amorphous calcium phosphate and fluoride on enamel lesions: an in vitro pH-cycling study.

PURPOSE: The purpose of this study was to show through enamel remineralization that a combination of casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) and fluoride is better than fluoride alone when the processing time for remineralization is short. METHODS: The bovine enamel slabs (n=28) were subjected to pH cycling for 4 days. Paste containing CPP-ACP and/or fluoride was applied for 30 minutes daily. The sections were observed using a polarizing microscope and microradiographs. RESULTS: In the groups treated solely with sodium fluoride (NaF) solution, tissue loss on the enamel surface was observed. On the other hand, in the groups that had been treated with a mixture of the NaF solution and CPP-ACP, the enamel surface was maintained. CONCLUSIONS: These results show that casein phosphopeptide-amorphous calcium phosphate-containing paste has the ability to maintain the enamel surface; the combined use of CPP-ACP paste and fluoride enhances this ability, thereby reducing demineralization.

Contribution of phosphoglucosamine mutase to the resistance of Streptococcus gordonii DL1 to polymorphonuclear leukocyte killing.

Phosphoglucosamine mutase (GlmM; EC 5.4.2.10) catalyzes the interconversion of glucosamine-6-phosphate to glucosamine-1-phosphate, an essential step in the biosynthetic pathway leading to the formation of the peptidoglycan precursor uridine 5'-diphospho-N-acetylglucosamine. We have recently identified the gene (glmM) encoding the enzyme of Streptococcus gordonii, an early colonizer on the human tooth and an important cause of infective endocarditis, and indicated that the glmM mutation in S. gordonii appears to influence bacterial cell growth, morphology, and sensitivity to penicillins. In the present study, we assessed whether the glmM mutation also affects escape from polymorphonuclear leukocyte (PMN)-dependent killing. Although no differences in attachment to human PMNs were observed between the glmM mutant and the wild-type S. gordonii, the glmM mutation resulted in increased sensitivity to PMN-dependent killing. Compared with the wild type, the glmM mutant induced increased superoxide anion production and lysozyme release by PMNs. Moreover, the glmM mutant is more sensitive to lysozyme, indicating that the GlmM may be required for synthesis of firm peptidoglycans for resistance to bacterial cell lysis. These findings suggest that the GlmM contributes to the resistance of S. gordonii to PMN-dependent killing. Enzymes such as GlmM could be novel drug targets for this organism.

Identification of the Streptococcus gordonii glmM gene encoding phosphoglucosamine mutase and its role in bacterial cell morphology, biofilm formation, and sensitivity to antibiotics.

Phosphoglucosamine mutase (EC 5.4.2.10) catalyzes the interconversion of glucosamine-6-phosphate into glucosamine-1-phosphate, an essential step in the biosynthetic pathway leading to the formation of peptidoglycan precursor uridine 5'-diphospho-N-acetylglucosamine. The gene (glmM) of Escherichia coli encoding the enzyme has been identified previously. We have now identified a glmM homolog in Streptococcus gordonii, an early colonizer on the human tooth and an important cause of infective endocarditis, and have confirmed that the gene encodes phosphoglucosamine mutase by assaying the enzymatic activity of the recombinant GlmM protein. Insertional glmM mutant of S. gordonii did not produce GlmM, and had a growth rate that was approximately half that of the wild type. Morphological analyses clearly indicated that the glmM mutation causes marked elongation of the streptococcal chains, enlargement of bacterial cells, and increased roughness of the bacterial cell surface. Furthermore, the glmM mutation reduces biofilm formation and increases sensitivity to penicillins relative to wild type. All of these phenotypic changes were also observed in a glmM deletion mutant, and were restored by the complementation with plasmid-borne glmM. These results suggest that, in S. gordonii, mutations in glmM appear to influence bacterial cell growth and morphology, biofilm formation, and sensitivity to penicillins.

Hsa, an adhesin of Streptococcus gordonii DL1, binds to alpha2-3-linked sialic acid on glycophorin A of the erythrocyte membrane.

Bacterial recognition of host sialic acid-containing receptors plays an important role in microbial colonization of the human oral cavity. The aggregation of human platelets by Streptococcus gordonii DL1 is implicated in the pathogenesis of infective endocarditis. In addition, we consider that hemagglutination of this organism may act as an additive factor to increase the severity of this disease. We previously reported that this interaction requires the bacterial expression of a 203-kDa protein (Hsa), which has sialic acid-binding activity. In the present study, we confirmed that erythrocyte surface sialoglycoproteins are the receptors for Hsa. We examined the effects of proteinase K, chymotrypsin, phospholipase C, and alpha(2-3) or alpha(2-3, 6, 8) neuraminidase on hemagglutination activity and found that the interaction occurs between Hsa and alpha2-3-linked sialic acid-containing proteins of erythrocytes. We expressed recombinant NR2, which is the putative binding domain of Hsa, fused with GST in Escherichia coli BL21. Dot-blot analysis demonstrated that GST-HsaNR2 binds both glycophorin A (GPA) and band 3. Moreover, GPA and a small amount of band 3 were detected by GST pull-down assays. These findings indicate that S. gordonii Hsa specifically binds to GPA and band 3, alpha2-3-linked sialic acid membrane glycoproteins.

Contribution of sialic acid-binding adhesin to pathogenesis of experimental endocarditis caused by Streptococcus gordonii DL1.

An insertional mutation in hsa, the gene encoding the sialic acid-binding adhesin of Streptococcus gordonii DL1, resulted in a significant reduction of the infection rate of the organism and an inflammatory reaction in the rat aortic valve with experimental endocarditis, suggesting that the adhesin contributes to the infectivity of the organism for heart valves.