Long-Term Mastication Changed Salivary Metabolomic Profiles.

Saliva is an ideal biofluid for monitoring oral and systemic health. Repeated mastication is a typical physical stimulus that improves salivary flow and oral hygiene. Recent metabolomic studies have shown the potential of salivary metabolomic components for various disease monitoring systems. Here, we evaluated the effect of long-term mastication on salivary metabolomic profiles. Young women with good oral hygiene (20.8 ± 0.3 years, n = 17) participated. They were prohibited from chewing gum during control periods (4 weeks each) and were instructed to chew a piece of gum base seven times a day for 10 min each time during the intervention period. Paired samples of unstimulated whole saliva collected on the last day of the control and intervention period were compared. Liquid chromatography−time-of-flight mass spectrometry successfully quantified 85 metabolites, of which 41 showed significant differences (p < 0.05, Wilcoxon paired test corrected by false discovery rate). Except for a few metabolites, such as citrate, most metabolites showed lower concentrations after the intervention. The pathways related to glycogenic amino acids, such as alanine, arginine, and glutamine, altered considerably. This study suggests that long-term mastication induces unstimulated salivary component-level changes.

Effects of Chewing Gum Base on Oral Hygiene and Mental Health: A Pilot Study.

The purpose of the present study was to clarify the long-term effects of frequent chewing of unflavored and odorless gum (hereafter, gum base) on oral hygiene and mental health. This single-arm study, which started with a 4-week control and ended with a 4-week intervention period, was conducted in two phases: one in 2017 and one in 2018. The participants comprised 36 dental hygiene students (17 in 2017, 19 in 2018). During the intervention period, all participants were required to chew a piece of gum base 7 times a day for 10 min each time. The unstimulated salivary flow rate and masticatory efficiency were measured and chewing number counted. Two questionnaires －the Profile of Mood States, second edition (POMS2) and the 30-item General Health Questionnaire (GHQ-30)－ were administered to assess mental health. In both phases, the unstimulated salivary flow rate showed a significant increase after the intervention period (p<0.05). In 2017, the GHQ-30 scores and masticatory efficiency showed a tendency toward a negative correlation after the intervention period (r=－0.4647, p=0.06). In 2018, a significant negative correlation was observed between chewing number and the POMS2 scores after the intervention period (r=－0.6296, p<0.01). These findings suggest that frequent chewing of gum base increases unstimulated salivary flow rate. However, no significant change was observed in the mental health.

Effects of 7S globulin 3 derived from the adzuki bean [Vigna angularis] on the CSP- and eDNA- dependent biofilm formation of Streptococcus mutans.

OBJECTIVE: Streptococcus mutans is a principal bacterium that forms pathogenic biofilm involved in the development of dental caries. S. mutans possesses a quorum sensing system (QS) stimulated by competence stimulating peptide (CSP), which is associated with bacteriocin production, genetic competency and biofilm formation. Inhibiting CSP-dependent QS is one of the aims leading to the inhibition of biofilm formation and is useful for establishing new prevention systems for dental caries. DESIGN: In this study, we selected adzuki bean [Vigna angularis] extract as a candidate component to inhibit CSP-dependent biofilm formation among various foods. To purify an inhibitory component from the adzuki extracts, we performed the salting-out method, two rounds of ion-exchange chromatography, and SDS and native PAGE. RESULTS: A primary protein band that inhibits CSP-dependent biofilm formation appeared at approximately 50 kDa and was identified as 7S globulin 3 (7S3), a major seed storage protein in adzuki bean. To determine the characteristics of 7S3 as an inhibitory component, aggregated proteins were extracted from the adzuki crude extracts at pH values lower than 6. The aggregated proteins inhibited CSP- and eDNA-dependent biofilm formation and showed 50 kDa band, which is identical with 7S3 in the purified sample. Moreover, 7S globulin 3 in the adzuki bean extract directly interacted with CSP at low pH conditions but not at neutral conditions, and inhibited CSP-dependent bacteriocin production. CONCLUSION: It was suggested that 7S3 might be a safe and useful material to prevent pathogenic activities in the biofilm formation of S. mutans.

Dental plaque development on a hydroxyapatite disk in young adults observed by using a barcoded pyrosequencing approach.

Dental plaque is a dynamic microbial biofilm ecosystem that comprises hundreds of species including difficult-to-cultivate bacteria. We observed the assembly of a plaque bacterial community through 16S rRNA gene analysis. Plaque samples that accumulated on a hydroxyapatite disk for 1, 2, 3, 4, 5, and 7 days with saliva on day 0 were collected from 19 young adults using a removable resin splint. Quantitative PCR analysis showed that the total bacterial amount gradually increased and reached a plateau on day 4. Barcoded pyrosequencing analysis revealed that the microbial richness and diversity particularly increased between days 5 and 7. A principal coordinate analysis plot based on unweighted UniFrac showed the community assembly in a time-related manner, which became increasingly similar to the salivary microbiota. Facultative anaerobic bacteria such as Streptococcus, Neisseria, Abiotrophia, Gemella, and Rothia were predominant in the plaque bacterial community in the earlier days, whereas obligate anaerobes, such as Porphyromonas, Fusobacterium, Prevotella, and Capnocytophaga showed increased dominance on later days. UniFrac analysis also demonstrated that dental caries experience had a significant effect on the assembly process. Our results reveal the development pattern of the plaque bacterial community as well as the inter-individual differences associated with dental caries experience.

Effect of xylitol gum on the level of oral mutans streptococci of preschoolers: block-randomised trial.

OBJECTIVES: To assess the influence of xylitol chewing gum consumption on mutans streptococci level of 3-4 years old Japanese preschoolers. METHODS: 248 participants were examined regarding caries-related factors at baseline and were followed up at 6, 9, and 12 months after the baseline: assessors were blinded, subjects were open labelled and blocked parallel randomised; 142 were selected to use xylitol gum for 3 months (from months 6 to 9) and 106 were controls. RESULTS: 161 participants were analysed (xylitol n = 76, control n = 85). Nineteen caries-related variables, including xylitol gum consumption, were analysed for any association with the main outcome, plaque mutans streptococci scores development within the intervention period, by logistic regression. Six showed statistically significant associations by univariate analysis (P < 0.05). However, only xylitol gum consumption remained a significant negative association (P < 0.05) by multiple analyses. Interestingly, over 10% xylitol group children experienced diarrhoea, which was larger than previous investigations. CONCLUSION: Xylitol gum is effective in avoiding increased plaque mutans streptococci in young children.

Remineralization effects of xylitol on demineralized enamel.

We morphologically determined the effects of xylitol on the remineralization of artificially demineralized enamel. The samples were demineralized and then immersed in a remineralizing solution with or without 20% xylitol at 37 degrees C for 2 weeks. Samples were observed using contact microradiography, a multipurpose image processor (MIP) and a high-resolution electron microscope (HRTEM). Contact microradiography indicated that remineralization occurred in the surface and deep layers of samples immersed in a non-xylitol solution. Samples immersed in a xylitol solution demonstrated less mineralization in the outer 10 microm of the outermost surface layers, but more mineralization in the middle and deep layers, than was observed in the non-xylitol samples. The MIP evaluation indicated that remineralization was more prominent in layers at depths of 50-60 microm in the xylitol samples than in the non-xylitol samples. Observation of the xylitol samples by HRTEM revealed crystals of various sizes and irregular shapes with unclear crystal angles in the outermost surface layers. In the middle layers, they had thickened and angles of crystals are clear. These results indicate that xylitol can induce remineralization of deeper layers of demineralized enamel by facilitating Ca2+ movement and accessibility.