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INTRODUCTION: Ammonia production via the arginine deiminase system (ADS) of oral bacteria can function to reduce the cariogenicity of oral biofilms by neutralizing glycolytic acids that cause tooth demineralization. OBJECTIVES: This cohort study investigated the relationship between ADS activity and bacterial profile changes of supragingival biofilms with caries experience among children over time. METHODS: A total of 79 children aged 2 to 7 y at baseline were assessed every 6 mo for a period of 18 mo. Children were grouped as caries free (CF), caries active with enamel lesions (CAE), or caries active with dentin lesions (CA). Supragingival plaque samples were collected from caries-free surfaces (PF) and from enamel (PE) and dentin (PD) lesions. Plaque ADS activity was measured by monitoring citrulline production from arginine and compared with ribosomal 16S rRNA–derived taxonomic profiles for the same samples. RESULTS: At baseline, 37% of the children were CF, 34% CAE, and 29% CA. At 18 mo, 26% were CF, 41% CAE, 23% CA, and 10% were caries experienced (new restorations but no caries activity). Throughout the study period, ADS activity was significantly higher in the CF group than the CA group (P < 0.0001), and ADS activity in the PF samples was significantly higher than in the PE and PD samples (P < 0.0001). Distance-based redundancy analysis showed that the bacterial communities could be differentiated when plaque samples are grouped into levels of high and low ADS activity. CONCLUSIONS: There is a positive correlation between caries activity and low arginolytic capacity of the supragingival oral biofilms of children and tooth surfaces over time. Measurements of arginine metabolism via ADS may be useful to differentiate the caries risk of individuals and tooth surfaces. KNOWLEDGE TRANSFER STATEMENT: Findings from this study support the development of new strategies for caries risk assessment and prevention based on modulation of the virulence of the oral microbiome through arginine metabolism in supragingival biofilms.

Oral Candida albicans has been detected in children with early childhood caries (ECC) and has demonstrated cariogenic traits in animal models of the disease. Conversely, other studies found no positive correlation between C. albicans and caries experience in children, while suggesting it may have protective effects as a commensal organism. Thus, this study aimed to examine whether oral C. albicans is associated with ECC. Seven electronic databases were searched. The data from eligible studies were extracted, and the risk of bias was evaluated. A fixed effects model (Mantel-Haenszel estimate) was used for meta-analysis, and the summary effect measure was calculated by odds ratio (OR) and 95% confidence interval (CI). Fifteen cross-sectional studies were included for the qualitative assessment and 9 studies for meta-analysis. Twelve studies revealed higher oral C. albicans prevalence in ECC children than in caries-free children, while 2 studies indicated an equivalent prevalence. A pooled estimate, with OR = 6.51 and 95% CI = 4.94-8.57, indicated a significantly higher ECC experience in children with oral C. albicans than those without C. albicans (p < 0.01). The odds of experiencing ECC in children with C. albicans versus children without C. albicans were 5.26 for salivary, 6.69 for plaque, and 6.3 for oral swab samples. This systematic review indicates that children with oral C. albicans have >5 times higher odds of having ECC compared to those without C. albicans. Further prospective cohort studies are needed to determine whether C. albicans could be a risk factor for ECC, and whether it is dependent on different sample sources (saliva/plaque).

Arginine metabolism via the arginine deiminase system (ADS) of oral bacteria generates ammonia, which can increase the pH of oral biofilms and decrease the risk for dental caries. Antagonistic interactions between ADS-positive and cariogenic bacteria in oral biofilms may be an important ecological determinant of caries. This study investigated the antagonistic potential and mechanisms of clinical isolates of arginolytic streptococci on and by Streptococcus mutans UA159, a well-characterized cariogenic human isolate. Low-passage isolates of Streptococcus gordonii, Streptococcus sanguinis, Streptococcus parasanguinis, Streptococcus australis, and Streptococcus cristatus inhibited the growth of S. mutans to various degrees when they were inoculated on growth media first or simultaneously with S. mutans. The antagonistic effects of arginolytic strains against S. mutans and the production of H2O2 by these strains were enhanced during growth in a less-rich medium or when galactose was substituted for glucose as the primary carbohydrate source. Pyruvate oxidase was the dominant pathway for H2O2 production by arginolytic strains, but lactate oxidase activity was also detected in some strains of S. gordonii and S. cristatus. UA159 inhibited the growth of all tested arginolytic strains when inoculated first, especially in aerobic conditions. However, the antagonistic effects of S. mutans on certain strains of S. gordonii and S. australis were not observed during anaerobic growth in the presence of arginine. Thus, arginolytic commensal streptococci may have a synergistically positive impact on the ecology of oral biofilms by moderating biofilm pH while antagonizing the growth and virulence of caries pathogens.

Background Alkali production via arginine deiminase system (ADS) of oral bacteria plays a significant role in oral ecology, pH homeostasis and inhibition of dental caries. ADS activity in dental plaque varies greatly between individuals, which may profoundly affect their susceptibility to caries. Objective To investigate the effect of arginine on the growth and biofilm formation of oral bacteria. Methods and results Polymicrobial dental biofilms derived from saliva were formed in a high-throughput active attachment biofilm model and L-arginine (Arg) was shown to reduce the colony forming units (CFU) counts of such biofilms grown for various periods or biofilms derived from saliva of subjects with different caries status. Arg hardly disturbed bacterial growth of Streptococcus mutans, Streptococcus sobrinus, Streptococcus sanguinis and Streptococcus gordonii in BHI medium, but only inhibited biofilm formation of S. mutans. Scanning electron microscope (SEM) showed S. mutans biofilms harboured fewer cells grown with Arg than that without Arg, even in the initial 2 h and 8 h phase. Confocal laser scanning microscope (CLSM) images of poly-microbial dental and S. mutans biofilms revealed the biofilms grown with Arg had lower exopolysaccharide (EPS)/bacteria ratios than those without Arg (P = 0.004, 0.002, respectively). Arg could significantly reduce the production of water-insoluble EPS in S. mutans biofilms (P < 0.001); however, quantitative real-time PCR (qRT-PCR) did not show significantly influence in gene expression of gtfB, gtfC or gtfD (P = 0.32, 0.06, 0.44 respectively). Conclusions Arg could reduce the biomass of poly-microbial dental biofilms and S. mutans biofilms, which may be due to the impact of Arg on water-insoluble EPS. Considering the contribution to pH homeostasis in dental biofilms, Arg may serve as an important agent keeping oral biofilms healthy thus prevent dental caries.

Objectives:Galla chinensiswater extract (GCE) has been demonstrated to inhibit dental caries by favorably shifting the demineralization/remineralization balance of enamel and inhibiting the biomass and acid formation of dental biofilm. The present study focused on the comparison of composition and anticaries effect ofGalla chinensisextracts with different isolation methods, aiming to improve the efficacy of caries prevention.Methods:The composition of water extract (GCE), ethanol extract (eGCE) and commercial tannic acid was compared. High performance liquid chromatography coupled to electrospray ionization-time of flight-mass spectrometry (HPLC-ESI-TOF-MS) analysis was used to analyze the main ingredients.In vitropH-cycling regime and polymicrobial biofilms model were used to assess the ability of differentGalla chinensisextracts to inhibit enamel demineralization, acid formation and biofilm formation.Results:All the GCE, eGCE and tannic acid contained a high level of total phenolics. HPLC-ESI-TOF-MS analysis showed that the main ingredients of GCE were gallic acid (GA), while eGCE mainly contained 4-7 galloylglucopyranoses (GGs) and tannic acid mainly contained 5-10 GGs. Furthermore, eGCE and tannic acid showed a better effect on inhibiting enamel demineralization, acid formation and biofilm formation compared to GCE.Conclusions:Galla chinensisextracts with higher tannin content were suggested to have higher potential to prevent dental caries.

Objective This study was designed to explore the effect of gallic acid (GA, one of the ingredients of chemical compounds from galla chinensis) on the morphology and growth of hydroxyapatite crystals. Methods The crystals was produced by mixing CaCl 2 and KH 2 PO 4 with or without GA (4 g/L) at room temperature for 3, 12, 24 h and 3, 7, 14 days. Subsequently, the micro-structure, morphology and composition of the crystals were investigated via SEM, XRD, ATR-FTIR and fluorescence microscopy. Results The mineral phase was hydroxyapatite in both groups after 14 days, but their processes and the morphology were completely different. The crystals from groups utilizing GA for 14 days were urchin-like, while loose needle-like crystals were observed in groups without GA. XRD results indicated that GA might limit the growth of the crystals, mainly on the 0 0 2 direction. The results of ATR-FTIR and fluorescence microscopy revealed that the unique structures might caused by the participation of GA during crystals formation. Conclusion GA might affect and participate into the formation of the hydroxyapatite, and regulate the morphology and structure of the crystals, to enhance the remineralization process.

To determine the chemical composition of Galla chinensis extract (GCE) by several analysis techniques and to compare the efficacy of GCE and its main component(s) in inhibition of enamel demineralization, for the development of future anticaries agents, main organic composition of GCE was qualitatively determined by liquid chromatography-time of flight-mass spectrometry (LC-TOF-MS) and quantified by high-performance liquid chromatography-diode array detector (HPLC-DAD). Inorganic ions were tested by inductively coupled plasma-atomic emission spectroscopy and F was especially measured by ion chromatography. Then, bovine enamel blocks were randomly divided into four treatment groups and were subjected to a pH-cycling regime for 12 times. Each cycle included 5-min applications with one of four treatments: 4 g⋅L(-1) GCE solution, 4 g⋅L(-1) gallic acid (GA) solution, 1 g⋅L(-1) NaF solution (positive control), deionized water (DDW, negative control), and then 60-min application in pH 5.0 acidic buffer and 5-min application in neutral buffer. Acidic buffers were retained for calcium analysis. The main organic composition of GCE were GA and its isomer, and, to a lesser extent, small molecule gallotannins. The content of GA in GCE was 71.3%±0.2% (w/w). Inorganic ions were present in various amounts, of which Ca was (136±2.82) µg⋅g(-1), and Zn was (6.8±0.1) µg⋅g(-1). No F was detected in GCE. In pH cycling, GA showed an effect similar to GCE in inhibiting enamel demineralization (P0.05). GA was found to be the main effective, demineralization inhibiting component of GCE and could be a promising agent for the development of anticaries agents.

Aim The aim of this survey was to compare Chinese natives and foreign inhabitants in Chengdu, China, with respect to: (1) attitudes towards dental appearance, (2) subjective orthodontic treatment need, and (3) the main factors influencing orthodontic treatment need. Methodology A total of 522 subjects, including 227 foreign inhabitants and 295 Chinese natives in Chengdu participated in the survey. A simple random sampling method was adopted and a face-to-face interview was conducted at some public sites using a questionnaire. Data was entered by two persons synchronously using Epidata 3.0, and SPSS 13.0 was used to analyze these data. Results 89.0% of foreign inhabitants were satisfied with their teeth compared to only 46.8% of Chinese natives. Females were more dissatisfied with their teeth than males. Chinese natives put improving appearance as the top priority (55.9%) for seeking orthodontic treatment; however, in foreign inhabitants, the main reason for seeking treatment was to improve masticatory function (44.1%), followed by ”to be pretty“ (35.2%). The importance of well-aligned teeth and self-perception of psycho-social impact of malocclusion were the same two main factors influencing subjective orthodontic treatment need (P

The purpose of this study is to detect the effect of gallic acid (GA) on hydroxyapatie crystal growth and find the mechanism of the regulation. We evaluated the morphology of HAP crystals grown under various amounts of GA (0, 0.05, 1, and 4 gL(-1)). Subsequently, the chemical composition, crystal size and the morphology were investigated via the energy-dispersive X-ray spectrometer, attenuated total fourier transform infrared spectroscopy, thermogravimetric analysis, X-ray diffraction analysis, transmission electron microscope and scanning electron microscopy. In all groups, the Ca/P ratio was closed to 1.67. In the absence of GA, crystals did not arrange, while in the presence of GA, crystals tended to form spherules. The size of the crystals decreased with the concentration of GA increased. These results indicated the role of GA on the growth and morphology of hydroxyapatite crystals, which might be the key mechanism for gallic acid regulating the mineralization.

Background. Gallic acid (GA) has been shown to inhibit demineralization and enhance remineralization of enamel; however, GA solution is highly acidic. This study was to investigate the stability of GA solutions at various pH and to examine the resultant effects on enamel demineralization. Methods. The stability of GA in H2O or in phosphate buffer at pH 5.5, pH 7.0 and pH 10.0 was evaluated qualitatively by ultraviolet absorption spectra and quantified by high performance liquid chromatography with diode array detection (HPLC-DAD). Then, bovine enamel blocks were subjected to a pH-cycling regime of 12 cycles. Each cycle included 5 min applications with one of the following treatments: 1 g/L NaF (positive control), 4 g/L GA in H2O or buffered at pH 5.5, pH 7.0 and pH 10.0 and buffers without GA at the same pH (negative control), followed by a 60 min application with pH 5.0 acidic buffers and a 5 min application with neutral buffers. The acidic buffers were analysed for dissolved calcium. Results. GA was stable in pure water and acidic condition, but was unstable in neutral and alkaline conditions, in which ultraviolet spectra changed and HPLC-DAD analysis revealed that most of the GA was degraded. All the GA groups significantly inhibited demineralization (p < 0.05) and there was no significant difference of the inhibition efficacy among different GA groups (p > 0.05). Conclusions. GA could inhibit enamel demineralization and the inhibition effect is not influenced by pH. GA could be a useful source as an anti-cariogenic agent for broad practical application.

A selected group of oral bacteria commonly associated with dental health is capable of producing alkali via the arginine deiminase system (ADS), which has a profound impact on the pH of human oral biofilms. An increased risk for dental caries has been associated with reduced ADS activity of the bacteria in oral biofilms. Arginolytic bacterial strains from dental plaque samples of caries-free and caries-active adults were isolated and characterized to investigate the basis for differences in plaque ADS activity between individuals. Fifty-six ADS-positive bacterial strains were identified by 16S rRNA gene sequencing, and their ADS activity levels were compared under standard growth conditions. The spectrum of bacterial ADS activity ranged from 45.2 to 688.0 units (mg protein)-1. Although Streptococcus sanguinis was the most prevalent species, other Streptococcus sp. were also represented. Biochemical assays carried out using 27 ADS-positive strains under conditions known to induce or repress ADS gene expression showed substantial variation in arginolytic activity in response to pH, oxygen and the availability of carbohydrate or arginine. This study reveals that the basis for the wide spectrum of arginolytic expression observed among clinical strains is, at least in part, attributable to differences in the regulation of the ADS within and between species. The results provide insights into the microbiological basis for intersubject differences in ADS activity in oral biofilms and enhance our understanding of dental caries as an ecologically driven disease in which arginine metabolism moderates plaque pH and promotes dental health.

Alkali production by oral bacteria in the oral cavity has been linked to protection against dental caries. The current study assessed various parameters associated with ammonium produced during arginine catabolism in dental biofilms. Polymicrobial biofilms were formed with saliva as the inoculum. The NH3 level and the pH of the spent medium were used to monitor and quantitate the bacterial reactions. The presence of sucrose, a low buffer capacity, and a low pH (≤ pH 4.5) were found to hamper alkali production from arginine. The rate of alkali production exhibited an optimum around pH 5.5. Biofilms were found to produce NH3 also from polypeptides and proteins in the medium. The biofilm age affected these processes. The experimental model proved valuable for the assessment of the collective bacterial reactions determining the overall pH outcome. This experimental approach could bridge the gap in our knowledge between pH-rise phenomena and caries susceptibility from clinical observations and studies performed on alkali-producing bacteria in well- controlled, though simplified, in vitro models. Analysis of our data supports the hypothesis that the initiation and progression of dental caries may be influenced by the relative rates of acid and base formation, which critically depend on the aforementioned parameters.

ObjectivesConsidering that Galla chinensis extract (GCE) solution has a low pH, which might dissolve dental enamel, we investigated the effects of elevation of pH on GCE stability, and on its anti-caries properties.DesignsStability of GCE solutions, either in H2O (pH less than 4.0) or when buffered at pH 5.5, 7.0 and 10.0, was assessed from UV-VIS spectra. Inhibition of enamel demineralization was determined in a pH-cycling set up, comprising treatments with either GCE solutions or negative control buffers and acid and neutral buffer immersions. Demineralization was assessed by calcium in the acetate buffers. To determine antimicrobial properties, polymicrobial biofilms were formed after saliva inoculation on glass surfaces which were treated after 48 h. Treatment output parameters were lactic acid formation and viability, the latter by colony forming unit (CFU) counts.ResultsAt pH 7.0 and higher GCE solutions changed colour and absorption spectra in UV-VIS, indicative of chemical changes. Regarding enamel demineralization, significant inhibitions (P < 0.05) were found for all GCE treatments when compared with corresponding controls. In polymicrobial biofilms, GCE reduced the acid production, compared with the negative controls (P < 0.05). However, this difference was only significant at the lower pH values.ConclusionsGCE solutions were unstable under neutral and alkaline conditions. pH did not significantly influence the inhibiting effect of GCE on enamel demineralization. However, GCE was not effective on polymicrobial biofilms at alkaline pH (8.5). To avoid enamel damage due to acidic treatment, GCE solutions should be used at about pH 5.5.

The performance of mastication is presumed to be influenced by afferent stimulation, but little information is available from awake, intact animals. Thirteen experiments were carried out with four miniature pigs. Electromyographic signals from the jaw muscles were recorded simultaneously with jaw movements during natural chewing of foods of differing hardnesses. Harder foods were found to be associated with higher activity levels of jaw-closing muscles and greater lateral deviation, but reduced jaw opening. The sensory supply from one side of the oral cavity was then removed by injected local anesthetic, and the recordings repeated. In addition to decreased activity levels, the jaw-closing muscles showed reduced ability to adjust to different foods. The frequency of mastication fell slightly, and the animals preferred to chew on the uninjected side. Lateral deviation was reduced, but at the same time the jaw-opening muscles usually became more active, and accordingly the jaw opened more widely. These findings provide new information on the role of oral sensory afferents in regulating natural chewing patterns. The decreased activities of the jaw-closing muscles are consistent with, although they do not demonstrate, the existence of a positive feedback from periodontal afferents.

Although the inactivation or removal of motoneurones and muscles are known to affect development and growth of the craniofacial skeleton and masticatory muscles, the effect of these on functional activity patterns is not clear. The goal of this study was to evaluate the immediate responses of masticatory muscle activities to temporary nerve blocks of individual jaw-closing muscles. Nineteen experiments were made with four miniature pigs (Sus scrofa). Electromyograms of the masticatory muscles and jaw movements were recorded during natural chewing. Then, the function of individual jaw-closing muscles was removed unilaterally by local anaesthesia, and the recording repeated. The results showed a general increase in the activities of the other jaw-closing muscles, particularly those that were synergistic with the blocked muscle in producing lateral movements. Furthermore, the lateral pterygoid showed stronger activity, and accordingly the lateral movement was increased. Digastric muscle activity and the magnitude of jaw opening also tended to increase. Thus, the response to loss of a muscle is a strong, immediate compensation by synergists.