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Objectives: Calcifying nanoparticles (CNPs), referred to as nanobacteria (NB), are recognized to be associated with ectopic calcification. This study aims to isolate and culture CNPs from the dental plaque of patients with periodontal disease and investigate their possible role in unravelling the aetiology of periodontal disease., Material and Methods: Supragingival and subgingival plaques were sampled from 30 periodontitis patients for CNPs isolation and culture. Alkaline phosphatase (ALP) content changes were tracked over time. Positive samples underwent thorough morphological identification via hematoxylin and eosin (HE) staining, Alizarin red S (ARS), and transmission electron microscopy (TEM). The chemical composition of CNPs analysis involved calcium (Ca) and phosphorus (P) content determination, Fourier transform infrared spectroscopy (FTIR), and X-ray diffraction (XRD)., Results: The subgingival plaque dental group exhibited a higher CNPs isolation rate at 36.67% (11/30) compared to the supragingival dental plaque group at 66.67% (20/30). ALP activity varied among the positive, negative and control groups. Morphological observation characterized the CNPs as round, oval, and ellipsoid particles with Ca deposits. Chemical analysis revealed the Ca/P ratio was 0.6753. Hydroxyl, methyl, carbonate, phosphate, hydrogen phosphate, and dihydrogen phosphate were detected by FTIR; the main chemical components detected by XRD were hydroxyapatite and tricalcium phosphate., Conclusion: CNPs were found in periodontitis-related dental plaque and exhibited the potential to develop calcified structures resembling dental calculus. However, the potential involvement of ALP in CNPs formation requires deeper exploration, as does the precise nature of its role and the interrelation with periodontitis demand a further comprehensive investigation., (© 2024 The Authors. Clinical and Experimental Dental Research published by John Wiley & Sons Ltd.)

Background & Aims: Oral nutritional supplements (ONS) administered to malnourished elders and other patients contain high-levels of carbohydrates that could be a potential risk factor for dental caries. This study aimed to evaluate the cariogenic potentials of ONS using intraoral plaque telemetry., Methods: Ten ONS were tested on five healthy volunteers (mean age: 76.8 ± 9.15 years). Participants were requested to refrain from performing oral hygiene 3-7 days prior to testing. The pH-value below the dental plaque on the tooth was measured while the ONS was being consumed. After neutralizing the participant's saliva, a control solution (10% sucrose) was administered and telemetry measurements were repeated. Mean relative cariogenicity (RC) was calculated for each ONS. ANOVA and post hoc tests were used for statistical analyses (p < 0.05)., Results: All ten ONS were potentially cariogenic on enamel with an overall RC of 0.519 ± 0.35 (Range: Min = 0.31 ± 0.16; Max = 1.00 ± 0.34). RC differed significantly between the ONS (p = 0.002). RC was lower in ONS that contained high-protein (p = 0.018). RC was not influenced by other factors such as readily consumable (p = 0.102), flavor (p = 0.869), consistency (p = 0.126), fiber containing (p = 0.134), style (p = 0.112), and age of plaque (p = 0.339)., Conclusions: The ONS administered to elders and malnourished patients are potentially cariogenic. It is imperative that the administration of ONS must be based on individual needs to potentiate a maximum benefit. Wherever possible, an attempt to limit the use of high-carbohydrate containing ONS must be practiced along with the adoption of suitable preventive measures to arrest the development and progression of caries., Competing Interests: Conflict of interest AS, IN, TA, MS: declare no conflicts of interest. FW: member of the board of Toothfriendly Switzerland., (Copyright © 2020 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Oral microbiome influences human health, specifically prediabetes and type 2 diabetes (Pre-DM/DM) and periodontal diseases (PDs), through complex microbial interactions. To explore these relations, we performed 16S rDNA sequencing, metabolomics, lipidomics, and proteomics analyses on supragingival dental plaque collected from individuals with Pre-DM/DM (n = 39), Pre-DM/DM and PD (n = 37), PD alone (n = 11), or neither (n = 10). We identified on average 2790 operational taxonomic units and 2025 microbial and host proteins per sample and quantified 110 metabolites and 415 lipids. Plaque samples from Pre-DM/DM patients contained higher abundance of Fusobacterium and Tannerella than plaques from metabolically healthy patients. Phosphatidylcholines, plasmenyl phosphatidylcholines, ceramides containing non-OH fatty acids, and host proteins related to actin filament rearrangement were elevated in plaques from PD versus non-PD samples. Cross-omic correlation analysis enabled the detection of a strong association between Lautropia and monomethyl phosphatidylethanolamine (PE-NMe), which is striking because synthesis of PE-NMe is uncommon in oral bacteria. Lipidomics analysis of in vitro cultures of Lautropia mirabilis confirmed the synthesis of PE-NMe by the bacteria. This comprehensive analysis revealed a novel microbial metabolic pathway and significant associations of host-derived proteins with PD., Competing Interests: Conflict of interest J. J. C. is a consultant for Thermo Fisher Scientific. The remaining authors declare no conflicts of interest related to this work., (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)

Dental plaque is a structurally organized biofilm which consists of diverse microbial colonies and extracellular matrix. Its composition may change when pathogenic microorganisms become dominating. Therefore, dental biofilm or plaque has been frequently investigated in the context of oral health and disease. Furthermore, its potential as an alternative matrix for analytical purposes has also been recognized in other disciplines like archeology, food sciences, and forensics. Thus, a careful in-depth characterization of dental plaque is worthwhile. Most of the conducted studies focused on the screening of microbial populations in dental plaque. Their lipid membranes, on the other hand, may significantly impact substance (metabolite) exchange within microbial colonies as well as xenobiotics uptake and incorporation into teeth. Under this umbrella, a comprehensive lipidomic profiling for determination of lipid compositions of in vivo dental plaque samples and of in vitro cultivated biofilm as surrogate matrix to be used for analytical purposes has been performed in this work. An untargeted lipidomics workflow utilizing a ultra-high-performance liquid chromatography (UHPLC)-quadrupole-time-of-flight (QTOF) platform together with comprehensive SWATH (sequential window acquisition of all theoretical fragment ion mass spectra) acquisition and compatible software (MS-DIAL) that comprises a vast lipid library has been adopted to establish an extensive lipidomic fingerprint of dental plaque. The main lipid components in dental plaque were identified as triacylglycerols, followed by cholesterol, cholesteryl esters as well as diacylglycerols, and various phospholipid classes. In vivo plaque is a rare matrix which is usually available in very low amounts. When higher quantities for specific research assays are required, efficient ways to produce an appropriate surrogate matrix are mandatory. A potential surrogate matrix substituting dental plaque was prepared by cultivation of in vitro biofilm from saliva and similarities and differences in the lipidomics profile to in vivo plaque were mapped by statistical evaluation post-analysis. It was discovered that most lipid classes were highly elevated in the in vitro biofilm samples, in particular diacylglycerols, phosphatidylglycerols, and phosphatidylethanolamines (PEs). Furthermore, an overall shift from even-chain lipid species to odd-chain lipids was observed in the cultivated biofilms. On the other hand, even-chain phosphatidylcholines (PCs), lysoPCs, cholesteryl esters, and cholesterol-sulfate were shown to be specifically increased in plaque samples. Graphical abstract.

Streptococcus mutans is an etiologic agent of human dental caries that forms dental plaque biofilms containing functional amyloids. Three amyloidogenic proteins, P1, WapA, and Smu&#95;63c were previously identified. C123 and AgA are naturally occurring amyloid-forming fragments of P1 and WapA, respectively. We determined that four amyloidophilic dyes, ThT, CDy11, BD-oligo, and MK-H4, differentiate C123, AgA, and Smu&#95;63c amyloid from monomers, but non-specific binding to bacterial cells in the absence of amyloid precludes their utility for identifying amyloid in biofilms. Congo red-induced birefringence is a more specific indicator of amyloid formation and differentiates biofilms formed by wild-type S. mutans from a triple ΔP1/WapA/Smu&#95;63c mutant with reduced biofilm forming capabilities. Amyloid accumulation is a late event, appearing in older S. mutans biofilms after 60 hours of growth. Amyloid derived from pure preparations of all three proteins is visualized by electron microscopy as mat-like structures. Typical amyloid fibers become evident following protease digestion to eliminate non-specific aggregates and monomers. Amyloid mats, similar in appearance to those reported in S. mutans biofilm extracellular matrices, are reconstituted by co-incubation of monomers and amyloid fibers. X-ray fiber diffraction of amyloid mats and fibers from all three proteins demonstrate patterns reflective of a cross-β amyloid structure.

PitA is the putative tip adhesin of the pilus islet 2 (PI-2)-encoded sortase-dependent pilus in the Gram-positive Streptococcus oralis, an opportunistic pathogen that often flourishes within the diseased human oral cavity. Early colonization by S. oralis and its interaction with Actinomyces oris seeds the development of oral biofilm or dental plaque. Here, the PI-2 pilus plays a vital role in mediating adherence to host surfaces and other bacteria. A recombinant form of the PitA adhesin has now been produced and crystallized. Owing to the large size (∼100 kDa), flexibility and complicated folding of PitA, obtaining diffraction-quality crystals has been a challenge. However, by the use of limited proteolysis with α-chymotrypsin, the diffraction quality of the PitA crystals was considerably enhanced to 2.16 Å resolution. These crystals belonged to space group P1, with unit-cell parameters a = 61.48, b = 70.87, c = 82.46 Å, α = 80.08, β = 87.02, γ = 87.70°. The anomalous signal from the terbium derivative of α-chymotrypsin-treated PitA crystals prepared with terbium crystallophore (Tb-Xo4) was sufficient to obtain an interpretable electron-density map via terbium SAD phasing.

Context: The comparison of fluoride levels in saliva and plaque following the use of conventional, 2800 and 5000 ppm dentifrices for different time intervals up to 24 hours has not been explored., Aim: The aim of the present study was to assess salivary and plaque fluoride levels at different time intervals following the use of high fluoride dentifrices., Study Design: This randomised control trial was conducted on sixty adolescents between the age group of 16 and 18 years divided into three groups A, B, and C., Intervention: Subjects were asked to brush the occlusal surfaces of posterior teeth with one of the dentifrices for 2 min. Unstimulated whole saliva and plaque samples were collected at different time intervals. Fluoride levels were determined using SPADNS method. Data was collected and statistically analyzed using SPSS 20 Inc. by mixed repeated measure ANOVA., Results: A significant difference in fluoride levels was observed at different time intervals in both saliva (p=0.048) and plaque (p=0.03). The variance was low with time and concentration of the dentifrice used in saliva, whereas the magnitude of treatment was large for time (>0.25) but small for (<0.09) concentration in plaque., Conclusion: A significant difference in fluoride levels was observed at different time intervals in both saliva and plaque among the three groups. There was a positive correlation between fluoride levels in saliva and plaque. Both high fluoride dentifrices were effective in maintaining higher intraoral fluoride levels up to 12 hours and 5000 ppm up to 24 hours compared to conventional dentifrice.

Aim: To clinically, biochemically, and microbiologically evaluate the influence of crown margins position on one-stage laser-microgrooved implants., Materials and Methods: Twenty-one-stage titanium implants with a laser-microgrooved collar surface, supporting screwed, single crown restorations, were placed in 20 partially edentulous patients and evaluated. Clinical parameters included modified plaque index, modified gingival index, peri-implant probing pocket depth, bleeding on probing, and distance between implant shoulder and mucosal margin. The parameters were recorded at baseline (crowns delivery) and at every 6-month recall visit, until the end of the 3 years follow-up period. At the same time intervals, radiographic marginal bone levels were assessed at the mesial and distal aspect of the implant sites. For biochemical analysis, the volume of the peri-implant sulcus fluid, and its levels of interleukin-1beta (IL-1β), interleukin-6 (IL-6), and of tumor necrosis factor-α, were utilized to evaluate the peri-implant health conditions at the end of the 3-year follow-up period. At the same time, microbiological analysis, including the concentration of five putative periodontal pathogens (Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, Treponema denticola, and Tannerella forsythensis), were assessed. The crown margins positions were classified into four groups (A = intracrevicular position >2 mm, B = intracrevicular position ≤2 mm/<1 mm, C = intracrevicular position ≤1 mm/<0 mm, and D = extracrevicular position), and the biochemical, and microbiological parameters were evaluated at 3 years., Results: No statistical differences of clinical and biochemical parameters were found between the four groups. In group A, compared to groups B, C, and D, a statistically significant higher concentration of putative periodontal pathogens was found., Conclusions: Results showed that the intracrevicular deeper position of the restoration margin does not influence the clinical and biochemical peri-implant parameters. The deeper position of the crown margin is associated with a greater amount of putative periodontal pathogenic microflora colonization., (© 2018 Wiley Periodicals, Inc.)

Through a correlative analytical approach encompassing backscattered electron scanning electron microscopy (BSE-SEM), energy dispersive X-ray spectroscopy (EDX), and micro-Raman spectroscopy, the composition of the mineralized biofilm around a dental implant, retrieved due to peri-implantitis, was investigated. The mineralized biofilm contains two morphologically distinct regions: (i) bacteria-containing zones (Bact+), characterized by aggregations of unmineralized and mineralized bacteria, and intermicrobial mineralization, and (ii) bacteria-free zones (Bact-), comprised mainly of randomly oriented mineral platelets. Intramicrobial mineralization, within Bact+, appears as smooth, solid mineral deposits resembling the morphologies of dental plaque bacteria. Bact- is associated with micrometer-sized Mg-rich mineral nodules. The Ca/P ratio of Bact+ is higher than Bact-. The inorganic phase of Bact+ is carbonated apatite (CHAp), while that of Bact- is predominantly octacalcium phosphate (OCP) and whitlockite (WL) inclusions. Compared with native bone, the inorganic phase of Bact+ (i.e., CHAp) exhibits higher mineral crystallinity, lower carbonate content, and lower Ca/P, C/Ca, Mg/Ca, and Mg/P ratios. The various CaPs found within the mineralized dental biofilm (CHAp, OCP, and WL) are related to the local presence/absence of bacteria. In combination with BSE-SEM and EDX, micro-Raman spectroscopy is a valuable analytical tool for nondestructive investigation of mineralized dental biofilm composition and development.

Background: Recent data support the implication of accelerated titanium dissolution products in peri-implantitis. It is unknown whether these dissolution products have an effect on the peri-implant microbiome, the target of existing peri-implantitis therapies., Purpose: This study assessed the relationship between the peri-implant microbiome, dissolved titanium levels, and peri-implantitis., Materials and Methods: Clinical, microbiome, and titanium data were collected from a periodontal population having implants in function for 10 years. Clinical examinations were performed, and submucosal plaque samples were collected from the deepest site per implant. An aliquot of the sample was used for 16S rRNA gene sequencing, with the remainder analyzed for titanium quantity using mass spectrometry. Sequences were clustered into taxonomic units at 97% minimum sequence similarity using the QIIME pipeline approach., Results: Fifteen implants were assessed. According to established case definitions, six had a diagnosis of peri-implantitis; nine were healthy. The genera Streptococcus, Prevotella and Haemophilus characterized peri-implant health. Peri-implantitis was associated with a marked increase in Veillonella. Quantities of dissolved titanium were identified in 40% of sites. Titanium presence was associated with peri-implant disease status (P = .02) and correlated to the first principal component of the microbiome (rho = 0.552) and its alpha-diversity (rho = -0.496). Canonical correlation analyses found that titanium levels, but not health or disease status of the implant, were significantly associated with the microbiota composition (P = .045)., Conclusions: These findings suggest an association between titanium dissolution products and peri-implantitis and support a role for these products in modifying the peri-implant microbiome structure and diversity., (© 2018 Wiley Periodicals, Inc.)

Background: To date, there has been no study identified that assessed peri-implant parameters and local levels of proinflammatory cytokines between waterpipe smokers (WS) and electronic cigarette users (VS). To evaluate the true effect of the risk indicator (WS and VS), a positive control (cigarette smokers [CS]) was added in the study design., Purpose: To compare clinical and radiographic peri-implant parameters and proinflammatory cytokine profile in the peri-implant sulcular fluid (PISF) among CS, WS, VS, and nonsmokers (NS)., Materials and Methods: Forty CS, WS, VS, and NS were included. Details regarding demographics, duration of implants, and habits were recorded. Clinical examination for periodontal and peri-implant plaque index (PI), bleeding on probing (BOP), and probing depth (PD ≥ 4 mm) were assessed. Radiographic bone loss (RBL) was assessed using digital periapical radiographs and viewed on a calibrated computer screen using an image analyzer. Levels of tumor necrosis factor-alpha (TNF-α), interleukin (IL)-6, and IL-1β in PISF were evaluated using enzyme-linked immunosorbent assay., Results: Mean peri-implant PI (P < .05), PD ≥ 4 mm (P < .05), and total RBL (P < .01) was significantly higher among CS, WS, and VS compared with NS. Statistical differences in BOP were observed in NS (P < .01) compared to CS, WS, and VS. CS and WS showed significantly higher PD ≥ 4 mm and RBL compared with VS (P < .05). Levels of TNF-α, IL-6, and IL-1β were significantly higher in CS, WS, and VS compared to NS. There were no statistical differences in the mean levels of all proinflammatory cytokines among individuals in CS and WS., Conclusion: Tobacco smoking is associated with poor peri-implant health. The effect of waterpipe smoking was of largely the same extent as that of cigarette smoking. Higher levels of inflammatory cytokines in tobacco smokers may propose increased peri-implant inflammatory process which could play a principal role in the progression of peri-implant tissue damage., (© 2018 Wiley Periodicals, Inc.)

Objectives: The objective of this study was to evaluate the association between salivary and plaque arginine levels/ADS activities with dental caries., Materials and Methods: A systematic search was performed as per PRISMA statement using PubMed, Scopus, Cochrane Library, and Web of Science. Published studies that investigated adults and children (P) with caries-active status (E) and caries-free status (C), whereby arginine levels/ADS activity (O) was measured in saliva/plaque to analyze exposure-outcome association compared to the control group were deemed eligible for inclusion. Quality assessment was performed using combined Newcastle-Ottawa Scale and Modified RTI Item Bank scale. Meta-analysis was performed for effect size, precision estimation, and subgroup effects analysis., Results: Of 233 records identified, seven (κ = 1.00) were included for qualitative synthesis (systematic review) and four for quantitative synthesis (meta-analysis). No specific bias could be identified in five studies assessed as per the Modified RTI Item Bank scale. Two studies received lower scores on the Newcastle Ottawa scale. Plaque ADS activity in adults (effect size = 0.93, p = 0.008), salivary ADS activity in adults and children (effect size = 0.85, p < 0.001), and salivary ADS activity in adults (effect size = 0.87, p < 0.001) identified a statistically significant effect size. Subgroup analysis demonstrated non-significant variance (Q value = 0.042, p = 0.838) between saliva and plaque ADS activities of adults., Conclusions: The results of this review suggest the salivary and plaque ADS activities appear to be promising caries risk indicators for adults, while results remain inconclusive in children., Clinical Relevance: Measuring ADS activities (saliva or plaque) can be a potential caries risk indicator in adults. The protocol was registered on PROSPERO database: CRD42017060701.

Alternative matrices play a major role in postmortem forensic toxicology, especially if common matrices (like body fluids or hair) are not available. Incorporation of illicit and medicinal drugs into non-mineralized dental biofilm (plaque) seems likely but has not been investigated so far. Analysis of plaque could therefore extend the spectrum of potentially used matrices in postmortem toxicology. For this reason, a rapid, simple and sensitive method for the extraction, determination and quantification of ten drugs of abuse from plaque using liquid chromatography-tandem mass spectrometry (LC-MS/MS) was developed and fully validated. Amphetamine, methamphetamine, 3,4-methylenedioxymethamphetamine (MDMA), 3,4-methylenedioxy-N-ethylamphetamine (MDEA), 3,4-methylenedioxyamphetamine (MDA), cocaine, benzoylecgonine, morphine, codeine and 6-acetylmorphine were extracted from 2mg of dried and powdered plaque via ultrasonication with acetonitrile. The extracts were analyzed on a triple-quadrupole linear ion trap mass spectrometer in scheduled multiple reaction monitoring mode (sMRM). The method was fully validated and proved accurate, precise, selective and specific with satisfactory linearity within the calibrated ranges. The lower limit of quantification was 10-15pgmg -1 for all compounds except for MDA (100pgmg -1 ) and amphetamine (200pgmg -1 ). The method has been successfully applied to three authentic postmortem samples with known drug history. Amphetamine, MDMA, cocaine, benzoylecgonine, morphine and codeine could be detected in these cases in concentrations ranging from 18pgmg -1 for cocaine to 1400pgmg -1 for amphetamine., (Copyright © 2017 Elsevier B.V. All rights reserved.)

There is limited evidence that arginine-containing fluoridated dentifrices (AFD) have a better anticaries effect than regular fluoridated dentifrices (FD), especially in subjects at a higher risk for caries development. This study aimed to assess the effect of AFD on enamel demineralization and on the microbial and biochemical compositions of biofilm formed under different frequencies of sucrose exposure. It consisted of an in situ split-mouth design, where 12 adult volunteers who used FD for at least 2 months prior to the beginning of this study wore acrylic palatal appliances containing 4 bovine enamel specimens (1 pair at each side of the appliance) during 2 phases of 14 days each. FD slurry (3×/day) and 20% sucrose solution (4× and 8×/day) were dripped on the specimens during the first experimental phase. The same volunteers then used AFD during a 2-month washout period, followed by a second experimental phase where the AFD slurry and sucrose solution were applied onto a new subset of specimens. The percentage of enamel surface hardness loss (%SHL), the lesion depth (LD), the integrated mineral loss (IML), microbial counts on biofilms, the biomass, and inorganic and insoluble extracellular polysaccharide (IEPS) biofilm concentrations were determined. Higher %SHL, biomass, and IEPS and lower fluoride values were found at sucrose 8×/day exposure. Lower IEPS were found in the presence of AFD compared to FD. Similar %SHL, LD, and IML values were found between FD and AFD, irrespectively of the cariogenic challenge. The results suggest that AFD have an anticaries effect similar to that of regular FD., (© 2018 S. Karger AG, Basel.)

Background: The pH strip method has been proposed for use in a clinical setting for the evaluation of dental plaque pH. The aim of this in-vivo study was to compare the reliability of the pH-indicator strip method to the microelectrode technique in plaque pH measurement., Methods: 136 subjects (7-9 years) were enrolled and, for each subject, measurements of plaque pH were made at two interproximal sites; a total of 272 sites were analyzed. Plaque pH was assessed in triplicate using pH-indicator strips (pH range: 4.0-7.0) and an iridium microelectrode with a diameter of 0.1 mm. The caries status of 544 proximal surfaces between two primary molars, in proximity at the plaque pH sites, was assessed. All measurements were performed before and 2, 5, 10, 15, 20, and 30 minutes after a mouth rinse with water solution of sucrose (10%). One-way ANOVA was performed to analyze statistically significant differences between the two techniques. Instrument reliability was measured by assessing the intraclass correlation coefficient., Results: Comparable, not statistically significant pH values (P values range from 0.98 to 1.00) were obtained by the two methods for AUC5.7, AUC6.2, maximum pH fall and minimum pH. High intraclass coefficients were recorded (Pearson's r=0.96, Yule's Q=0.99), implying an almost perfect association when the measurements were grouped as a carious lesion being present or not on the surface near the site of measurement., Conclusions: The use of the pH strip method has high validity and may replace the traditional microtouch electrode technique for clinical use.

Background: An ability to predict the response to conventional non-surgical treatment of a periodontal site would be advantageous. However, biomarkers or tests devised to achieve this have lacked sensitivity. The aim of this study is to assess the ability of a novel combination of biomarkers to predict treatment outcome of patients with chronic periodontitis., Methods: Gingival crevicular fluid (GCF) and subgingival plaque were collected from 77 patients at three representative sites, one healthy (probing depth [PD] ≤3 mm) and two diseased (PD ≥6 mm), at baseline and at 3 and 6 months after treatment. Patients received standard non-surgical periodontal treatment at each time point as appropriate. The outcome measure was improvement in probing depth of ≥2 mm. Concentrations of active enzymes (matrix metalloproteinase [MMP]-8, elastase, and sialidase) in GCF and subgingival plaque levels of Porphyromonas gingivalis, Tannerella forsythia, and Fusobacterium nucleatum were analyzed for prediction of the outcome measure., Results: Using threshold values of MMP-8 (94 ng/μL), elastase (33 ng/μL), sialidase (23 ng/μL), and levels of P. gingivalis (0.23%) and T. forsythia (0.35%), receiver operating characteristic curves analysis demonstrated that these biomarkers at baseline could differentiate healthy from diseased sites (sensitivity and specificity ≥77%). Furthermore, logistic regression showed that this combination of these biomarkers at baseline provided accurate predictions of treatment outcome (≥92%)., Conclusion: The "fingerprint" of GCF enzymes and bacteria described here offers a way to predict the outcome of non-surgical periodontal treatment on a site-specific basis.

Objectives: To compare the fluoridating potential of selected European toothpastes using a combination of enamel, dentin, and plaque in vitromodels., Methods: Four in vitromodels were included: 1) Enamel Fluoride (F) Uptake (EFU); 2) Dentin F Uptake (DFU); 3) Enamel Solubility Reduction (ESR); and 4) Plaque F Uptake (PFU). A core set of marketed products was included in all studies, plus a standard toothpaste (1100 ppm F as NaF/silica) and placebo control (the PFU study did not include a placebo control). Test dentifrices: [A] Fluocaril® Bi-Fluoré 250 (1500 ppm F as NaF+1000 ppm F as SMFP); [B] LacerAnticaries (2500 ppm F as SMFP); [C] Elmex® Caries Professional™ (1450 ppm F as SMFP+1.5% arginine); [D] Colgate® Triple Action (1450 ppm F as SMFP); [E] Placebo (0 ppm F); and [F] standard toothpaste (1100 ppm as NaF/silica). In all studies (EFU, DFU, ESR, and PFU), assessments were compared for each pair using the Tukey-Kramer HSD test (p < 0.05)., Results: In all studies of fluoride uptake, the Fluocaril dentifrice [A] provided the greatest numerical benefit, regardless of the substrate. Statistical groupings were EFU: A > F = B = C = D > E; DFU: A > F = B = C = D = E; PFU: A = B > F = C = D). In demineralization prevention, the Fluocaril dentifrice [A] also provided the greatest benefit (ESR: A = F = C = B = D > E). In all studies that included a placebo control, all of the F-containing dentifrices performed better than the placebo control., Conclusions: While these results demonstrate that all of the marketed products tested provide effective anticaries benefits, the Fluocaril Bi-Fluoré 250 dentifrice consistently delivered unsurpassed performance. It delivered the highest level of F to plaque, provided greater measures of efficacy in both remineralization and inhibition of demineralization, and delivered substantial improvement in fluoridation of dentin, suggesting the potential for delivering both coronal and root caries benefits., Competing Interests: All authors are employees of Procter & Gamble.

Background: Peri-implantitis represents a disruption of the biocompatible interface between the titanium dioxide layer of the implant surface and the peri-implant tissues. Increasing preclinical data suggest that peri-implantitis microbiota not only triggers an inflammatory immune response but also causes electrochemical alterations of the titanium surfaces, i.e., corrosion, that aggravate this inflammatory response. Thus, it was hypothesized that there is an association between dissolution of titanium from dental implants, which suggests corrosion, and peri-implantitis in humans. The objective of this study is to compare levels of dissolved titanium in submucosal plaque collected from healthy implants and implants with peri-implantitis., Methods: Submucosal plaque from 20 implants with peri-implantitis and 20 healthy implants was collected with sterile curets from 30 participants. Levels of titanium were quantified using inductively coupled plasma mass spectrometry and normalized for mass of bacterial DNA per sample to exclude confounding by varying amounts of plaque per site. Statistical analysis was performed using generalized estimated equations to adjust for clustering of implants per participant., Results: Implants with peri-implantitis harbored significantly higher mean levels of titanium (0.85 ± 2.47) versus healthy implants (0.07 ± 0.19) after adjusting for amount of plaque collected per site (P = 0.033)., Conclusions: Greater levels of dissolved titanium were detected in submucosal plaque around implants with peri-implantitis compared with healthy implants, indicating an association between titanium dissolution and peri-implantitis. Factors triggering titanium dissolution, as well as the role of titanium corrosion in the peri-implant inflammatory process, warrant further investigation.

Objective: Caries risk assessment is an important component of clinical practice. The goal of this project was to evaluate the diagnostic performance of two commercially available kits (the Saliva-Check mutans and the Plaque-Check pH kit; GC-America) in distinguishing caries-active from caries-free individuals. The effect of following instructions not to eat, drink, or perform oral hygiene, prior to testing, was also investigated on the effectiveness of these kits., Method and Materials: The subjects included 26 caries-affected children and 20 caries-free adults. Plaque and saliva samples were collected for analysis by the two kits following the manufacturer's instructions. For 63 additional subjects, instructions not to eat, drink, or practice oral hygiene were omitted prior to sample collection. The sensitivity and specificity in distinguishing caries-free from caries-active subjects was calculated for both kits., Results: The sensitivity (88%) and specificity (90%) of the Saliva-Check mutans kits were satisfactory in the subjects refraining from eating etc. In contrast, both the sensitivity and the specificity of this kit were reduced when subjects were tested without use instructions. When tested in subjects that followed instructions, the pH kit's sensitivity and specificity were 72% and 55% respectively. This modest diagnostic performance was reduced when pH values were measured in subjects not following use instructions., Conclusion: These kits, particularly the Saliva-Check mutans kits, can contribute to patient education by documenting microbial caries risk factors. Though difficult to implement in practice, subjects should refrain from eating or other activities that disturb the oral environment prior to sample collection.

Objectives: There is some evidence that water rinsing immediately after topical fluoride therapy has the potential to reduce the effectiveness of fluoride. The aim was to determine if covering fluoridated teeth with a layer of mousse containing CPP-ACP could prevent the adverse effect of rinsing on fluoride and consequently its buffering effect on dental plaque pH during cariogenic challenge., Study Design: This randomized, controlled, crossover, in situ study was conducted on 25 participants. The participants were subjected to acidulated phosphate fluoride (APF) application followed by five treatment protocols: (1) water rinsing after 30 minutes (APF-30) or (2) immediate water rinsing (APF-0); (3) using CPP-ACP immediately before water rinsing (F-CPP-ACP); and two control groups: (4) no fluoride therapy (No-F) and (5) using CPP-ACP and immediate water rinsing (CPP-ACP-0). After 48 hours, teeth were rinsed with 10% sucrose solution and plaque pH was measured before and after 5, 10, 15, 20 and 30 minutes., Results: The least pH changes, the lowest pH drop, and the quickest pH recovery were found in the APF-30 and F-CPP-ACP groups. APF-0 ranked in the middle and the highest values were in the control groups., Conclusions: The results show that in the case using CPP-ACP on fluoridated teeth, water rinsing immediately after topical fluoride therapy did not seem to influence the inhibitory effect of fluoride on plaque acidity.

Fetuin-A is a potent inhibitor of calcium-phosphate precipitation and of the calcification process, therefore it can also be related with dental calculus. Thus, we aimed to investigate a possible relationship between fetuin-A gene polymorphism and the presence of dental calculus. A possible relationship between serum, saliva and gingival crevicular fluid (GCF) levels of fetuin-A was also investigated. Fetuin-A c.742C > T and c.766C > G polymorphisms were investigated in 103 patients with or without dental calculus. Additionally, serum, saliva and GCF fetuin-A levels of patients were compared according to dental calculus presence. A significant difference was not observed in the distribution of the fetuin-A c.742C > T and c.766C > G polymorphisms between patients with or without dental calculus. Saliva and GCF fetuin-A concentrations of patients with dental calculus were statistically higher than those without dental calculus (P=0.001, P=0.036 respectively). According to our results, fetuin-A c.742C > T and c.766C > G polymorphisms were not associated with presence of dental calculus. However, higher GCF and saliva fetuin-A levels were detected in patients with dental calculus than in patients without dental calculus, which may result from an adaptive mechanism to inhibit mineral precipitation and eventually calculus formation.

Objective: To determine the effect of the mode of drinking fluoridated milk on salivary and plaque fluoride concentrations., Methods: Fluoridated milk was ingested by 32 children in three ways: (a) directly from the container (1.0 and 5.0 mg F/litre), (b) through a straw with the tip between the lips (5.0 mg F/litre), and (c) with the tip deep in the oral cavity (5.0 mg F/litre). Saliva was collected at baseline and 2, 15, and 40 min and plaque at baseline and 20 min after drinking. Fluoride concentrations were determined using the electrode after HMDS-facilitated diffusion., Results: The mode of drinking did not affect fluoride concentrations in saliva or plaque. The average 2-min salivary concentrations were 65 ng F/mL for the 1.0 mg F/litre group and 276 ng F/mL for the three 5.0 mg F/litre groups (P < 0.01). The average of the 15- and 40-min salivary concentrations was 22 ng F/mL for the 1.0 mg F/litre group and 41 ng F/mL for the 5.0 mg F/litre groups (P < 0.01). Plaque concentrations showed the same patterns as in saliva, that is, they were higher in the three 5.0 mg F/litre groups than in the 1.0 mg F/litre group and the differences among the 5.0 mg F/litre groups were not statistically significant., Conclusion: Salivary and plaque fluoride concentrations were independent of the mode of drinking but directly related to milk fluoride concentrations., (© 2015 BSPD, IAPD and John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

This review will discuss the importance of Zinc in the maintenance of oral health. Zinc (Zn) is a trace element of valuable importance. In the oral cavity, it is naturally present at various sites such as dental plaque, dental hard tissues and saliva. It is proven to be effective against common prevalent oral health problems such as dental caries, gingivitis, periodontitis and malodour. It is being used in various oral health care products to control the formation of dental plaque and inhibiting the formation of dental calculus. It has the potential to sustain and maintain its elevated concentrations for a longer time particularly in the dental plaque and saliva on delivery from the mouth rinses and toothpastes. It has been reported that low concentrations of zinc have the capability to reduce dissolution and promote remineralization under caries simulating conditions. Most importantly low Zn2+ levels in the serum are useful as a tumour marker. Thus taking a note of its potentials, it can be concluded that zinc is a precious element for the maintenance of oral health.

The aim of this study was to determine whether the degree of tongue and interdental plaque can be used to assess oral malodor by quantifying their fluorescence as detected using quantitative light-induced fluorescence (QLF) technology. Ninety-nine subjects who complained of oral malodor were included. The level of oral malodor was quantified using the organoleptic score (OLS) and the concentration of volatile sulfur compounds (VSCs). The fluorescence properties of tongue and interdental plaque were quantified as scores calculated by multiplying the intensity and area of fluorescence in QLF-digital images, and the combined plaque fluorescence (CPF) score was obtained by summing the scores for the two regions. The associations of the scores with malodor levels and the diagnostic accuracy of the CPF score were analyzed. The two plaque fluorescence scores and their combined score differed significantly with the level of oral malodor (p<0.001). The CPF score was moderately correlated with OLS (r=0.64) and VSC levels (r=0.54), and its area under the receiver operating characteristic curve was 0.77 for identifying subjects with definite oral malodor (OLS≥2). In conclusion, plaque fluorescence from tongue and interdental sites as detected using QLF technology can be used to assess the level of oral malodor.

Background: This study explores the association of disabilities and oral health. The aim of the study was to assess the salivary and plaque pH and oral health status of children with and without disabilities., Materials and Methods: A total of 100 schoolchildren (50 with disabilities and 50 without disabilities) were examined from 9 to 15 years age group. Saliva and plaque pH analysis were done to both the groups. Clinical data were collected on periodontal status, dental caries using WHO criteria. pH values of different groups, difference between the means were calculated using independent t-test, and frequency distribution was analyzed using Chi-square test. Statistical significance, P value was set at 0.05., Results: Mean plaque and salivary pH scores were lesser (5.73 and 5.67) in children with intellectual disabilities (IDs) (P< 0.001). Subjects with disabilities had also statistically significant higher CPI scores and decayed, missing, and filled scores than their healthy counterparts (P< 0.001)., Conclusion: There is a statistically significant difference in plaque and salivary pH among children with and without ID with lower plaque and salivary pH among children with ID. In addition to this, the oral health was also more compromised in children with ID, which confirms a need for preventive treatment for these children.

Cavitation in teeth results from a pathogenic process termed dental caries that has occurred on the tooth surface for weeks or even years. Accumulation of dental plaque (biofilm) on the tooth is usually the first manifestation of the disease. Although acid production is the immediate and proximal cause of dissolution of teeth; it is the milieu within which the acid is formed that should be of primary concern. Focusing on the 'critical pH' has detracted attention from the more biological aspects (biofilm formation) of dental caries. Dental caries is unique; it is a biological process occurring on essentially an inert surface. Investigation of the multitude of interactions occurring in plaque ranging from enamel interfaces to surfaces of bacteria and matrices poses challenges worthy of the best scientific minds. The mouth clearly offers unique opportunities to investigate the multi facets of biofilm formation in vivo, generating data that have relevance way beyond the mouth. Prevention of this ubiquitous disease, dental caries, continues to present serious challenges. The public health benefits of fluoride delivered in its various formats are well recognized. Nevertheless, additional preventive approaches are required. Overcoming the rapid clearance of agents from the mouth is particularly challenging. Building on the polymerizing capacity of glucosyltransferases it may be possible to incorporate a therapeutic agent into the matrix plaque, thereby delivering therapeutic agents precisely to where they are needed., (© 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Aim: To systematically evaluate the cariogenic potential of various commercially available infant formulas., Materials and Methods: A literature search was conducted using Pubmed and Scopus databases for articles published between 1966 and November 2014. Reference lists of all eligible studies were searched. Only human studies were included. Data extraction and risk of bias assessments were performed., Results: Seven of the 83 articles identified were included in this review, of which six studies employed plaque harvesting methods, while one study utilised an intra-oral cariogenicity/in situ model. Three studies compared milk-based formulas (MBFs) and soy-based formulas (SBFs), two compared protein hydrolysate formulas (PHFs) with MBFs and SBFs, four compared formulas with various types of sugar, and two studies compared formulas with varying casein content. Based on a single study, SBFs were significantly more cariogenic than MBFs. Formulas containing only non-milk extrinsic sugars (NMES) and those containing lactose + NMES were found to be significantly more cariogenic than formulas containing only lactose. No significant correlation was found between cariogenicity and casein content in infant formula. The results of studies comparing PHFs with MBFs and SBFs were contradictory. Risk of bias assessment revealed that five studies were at moderate risk of bias, and two were assessed to be at high risk of bias., Conclusion: The result for cariogenicity of various types of infant formulas remains inconclusive, thus no concrete recommendations can be made. Further well-designed studies are needed to clarify the effect of casein content on cariogenicity.

Gingival recession is a common clinical problem that results in esthetic deficiencies and poor plaque control and predominantly occurs in aged patients. In order to restore the cervical region, ideal biomaterials should possess the ability to stimulate proliferation and osteogenesis/cementogenesis of human gingival fibroblasts (HGF) and have a strong antibiofilm effect. The aim of the present study was to investigate the interactions of HGF and oral multispecies biofilms with Ca, Mg and Si-containing bredigite (BRT, Ca7MgSi4O16) bioceramics. BRT extract induced osteogenic/cementogenic differentiation of HGF and its inhibition of plaque biofilm formation were systematically studied. BRT extract in concentrations lower than <200 mg mL(-1) presented high biocompatibility to HGF cells in 3 days. Ion extracts from BRT also stimulated a series of bone-related gene and protein expressions in HGF cells. Furthermore, BRT extract significantly inhibited oral multispecies plaque biofilm growth on its surface and contributed to over 30% bacterial cell death without additional antibacterial agents in two weeks. A planktonic killing test showed that BRT suppressed 98% plaque bacterial growth compared to blank control in 3 days. The results also revealed that BRT extract has an osteostimulation effect on HGF. The suppression effect on plaque biofilms suggested that BRT might be used as a bioactive material for cervical restoration and that the synergistic effect of bioactive ions, such as Ca, Mg and Si ions, played an important role in the design and construction of bifunctional biomaterials in combination with tissue regeneration and antibiofilm activity.

Background: The etiology of dental caries is multifactorial, but frequent consumption of free sugars, notably sucrose, appears to be a major factor driving the supragingival microbiota in the direction of dysbiosis. Recent 16S rRNA-based studies indicated that caries-associated communities were less diverse than healthy supragingival plaque but still displayed considerable taxonomic diversity between individuals. Metagenomic studies likewise have found that healthy oral sites from different people were broadly similar with respect to gene function, even though there was an extensive individual variation in their taxonomic profiles. That pattern may also extend to dysbiotic communities. In that case, shifts in community-wide protein relative abundance might provide better biomarkers of dysbiosis that can be achieved through taxonomy alone., Results: In this study, we used a paired oral microcosm biofilm model of dental caries to investigate differences in community composition and protein relative abundance in the presence and absence of sucrose. This approach provided large quantities of protein, which facilitated deep metaproteomic analysis. Community composition was evaluated using 16S rRNA sequencing and metaproteomic approaches. Although taxonomic diversity was reduced by sucrose pulsing, considerable inter-subject variation in community composition remained. By contrast, functional analysis using the SEED ontology found that sucrose induced changes in protein relative abundance patterns for pathways involving glycolysis, lactate production, aciduricity, and ammonia/glutamate metabolism that were conserved across taxonomically diverse dysbiotic oral microcosm biofilm communities., Conclusions: Our findings support the concept of using function-based changes in protein relative abundance as indicators of dysbiosis. Our microcosm model cannot replicate all aspects of the oral environment, but the deep level of metaproteomic analysis it allows makes it suitable for discovering which proteins are most consistently abundant during dysbiosis. It then may be possible to define biomarkers that could be used to detect at-risk tooth surfaces before the development of overt carious lesions.

Objectives: To investigate the neutralising effects of subsequent urea rinse on sucrose-induced decrease in plaque pH with interdental plaque pH telemetry., Method: Six participants wearing partial lower prostheses which incorporated a miniature glass pH electrode were included. After 5 or 6 days of plaque accumulation on the tip of the electrode, the subjects rinsed with a 15 ml 10% sucrose solution, followed by no subsequent rinsing or rinsing with either 15 ml of water, 0.25%, 0.50% or 1.00% urea solution, for 2 min. The plaque pH was continuously recorded for 120 min., Results: Without subsequent rinsing, the plaque pH decreased at 10 min to 4.39 and stayed below the critical pH of 5.7 for 80 min following a sucrose rinse. Subsequent water rinse showed little effects on the sucrose-induced decrease in plaque pH, whereas subsequent urea rinses all immediately and effectively neutralised sucrose-induced decrease in plaque pH, and remained above the critical pH of 5.7 until the end of data collection., Conclusion: Urea rinse could effectively counteract the pH fall following sucrose administration. These results strongly suggested that the regular use of low-concentration urea rinse after carbohydrate consumption may help prevent caries.

Objectives: This study aimed to evaluate saliva and plaque as indicators of intraoral fluoride (F) levels after the use of conventional and high-fluoride dentifrices., Materials and Methods: Subjects were randomly assigned to brush their teeth with conventional (1000 ppm F), high-fluoride (5000 ppm F), and placebo dentifrices (fluoride free) for 10 days, following a double-blind, crossover protocol. Saliva and plaque samples were collected on the morning of the 5(th) and 10th days, respectively at 1 and 12 h after brushing, and analyzed with an ion-selective electrode after HMDS-facilitated diffusion. Data were analyzed by two-way repeated measures ANOVA, Tukey's test and Spearman's correlation coefficient (p < 0.05)., Results: Plaque and salivary F levels were significantly increased after the use of conventional and high-fluoride dentifrices when compared to values obtained for placebo, except plaque 12 h after the use of conventional dentifrice. A positive and significant correlation was found between fluoride concentrations in plaque and saliva for both times of sample collection., Conclusion: Both indicators assessed were able to detect significant differences among treatments and between times after brushing. The use of a high-fluoride dentifrice is able to significantly increase intraoral fluoride levels throughout the day, being therefore a useful therapy for patients at high caries risk., Clinical Relevance: A dentifrice with high fluoride concentration could be regarded as a useful therapy of F delivery for high caries-risk patients, since intraoral F levels were sustained throughout most of the day after using this formulation.

Objective: To assess the efficacy of an experimental 1450 ppm fluoride as sodium monofluorophosphate (SMFP)/zinc-based dentifrice (Test), a clinically proven 1450 ppm fluoride as SMFP/zinc-based dentifrice (Positive Control), and a 1000 ppm fluoride as SMFP and 450 ppm fluoride as sodium fluoride (NaF) dentifrice (Negative Control) in delivering 12-hour anti-tartar benefits as measured by calcium buildup after a single product brushing., Methods: Fourteen (14) adult subjects completed this single-center, double-blind, randomized, cross-over clinical study conducted in Piscataway, New Jersey. An intra-oral appliance was custom-made for each subject as a surface on which the anti-calculus agents' inhibition of early calculus formation could be analyzed. After brushing with their assigned toothpaste, each subject wore their respective appliance for 12 hours overnight. When the appliance was removed, it was washed, suspended in 0.1 MHCL to release Ca2+ from deposits, and analyzed by Inductively Coupled Plasma (ICP) for deposited calcium. There was a one-week washout period between each product use. Statistical analyses were performed on the mean level of calcium (μg). Comparisons of the treatment groups with respect to 12-hour post-brushing (overnight) calcium levels were performed via a two-way analysis of variance (ANOVA), with subjects and products as factors in the ANOVA model. Post-ANOVA pair-wise comparisons of the study treatments were performed using the Tukey's test for multiple comparisons. All statistical tests of hypothesis were two-sided, and employed a level of significance of α = 0.05., Results: At the 12-hour post-brushing examination, the mean level of calcium buildup was 0.46 μg for subjects assigned to the experimental Test dentifrice, 0.41 μg for subjects assigned to the Positive Control dentifrice, and 2.47 μg for subjects assigned to the Negative Control dentifrice. After 12 hours post-brushing, subjects assigned to the experimental Test dentifrice and Positive Control dentifrice both exhibited statistically significantly (p = 0.005 and p = 0.004, respectively) lower levels of calcium buildup as compared to the subjects assigned to the Negative Control dentifrice. No statistically significant (p = 0.996) difference was observed between the experimental Test dentifrice and Positive Control dentifrice with respect to tartar control efficacy after 12 hours post-brushing. No adverse effects of the hard or soft oral tissue were observed by the clinical investigators or reported by the subjects., Conclusion: Both the experimental Test dentifrice (1450 ppm fluoride as SMFP/zinc-based dentifrice) and the Positive Control dentifrice (1450 ppm fluoride as SMFP/zinc-based dentifrice) provided significantly greater 12-hour tartar control as measured by calcium buildup, as compared to the Negative Control dentifrice containing 1000 ppm fluoride as SMFP and 450 ppm fluoride as NaF.

Increasing the concentration of free fluoride in oral fluids is an important goal in the use of topical fluoride agents. Although sodium lauryl sulfate (SLS) is a common dentifrice ingredient, the influence of this ion on plaque fluid and salivary fluid fluoride has not been examined. The purpose of this study was to investigate the effect of SLS on these parameters and to examine the effect of this ion on total (or whole) plaque fluoride, an important source of plaque fluid fluoride after a sufficient interval following fluoride administration, and on total salivary fluoride, a parameter often used as a surrogate measure of salivary fluid fluoride. Ten subjects accumulated plaque for 48 h before rinsing with a 12 mmol/l NaF (228 µg/g F) rinse containing or not containing 0.5% (w/w) SLS. SLS had no statistically significant effect on total plaque and total saliva fluoride but significantly increased salivary fluid and plaque fluid fluoride (by 147 and 205%, respectively). These results suggest that the nonfluoride components of topical agents can be manipulated to improve the fluoride release characteristics from oral fluoride reservoirs and that statistically significant change may be observed in plaque fluid and salivary fluid fluoride concentrations that may not be observed in total plaque and total saliva fluoride concentrations.

The aim of this study was to investigate the effects of the Swedish moist powder tobacco product known as "snus" on dental caries and to measure the pH fall in dental plaque. The subjects comprised male and female adults between 26 and 62 years of age (n = 102), all habitual snus users for ≥ 10 years. The control group (n = 101) consisted of similar individuals in terms of gender, age and educational level but with no tobacco use for ≥ 10 years. A clinical and radiographic examination and a questionnaire were completed. The pH fall after a sucrose rinse was estimated in situ in 10 randomly selected subjects per group. The salivary secretion rate was higher in snus users than non-users (2.5 vs 2.2 ml/min, p = 0.005).There was no statistically significant difference regarding salivary buffer capacity. No differences were found between the two groups in terms of the plaque index, primary or secondary enamel and dentine caries, DFS and salivary counts of mutans streptococci or lactobacilli. The pH fall was somewhat more pronounced among non-users compared with snus users (NS). Snus users had a lower intake of snacks between meals and a less frequent intake of cookies (p = 0.000). Furthermore, snus users had a mean gingival index (± SD) for the whole dentition of 20.4 ± 18.2, while the index for non-users was 14.4 ± 13.9 (p = 0.09); the corresponding values for teeth 13-23 were 14.9 ± 20.6 and 7.7 ± 11.9 respectively (p = 0.003). To conclude, this clinical study revealed no statistically significant differences in caries prevalence between snus users and non-users and only minor differences regarding different caries associated factors.

Objectives: This study investigated the relationship between urease and arginine deiminase system (ADS) activities and dental caries through a cross-sectional study., Material and Methods: Urease and ADS activities were measured in saliva and plaque samples from 10 caries-free subjects and 13 caries-active. Urease activity was obtained from the ammonia produced by incubation of plaque and saliva samples in urea. ADS activity was obtained from the ammonia generated by the arginine-HCl and Tris-maleate buffer. Specific activity was defined as micromoles of ammonia per minute per milligram of protein. Shapiro-Wilk statistical test was used to analyze the distribution of the data, and Mann-Whitney test was used to determine the significance of the data., Results: The specific urease activity in saliva and plaque was significantly higher in individuals with low DMFT scores. ADS activity in saliva (6.050 vs 1.350, p=0.0154) and plaque (8.830 vs 1.210, p=0.025) was also higher in individuals with low DMFT scores., Conclusions: Caries-free subjects had a higher ammonia generation activity by urease and arginine deiminase system for both saliva and plaque samples than low caries-active subjects. High levels of alkali production in oral environment were related to caries-free subjects.

Objective: This study compares pH and microbiological profile of dental plaque in children and adults of a low caries population., Material and Methods: Thirty-nine children, 12-14 years of age and 45 adults between 20 and 39 years of age in 5 Karen villages of the Tak province, Northern Thailand were examined for plaque, calculus, caries (DMFT) and pH measurements in resting plaque and after a sucrose rinse. Information on dietary and oral hygiene habits was obtained through interviews using a fixed questionnaire. Microbiological profile of plaque samples was analyzed with DNA-DNA checkerboard technique., Results: Mean DMFT was 0.77 ± 1.56 and 87% of the adults and 67% of the children were caries free (p < 0.05). The mean resting pH was for both age groups in the range of 7.0-7.1 and significantly higher than a Swedish caries free reference group. Karen adult men had significantly lower pH minimum than females and children (p < 0.05). Supragingival plaque samples showed high levels of low acidogenic and anaerobic species, which dominated over strong acid producers such as streptococci., Conclusion: The study indicates that the Karen children and adults has a plaque physiology and microbiology predominating by low acidogenic anaerobes, which in addition to the low sucrose intake explains the low caries prevalence in this population., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Purpose: To evaluate the clinical effect on plaque metabolism of a dentifrice containing 1.5% arginine, an insoluble calcium compound and 1,450 ppm fluoride compared to a commercially available dentifrice containing 1,450 ppm fluoride in a silica base., Methods: A 12-week, parallel, randomized, double-blind study using 48 subjects was conducted at the Colgate-Palmolive Technology Center (Piscataway, NJ, USA). One group used a test dentifrice containing 1.5% arginine, an insoluble calcium compound and 1,450 ppm fluoride as sodium monofluorophosphate (MFP), and the other group used a commercial silica dentifrice with 1,450 ppm fluoride as sodium fluoride (NaF) as a control. Plaque metabolism analyses were conducted at baseline and after 2, 4, 6, 8, and 12 weeks of assigned product use. The plaque analyses included pH measurements before and after a sucrose rinse, ammonia production and lactic acid production., Results: Subjects using the test dentifrice had significantly higher plaque pH values before (P< or = 0.01) and after (P< or = 0.045) a sucrose challenge than those using the commercially available control dentifrice. Subjects using the test dentifrice also produced higher levels of ammonia and lower levels of lactic acid compared to subjects using the control dentifrice.

Alkali production by oral bacteria via the arginine deiminase system (ADS) increases the pH of oral biofilms and reduces the risk for development of carious lesions. This study tested the hypothesis that increased availability of arginine in the oral environment through an exogenous source enhances the ADS activity levels in saliva and dental plaque. Saliva and supra-gingival plaque samples were collected from 19 caries-free (CF) individuals (DMFT = 0) and 19 caries-active (CA) individuals (DMFT ≥ 2) before and after treatment, which comprised the use of a fluoride-free toothpaste containing 1.5% arginine, or a regular fluoride-containing toothpaste twice daily for 4 weeks. ADS activity was measured by quantification of ammonia produced from arginine by oral samples at baseline, after washout period, 4 weeks of treatment, and 2 weeks post-treatment. Higher ADS activity levels were observed in plaque samples from CF compared to those of CA individuals (P = 0.048) at baseline. The use of the arginine toothpaste significantly increased ADS activity in plaque of CA individuals (P = 0.026). The plaque microbial profiles of CA treated with the arginine toothpaste showed a shift in bacterial composition to a healthier community, more similar to that of CF individuals. Thus, an anti-caries effect may be expected from arginine-containing formulations due in large part to the enhancement of ADS activity levels and potential favorable modification to the composition of the oral microbiome., (© 2013 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

This single-blind, randomized, crossover study aimed at assessing the long-term fluoride concentrations in saliva and in dental biofilm after different home-use fluoride treatments. The study volunteers (n = 38) were residents of an area with fluoridated drinking water. They were administered four treatments, each of which lasted for one week: twice-daily placebo dentifrice, twice-daily fluoride dentifrice, twice-daily fluoride dentifrice and once-daily fluoride mouthrinse, and thrice-daily fluoride dentifrice. At the end of each treatment period, samples of unstimulated saliva and dental biofilm were collected 8 h after the last oral hygiene procedure. Fluoride concentrations in saliva and dental biofilm were analyzed using a specific electrode. The fluoride concentrations in saliva and dental biofilm 8 h after the last use of fluoride products did not differ among treatments. The results of this study suggest that treatments with home-use fluoride products have no long-term effect on fluoride concentrations in saliva and in dental biofilm of residents of an area with a fluoridated water supply.

The purpose of this study is to determine whether a calcium (Ca) prerinse used before a 228 µg/g (ppm) fluoride (F) rinse would induce the formation of 'calcium fluoride-like' (CaF2-like) deposits in human dental plaque. Sixty minutes after the use of the Ca prerinse/F rinse, plaque samples were collected from 10 volunteers, homogenized, and split into 2 aliquots. The plaque mass from one aliquot was then extracted with a 'plaque-like' solution that extracted all the CaF2-like deposits. The total F in both aliquots was then determined and compared. The results demonstrated that, as in previous studies, the Ca prerinse induced large increases in plaque fluid and total plaque F. However, unlike previous results without the Ca prerinse, 30% of the plaque F deposits were CaF2 or CaF2-like. Given that maintaining an elevated F concentration in the vicinity of a developing lesion may play an important role in the cariostatic effect of this ion, and the potential advantages of CaF2-like deposits as an F source, these results suggest that a Ca prerinse may increase the cariostatic effect of topical agents.

The aim of this study was to evaluate the effect of a commercial lactose-containing stevioside sweetener on biofilm acidogenicity in vivo. Nine volunteers refrained from brushing their teeth for 3 days in five phases. On the 4th day of each phase, the pH of the biofilm was measured by the "Strip method". Interproximal plaque pH was measured before and up to 60 minutes after a 10 mL mouthrinse for 1 minute with the test solutions: I - sweetener with 93% lactose and 7% stevioside; II - sweetener with 6.8% saccharin, 13.6% cyclamate, and 0.82% stevioside; III - 18% sucrose solution (positive control); IV - mineral water (negative control); and V- 93% lactose solution. The results revealed that the most pronounced pH fall was found with sucrose (positive control), followed by the 93% lactose solution, the sweetener with lactose + stevioside, the sweetener with saccharin + cyclamate + stevioside, and finally water (negative control). According to the area under the curve, the two sweeteners containing stevioside were significantly different, and the sweetener with lactose + stevioside was significantly different from water but not from sucrose. The critical pH for dentin demineralization (pH ≤ 6.5) was reached by all volunteers after rinsing with sucrose solution, lactose solution, and the stevioside + lactose sweetener. Analysis of the data suggests that lactose-containing stevioside sweeteners may be cariogenic, especially to dentin.

Purpose: This double-blind, crossover study evaluated whole plaque fluoride concentration (F), as well as whole plaque calcium concentrations (Ca) after brushing with a placebo (PD - fluoride free), low-fluoride (LFD, 513 microg F/g) and conventional (CD, 1,072 microg F/g) dentifrices., Methods: Children (n=20) were randomly assigned to brush twice daily with one of the dentifrices, during 7 days. On the 7th day, samples were collected at 1 and 12 hours after brushing. F and Ca were analyzed with an ion-selective electrode and with the Arsenazo III method, respectively. Data were analyzed by ANOVA, Tukey's test and by Pearson correlation coefficient (P< 0.05)., Results: The use of the fluoridated dentifrices significantly increased plaque [F]s 1 hour after brushing when compared to PD, returning to baseline levels 12 hours after. Positive and significant correlations were found between plaque [F] and (Ca) under most of the conditions evaluated. The mean increase in plaque [F] observed 1 hour after brushing with the CD were only about 47% higher than those obtained for the LFD. The use of a LFD promotes proportionally higher increases in plaque [F] when compared to a CD. Plaque F concentrations were also shown to be dependent on plaque Ca concentrations.

Objective: The objective of the present paper is to report results from oral biologic studies carried out in connection with a caries study., Methods: Samples of whole-mouth saliva and dental plaque were collected from initially 7- to 8-year-old subjects who participated in a 3-year school-based programme investigating the effect of the consumption of polyol-containing candies on caries rates. The subjects were randomized in three cohorts, consumed erythritol, xylitol, or sorbitol candies. The daily polyol consumption from the candies was approximately 7.5 g., Results: A significant reduction in dental plaque weight from baseline (p<0.05) occurred in the erythritol group during almost all intervention years while no changes were found in xylitol and sorbitol groups. Usage of polyol candies had no significant or consistent effect on the levels of plaque protein, glucose, glycerol, or calcium, determined yearly in connection with caries examinations. After three years, the plaque of erythritol-receiving subjects contained significantly (p<0.05) lower levels of acetic acid and propionic acid than that of subjects receiving xylitol or sorbitol. Lactic acid levels partly followed the same pattern. The consumption of erythritol was generally associated with significantly (p<0.05) lower counts of salivary and plaque mutans streptococci compared with the other groups. There was no change in salivary Lactobacillus levels., Conclusion: Three-year consumption of erythritol-containing candies by initially 7- to 8-year old children was associated with reduced plaque growth, lower levels of plaque acetic acid and propionic acid, and reduced oral counts of mutans streptococci compared with the consumption of xylitol or sorbitol candies., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Objectives: To compare the effect of 40 seconds versus 2 minutes brushing on saliva and dental biofilm fluid fluoride in children ages 4-5 years over 1 hour., Design: This was a single-blind, cross-over, randomised, two-period clinical study in healthy children. Three days before the start of each treatment subjects received a thorough brushing and then refrained from all oral hygiene procedures. At treatment visits, after collecting baseline biofilm and saliva samples, staff brushed the occlusal surfaces of the subject's posterior teeth with a pea-sized amount (0.25 g) of NaF/silica toothpaste for the randomised time. Samples were taken at 5 minutes, 15 minutes, 30 minutes and 60 minutes after brushing and analysed for fluoride using a microanalytical methodology. There was a minimum 4-day washout period between treatments., Results: Log changes from baseline biofilm fluid and saliva fluoride were statistically significant (P < 0.05) for both brushing times at all post-brushing time-points [except 60 minutes saliva where P = 0.06 (t-test)]. Statistically significantly greater ln-AUC (area under the curve) was found for biofilm fluid and salivary fluoride after brushing for 2 minutes compared with brushing for 40 seconds over the 1-hour test period. There was a statistically significantly higher concentration of fluoride in the log change from baseline saliva levels after 5, 15, 30 and 60 minutes for the 2-minute brushing time compared with 40 seconds brushing time. There was no statistically significant difference in concentration of log change from baseline fluoride levels in biofilm fluid at each individual time-point (5, 15, 30 and 60 minutes) for the 2-minute brushing time compared with the 40-second brushing time, but significant differences were observed for 15, 30 and 60 minutes in favour of 2-minute brushing time when log biofilm fluid value was analysed., Conclusion: The findings provide further evidence for the benefits of increased duration of brushing with respect to fluoride delivery., (© 2013 FDI World Dental Federation.)

Aim: The effects on plaque parameters of sugar free chewing-gums (CG) sweetened with either maltitol or xylitol were assessed to better understand the role polyols can play in dental caries prevention., Materials and Methods: A double-blind, parallel, randomised, controlled study was conducted in China. Subjects (N = 258, age = 13 to 15 years-old) were divided into 4 groups: 2 receiving polyols CG, containing respectively maltitol or xylitol, a group receiving gum base (placebo) and a negative control group not receiving any gum. CG were chewed for 30 days. This corresponds to a 10 g consumption of polyol per day. Plaque parameters (growth, pH, bacteria and insoluble glucans) were evaluated throughout the experimental period., Results: All parameters studied were significantly modified with gum base compared to no-gum: plaque pH increased; plaque growth, bacteria (S. mutans, S. sobrinus, A. viscosus and Lactobacillus) and insoluble glucans decreased. Maltitol and xylitol CG led similarly to a higher plaque pH (AUC, p⋜0.05) on short (at baseline after the first CG consumption) and long term (after 4 weeks of daily CG consumption), with or without saliva stimulation compared to both control and placebo groups. They led to a decrease in plaque growth (p=0.02) over the experimental period compared to controls. Moreover, they significantly reduced the concentration of 4 cariogenic bacteria species (p⋜0.05) in dental plaque compared to gum base., Conclusion: Sugar free CG sweetened with either maltitol or xylitol can similarly reduce plaque acidogenicity compared to gum base through a decrease in oral bacteria presence. The use of a gum base placebo allowed to isolate effects on parameters involved in dental caries development specific to maltitol and xylitol, and to show these effects were similar.

Unlabelled: This article discusses the possible cariostatic mechanisms of the action of fluoride. In the past, fluoride inhibition of caries was ascribed to reduced solubility of enamel due to incorporation of fluoride (F-) into the enamel minerals. The present evidence from clinical and laboratory studies suggests that the caries-preventive mode of action of fluoride is mainly topical. There is convincing evidence that fluoride has a major effect on demineralisation and remineralisation of dental hard tissue. The source of this fluoride could either be fluorapatite (formed due to the incorporation of fluoride into enamel) or calcium fluoride (CaF2)-like precipitates, which are formed on the enamel and in the plaque after application of topical fluoride. Calcium fluoride deposits are protected from rapid dissolution by a phosphate -protein coating of salivary origin. At lower pH, the coating is lost and an increased dissolution rate of calcium fluoride occurs. The CaF2, therefore, act as an efficient source of free fluoride ions during the cariogenic challenge. The current evidence indicates that fluoride has a direct and indirect effect on bacterial cells, although the in vivo implications of this are still not clear., Conclusion: A better understanding of the mechanisms of the action of fluoride is very important for caries prevention and control. The effectiveness of fluoride as a cariostatic agent depends on the availability of free fluoride in plaque during cariogenic challenge, i.e. during acid production. Thus, a constant supply of low levels of fluoride in biofilm/saliva/dental interference is considered the most beneficial in preventing dental caries., (Copyright © 2013 by Academy of Sciences and Arts of Bosnia and Herzegovina.)

Aim: The aim of this study was to investigate oral rehydration therapy (ORT) products and their effect on plaque pH under normal and dry mouth conditions., Design: Three commercial oral rehydration therapy products, prepared according to the manufacturers' instruction, plus a 10% glucose solution, which served as control, were tested in 10 healthy subjects (mean age 25 years): (1) Electral (Arrow Pharmaceuticals), (2) Resorb Junior (Nestlé) and (3) Vätskeersättning (Semper). pH was measured in the maxillary premolar region in situ with the so-called microtouch method. The area under the pH curve was calculated. Their carbohydrate content was also analysed., Results: The oral rehydration therapy products gave the same low pH values in plaque as the glucose solution. Dry mouth condition, obtained by injection of methylscopolamine nitrate in the labial sulcus, resulted in an overall more pronounced fall in pH and in a delay in the pH recovery compared with normal salivary conditions. The chemical analyses showed that all three test-products contained glucose and starch as the main carbohydrates., Conclusions: Rinsing with oral rehydration therapy products leads to a greater pH decrease in dental plaque during low compared with normal salivary conditions. As diarrhoea may result in dehydration and thereby in dry mouth, we believe that ORT products have a high cariogenic potential., (© 2013 FDI World Dental Federation.)

Unlabelled: This research paper primarily focuses on the importance of use of xylitol among school children., Purpose: The purpose of this paper is to evaluate the salivary and dental plaque pH changes after consumption of sugared and sugar-free (xylitol) chewing gums in children., Materials and Methods: A total of 30 school children were selected for this study and were divided into two equal groups and given both chewing gums for the experiment., Results: Children consuming the sugar-free (xylitol) chewing gum showed a marked increase in the pH of saliva and plaque when compared to their counterpart. All these values had a significant difference of P ≤ 0.0001., Conclusion: Xylitol is a safe all-natural sweetener which helps to reduce tooth decay. It plays a unique role in preventive strategies for better health.

This study explored differences in dental biofilm solids and fluid fluoride after ingestion of NaF or Na2FPO3 in milk or non-fluoridated milk. Eighteen volunteers ingested 1 mg fluoride in 200 mL of milk or 200 mL of non-fluoridated milk. Biofilm samples were collected at baseline, 30, 60, and 240 min and biofilm solids and fluid were micro-analyzed for fluoride. Analysis of variance was performed and the total delivery, retention, and clearance of fluoride to biofilm solids and fluid were estimated as the area under the curve between 0 and 240 min. No statistically significant differences were found for baseline values. Biofilm fluid fluoride values ranged from 0.11 ± 0.05 to 0.21 ± 0.08 µg F/mL while biofilm solid values ranged from 0.62 ± 0.39 to 1.75 ± 1.16 µg F/g. Biofilm fluid values were significantly lower at 60 min for Na2FPO3 in milk. Clearance profiles for biofilm fluid diverged after the initial 60 min. Na2FPO3 had a smaller area under the curve from 60 to 240 min than NaF. Results of this study indicate that the release and clearance of fluoride in biofilm vary among fluoridation compounds and that the concentrations in biofilm solids and fluid also vary and are not correlated to each other in many cases.