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Journal of Dentistry Jul 2024The aim of this work was to evaluate the antibiofilm and anticaries properties of the association of arginine (Arg) with calcium glycerophosphate (CaGP) and fluoride (F).
OBJECTIVE
The aim of this work was to evaluate the antibiofilm and anticaries properties of the association of arginine (Arg) with calcium glycerophosphate (CaGP) and fluoride (F).
METHODS
An active attachment, polymicrobial biofilm model obtained from saliva and bovine teeth discs were used. After the initial biofilm growth period, the enamel discs were transferred to culture medium. The treatment solutions were added to the culture media to achieve the desired final concentration. The following groups were used: negative control (Control); F (110 ppm F); CaGP (0.05 %); Arg (0.8 %) and their associations (F + CaGP; Arg + F; Arg + CaGP; Arg +F + CaGP). The following analyses were carried out: bacterial viability (total bacteria, aciduric bacteria and mutans streptococci), pH assessment of the spent culture medium, dry weight quantification, evaluation of surface hardness loss (%SH) and subsurface mineral content. Normality and homoscedasticity were tested (Shapiro-Wilk and Levene's test) and the following tests were applied: two-way ANOVA (acidogenicity), Kruskall-Wallis (microbial viability) and one way ANOVA (dry weight, %SH, mineral content).
RESULTS
The association Arg + F + CaGP resulted in the lowest surface hardness loss in tooth enamel (-10.9 ± 2.3 %; p < 0.05). Arg +F + CaGP exhibited highest values of subsurface mineral content (10.1 ± 2.9 g/cm) in comparison to Control and F (p < 0.05). In comparison to Control and F, Arg +F + CaGP promoted the highest reduction in aciduric bacteria and mutans streptococci (5.7 ± 0.4; 4.4 ± 0.5 logCFU/mL, p < 0.05).
CONCLUSIONS
The Arg-F-Ca association demonstrated to be the most effective combination in protecting the loss of surface hardness and subsurface mineral content, in addition to controlling important virulence factors of the cariogenic biofilm.
CLINICAL SIGNIFICANCE
Our findings provide evidence that the Arg-F-Ca association showed an additive effect, particularly concerning protection against enamel demineralization. The combination of these compounds may be a strategy for patients at high risk of caries.
Topics: Arginine; Biofilms; Cattle; Animals; Dental Enamel; Streptococcus mutans; Fluorides; Glycerophosphates; Cariostatic Agents; Saliva; Hydrogen-Ion Concentration; Dental Caries; Microbial Viability; Hardness; Humans; Tooth Demineralization; Surface Properties
PubMed: 38714243
DOI: 10.1016/j.jdent.2024.105039 -
Carbohydrate Polymers Aug 2024Water-insoluble α-glucans synthesized from sucrose by glucansucrases from Streptococcus spp. are essential in dental plaque and caries formation. Because limited...
Water-insoluble α-glucans synthesized from sucrose by glucansucrases from Streptococcus spp. are essential in dental plaque and caries formation. Because limited information is available on the fine structure of these biopolymers, we analyzed the structures of unmodified glucans produced by five recombinant Streptococcus (S.) mutans DSM 20523 and S. salivarius DSM 20560 glucansucrases in detail. A combination of methylation analysis, endo-dextranase and endo-mutanase hydrolyses, and HPSEC-RI was used. Furthermore, crystal-like regions were analyzed by using XRD and C MAS NMR spectroscopy. Our results showed that the glucan structures were highly diverse: Two glucans with 1,3- and 1,6-linkages were characterized in detail besides an almost exclusively 1,3-linked and a linear 1,6-linked glucan. Furthermore, one glucan contained 1,3-, 1,4-, and 1,6-linkages and thus had an unusual, not yet described structure. It was demonstrated that the glucans had a varying structural architecture by using partial enzymatic hydrolyses. Furthermore, crystal-like regions formed by 1,3-glucopyranose units were observed for the two 1,3- and 1,6-linked glucans and the linear 1,3-linked glucan. 1,6-linked regions were mobile and not involved in the crystal-like areas. Altogether, our results broaden the knowledge of the structure of water-insoluble α-glucans from Streptococcus spp.
Topics: Glucans; Water; Glycosyltransferases; Streptococcus; Solubility; Streptococcus mutans
PubMed: 38710558
DOI: 10.1016/j.carbpol.2024.122164 -
Journal of Visualized Experiments : JoVE Apr 2024Most in vitro models lack the capacity to fully probe bacterial phenotypes emerging from the complex interactions observed in real-life environments. This is...
Most in vitro models lack the capacity to fully probe bacterial phenotypes emerging from the complex interactions observed in real-life environments. This is particularly true in the context of hard-to-treat, chronic, and polymicrobial biofilm-based infections detected in the airways of individuals living with cystic fibrosis (CF), a multiorgan genetic disease. While multiple microbiome studies have defined the microbial compositions detected in the airway of people with CF (pwCF), no in vitro models thus far have fully integrated critical CF-relevant lung features. Therefore, a significant knowledge gap exists in the capacity to investigate the mechanisms driving the pathogenesis of mixed species CF lung infections. Here, we describe a recently developed four-species microbial community model, including Pseudomonas aeruginosa, Staphylococcus aureus, Streptococcus sanguinis, and Prevotella melaninogenica grown in CF-like conditions. Through the utilization of this system, clinically relevant phenotypes such as antimicrobial recalcitrance of several pathogens were observed and explored at the molecular level. The usefulness of this in vitro model resides in its standardized workflow that can facilitate the study of interspecies interactions in the context of chronic CF lung infections.
Topics: Cystic Fibrosis; Biofilms; Humans; Phenotype; Pseudomonas aeruginosa; Staphylococcus aureus; Microbiota; Streptococcus sanguis; Prevotella melaninogenica
PubMed: 38709077
DOI: 10.3791/66785 -
Journal of Microbiological Methods Jun 2024Methylation analysis was performed on methylated alditol acetate standards and Streptococcus mutans extracellular polymeric substances (EPS) produced from wild-type and...
Revisiting gas-chromatography/mass-spectrometry molar response factors for quantitative analysis (FID or TIC) of glycosidic linkages in polysaccharides produced by oral bacterial biofilms.
Methylation analysis was performed on methylated alditol acetate standards and Streptococcus mutans extracellular polymeric substances (EPS) produced from wild-type and Gtf knockout strains (∆GtfB, ∆GtfB, and ∆GtfD). The methylated alditol acetate standards were representative of glycosidic linkages found in S. mutans EPS and were used to calibrate the GC-MS system for an FID detector and MS (TIC) and produce molar response factor, a necessary step in quantitative analysis. FID response factors were consistent with literature values (Sweet et al., 1975) and found to be the superior option for quantitative results, although the TIC response factors now give researchers without access to an FID detector a needed option for molar response factor correction. The GC-MS analysis is then used to deliver the ratio of the linkage types within a biofilm.
Topics: Biofilms; Streptococcus mutans; Gas Chromatography-Mass Spectrometry; Polysaccharides, Bacterial; Glycosides; Methylation; Extracellular Polymeric Substance Matrix; Polysaccharides
PubMed: 38704038
DOI: 10.1016/j.mimet.2024.106942 -
BMC Oral Health May 2024To evaluate the antibacterial effectiveness of a combination of ε-poly-L-lysine (ε-PL), funme peptide (FP) as well as domiphen against oral pathogens, and assess the... (Randomized Controlled Trial)
Randomized Controlled Trial
OBJECTIVE
To evaluate the antibacterial effectiveness of a combination of ε-poly-L-lysine (ε-PL), funme peptide (FP) as well as domiphen against oral pathogens, and assess the efficacy of a BOP® mouthwash supplemented with this combination in reducing halitosis and supragingival plaque in a clinical trial.
MATERIALS AND METHODS
The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of the compound against Fusobacterium nucleatum, Porphyromonas gingivalis, Streptococcus mutans, and Aggregatibacter actinomycetemcomitans were determined by the gradient dilution method. Subsequently, the CCK-8 assay was used to detect the toxicity of mouthwash on human gingival fibroblastst, and the effectiveness in reducing halitosis and supragingival plaque of the mouthwash supplemented with the combination was analyzed by a randomized, double-blind, parallel-controlled clinical trial.
RESULTS
The combination exhibited significant inhibitory effects on tested oral pathogens with the MIC < 1.56% (v/v) and the MBC < 3.13% (v/v), and the mouthwash containing this combination did not inhibit the viability of human gingival fibroblasts at the test concentrations. The clinical trial showed that the test group displayed notably lower volatile sulfur compounds (VSCs) at 0, 10, 24 h, and 7 d post-mouthwash (P < 0.05), compared with the baseline. After 7 days, the VSC levels of the and control groups were reduced by 50.27% and 32.12%, respectively, and notably cutting severe halitosis by 57.03% in the test group. Additionally, the Plaque Index (PLI) of the test and control group decreased by 54.55% and 8.38%, respectively, and there was a significant difference in PLI between the two groups after 7 days (P < 0.01).
CONCLUSIONS
The combination of ε-PL, FP and domiphen demonstrated potent inhibitory and bactericidal effects against the tested oral pathogens, and the newly formulated mouthwash added with the combination exhibited anti-dental plaque and anti-halitosis properties in a clinical trial and was safe.
TRIAL REGISTRATION
The randomized controlled clinical trial was registered on Chinese Clinical Trial Registry (No. ChiCTR2300073816, Date: 21/07/2023).
Topics: Humans; Halitosis; Mouthwashes; Dental Plaque; Double-Blind Method; Male; Female; Polylysine; Adult; Microbial Sensitivity Tests; Young Adult; Anti-Bacterial Agents; Porphyromonas gingivalis; Fusobacterium nucleatum; Fibroblasts; Peptides; Aggregatibacter actinomycetemcomitans; Streptococcus mutans
PubMed: 38702623
DOI: 10.1186/s12903-024-04255-0 -
BMC Microbiology May 2024Fluoride-resistant Streptococcus mutans (S. mutans) strains have developed due to the wide use of fluoride in dental caries prevention. However, the metabolomics of...
BACKGROUND
Fluoride-resistant Streptococcus mutans (S. mutans) strains have developed due to the wide use of fluoride in dental caries prevention. However, the metabolomics of fluoride-resistant S. mutans remains unclear.
OBJECTIVE
This study aimed to identify metabolites that discriminate fluoride-resistant from wild-type S. mutans.
MATERIALS AND METHODS
Cell supernatants from fluoride-resistant and wild-type S. mutans were collected and analyzed by liquid chromatography-mass spectrometry. Principal components analysis and partial least-squares discriminant analysis were performed for the statistical analysis by variable influence on projection (VIP > 2.0) and p value (Mann-Whitney test, p < 0.05). Metabolites were assessed qualitatively using the Human Metabolome Database version 2.0 ( http://www.hmdb.ca ), or Kyoto Encyclopedia of Genes and Genomes ( http://www.kegg.jp ), and Metaboanalyst 6.0 ( https://www.metaboanalyst.ca ).
RESULTS
Fourteen metabolites differed significantly between fluoride-resistant and wild-type strains in the early log phase. Among these metabolites, 5 were identified. There were 32 differential metabolites between the two strains in the stationary phase, 13 of which were identified. The pyrimidine metabolism for S. mutans FR was matched with the metabolic pathway.
CONCLUSIONS
The fructose-1,6-bisphosphate concentration increased in fluoride-resistant strains under acidic conditions, suggesting enhanced acidogenicity and acid tolerance. This metabolite may be a promising target for elucidating the cariogenic and fluoride resistant mechanisms of S. mutans.
Topics: Streptococcus mutans; Metabolomics; Fluorides; Drug Resistance, Bacterial; Fructosediphosphates; Humans; Metabolome; Dental Caries; Chromatography, Liquid
PubMed: 38702601
DOI: 10.1186/s12866-024-03310-8 -
Archives of Oral Biology Aug 2024The effectiveness of supragingival dental biofilm control during orthodontic treatment and changes in the bacterial profile were analyzed. (Randomized Controlled Trial)
Randomized Controlled Trial
OBJECTIVE
The effectiveness of supragingival dental biofilm control during orthodontic treatment and changes in the bacterial profile were analyzed.
DESIGN
Sixty-four participants aged 12-22 years (57% female) were included in the study. Participants underwent orthodontic treatment with fixed appliances and were randomly assigned to one of the three groups, which during a period of one month: (I) used chlorhexidine digluconate (CHX), (II) used high concentration of fluoride (F) gel and (III) performed standard oral hygiene. The plaque and gingivitis index, pH of biofilm and white spot lesions (WSL) were assessed. Changes of the bacteria in the biofilm were analyzed by the quantitative polymerase chain reaction RESULTS: Increase in the plaque index, pH of biofilm, and WSL was observed during orthodontic treatment with standard oral hygiene. Large interindividual variability was present, and the effects of one-month use of fluorides and CHX on clinical parameters were not significant. Despite standard hygiene the abundance of studied biofilm bacteria increased - the most Streptoccocus mutans (14.2x) and S. salivarius (3.3x), moderate Veillonella parvula (3x) and the least S. sobrinus (2.3x) and Agregatibacter actinomycetemcomitans (1.9x). The use of CHX reduced S. sobrinus (2.2x) and A. actinomycetemcomitans (1.9x). Fluoride use reduced A. actinomycetemcomitans (1.3x) and S. sobrinus (1.2x). Fluorides better controlled S. mutans than CHX.
CONCLUSION
Bacterial biomass in supragingival biofilm increased during treatment with metal orthodontic appliances, with greater increase in cariogenic bacteria than periopathogens. Fluoride controlled S. mutans, while CHX S. sobrinus and A. actinomycetemcomitans.
Topics: Humans; Biofilms; Female; Adolescent; Chlorhexidine; Child; Male; Orthodontic Appliances, Fixed; Young Adult; Fluorides; Dental Plaque Index; Oral Hygiene; Dental Plaque; Hydrogen-Ion Concentration; Streptococcus mutans; Gingivitis; Anti-Infective Agents, Local; Polymerase Chain Reaction; Dental Caries
PubMed: 38701663
DOI: 10.1016/j.archoralbio.2024.105984 -
Journal of Dentistry Jul 2024To evaluate the mechanical, wear, antibacterial properties, and biocompatibility of injectable composite materials.
OBJECTIVES
To evaluate the mechanical, wear, antibacterial properties, and biocompatibility of injectable composite materials.
METHODS
Two injectable composite resins (GU and BI), one flowable composite resin (FS), and one flowable compomer (DF), in A2 shade, were tested. Mechanical properties were tested via three-point bending test immediately after preparation and after 1-day, 7-day, 14-day, and 30-day water storage. Under water-PMMA slurry immersion, specimens were subjected to a 3-body wear test (10,000 cycles) against stainless steel balls, while the roughness, wear depth, and volume loss were recorded. After 1-day and 3-day MC3T3-E1 cell culture, cell viability was evaluated with CCK-8 test kits, while the cell morphology was observed under CLSM and SEM. Antibacterial properties on S. mutans were assessed via CFU counting, CLSM, and SEM observation. SPSS 26.0 was used for statistical analysis (α = 0.05).
RESULTS
The mechanical properties were material-dependent and sensitive to water storage. Flexural strength ranked GU > FS > BI > DF at all testing levels. Three nanocomposites had better wear properties than DF. No significant difference on 1-day cell viability was found, but DF showed significantly lower cell proliferation than nanocomposites on 3-day assessment. GU and FS had more favourable cell adhesion and morphology. CFU counting revealed no significant difference, while FS presented a slightly thicker biofilm and BI showed relatively lower bacteria density.
CONCLUSIONS
Injectable nanocomposites outperformed the compomer regarding mechanical properties, wear resistance, and biocompatibility. The tested materials presented comparable antibacterial behaviours. Flowable resin-based composites' performances are affected by multiple factors, and their compositions can be attributed.
CLINICAL SIGNIFICANCE
A profound understanding of the mechanical, wear, and biological properties of the restorative material is imperative for the clinical success of dental restorations. The current study demonstrated superior properties of highly filled injectable composite resins, which imply their wider indications and better long-term clinical performances.
Topics: Composite Resins; Materials Testing; Anti-Bacterial Agents; Streptococcus mutans; Biocompatible Materials; Mice; Animals; Cell Survival; Surface Properties; Dental Materials; Compomers; Water; Nanocomposites; Flexural Strength; Injections; Polymethyl Methacrylate
PubMed: 38697507
DOI: 10.1016/j.jdent.2024.105025 -
Photodiagnosis and Photodynamic Therapy Jun 2024To assess the impact of various cavity disinfectants PC-PDT (Phycocyanin activated by Photodynamic therapy), PC@AgNPs-PDT (Phycocyanin and silver nanoparticles activated...
Phycocyanin-loaded silver nanoparticles activated with photodynamic therapy and Nd: YAG laser for caries-affected dentin disinfection: Impact on Streptococcus mutans survival rate and shear bond strength to the tooth-colored restorative material.
To assess the impact of various cavity disinfectants PC-PDT (Phycocyanin activated by Photodynamic therapy), PC@AgNPs-PDT (Phycocyanin and silver nanoparticles activated by PDT), and Nd: YAG laser on the survival rate of S.mutans and the bond integrity of composite restoration METHODS: Sixty human mandibular molars that scored 4 and 5 based on ICDAS criteria were included. The infected dentin was removed while the CAD was preserved based on visual, tactile, and staining assessment. S.mutans were cultured on the CAD of twenty samples. All the specimens were indiscriminately distributed into four groups based on cavity disinfection (n=20 each includes n = 5 each group incubated with S.mutans) Group 1: CHX, Group 2: Nd:YAG laser, Group 3: PC-PDT and Group 4: PC@AgNPs-PDT. S.mutans survival rate was assessed for each group(n = 5). Forty samples underwent composite bonding for SBS and failure mode assessment using universal testing machine (UTM) and stereomicroscope. The calculations for the mean and standard deviation (SD) and their comparison among different groups were performed using a one-way analysis of variance (ANOVA) and the Tukey post hoc test (p ≤ 0.05) RESULTS: CAD surface treated disinfected with PC@AgNPs-PDT yielded the lowest survival rates (0.13 ± 0.05 CFU/ml) and highest SBS (17.23 ± 1.45 MPa). Group 1 (CHX) unveiled the highest survival rate of S.mutans (0.33 ± 0.12 CFU/ml). However, Group 2 (Nd:YAG Laser) (11.87 ± 0.67 MPa) presented the lowest SBS CONCLUSION: The combination of Phycocyanin loaded with silver nanoparticles and activated with Photodynamic therapy demonstrates the highest antimicrobial potential and bond strength of composite restorations.
Topics: Photochemotherapy; Silver; Humans; Streptococcus mutans; Metal Nanoparticles; Phycocyanin; Photosensitizing Agents; Lasers, Solid-State; Dental Caries; Dentin; Disinfection; Shear Strength; Molar
PubMed: 38697450
DOI: 10.1016/j.pdpdt.2024.104108 -
Journal of Applied Oral Science :... 2024Specific products containing natural resources can contribute to the innovation of complete denture hygiene.
UNLABELLED
Specific products containing natural resources can contribute to the innovation of complete denture hygiene.
OBJECTIVE
To conduct an in vitro evaluation of experimental dentifrices containing essential oils of Bowdichia virgilioides Kunth (BvK), Copaifera officinalis (Co), Eucalyptus citriodora (Ec), Melaleuca alternifolia (Ma) and Pinus strobus (Ps) at 1%.
METHODOLOGY
The variables evaluated were organoleptic and physicochemical characteristics, abrasiveness (mechanical brushing machine) simulating 2.5 years, and microbial load (Colony Forming Units - CFU/mL), metabolic activity (XTT assay) and cell viability (Live/Dead® BacLight™ kit) of the multispecies biofilm (Streptococcus mutans: Sm, Staphylococcus aureus: Sa, Candida albicans: Ca and Candida glabrata: Cg). Specimens of heat-polymerized acrylic resins (n=256) (n=96 specimens for abrasiveness, n=72 for microbial load count, n=72 for biofilm metabolic activity, n=16 for cell viability and total biofilm quantification) with formed biofilm were divided into eight groups for manual brushing (20 seconds) with a dental brush and distilled water (NC: negative control), Trihydral (PC: positive control), placebo (Pl), BvK, Co, Ec, Ma or Ps. After brushing, the specimens were washed with PBS and immersed in Letheen Broth medium, and the suspension was sown in solid specific medium. The organoleptic characteristics were presented by descriptive analysis. The values of density, pH, consistency and viscosity were presented in a table. The data were analyzed with the Wald test in a generalized linear model, followed by the Kruskal-Wallis test, Dunn's test (mass change) and the Bonferroni test (UFC and XTT). The Wald test in Generalized Estimating Equations and the Bonferroni test were used to analyze cell viability.
RESULTS
All dentifrices showed stable organoleptic characteristics and adequate physicochemical properties. CN, Ec, Ps, Pl and PC showed low abrasiveness. There was a significant difference between the groups (p<0.001) for microbial load, metabolic activity and biofilm viability.
CONCLUSIONS
It was concluded that the BvK, Ec and Ps dentifrices are useful for cleaning complete dentures, as they have antimicrobial activity against biofilm. The dentifrices containing Bowdichia virgilioides Kunth showed medium abrasiveness and should be used with caution.
Topics: Biofilms; Dentifrices; Oils, Volatile; Denture, Complete; Materials Testing; Time Factors; Reproducibility of Results; Toothbrushing; Colony Count, Microbial; Staphylococcus aureus; Statistics, Nonparametric; Streptococcus mutans; Analysis of Variance; Microbial Viability; Candida albicans; Reference Values; Acrylic Resins
PubMed: 38695444
DOI: 10.1590/1678-7757-2023-0397