Relationship between the IgA antibody response against Streptococcus mutans GbpB and severity of dental caries in childhood

ObjectiveExplore the associations between the severity of dental caries in childhood, mutans streptococci (MS) levels and IgA antibody response against Streptococcus mutans GbpB. Moreover, other caries-related etiological factors were also investigated.Design36–60 month-old children were grouped into Caries-Free (CF, n = 19), Early Childhood Caries (ECC, n = 17) and Severe Early Childhood Caries (S-ECC, n = 21). Data from socio-economic-cultural status, oral hygiene habits and dietary patterns were obtained from a questionnaire and a food-frequency diary filled out by parents. Saliva was collected from children for microbiological analysis and detection of salivary IgA antibody reactive with S. mutans GbpB in western blot.ResultsS-ECC children had reduced family income compared to those with ECC and CF. There was difference between CF and caries groups (ECC and S-ECC) in MS counts. Positive correlations between salivary IgA antibody response against GbpB and MS counts were found when the entire population was evaluated. When children with high MS counts were compared, S-ECC group showed significantly lower IgA antibody levels to GbpB compared to CF group. This finding was not observed for the ECC group.ConclusionsThis study suggests that children with S-ECC have reduced salivary IgA immune responses to S. mutans GbpB, potentially compromising their ability to modify MS infection and its cariogenic potential. Furthermore, a reduced family income and high levels of MS were also associated with S-ECC.     

Antimicrobial peptides in saliva of children with severe early childhood caries

ObjectiveControversies exist regarding the relationship between the concentrations of antimicrobial peptides (AMPs) and presence of dental caries in children. Thus, the aim of this study was to examine levels of AMPs in saliva of caries-free (CF), early childhood caries (ECC) and severe early childhood caries (S-ECC) children to determine if the levels of these salivary peptides individually or in combinations were related to caries severity and mutans streptococci levels.Design36 to 60 month-old children were selected to participate in this study. Children were grouped into CF group (n = 29), ECC group (n = 25) and S-ECC group (n = 29). Saliva was collected from children for microbiological analysis by culture. Salivary concentrations of cathelicidin LL-37, human β-defensin 2 (hBD-2), human β-defensin 3 (hBD-3) and histatin-5 (HTN-5) were determined by ELISA.ResultsSalivary concentrations of AMPs did not differ among CF, ECC and S-ECC groups. Data showed positive correlations between mutans streptococci levels and salivary hBD-2 or HTN-5. Positive correlations were found between hBD-2, hBD-3, LL-37 and HTN-5. Combinations among AMPs, mainly LL-37, were positively associated with caries levels.ConclusionsSalivary concentrations of AMPs individually were not associated with the severity of early childhood caries. The stimulus of caries appears to trigger a biological response, however, with a combination of these peptides.     

Quantitative analysis of biofilm bacteria according to different stages of early childhood caries

ObjectiveMonitoring selected key species related to health or disease may facilitate caries risk assessment and discovery of novel ecological preventive and therapeutic approaches. This study aimed at quantifying Actinomyces naeslundii, Bifidobacterium spp., Lactobacillus acidophilus, Lactobacillus casei group, Streptococcus gordonii, Mitis group and Streptococcus mutans by quantitative polymerase chain reaction (qPCR) in dental biofilm from Brazilian children with different stages of early childhood caries (ECC).DesignSeventy-five preschool children were clinically evaluated by ICDAS criteria and divided into groups: caries-free (CF; n = 20), enamel caries lesions (ECL; n = 17) and dentine caries lesions (DCL; n = 38). Plaque samples from all children were collected for detection and quantification of the selected bacteria.ResultsL. acidophilus and L. casei group were absent in almost all plaque samples. No differences in relative proportions of A. naeslundii, Mitis group and S. gordonii were observed in any stage of caries. However, S. mutans and Bifidobacterium spp. were present at higher concentrations in the biofilm of children with DCL (p < 0.001). Multivariate analysis showed that S. mutans and Bifidobacterium spp. were strongly associated with biofilm in children with DCL.ConclusionDifferences were observed in the proportion of acidogenic and aciduric bacteria with dental caries progression. The data indicate that S. mutans and Bifidobacterium spp. in dental biofilm may be involved in some progression processes for ECC.     

PCR detection of Scardovia wiggsiae in combination with Streptococcus mutans for early childhood caries‐risk prediction

The microbial factor is an important determinant in caries risk assessment. This study aimed to use detection, by PCR, of Scardovia wiggsiae, in combination with Streptococcus mutans, for the accurate prediction of caries risk in children. Detection of Lactobacillus, as a caries‐specific species, was also performed. Dental plaque, as well as infected dentine when available, was collected from children who were caries‐free (n = 30) or diagnosed with early childhood caries (n = 30), and the prevalence and abundance of S. wiggsiae and S. mutans were estimated using quantitative PCR. Lactobacillus was amplified by Lactobacillus genus‐specific primers and then sequenced. Both S. wiggsiae and S. mutans were concurrently detected in 19 children diagnosed with early childhood caries, but in none of the caries‐free children. The positive predictive value was 1 in children with S. wiggsiae‐ and S. mutans‐positive test results, compared with 0.58 when only S. mutans was detected and 0.9 when only S. wiggsiae was detected. The abundance of S. wiggsiae and S. mutans in infected dentine was higher than that in dental plaque from children. Diverse Lactobacillus species were observed in dental plaque but none appeared to be caries‐specific. In conclusion, the detection of S. wiggsiae in combination with S. mutans improves the positive predictive value and the specificity of the test.     

Increased activity of the antioxidants systems modulate the oxidative stress in saliva of toddlers with early childhood caries

ObjectiveThis study aimed to evaluate the oxidative stress levels and the enzymatic and non-enzymatic antioxidant systems in saliva of toddlers with severe early childhood caries (S-ECC).DesignUnstimulated saliva samples were collected at the morning from 0 to 3 year-old S-ECC (n = 30) or caries-free (CF) children (n = 30/group) for evaluation of oxidative stress (OS) and total antioxidant capacity (TAC), which were measured by the ferric reducing antioxidant power (FRAP) assay, as well as to assess the activity of enzymatic (superoxide dismutase, SOD) and non-enzymatic (uric acid, UA) antioxidant systems, respectively. Data were analyzed by Student’s t-test (p < 0.05).ResultsSignificantly higher protein levels were observed in saliva of S-ECC children (0.083 mg/mL) than in the CF group (0.070 mg/mL). Oxidative damage was significantly lower in saliva of S-ECC children (0.0019 μmol/L/mg protein) than in CF children (0.0039 μmol/L/mg protein), while salivary TAC (61.5 μmol/L), SOD activity (36.6 UE/mL) and uric acid (7.05 mg/mL) were significantly higher in saliva of S-ECC when compared to the CF group (49.1 μmol/L, 26.8 UE/mL and 5.02 mg/mL, respectively for TAC, SOD and UA).ConclusionOxidative stress levels were significantly lower in saliva of S-ECC children, what might be associated with the increased activity of salivary enzymatic (SOD) and non-enzymatic (uric acid) antioxidant systems.     

Detection of mutans streptococci in plaque samples from Mongolian preschool and school children

Mutans streptococci, in particular Streptococcus mutans and Streptococcus sobrinus, are generally considered to be the principal microbial pathogen of dental caries. The objective of the study was to isolate S. mutans and S. sobrinus, identify them by PCR, and to compare their presence with the caries status and caries risk in Mongolian preschool and school children. Forty one preschool children aged 3–5 years and 40 school children aged 12–15 years were enrolled in this study. As assessed using Cariostat test, 75.6% of preschool children had high caries risk and 37.5% of school children had high caries risk. In preschool children, the prevalence of S. mutans and S. sobrinus were 100% and 36.6%, respectively; 63.4% were positive for S. mutans alone and 36.6% were positive for both S. mutans and S. sobrinus. In school children, the prevalence of S. mutans and S. sobrinus were 100% and 25.0%, respectively; 75.0% carried S. mutans alone and 25.0% had both S. mutans and S. sobrinus. The percentage of children positive for both S. mutans and S. sobrinus in the high caries risk group were significantly higher than those in the low risk group of either preschool (42.0% vs. 10.0%, P< 0.001) or school children (46.6% vs. 12.0%, P<0.001). Moreover, the caries status of children positive for both S. mutans and S. sobrinus were significantly higher than those positive for S. mutans alone (P< 0.01 for preschool children, and P< 0.05 for school children).     

Presence of Streptococcus mutans or Streptococcus sobrinus in Cariostat®-inoculated plaque samples from Japanese mother-child pairs

The aim of this study was to determine the presence of Streptococcus mutans or Streptococcus sobrinus in Cariostat-inoculated plaque samples obtained from Japanese mother-child pairs through a conventional PCR technique and to establish the presence of these bacteria and caries risk. Oral examination and caries risk assessment using the Cariostat^® were carried out on 168 children, aged 6–31 months, and their mothers. The presence of S. mutans and S. sobrinus in Cariostat-inoculated plaque samples was checked through PCR and tested for relevance with caries risk. A significant correlation (P < 0.001) was found between caries risk of mothers and presence of S. mutans or S. sobrinus in plaque samples from their children in the 19–31-month-old age range. However, no significant relationship found between the presence of either strain in the plaque of younger children (6–18 months) and caries risk of mothers. Likewise, high caries risk was seen in 49.1% of the 19–31-month-old children of highrisk mothers (P < 0.001) and 27% of the 6–18-month-old children of high-risk mothers (P < 0.05). The effectiveness of the Cariostat method for prediction of caries risk can be improved by detecting the presence of S. mutans and S. sobrinus in plaque samples obtained from mothers and their children through conventional PCR techniques.     

Scardovia wiggsiae and its potential role as a caries pathogen

Background

Streptococcus mutans has been strongly associated with dental caries but caries also occurs in its absence. Association of a new species, Scardovia wiggsiae with childhood caries suggests this could be a new caries pathogen.

Risk behaviors and their association with presence of S.mutans or S.sobrinus and caries activity in 18-month-old Japanese children

The purpose of this study was to investigate risk behaviors associated with the presence of S. mutans or S. sobrinus and caries activity. The subjects were 448 mother-child pairs who underwent dental health examinations between February 2004 and November 2004 when the children were 18 months old. Caries activity was assessed by the Cariostat test. The presence of S. mutans and S. sobrinus was detected using PCR techniques. Questionnaires regarding risk behaviors were completed by the mothers. A statistically significant correlation was found for the detection of S. mutans and/or S. sobrinus in children and mothers (P < 0.01). High-risk mothers were more likely to have high-risk children (P < 0.001). In children in whom bacteria were detected, breast-feeding was ranked as the most important risk factor (P < 0.01), followed by eating snacks while playing (P < 0.01), getting snacks from neighbors (P < 0.05), being cared for by grandparents (P < 0.05) and pre-chewing of children's food by mothers (P < 0.05). In children with high caries risk, breast-feeding and pre-chewing were the most important risk factors (P < 0.01), followed by taking meals at irregular intervals and mothers not attending maternity classes (P < 0.05).     

Effect of arginine on the growth and biofilm formation of oral bacteria

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

Effects of S. mutans gene-modification and antibacterial monomer dimethylaminohexadecyl methacrylate on biofilm growth and acid production

ObjectivesAntibacterial quaternary ammonium monomers (QAMs) are used in resins. The rnc gene in Streptococcus mutans (S. mutans) plays a key role in resisting antibiotics. The objectives of this study were to investigate for the first time: (1) the effects of rnc deletion on S. mutans biofilms and acid production; (2) the combined effects of rnc deletion with dimethylaminohexadecyl methacrylate (DMAHDM) on biofilm-inhibition efficacy.MethodsParent S. mutans strain UA159 (ATCC 700610) and the rnc-deleted S. mutans were used. Bacterial growth, minimum inhibitory concentration (MIC), and minimal bactericidal concentration (MBC) were measured to analyze the bacterial susceptibility of the parent and rnc-deleted S. mutans against DMAHDM, with the gold-standard chlorhexidine (CHX) as control. Biofilm biomass, polysaccharide and lactic acid production were measured.ResultsThe drug-susceptibility of the rnc-deleted S. mutans to DMAHDM or CHX was 2-fold higher than parent S. mutans. The drug-susceptibility did not increase after 10 passages (p < 0.05). Deleting the rnc gene increased the biofilm susceptibility to DMAHDM or CHX by 2-fold. The rnc-deletion in S. mutans reduced biofilm biomass, polysaccharide and lactic acid production, even at no drugs. DMAHDM was nearly 40 % more potent than the gold-standard CHX. The combination of rnc deletion + DMAHDM treatment achieved the greatest reduction in biofilm biomass, polysaccharide synthesis, and lactic acid production.SignificanceGene modification by deleting the rnc in S. mutans reduced the biofilm growth and acid production, and the rnc deletion + DMAHDM method showed the greatest biofilm-inhibition efficacy, for the first time. The dual strategy of antibacterial monomer + bacterial gene modification shows great potential to control biofilms and inhibit caries.     

Antimicrobial and antibiofilm action of Casbane Diterpene from Croton nepetaefolius against oral bacteria

ObjectiveThe antibacterial activity of Casbane Diterpene (CD) was evaluated in vitro against Streptococcus oralis, S. mutans, S. salivarius, S. sobrinus, S. mitis and S. sanguinis. The viability of planktonic cells was analysed by susceptibility tests (MIC and MBC) and antibiofilm action was assayed.MethodsThe minimal inhibitory and bactericidal concentrations (MIC and MBC) of oral Streptococcus were evaluated through microdilution tests. To assay antibiofilm activity, biofilms were generated on 96-wells polystyrene plates under the presence of CD and quantified by a crystal violet technique and colonies forming units counting.ResultsThe CD isolated from Croton nepetaefolius showed antimicrobial effect on planktonic forms and biofilms of oral pathogens, with MIC values of 62.5 μg/mL against Streptococcus oralis and values between 125 and 500 μg/mL against S. mutans, S. salivarius, S. sobrinus, S. mitis and S. sanguinis. CD showed an inhibitory effect on S. mutans biofilm formation at 250 μg/mL, and a decrease on viable cell of 94.28% compared to the normal biofilm growth.ConclusionsThe compound CD can be considered as a promising molecule for the treatment against oral pathogens responsible for dental biofilm.     

Effectiveness of Salvadora persica extracts against common oral pathogens

Objective

The purpose of this study was to evaluate the antibacterial activity of ethanol and hexane extracts of Salvadora persica against common oral pathogens.

Risk behaviors and its association with caries activity and dental caries in Japanese children

The aim of this study was to assess risk behaviors and its association with caries activity and dental caries in Japanese children. The subjects were 392 young Japanese children who underwent dental health check-up at 18, 30 and 42 months of age. Oral examinations, Cariostat tests and dental health questionnaires were carried out at each time. The caries prevalence of children was 1.5% at 18 months, 9.9% at 30 months and 28.1% at 42 months. Caries activity based on the Cariostat scores of children was correlated with caries status (caries-free/carious) at 42 months. In children with caries during each examination period at 42 months, eating snacks while playing was ranked as the most important behavioral risk (P < 0.001), followed by breast-feeding (P < 0.01), non setting of time for snacks (P < 0.05) and frequency of snacks (P < 0.05) at 18 months old; non brushing by mother (P < 0.05) and eating snacks while playing (P < 0.05) were ranked highest at 30 months old. In addition, eating snacks while playing (P < 0.001) at 42 months old was the only a significant factor for children with caries. Caries activity and risk behaviors were associated with caries experience at different age periods of childhood.     

Effect of bovine lactoferrin on enamel demineralization and acid fermentation by Streptococcus mutans

The purpose of this study was to evaluate the effects of bovine lactoferrin on acid fermentation and enamel demineralization using Streptococcus mutans in a culture system and an artificial mouth model system. The antibacterial activity of bovine lactoferrin (bLF) against S. mutans was analyzed by a radial diffusion assay. In the culture system, the effect of bLF on the synthesis and adherence of water insoluble glucan (WIG) and the adherence of S. mutans to a glass surface was examined by a batch culture. In the artificial mouth model system, cell suspension of S. mutans, heart infusion broth supplemented with sucrose, and PBS or lactoferrin solution were supplied separately and constantly for 21 hours. The following parameters were determined for evaluation: the amount of artificial biofilm, the changes in pH underneath the biofilm; and the changes in enamel microhardness measured by a Vicker's hardness tester. The antibacterial activity of bLF against S. mutans was observed. The amounts of bacterial cells in the total adherent fractions were inhibited by bLF in a dose dependent manner. The amounts of WIG in a firm-adherent fraction were significantly inhibited by 0.1–1.0% bLF. The changes in microhardness on enamel slabs in the bLF group (2.4 ± 0.8) showed significantly less hardness reduction than those in the control group (22.3 ± 2.5) (P < 0.001). The artificial biofilm accumulation was not reduced by bLF. The results of this study suggest that bLF might have inhibitory effects against acid fermentation and demineralization of enamel by S. mutans.     

Influence of sucrose and xylitol on an early Streptococcus mutans biofilm in a dental simulator

ObjectivesIn vitro methods to study dental biofilms are useful in finding ways to support a healthy microbial balance in the oral cavity. The effects of sucrose, xylitol, and their combination on three strains of Streptococcus mutans and one strain of Streptococcus sobrinus were studied using a dental simulator.MethodsA simulator was used to mimic the oral cavity environment. It provided a continuous-flow system using artificial saliva (AS), constant temperature, mixing, and hydroxyapatite (HA) surface in which the influence of xylitol was studied. The quantities of planktonic and adhered bacteria were measured by real-time qPCR.ResultsCompared against the untreated AS, adding 1% sucrose increased the bacterial colonization of HA (p < 0.0001) whereas 2% xylitol decreased it (p < 0.05), with the exception of clinical S. mutans isolate 117. The combination of xylitol and sucrose decreased the bacterial quantities within the AS and the colonization on the HA by clinical S. mutans isolate 2366 was reduced (p < 0.05). Increasing the concentration (2%–5%) of xylitol caused a reduction in bacterial counts even in the presence of sucrose.ConclusionsThe continuous-culture biofilm model showed that within a young biofilm, sucrose significantly promotes whereas xylitol reduces bacterial colonization and proliferation. The results indicate that xylitol affects the ability of certain S. mutans strains to adhere to the HA. Clinical studies have also shown that xylitol consumption decreases caries incidence and reduces the amount of plaque. This study contributes to the understanding of the mechanism behind these clinical observations.     

Influence of a Brazilian wild green propolis on the enamel mineral loss and Streptococcus mutans’ count in dental biofilm

ObjectiveThis study investigated the anti-demineralizing and antibacterial effects of a propolis ethanolic extract (EEP) against Streptococcus mutans dental biofilm.DesignBlocks of sound bovine enamel (n = 24) were fixed on polystyrene plates. S. mutans inoculum (ATCC 25175) and culture media were added (48 h–37 °C) to form biofilm. Blocks with biofilm received daily treatment (30 μL/1 min), for 5 days, as following: G1 (EEP 33.3%); G2 (chlorhexidine digluconate 0.12%); G3 (ethanol 80%); and G4 (Milli-Q water). G5 and G6 were blocks without biofilm that received only EEP and Milli-Q water, respectively. Final surface hardness was evaluated and the percentage of hardness loss (%HL) was calculated. The EEP extract pH and total solids were determined. S. mutans count was expressed by log₁₀ scale of Colony-Forming Units (CFU/mL). One way ANOVA was used to compare results which differed at a 95% significance level.ResultsG2 presented the lowest average %HL value (68.44% ± 12.98) (p = 0.010), while G4 presented the highest (90.49% ± 5.38%HL) (p = 0.007). G1 showed %HL (84.41% ± 2.77) similar to G3 (87.80% ± 6.89) (p = 0.477). Groups G5 and G6 presented %HL = 16.11% ± 7.92 and 20.55% ± 10.65; respectively (p = 0.952). G1 and G4 differed as regards to S. mutans count: 7.26 ± 0.08 and 8.29 ± 0.17 CFU/mL, respectively (p = 0.001). The lowest bacterial count was observed in chlorhexidine group (G2 = 6.79 ± 0.10 CFU/mL) (p = 0.043). There was no difference between S. mutans count of G3 and G4 (p = 0.435). The EEP showed pH 4.8 and total soluble solids content = 25.9 Brix.ConclusionThe EEP seems to be a potent antibacterial substance against S. mutans dental biofilm, but presented no inhibitory action on the de-remineralization of caries process.     

Inhibition of Streptococcus mutans biofilm formation using extracts from Assam tea compared to green tea

ObjectiveStreptococcus mutans, a gram-positive oral bacterium, has been identified as one of the principal etiological agents of human dental caries. To clarify the nature of the difference anti-biofilm effect against S. mutans between Assam tea from Camellia sinensis var. assamica, partially fermented, and green tea from Camellia sinensis, non-fermented, active agents from the teas were purified.MethodsEffects of Assam tea and green tea samples on biofilm were assessed by using the conventional titer plate method and the human saliva-coated hydroxyapatite discs. The purification and identification of inhibitors were performed by using ultrafiltration with centrifugal filter devices and high performance liquid chromatography.ResultsAssam tea has stronger biofilm inhibition activity against S. mutans than green tea. A substance of <10 kDa in mass in Assam tea had a high concentration of galloylated catechins and a stronger biofilm inhibiting activity than green tea. In contrast, substances >10 kDa in mass from green tea included higher concentrations of polysaccharides composed of galacturonic acid, such as pectin, that enhance biofilm formation.ConclusionsThe higher concentrations of galloylated catechins in Assam tea may assist in prevention of dental caries, whereas in green tea, this mode of inhibition was likely offset by the presence of pectin. Purification of catechins in partially fermented Assam tea with lower-molecular-weight polysaccharide than pectin may be useful for developing oral care products such as toothpaste and oral care gel pastes.     

Resin composite blocks for dental CAD/CAM applications reduce biofilm formation in vitro

ObjectivesModern dentistry is increasingly focusing on digital procedures, including CAD/CAM technologies. New materials have to resist in a demanding environment that includes secondary caries occurrence. The current study hypothesized that the microbiological behavior of different RBCs for CAD/CAM applications is better than that of their counterparts for direct restorations due to differences in the surface characteristics.MethodsBoth direct and CAD/CAM RBCs were tested. Specimens were obtained from each group, polished, cleaned, stored in artificial saliva (1 w), then sterilized under UV (24 h). Specimens’ surface was assessed using profilometry, scanning electron microscopy, energy-dispersive X-ray spectroscopy, and X-ray diffraction; resin/filler content was assessed using thermogravimetry. After pre-incubation with sterile human saliva (24 h), the microbiological behavior of the materials was assessed using four models: Streptococcus mutans adherence (2 h), S. mutans biofilm formation in an orbital shaking bioreactor (24 h), S. mutans biofilm formation in a continuous-flow bioreactor simulating shear forces (24 h), and mixed-plaque formation in the bioreactor (24 h). The viable biomass adhering to the specimens’ surfaces was measured using a tetrazolium dye-based test. Statistical analysis included verification of normality of distribution and homoscedasticity, then Oneway ANOVA and Tukey's test (α = 5%).ResultsWhen using the bioreactor setup, CAD/CAM RBCs generally yielded lower S. mutans and mixed-plaque biofilm formation compared to direct RBCs. This difference was not evidenced in the first two microbiological models. Differences in manufacturing and curing processes rather than in materials’ surface roughness and composition could explain these results.SignificanceCAD/CAM RBCs are promising materials from a microbiological point of view, featuring reduced biofilm formation on their surfaces when shear conditions similar to in vivo ones are present.     

Curcumin reduces Streptococcus mutans biofilm formation by inhibiting sortase A activity

BackgroundSortase A is an enzyme responsible for the covalent attachment of Pac proteins to the cell wall in Streptococcus mutans. It has been shown to play a role in modulating the surface properties and the biofilm formation and influence the cariogenicity of S. mutans. Curcumin, an active ingredient of turmeric, was reported to be an inhibitor for Staphylococcus aureus sortase A. The aim of this study was to investigate the inhibitory ability of curcumin against S. mutans sortase A and the effect of curcumin for biofilm formation.MethodsThe antimicrobial activity of the curcumin to the S. mutans and inhibitory ability of the curcumin against the purified sortase A in vitro were detected. Western-blot and real-time PCR were used to analysis the sortase A mediated Pac protein changes when the S. mutans was cultured with curcumin. The curcumin on the S. mutans biofilm formation was determined by biofilm formation analysis.ResultsCurcumin can inhibit purified S. mutans sortase A with a half-maximal inhibitory concentration (IC₅₀) of (10.2 ± 0.7) μmol/l, which is lower than minimum inhibitory concentration (MIC) of 175 μmol/l. Curcumin (15 μmol/l) was found to release the Pac protein to the supernatant and reduce S. mutans biofilm formation.ConclusionsThese results indicated that curcumin is an S. mutans sortase A inhibitor and has promising anti-caries characteristics through an anti-adhesion-mediated mechanism.