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.     

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.     

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.     

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.     

De novo synthetic short antimicrobial peptides against cariogenic bacteria

ObjectiveAntimicrobial peptides (AMPs) have shown the ability to inhibit planktonic bacteria and biofilms. The objectives of this study were to de novo design and synthesize a series of cationic, amphipathic α-helical AMPs that would be shorter, less cytotoxic, and more potent than existing AMPs against cariogenic bacteria.DesignThree short AMPs (GH8, GLLWHLLH-NH₂; GH12, GLLWHLLHHLLH-NH₂; and GH16, GLLWHLLHHLLHLLHH-NH₂) were designed, synthesized and characterized structurally. Minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) against eight major cariogenic bacteria were tested to select the most promising peptide. Scanning electron microscopy (SEM) was used to observe the bacterial membrane after treatment with selected peptides. The bactericidal kinetics, effects on biofilm and cytotoxity were further investigated.ResultsOf the three AMPs, GH12 had the most balanced structural parameters and a high content of α-helical structure. GH12 had a MIC of 4.0-8.0 μg/mL and MBC of 8.0-32.0 μg/mL. The corresponding values for the other two AMPs were 2- to 64- fold higher. In time-kill assays, GH12 killed all bacterial strains within 60 min at 4- fold MBC. SEM observed lysis and pore formation of the cytomembrane after treatment with GH12. 8.0 μg/mL GH12 inhibited Streptococcus mutans biofilm formation. Confocal laser scanning microscopy showed that GH12 effectively reduced the biomass of 1-day-old S. mutans biofilm. Cytotoxicity assays indicated that GH12 showed little toxic effect on the viability of human gingival fibroblasts.ConclusionThese results indicate that GH12 shows antimicrobial activity against cariogenic bacteria and biofilms in vitro.     

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.     

Comparative effect of a stannous fluoride toothpaste and a sodium fluoride toothpaste on a multispecies biofilm

ObjectivesThis paper aimed to compare the mode of action of a stannous fluoride-containing toothpaste with a conventional sodium fluoride-containing toothpaste on anti-biofilm properties.MethodsA three-species biofilm model that consists of Streptococcus mutans, Streptococcus sanguinis and Porphyromonas gingivalis was established to compare the anti-biofilm properties of a stannous fluoride-containing toothpaste (CPH), a conventional sodium fluoride-containing toothpaste (CCP) and a negative control (PBS). The 48 h biofilms were subjected to two-minute episodes of treatment with test agents twice a day for 5 consecutive days. Crystal violet staining and XTT assays were used to evaluate the biomass and viability of the treated biofilm. Live/dead staining and bacteria/extracellular polysaccharides (EPS) double-staining were used to visualize the biofilm structure and to quantify microbial/extracellular components of the treated biofilms. Species-specific fluorescent in situ hybridization and quantitative polymerase chain reaction (qPCR) were used to analyze microbial composition of the biofilms after treatment.ResultsThe biomass and viability of the biofilms were significantly reduced after CPH toothpaste treatment. The inhibitory effect was further confirmed by the live/dead staining. The EPS amounts of the three-species biofilm were significantly reduced by CCP and CPH treatments, and CPH toothpaste demonstrated significant inhibition on EPS production. More importantly, CPH toothpaste significantly suppressed S. mutans and P. gingvalis, and enriched S. sanguinis in the three-species biofilm. In all experiments CPH had a significantly greater effect than CCP (p < 0.05) and CCP had a greater effect than PBS (p < 0.05).ConclusionsStannous fluoride-containing toothpaste not only showed better inhibitory effect against oral microbial biofilm, but was also able to modulate microbial composition within multi-species biofilm compared with conventional sodium fluoride-containing toothpaste.     

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.     

Clotrimazole and econazole inhibit Streptococcus mutans biofilm and virulence in vitro

Objective: The aim of this study was to determine the inhibitory effect of eight antifungal drugs on S. mutans growth, biofilm formation and virulence factors.MethodsThe actions of antifungal drugs on S. mutans were determined by recovery plates and survival kinetic curves. Biofilms were observed by scanning electron microscopy and the viable cells were recovered on BHI plates, meanwhile biofilms were stained by BacLight live/dead kit to investigate the biofilm viability. Bacteria/extracellular polysaccharides staining assays were performed to determine the EPS production of S. mutans biofilms. Acidogenicity and acidurity of S. mutans were determined using pH drop and acid tolerance assays, and the expression of ldh gene was evaluated using qPCR.ResultsWe found that clotrimazole (CTR) and econazole (ECO) showed antibacterial activities on S. mutans UA159 and S. mutans clinical isolates at 12.5 and 25 mg/L, respectively. CTR and ECO could also inhibit S. mutans biofilm formation and reduce the viability of preformed biofilm. CTR and ECO affected the live/dead ratio and the EPS/bacteria ratio of S. mutans biofilms. CTR and ECO also inhibited the pH drop, lactate acid production, and acid tolerance. The abilities of CTR and ECO to inhibit S. mutans ldh expression were also confirmed.ConclusionsWe found that two antifungal azoles, CTR and ECO, had the abilities to inhibit the growth and biofilm formation of S. mutans and more importantly, they could also inhibit the virulence factors of S. mutans.     

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.     

Nano-calcium phosphate and dimethylaminohexadecyl methacrylate adhesive for dentin remineralization in a biofilm-challenged environment

ObjectiveDentin remineralization at the bonded interface would protect it from external risk factors, therefore, would enhance the longevity of restoration and combat secondary caries. Dental biofilm, as one of the critical biological factors in caries formation, should not be neglected in the assessment of caries preventive agents. In this work, the remineralization effectiveness of demineralized human dentin in a multi-species dental biofilm environment via an adhesive containing nanoparticles of amorphous calcium phosphate (NACP) and dimethylaminohexadecyl methacrylate (DMAHDM) was investigated.MethodsDentin demineralization was promoted by subjecting samples to a three-species acidic biofilm containing Streptococcus mutans, Streptococcus sanguinis, Streptococcus gordonii for 24 h. Samples were divided into a control group, a DMAHDM adhesive group, an NACP group, and an NACP + DMAHDM adhesive group. A bonded model containing a control-bonded group, a DMAHDM-bonded group, an NACP-bonded group, and an NACP + DMAHDM-bonded group was also included in this study. All samples were subjected to a remineralization protocol consisting of 4-h exposure per 24-h period in brain heart infusion broth plus 1% sucrose (BHIS) followed by immersion in artificial saliva for the remaining period. The pH of BHIS after 4-h immersion was measured every other day. After 14 days, the biofilm was assessed for colony-forming unit (CFU) count, lactic acid production, live/dead staining, and calcium and phosphate content. The mineral changes in the demineralized dentin samples were analyzed by transverse microradiography.ResultsThe in vitro experiment results showed that the NACP + DMAHDM adhesive effectively achieved acid neutralization, decreased biofilm colony-forming unit (CFU) count, decreased biofilm lactic acid production, and increased biofilm calcium and phosphate content. The NACP + DMAHDM adhesive group had higher remineralization value than the NACP or DMAHDM alone adhesive group.SignificanceThe NACP + DMAHDM adhesive was effective in remineralizing dentin lesion in a biofilm model. It is promising to use NACP + DMAHDM adhesive to protect bonded interface, inhibit secondary caries, and prolong the longevity of restoration.     

Effect of different pre-irradiation times on curcumin-mediated photodynamic therapy against planktonic cultures and biofilms of Candida spp

ObjectivesThe aim of this study was to evaluate the effects of pre-irradiation time (PIT) on curcumin (Cur)-mediated photodynamic therapy (PDT) against planktonic and biofilm cultures of reference strains of Candida albicans, Candida glabrata and Candida dubliniensis.Materials and methodsSuspensions and biofilms of Candida species were maintained in contact with different concentrations of Cur for time intervals of 1, 5, 10 and 20 min before irradiation and LED (light emitting diode) activation. Additional samples were treated only with Cur, without illumination, or only with light, without Cur. Control samples received neither light nor Cur. After PDT, suspensions were plated on Sabouraud Dextrose Agar, while biofilm results were obtained using the XTT-salt reduction method. Confocal Laser Scanning Microscopy (CLSM) observations were performed to supply a better understanding of Cur penetration through the biofilms after 5 and 20 min of contact with the cultures.ResultsDifferent PITs showed no statistical differences in Cur-mediated PDT of Candida spp. cell suspensions. There was complete inactivation of the three Candida species with the association of 20.0 μM Cur after 5, 10 and 20 min of PIT. Biofilm cultures showed significant reduction in cell viability after PDT. In general, the three Candida species evaluated in this study suffered higher reductions in cell viability with the association of 40.0 μM Cur and 20 min of PIT. Additionally, CLSM observations showed different intensities of fluorescence emissions after 5 and 20 min of incubation.ConclusionPhotoinactivation of planktonic cultures was not PIT-dependent. PIT-dependence of the biofilm cultures differed among the species evaluated. Also, CLSM observations confirmed the need of higher time intervals for the Cur to penetrate biofilm structures.     

Bioactive resin-based composite with surface pre-reacted glass-ionomer filler and zwitterionic material to prevent the formation of multi-species biofilm

ObjectiveThis study evaluated the synergetic effect between surface pre-reacted glass-ionomer (SPRG) filler and 2-methacryloyloxyethyl phosphorylcholine (MPC), for inhibiting multi-species biofilm formation, while maintaining or even improving the original beneficial features of SPRG-filled resin-based composite (RBC).MethodsMPC (1.5–10 wt%) was incorporated into commercial SPRG-filled RBC. Then, the inherent properties of RBC, and ion release and acid-neutralising properties associated with SPRG were investigated. Further, protein adsorptions and bacterial adhesion and viability on the SPRG-filled RBC surfaces were studied using four kinds of oral bacteria; Streptococcus mutans, Actinomyces naeslundii, Veillonella parvula, and Porphyromonas gingivalis. Finally, the thickness and biomass of the human saliva-derived biofilm model cultured on test and control samples were analysed.ResultsAddition of MPC content resulted in decreased flexural strength and wettability of SPRG-filled RBC. SPRG-filled RBC released significantly higher amounts of multiple ions as contents of MPC increased. Meanwhile, SPRG-filled RBC with 5-wt% MPC significantly improved acid-neutralising properties than those of other test and control samples (P < 0.001). SPRG-filled RBC with 3 wt% MPC significantly reduced the amount of adsorbed bovine serum albumin and proteins from the brain heart infusion medium as compared to the control (P < 0.01). A similar trend was observed in the attachment of four types of bacteria and multi-species biofilm (P < 0.01).SignificanceDespite limitation in terms of deteriorations of some physical properties, addition of 3% MPC to SPRG-filled RBC leads to inhibition of the attachment of multi-species bacteria on its surface, as well as inhibition of biofilm growth. Moreover, the original important bioactive features of SPRG-filled RBC such as ion release and acid neutralisations are either maintained or improved upon adding MPC.     

Antibacterial effect of bactericide immobilized in resin matrix

ObjectiveBiomaterials with anti-microbial properties are highly desirable in the oral cavity. Ideally, bactericidal molecules should be immobilized within the biomaterial to avoid unwanted side-effects against surrounding tissues. They may then however loose much of their antibacterial efficiency. The aim of this study was to investigate how much antibacterial effect an immobilized bactericidal molecule still has against oral bacteria.MethodsExperimental resins containing 0, 1 and 3% cetylpyridinium chloride (CPC) were polymerized, and the bacteriostatic and bactericidal effects against Streptococcus mutans were determined. Adherent S. mutans on HAp was quantitatively determined using FE-SEM and living cells of S. mutans were quantified using real-time RT-PCR. The amount of CPC released from the 0%-, 1%- and 3%-CPC resin sample into water was spectrometrically quantified using a UV–vis recording spectrophotometer.ResultsUV spectrometry revealed that less than 0.11 ppm of CPC was released from the resin into water for all specimens, which is lower than the minimal concentration generally needed to inhibit biofilm formation. Growth of S. mutans was significantly inhibited on the surface of the 3%-CPC-containing resin coating, although no inhibitory effect was observed on bacteria that were not in contact with its surface. When immersed in water, the antibacterial capability of 3%-CPC resin lasted for 7 days, as compared to resin that did not contain CPC.SignificanceThese results demonstrated that the bactericidal molecule still possessed significant contact bacteriostatic activity when it was immobilized in the resin matrix.     

Antimicrobial activity of nanoemulsion on cariogenic planktonic and biofilm organisms

IntroductionNanoemulsions (NE) are a unique class of disinfectants produced by mixing a water immiscible liquid phase into an aqueous phase under high shear forces. NE have antimicrobial properties and are also effective anti-biofilm agents.Materials and methodsThe effectiveness of nanoemulsion and its components was determined against Streptococcus mutans and Lactobacillus casei by live/dead staining. In vitro antimicrobial effectiveness of nanoemulsion against planktonic S. mutans, L. casei, Actinomyces viscosus, Candida albicans and mixed culture was determined by a serial dilution technique to obtain minimum inhibitory concentration and minimum bactericidal concentration (MIC/MBC). In addition, efficacy was investigated by kinetics of killing, adherence and biofilm assays.ResultsCompared to its components, nanoemulsion showed notable antimicrobial activity against biofilm organisms, up to 83.0% kill within 1 min. NE dilutions ranging from 243 to 19683 were effective against planktonic S. mutans, L. casei, A. viscosus, C. albicans and mixed culture of these four strains as shown through MIC/MBC assays. NE showed antimicrobial activity against planktonic cells at high dilutions, confirmed by time kill studies. The level of adhesion on glass surface was reduced by 94.2–99.5% in nanoemulsion treated groups (p < 0.001). 4-Day-old S. mutans, L. casei, A. viscosus, C. albicans and mixed cultures biofilms treated with NE showed reductions of bacterial counts with decreasing dilutions (p < 0.001).ConclusionThese results suggest that nanoemulsion has effective anti-cariogenic activity against cariogenic microorganisms and may be a useful medication in the prevention of caries.     

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.     

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).     

Comparative effectiveness of NiCl2, Ni- and NiO-NPs in controlling oral bacterial growth and biofilm formation on oral surfaces

ObjectiveOral ailments are often treated with antibiotics, which are rendered ineffective as bacteria continue to develop resistance against them. It has been suggested that the nanoparticles (NPs) approach may provide a safer and viable alternative to traditional antibacterial agents. Therefore, nickel (Ni)- and nickel oxide (NiO)-NPs were synthesized, characterized and assessed for their efficacy in reducing oral bacterial load in vitro. Also, the effects of bulk compound NiCl₂ (Ni ions), along with the Ni- and NiO-NPs on bacterial exopolysaccharide (EPS) production and biofilm formation on the surface of artificial teeth, and acrylic dentures, were investigated.MethodsTotal bacteria from a healthy male were collected and adjusted to 4 × 109 cells/ml for all the tests. Effect of the NPs on growth, biofilm formation, EPS production and acid production from glucose was tested using standard protocols.ResultsData revealed that the Ni-NPs (average size 41.23 nm) exhibited an IC50 value of 73.37 μg/ml against total oral bacteria. While, NiO-NPs (average size 35.67 nm) were found less effective with much higher IC50 value of 197.18 μg/ml. Indeed, the Ni ions exhibited greater biocidal activity with an IC50 value of 70 μg/ml. Similar results were obtained with biofilm inhibition on the surfaces of dental prostheses. The results explicitly suggested the effectiveness of tested Ni compounds on the growth of oral bacteria and biofilm formation in the order as NiCl₂ > Ni-NPs > NiO-NPs.ConclusionThe results elucidated that Ni-NPs could serve as effective nanoantibiotics against oral bacteria.