Multiple biosurfactant production by Aureobasidium pullulans strain YTP6-14 in aqueous and heavy oil layers.

Aureobasidium pullulans YTP6-14 was demonstrated to be an excellent multiple biosurfactant producer utilizing cheap carbon sources available in Thailand, including glycerol and cassava flour hydrolysate. A. pullulans YTP6-14 maximally produced 1.81 g/l biosurfactant in an aqueous layer (BS-AQ) in a medium containing glycerol, and 7.37 or 6.37 g/l biosurfactant in a heavy oil layer (BS-HO) in cassava flour hydrolysate or a glucose containing medium, respectively. Each BS-AQ and BS-HO had critical micelle concentration values of 41.32 mg/l and 13.51 mg/l, and both biosurfactants formed a stable food oil emulsion and reduced the amount of biofilms formed by Streptococcus sobrinus and Streptococcus mutans. BS-AQ and BS-HO were mainly composed of liamocins or exophilins and massoia lactone, respectively.

Tea extracts differentially inhibit Streptococcus mutans and Streptococcus sobrinus biofilm colonization depending on the steeping temperature.

This study aimed to evaluate the effects of tea extracts on oral biofilm colonization depending on steeping temperature. S. mutans and S. sobrinus were cultured and treated with green or black tea extracts prepared under different steeping conditions. Biofilm formation, glucosyltransferase (GTF) levels, bacterial growth, and acidogenicity were evaluated. Biofilms were also assessed by gas chromatography-mass spectrometry and confocal laser scanning microscopy. All extracts with hot steeping showed higher inhibitory effects on biofilm formation and cell viability and lower GTF levels compared with those with cold steeping (p < 0.05). Hot steeping significantly reduced bacterial growth (p < 0.05) and maintained the pH. Catechins were only identified from hot steeping extracts. Within the limits of this study, extracts with cold steeping showed lower inhibitory effects on oral biofilms. The different effects between steeping extracts may be attributed to the difference in catechins released from tea extracts under the different steep conditions.

Isolation and Bacteriocin-Related Typing of Streptococcus dentisani.

Streptococcus oralis subspecies dentisani is explored as an anti-cariogenic probiotic. Here, subjecting freshly stimulated saliva samples of 35 healthy volunteers, six epidemiologically unrelated and two related strains were isolated (prevalence around 20%) applying a newly developed three-step procedure. Furthermore, the probiotic strain S. dentisani 7746 (AB-Dentisanium®) was tested under a variety of environmental conditions for its inhibitory effect on six S. mutans, two S. sobrinus, 15 other oral or intestinal streptococci, 15 S. dentisani strains, and six representatives of other species including periodontopathogens. All except one of the S. mutans strains were inhibited by 7746 colonies or culture supernatant concentrate but only if either the test cell number was low or the producer or its bacteriocin concentration, respectively, was high. S. sanguinis OMI 332, S. salivarius OMI 315, S. parasanguinis OMI 335, S. vestibularis OMI 238, and the intestinal S. dysgalactiae OMI 339 were not inhibited, while the other 10 streptococcal strains (especially S. oralis OMI 334 and intestinal S. gallolyticus OMI 326) showed a certain degree of inhibition. From the panel of other bacterial species only Aggregatibacter actinomycetemcomitans was slightly inhibited. With the exception of OMI 285 and OMI 291 that possessed a 7746 bacteriocin-like gene cluster, all S. dentisani strains and especially type strain 7747T were strongly inhibited by 7746. In conclusion, probiotic strain 7746 might antagonize the initiation and progression of dental caries by reducing S. mutans if not too abundant. S. dentisani strains inhibit each other, but strains with similar bacteriocin-related gene clusters, including immunity genes, are able to co-exist due to cross-resistance. In addition, development of resistance and adaptation to 7746-bacteriocins was observed during our study and needs attention. Hence, mechanisms underlying such processes need to be further investigated using omics-approaches. On the manufacturing level, probiotic strains should be continuously tested for function. Further clinical studies investigating inhibition of S. mutans by AB-Dentisanium® are required that should also monitor the impact on the oral microbiome composition including resident S. dentisani strains.

Effects of UVB and UVC irradiation on cariogenic bacteria in vitro.

The aim of this study was to evaluate the bactericidal effect of ultraviolet (UV) light at 265-nm (UVC) and 310-nm (UVB) wavelengths from a newly developed UV light-emitting diode (LED) device against cariogenic bacteria in vitro. Suspensions of Streptococcus mutans (S. mutans) and Streptococcus sobrinus were directly irradiated by UVB or UVC for 2.5 min or 5 min. Numbers of colonies were counted and calculated as colony forming units (CFU) per milliliter. Fluorescence microscopy (FM) and optical density measurements at 490 nm (OD490) were also taken after irradiation. In addition, the bactericidal effects of irradiation against S. mutans under 0.5 mm-thick dentin were compared using culture tests and OD490 measurements. Direct UV-LED irradiation with both UVB and UVC showed strong bactericidal effects. UVB showed superior bactericidal effect through 0.5-mm-thick dentin than did UVC, especially after demineralization. These results suggest that UVB irradiation could be utilized for the prevention and management of dental caries.

Preparation of fluoride-loaded microcapsules for anticariogenic bacterial growth using a coaxial ultrasonic atomizer.

A new method to deliver fluoride using biodegradable poly(lactic-co-glycolic acid) microcapsules to suppress cariogenic bacterial growth during orthodontic treatment was investigated. A coaxial ultrasonic atomizer was used to encapsulate KF solution into microcapsules. The orthodontic adhesive resin disk containing fluoride loaded microcapsules (DFLM) was prepared by LED light curing. The microstructure of microcapsules, successful loading of KF, fracture strength, and shear bonding strength were assessed by FE-SEM, confocal laser scanning microscope, and general purpose testing machine, respectively. Fluoride release from the DFLM in phosphate buffered saline and pH changes were measured after different periods of soaking time. Antibacterial activity of the DFLM was assessed in tryptic soy broth containing mutant streptococci. The starting inoculum and the orthodontic resin disk containing microcapsules not loaded with KF were used as negative and positive controls, respectively. As results, the cumulative amount of KF after 49 days was about 85% of the initial amount of fluoride contained in the microcapsules. The fracture and shear bonding strengths of the orthodontic resin disks with and without the microcapsules were similar to each other. The DFLM showed lower bacterial growth than the control groups, whereas no statistically significant differences were found between the negative and positive controls. It can be concluded that the microcapsules loaded with fluoride prepared by a coaxial ultrasonic atomizer have good potential for application as an antibacterial agent due to their excellent cariogenic antibacterial activity when incorporated into orthodontic adhesive resin. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 31-39, 2018.

Antimicrobial and biological activity of leachate from light curable pulp capping materials.

OBJECTIVES: Characterization of a number of pulp capping materials and assessment of the leachate for elemental composition, antimicrobial activity and cell proliferation and expression. METHODOLOGY: Three experimental light curable pulp-capping materials, Theracal and Biodentine were characterized by scanning electron microscopy, energy dispersive spectroscopy and X-ray diffraction. The elemental composition of the leachate formed after 24h was assessed by inductively coupled plasma (ICP). The antimicrobial activity of the leachate was determined by the minimum inhibitory concentration (MIC) against multispecies suspensions of Streptococcus mutans ATCC 25175, Streptococcus gordonii ATCC 33478 and Streptococcus sobrinus ATCC 33399. Cell proliferation and cell metabolic function over the material leachate was assessed by an indirect contact test using 3-(4,5 dimethylthiazolyl-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. RESULTS: The hydration behavior of the test materials varied with Biodentine being the most reactive and releasing the highest amount of calcium ions in solution. All materials tested except the unfilled resin exhibited depletion of phosphate ions from the solution indicating interaction of the materials with the media. Regardless the different material characteristics, there was a similar antimicrobial activity and cellular activity. All the materials exhibited no antimicrobial activity and were initially cytotoxic with cell metabolic function improving after 3days. CONCLUSIONS: The development of light curable tricalcium silicate-based pulp capping materials is important to improve the bonding to the final resin restoration. Testing of both antimicrobial activity and biological behavior is critical for material development. The experimental light curable materials exhibited promising biological properties but require further development to enhance the antimicrobial characteristics.

Compositional differences in multi-species biofilms formed on various orthodontic adhesives.

OBJECTIVES: To investigate the aspects of multi-species biofilm formation on various orthodontic adhesives with different surface characteristics. METHODS: Multi-species biofilms using 13 bacterial species were grown on the surfaces of composite, compomer, and resin-modified glass-ionomer cement (RMGI). The changes in Streptococcus mutans (Sm), Streptococcus sobrinus (Ss), Porphyromonas gingivalis (Pg), Aggregatibacter actinomycetemcomitans (Aa), and total bacteria were determined at day 1 (T1) and day 4 (T2) using real-time polymerase chain reaction. Surface roughness (SR), surface free energy (SFE), and surface texture were analyzed to explain the differences in bacterial compositions among the adhesives. Repeated measures analysis of variance was used to determine time-related changes in bacterial compositions with respect to adhesive type. The Kruskal-Wallis test was used to determine differences in SR and SFE among the adhesives. RESULTS: There were no significant differences in the adhesion of total bacteria among the adhesives; however, the adhesion of Sm, Ss, and Pg was higher to RMGI than the other adhesives. The amount of Sm, Ss, and total bacteria increased from T1 to T2, while Pg and Aa decreased from T1 to T2. RMGI showed a rougher surface relative to composite or compomer due to the presence of micro-pores and/or flaws. Compomer had the greatest SFE followed by RMGI and composite. Interestingly, SR differences were about 10 times greater than SFE differences among the adhesives. CONCLUSIONS: Considering the greater differences in SR than SFE among the adhesives, the rougher surface of RMGI may cause greater adhesion of Sm, Ss, and Pg.

Antimicrobial effect of the extracts from Hypericum perforatum against oral bacteria and biofilm formation.

CONTEXT: One traditional medicines, Hypericum perforatum L. (Hypericaceae), possesses several beneficial effects against depression, ulcers, dyspepsia, abdominal pains, burns, bacterial infections, migraine headaches, and sciatica. OBJECTIVE: The present study investigates the antimicrobial activity of the ethanol extract (HP-EtOH) of H. perforatum and its sub-extracts, namely n-hexane (HP-hexane), chloroform (HP-CHCl3), ethyl acetate (HP-EtOAc), n-butanol (HP-n-BuOH), and water (HP-H2O) extracts, against Streptococcus mutans, S. sobrinus, Lactobacillus plantarum, and Enterococcus faecalis. MATERIALS AND METHODS: For the evaluation of the antimicrobial activity, flowering aerial parts of H. perforatum were extracted with EtOH and then this extract was fractionated to obtain five sub-extracts in different polarities. Antimicrobial activities of HP-EtOH and its sub-extracts against Streptococcus mutans, S. sobrinus, L. plantarum, and E. faecalis were assessed by using colorimetric micro-well dilution at concentration ranges of 64-0.5 µg/ml as well as resazurin microplate and modified microtiter-plate assays between the ranges of 100 and 0.78125 µg/ml. RESULTS: According to the results of the present study, HP-H2O sub-extract displayed strong antibacterial activity (MIC values 8 µg/mL) against S. sobrinus and L. plantarum, and exerted moderate activity against S. mutans and E. faecalis at 32 and 16 µg/mL concentrations, respectively. Other sub-extracts also demonstrated antimicrobial activity against S. sobrinus at a concentration of 16 µg/mL. HP-EtOAc and HP-n-BuOH showed antimicrobial activity against L. plantarum and HP-EtOAc and HP-H2O were also active against E. faecalis at the same concentrations (16 µg/mL). CONCLUSION: According to the results, we suggest that H. perforatum could be employed as a natural antibacterial agent in oral care products.

Application of chimeric glucanase comprising mutanase and dextranase for prevention of dental biofilm formation.

Water-insoluble glucan (WIG) produced by mutans streptococci, an important cariogenic pathogen, plays an important role in the formation of dental biofilm and adhesion of biofilm to tooth surfaces. Glucanohydrolases, such as mutanase (α-1,3-glucanase) and dextranase (α-1,6-glucanase), are able to hydrolyze WIG. The purposes of this study were to construct bi-functional chimeric glucanase, composed of mutanase and dextranase, and to examine the effects of this chimeric glucanase on the formation and decomposition of biofilm. The mutanase gene from Paenibacillus humicus NA1123 and the dextranase gene from Streptococcus mutans ATCC 25175 were cloned and ligated into a pE-SUMOstar Amp plasmid vector. The resultant his-tagged fusion chimeric glucanase was expressed in Escherichia coli BL21 (DE3) and partially purified. The effects of chimeric glucanase on the formation and decomposition of biofilm formed on a glass surface by Streptococcus sobrinus 6715 glucosyltransferases were then examined. This biofilm was fractionated into firmly adherent, loosely adherent, and non-adherent WIG fractions. Amounts of WIG in each fraction were determined by a phenol-sulfuric acid method, and reducing sugars were quantified by the Somogyi-Nelson method. Chimeric glucanase reduced the formation of the total amount of WIG in a dose-dependent manner, and significant reductions of WIG in the adherent fraction were observed. Moreover, the chimeric glucanase was able to decompose biofilm, being 4.1 times more effective at glucan inhibition of biofilm formation than a mixture of dextranase and mutanase. These results suggest that the chimeric glucanase is useful for prevention of dental biofilm formation.

In vitro study of antigrowth capacity and antiacid capacity on Sreptococcus sobrinus 6715 of sorghum procyanidin dimers.

The bacteria grow in oral cavity and product acids, which could induce dental caries. In this study, in order to obtain the relationship between procyanidin dimers from sorghum episperm (sorghum procyanidins, SPC) and its anticaries effect. The extract of SPC purified by macroporous resin was divided into three parts by gel chromatography, marked as GPC-1, GPC-2, and GPC-3 in order. The ESI-MS and MS/MS analysis indicated that the main composition of GPC-2 was procyanidin dimers. In addition, the capacities of antigrowth and antiacid on Sreptococcus sobrinus 6715 were analysised to investigate the effect of SPC dimers in protecting against dental caries. The results indicated that the minimum inhibitory concentration (MIC) of SPC dimers was 16 mg/mL. Furthermore, the SPC dimers had notable preventive effect < against the acid production of Sreptococcus sobrinus 6715 compared with the control group, which suppressed in a dose-dependent manner by pH decline. These findings indicated that SPC dimers had potential to be used as anticaries preventive medicine due to its strong capacity of antigrowth and antiacid.

Efficacy of three hygienic protocols in reducing biofilm adherence to removable thermoplastic appliance.

OBJECTIVES: To examine the ability of a removable thermoplastic appliance (RTA) to adsorb hygienic solutions and inhibit bacterial growth and to examine the efficacy of three hygiene protocols in reducing bacterial biofilm adherence to RTA. MATERIALS AND METHODS: Solution adsorption and bacterial growth inhibition were examined in vitro using paper vs RTA discs. Subsequently, 11 patients treated with RTA (mean age, 29.1 ± 4.7 years) were assigned into a sequence of three hygiene protocols: regular RTA brushing (baseline), immersion RTAs in chlorhexidine mouthwash (CHX), and using a vibrating bath with cleaning solution (VBC). For each patient, 12 upper RTAs were examined (2 baseline RTAs, 5 CHX RTAs, and 5 VBC RTAs), for a total of 132 RTAs. All RTAs were stained with gentian violet, and biofilm presence was measured using a photodensitometer. RESULTS: The RTA discs did not adsorb CHX or cleaning solution. The later agent did not show antibacterial features. Baseline RTAs showed significant biofilm adherence (P < .001) on the posterior palatal side of the aligner and on the anterior incisal edge. CHX and VBC hygienic protocols significantly (P < .001) reduced baseline biofilm adherence by 16% and 50%, respectively. Hygienic improvement was maintained over 140 days when CHX and VBC were used. However, VBC was three times more efficient than CHX. CONCLUSIONS: This study highly recommends the use of a VBC protocol. Biofilm deposits on the RTA, especially on incisal edges and attachment dimples, could lead to inadequate tooth/RTA and attachment/RTA overlap and consequently impair tooth alignment.

Effects of Withania somnifera on the growth and virulence properties of Streptococcus mutans and Streptococcus sobrinus at sub-MIC levels.

The aim of this study was to evaluate the effect of the methanol extract of Withania somnifera (MEW) on the growth and virulence properties of Streptococcus mutans and Streptococcus sobrinus at sub-minimum inhibitory concentration (MIC) levels and to identify the main components of MEW. First, antibacterial activity of MEW against oral bacteria was determined using a micro-dilution method. Then, the effect of MEW on the growth of S. mutans and S. sobrinus was investigated at sub-MIC levels. To test the effect of MEW on the virulence properties of S. mutans and S. sobrinus, assays for acid production, acid tolerance, and biofilm formation were performed at sub-MIC levels. A GC-MS analysis for the main components of MEW was also carried out. MEW showed a broad antibacterial range against oral bacteria (MIC: 0.125-2 mg/mL). At sub-MIC levels, MEW dose-dependently increased doubling times of S. mutans and S. sobrinus up to 258% and 400%, respectively. Furthermore, MEW inhibited acid production, acid tolerance, and biofilm formation of S. mutans and S. sobrinus at sub-MIC levels. The GC-MS analysis revealed the presence of mono- and disaccharides, sugar alcohols, and organic acids as main components. These data suggest that MEW might be useful for restraining physiological activities of cariogenic bacteria.

Quantitative analysis of S. mutans and S. sobrinus cultivated independently and adhered to polished orthodontic composite resins.

UNLABELLED: In orthodontics, fixed appliances placed in the oral cavity are colonized by microorganisms. OBJECTIVE: The purpose of this study was to quantitatively determine the independent bacterial colonization of S. mutans and S. sobrinus in orthodontic composite resins. MATERIAL AND METHODS: Seven orthodontic composite adhesives for bonding brackets were selected and classified into 14 groups; (GIm, GIs) Enlight, (GIIm, GIIs) Grengloo, (GIIIm, GIIIs) Kurasper F, (GIVm, GIVs) BeautyOrtho Bond, (GVm, GVs) Transbond CC, (GVIm, GVIs) Turbo Bond II, (GVIIm, GVIIs) Blugloo. 60 blocks of 4x4x1 mm of each orthodontic composite resin were made (total 420 blocks), and gently polished with sand-paper and ultrasonically cleaned. S. mutans and S. sobrinus were independently cultivated. For the quantitative analysis, a radioactive marker was used to codify the bacteria (³H) adhered to the surface of the materials. The blocks were submerged in a solution with microorganisms previously radiolabeled and separated (210 blocks for S. mutans and 210 blocks for S. sobrinus) for 2 hours at 37 ºC. Next, the blocks were placed in a combustion system, to capture the residues and measure the radiation. The statistical analysis was calculated with the ANOVA test (Sheffè post-hoc). RESULTS: Significant differences of bacterial adhesion were found amongst the groups. In the GIm and GIs the significant lowest scores for both microorganisms were shown; in contrast, the values of GVII for both bacteria were significantly the highest. CONCLUSIONS: This study showed that the orthodontic composite resin evaluated in the GIm and GIs, obtained the lowest adherence of S. mutans and S. sobrinus, which may reduce the enamel demineralization and the risk of white spot lesion formation.

Contact-killing of adhering streptococci by a quaternary ammonium compound incorporated in an acrylic resin.

PURPOSE: Acrylates for bonding of joint prostheses and stainless-steel brackets in orthopedics and orthodontics are prone to bacterial adhesion and biofilm formation, respectively, leading to serious infectious complications. Here we describe the preparation of a contact-killing acrylic resin by incorporation of [3-(methacryloylamino)propyl] trimethylammonium chloride (MAPTAC). METHODS: Physicochemical properties of the acrylates with and without MAPTAC incorporated were determined with X-ray photoelectron spectroscopy and water contact angles. The bond-strength of the acrylate with different percentages of MAPTAC was determined in a shear mode. The efficacy in contact-killing of the acrylate with MAPTAC incorporated with and without an adsorbed salivary coating was evaluated for various oral streptococcal strains. Cytotoxicity was tested against human skin fibroblasts. RESULTS: Acrylates with 16 wt% and 20 wt% incorporated MAPTAC showed strong contact-killing of various oral streptococcal strains up to challenge concentrations of 109 mL-1 within 15 min, with no elution of antimicrobial polymers. Contact-killing reduced after coating with a salivary conditioning film, but still remained significant up to a challenge concentration of 105 mL-1. No cytotoxicity of acrylate with incorporated MAPTAC was observed toward human skin fibroblasts. The bond strengths of stainless-steel brackets fixed to etched enamel through the resin (12 ± 3 MPa) decreased with increasing amounts MAPTAC to half of the original value when 20 wt% of MAPTAC was incorporated, which remained within a clinically acceptable range. CONCLUSIONS: These results suggest that MAPTAC can be effectively incorporated in orthodontic resin to provide long-term bactericidal activity against oral bacteria, with potential application in orthopedics.

Quantitative analysis of S. mutans and S. sobrinus cultivated independently and adhered to polished orthodontic composite resins

In Orthodontics, fixed appliances placed in the oral cavity are colonized by microorganisms. OBJECTIVE: The purpose of this study was to quantitatively determine the independent bacterial colonization of S. mutans and S. sobrinus in orthodontic composite resins. MATERIAL AND METHODS: Seven orthodontic composite adhesives for bonding brackets were selected and classified into 14 groups; (GIm, GIs) Enlight, (GIIm, GIIs) Grengloo, (GIIIm, GIIIs) Kurasper F, (GIVm, GIVs) BeautyOrtho Bond, (GVm, GVs) Transbond CC, (GVIm, GVIs) Turbo Bond II, (GVIIm, GVIIs) Blugloo. 60 blocks of 4x4x1 mm of each orthodontic composite resin were made (total 420 blocks), and gently polished with sand-paper and ultrasonically cleaned. S. mutans and S. sobrinus were independently cultivated. For the quantitative analysis, a radioactive marker was used to codify the bacteria (³H) adhered to the surface of the materials. The blocks were submerged in a solution with microorganisms previously radiolabeled and separated (210 blocks for S. mutans and 210 blocks for S. sobrinus) for 2 hours at 37ºC. Next, the blocks were placed in a combustion system, to capture the residues and measure the radiation. The statistical analysis was calculated with the ANOVA test (Sheffè post-hoc). RESULTS: Significant differences of bacterial adhesion were found amongst the groups. In the GIm and GIs the significant lowest scores for both microorganisms were shown; in contrast, the values of GVII for both bacteria were significantly the highest. CONCLUSIONS: This study showed that the orthodontic composite resin evaluated in the GIm and GIs, obtained the lowest adherence of S. mutans and S. sobrinus, which may reduce the enamel demineralization and the risk of white spot lesion formation.

Effects of two fluoride varnishes and one fluoride/chlorhexidine varnish on Streptococcus mutans and Streptococcus sobrinus biofilm formation in vitro.

AIMS: The aim of this study was to evaluate and to compare the effect of two fluoride varnishes and one fluoride/chlorhexidine varnish on Streptococcus mutans and Streptococcus sobrinus biofilm formation, in vitro. STUDY DESIGN: Standard acrylic discs were prepared and divided into groups based on the varnish applied to the disc surface: Fluor Protector, Bifluoride 12, and Fluor Protector + Cervitec (1:1). Untreated discs served as controls. In the study groups, biofilms of S. mutans and S. sobrinus were formed over 24 h, 48 h, and 5 days. The fluoride concentrations in the monospecies biofilms and viable counts of S. mutans and S. sobrinus were investigated. RESULTS: In all study groups, a statistically significant increase in the viable number of S. mutans and S. sobrinus cells was observed between 24 h and 5 days. In both monospecies biofilms, the greatest antibacterial efficacy was detected in the Fluor Protector and Fluor Protector + Cervitec groups at 24 h. For all groups, the amount of fluoride released was highest during the first 24 h, followed by a significant decrease over the next 4 days. A negative correlation was detected between fluoride concentration and antibacterial effect in those groups with biofilms containing both species. Despite the release of high levels of fluoride, the greatest number of viable S. mutans and S. sobrinus cells was detected in the Bifluoride 12 group. STATISTICS: The data were analyzed using GraphPad Prism software (ver. 3). CONCLUSIONS: The Fluor Protector + Cervitec varnish exerted prolonged antibacterial effects on S. mutans and S. sobrinus biofilms compared to the other varnishes tested.

Inhibitory properties of Coffea canephora extract against oral bacteria and its effect on demineralisation of deciduous teeth.

OBJECTIVES: The antibacterial activity of Coffea canephora extract was evaluated in vitro against Streptococcus mutans and Streptococcus sobrinus. The viability of planktonic cells was analysed by susceptibility tests (MIC and MBC) and time-kill assays. The effect of the extract on dental demineralisation was also investigated. METHODS: Primary 1st molar fragments (n=24) were inoculated with a saliva pool and sustained in a multiple plaque growth system for 10 days to form biofilm. The biofilm was treated with light roasted C. canephora extract at 20%, Milli-Q water (negative control) and chlorhexidine (positive control) once a day, during a week. Blank controls comprised fragments without treatment. Biofilm pH was monitored in the last day of treatment. Changes in tooth mineralisation were assessed by cross-sectional microhardness (CSMH) test. RESULTS: MIC and MBC for S. mutans were 7±2 mg/mL and 160±0 mg/mL, respectively, showing no activity for S. sobrinus. The extract produced a 4-log reduction in the number of colonies of S. mutans after 3-h treatment (p<0.05) with undiluted extract (20%) and MBC concentration (16%). There was no difference among negative/blank controls and coffee plaque pH. Differences between CSMH values of dental fragments subjected to the coffee extract and to chlorhexidine were not significant. At depths up to 30 µm from the enamel surface, coffee extract and chlorhexidine promoted higher CSMH values when compared to blank/negative controls (p<0.05). CONCLUSION: Our data suggest that light roasted C. canephora extract is beneficial as an anticariogenic substance.

Antibacterial effects of 4-META/MMA-TBB resin containing chlorhexidine.

This study evaluated the antibacterial effects of 4-acryloyloxyethyl trimellitate anhydride/methyl methacrylate-tri-n-butylborane (4-META/MMA-TBB) resin containing chlorhexidine (CHX) digluconate. The CHX was incorporated into the resins at varying concentrations from 0.0 (control) to 3.0%. The antibacterial effect of each resin against seven microorganisms was examined using agar diffusion tests. Growth inhibition of two streptococci was also tested in brain heart infusion (BHI) liquid broth containing each resin. Release of CHX was analyzed using HPLC, and enamel and dentin bond strengths of each resin were measured. In the agar diffusion tests, growth of all the microorganisms was inhibited by the resin specimens containing CHX at 1.0% or higher except for Enterococcus faecalis. There was greater sensitivity in Streptococcus mutans and Streptococcus sobrinus than in the other bacteria tested. Complete growth inhibition of these two streptococci was observed in BHI liquid broth containing 1.0-1.5% or greater CHX-incorporated resin. According to HPLC analysis, 1.0% and 1.5% CHX-containing resins released means of 3.63 and 8.59 microg/mL of CHX, respectively. Specimens with 0.5-1.5% and 0.5-2.0% CHX exhibited no significant reduction in enamel and dentin bond strengths, respectively, when compared to the control (p > 0.05). This in vitro study suggested that incorporation of 1.0-1.5% CHX digluconate into the 4-META resin is optimal in terms of antibacterial effects and bond strength to the tooth.

Final pH affects the interference capacity of naturally occurring oral Lactobacillus strains against mutans streptococci.

OBJECTIVES: To establish the effects of three factors: previous caries experience; colonization of Streptococcus mutans; and final pH on autologous lactobacilli-mediated inhibition against a panel of mutans streptococci in young subjects with different caries experiences. DESIGN: The inhibition capacity was determined by the use of the agar overlay method and the final pH in culture medium was measured after 20 h. Using a logistic regression model, the risk of having an incomplete lactobacilli-mediated inhibition was calculated. RESULTS: All three factors significantly influenced the interference outcome in the order; final pH of the Lactobacillus strains, oral colonization of autologous S. mutans and caries experience. A high risk occurred at a lower pH and at a wider pH range for individuals with previous caries experience and autologous colonization of S. mutans compared with caries-free subjects who were not colonized. At a final pH of 4.0, this risk was approximately eight times higher than that of the latter group. Two mutans Streptococcus strains in the test panel demonstrated high individual predictive values of inhibition mediated by oral lactobacilli. CONCLUSIONS: Generation of a low pH either directly via organic acid production and/or production of bacteriocins or metabolites at a low pH may promote mutans Streptococcus growth inhibition, in vitro. Furthermore, a shift of pH range for the risk of incomplete inhibition of mutans streptococci suggests a less effective inhibition at a wider pH range for naturally occurring lactobacilli from individuals with earlier caries experience containing own S. mutans.

In vitro antimicrobial effect of bacteriocin PsVP-10 in combination with chlorhexidine and triclosan against Streptococcus mutans and Streptococcus sobrinus strains.

OBJECTIVES: To determine the minimal inhibitory concentrations (MICs) of bacteriocin PsVP-10, chlorhexidine and triclosan on S. mutans and S. sobrinus and to study the potential synergistic combination between these antimicrobial and the bacteriocin PsVP-10. DESIGN: Were determined MICs of bacteriocin PsVP-10, triclosan and chlorhexidine on strains of S. mutans and S. sobrinus, which formed a biofilm or did not form a biofilm. In addition, the synergistic effect was analysed by the determination of respective fractionary inhibitory concentrations (FICs) between bacteriocin PsVP-10 plus chlorhexidine and bacteriocin PsVP-10 plus triclosan. RESULTS: MICs of three antimicrobials used were higher in those bacterial strains, which formed a biofilm. An interesting synergistic effect on both studied species was observed when bacteriocin and chlorhexidine were combined. A slighter synergy was determined for the combination bacteriocin PsVP-10 and triclosan. CONCLUSIONS: The results showed that the combination of chlorhexidine bacteriocin PsVP-10 could reduce the number of cariogenic bacteria for in vitro studies. In the future this synergistic combination could be an alternative to antimicrobial therapy against S. mutans or S. sobrinus.