Effect of desensitising paste containing 8% arginine and calcium carbonate on biofilm formation of Streptococcus mutans in vitro

Objectives

To evaluate the influence of desensitising paste containing 8% arginine and calcium carbonate (Ar-Ca) on biofilm formation on dentine.

Methods

Dentine discs were cut from extracted third molars and divided into the following three groups: no treatment, pumice treatment and Ar-Ca treatment. Surface topography and roughness were examined using scanning electron microscopy (SEM) and non-contact 3D surface profiler. After sterilisation, samples were incubated with Streptococcus mutans (S. mutans) for 4 h, 24 h and 72 h. Bacterial adhesion and biofilm formation were analysed using SEM, whereas MTT and lactic acid production assays were used to analyse the metabolic activity of S. mutans.

Results

After polishing with either pumice or Ar-Ca, the surfaces of the samples became smoother than in the control group. The Ra values of the three experimental groups decreased significantly to 0.43 μm, 0.3 μm and 0.26 μm, respectively. Compared to the control group, fewer bacteria adhered to the dentine surface in the Ar-Ca group, while biofilm thickness decreased significantly for both groups after incubating for 24 h and 72 h. MTT and lactic acid production levers also showed a significant reduction in the Ar-Ca group.

Conclusions

Ar-Ca appears to present antibiofilm efficacy and may provide a promising approach to combat bacterial infection in hypersensitive dentinal lesions.

Clinical significance

As a clinical application of desensitising polishing paste, the paste containing 8% arginine and calcium carbonate could also inhibit the biofilm formation effectively.     

Substantivity of a single chlorhexidine mouthwash on salivary flora: Influence of intrinsic and extrinsic factors

Objectives

To analyse the influence of intrinsic and extrinsic factors on the in vivo antimicrobial activity of a chlorhexidine (CHX) digluconate mouthwash on the salivary flora up to 7 h after its application, using epifluorescence microscopy.

Methods

Ten volunteers performed the following mouthwashes: 0.12% CHX (10 ml/30 s, 15 ml/30 s and 10 ml/1 min); 0.2% CHX (10 ml/30 s, 15 ml/30 s and 10 ml/1 min); 0.2% CHX (10 ml/30 s) plus different daily activities (eating, drinking, chewing or smoking).

Results

On comparing 0.12% CHX (10 ml versus 15 ml), the greatest differences in bacterial viability were detected at 1 h and 3 h. On comparing 0.12% CHX (30 s versus 1 min) the greatest differences in viability were detected at 1 h, 3 h, and 5 h; and with 0.2% CHX (30 s versus 1 min), at 5 h and 7 h. On comparing 0.12% CHX (15 ml) versus 0.2% CHX (10 ml) and 0.12% CHX (1 min) versus 0.2% CHX (30 s), the percentage of viable bacteria was higher with the 0.12% concentration. On comparing 0.2% CHX versus 0.2% CHX plus daily activities, the higher differences were detected after eating and chewing, followed by drinking.

Conclusion

An increase in the volume of 0.12% or 0.2% CHX mouthwashes does not affect the duration of antimicrobial activity in saliva, whereas increasing the duration produces a marked increase in substantivity. Substantivity was greater with 0.2% CHX than 0.12% CHX. Eating, chewing or drinking significantly reduces the 0.2% CHX substantivity in saliva.     

The preventive effect of grape seed extract on artificial enamel caries progression in a microbial biofilm-induced caries model

Objectives

The aim of this study was to evaluate the effect of grape seed extract (GSE) on enamel caries lesion formation in an in vitro Streptococcus mutans biofilm model.

Methods

Enamel fragments were prepared from bovine incisors and divided into six treatment groups (n = 12): inoculated Brain Heart Infusion with 1% sucrose (BHIS), 1 mg/mL GSE, 2 mg/mL GSE, 3 mg/mL GSE, 10 ppm fluoride as NaF, and uninoculated BHIS. For biofilm formation, tooth fragments were incubated anaerobically in polystyrene 6-well tissue culture plates containing BHIS, the respective agents, and S. mutans (1 × 10⁵ CFU/mL) for 24 h at 37 °C. Culture medium was replaced with fresh BHIS and respective agents daily over a 7-day period. Following caries lesion formation, lesion depth (LD) and relative optical density (ROD) were determined by polarized light microscopy (PLM) and confocal laser scanning microscopy (CLSM), respectively, to evaluate lesion progression.

Results

LDs of the 2 mg/mL GSE group (122.86 ± 13.41 μm) and the 3 mg/mL GSE group (111.92 ± 11.39 μm) were significantly smaller than those of the 1 mg/mL GSE (198.33 ± 17.70 μm) and control groups (210.86 ± 15.50 μm) (p < 0.05). Compared with the 2 mg/mL and 3 mg/mL groups, the control and 1 mg/mL GSE groups showed significantly lower ROD values when depth was less than 200 μm, indicating greater mineral loss.

Conclusions

Dose-dependent GSE inhibits in vitro enamel caries formation due to its ability to suppress growth of S. mutans and the formation of bio?lm.

Clinical signi?cance

Grape seed extract may be a novel virulence-targeted natural antimicrobial agent for caries prevention.     

Inhibition of microbial adhesion to plastic surface and human buccal epithelial cells by Rhodomyrtus tomentosa leaf extract

Objective

The adherence of oral pathogenic microorganisms to host tissues is the initial step for successful process of oral diseases. This study aimed to determine the effect of the Rhodomyrtus tomentosa leaf extract and rhodomyrtone, an antibacterial compound from R. tomentosa leaf, on adhesion of some oral pathogens to polystyrene plastic surface and human buccal epithelial cells.

Methods

The minimum inhibitory concentration (MIC) was evaluated using broth microdilution method. The microbial adhesion to the plastic surface and buccal cells was determined using microtiter plate method and microscopy technique.

Results

The ethanol extract of leaf demonstrated antibacterial activity against oral microorganisms including Staphylococcus aureus ATCC 25923, Streptococcus mutans (clinical isolate), and Candida albicans ATCC 90028 with the MIC values of 31.25, 15.62, and 1000 μg/ml, respectively. Rhodomyrtone displayed activity with the MIC values of 0.78 and 0.39 μg/ml against S. aureus ATCC 25923 and S. mutans, respectively. The MIC value of the compound against C. albicans ATCC 90028 was more than 100 μg/ml which was the highest test concentration. All pathogenic microorganisms treated with the extract and rhodomyrtone at their subinhibitory concentrations resulted in a decrease in their adherence ability to both plastic surface and buccal cells.

Conclusion

It is suggested that R. tomentosa extract and rhodomyrtone may be useful in therapy or as prophylaxis in infections involving oral pathogens.     

Effect of charge density of bonding agent containing a new quaternary ammonium methacrylate on antibacterial and bonding properties

Objective

Quaternary amine charge density is important because when the negatively charged bacteria contact the positive quaternary amine charge, the electric balance is disturbed and the bacterium could be disrupted. There has been no report on the effects of charge density on the antibacterial efficacy of dental bonding agents. The objective of this study was to synthesize a new quaternary ammonium methacrylate, and investigate the effects of charge density of bonding agent on bacteria early-attachment, biofilm colony-forming units (CFU) and dentin bond strength.

Methods

Dimethylaminododecyl methacrylate (DMAHDM) with an alkyl chain length of 16 was synthesized and mixed into Scotchbond Multi-Purpose adhesive and primer (SBMP) at mass fractions of 0%, 2.5%, 5%, 7.5%, and 10%. A microtensile dentin bond test was performed. The density of quaternary ammonium groups was measured using a fluorescein dye method. Streptococcus mutans (S. mutans) early-attachment was examined at 4 h, and biofilm colony-forming units (CFU) were measured at 2 days.

Results

All groups had similar microtensile bonding strengths (mean ± sd; n = 40) of about 60 MPa (p > 0.1). Quaternary amine charge density of bonding agents monotonically increased with increasing DMAHDM mass fraction. Bacteria early-attachment coverage greatly decreased with increasing DMAHDM content in the resin. Biofilm CFU at 10% DMAHDM was reduced by more than 4 log, compared to SBMP control. Charge density of bonding agent was inversely proportional to bacteria early-attachment coverage and biofilm CFU.

Significance

Increasing the quaternary amine charge density of dentin bonding agent resin was shown to greatly reduce S. mutans attachment and decrease biofilm CFU by four orders of magnitude, without compromising the dentin bond strength. The new DMAHDM is promising for use in bonding agents and other antibacterial restorative materials to inhibit caries.     

An in vitro study of alginate oligomer therapies on oral biofilms

Objectives

The in vitro effect of a novel, oligosaccharide nanomedicine OligoG against oral pathogen-related biofilms, both alone and in the presence of the conventional anti-bacterial agent triclosan, was evaluated.

Methods

The effect of OligoG ± triclosan was assessed against established Streptococcus mutans and Porphyromonas gingivalis biofilms by bacterial counts and image analysis using LIVE/DEAD^® staining and atomic force microscopy (AFM). The effect of triclosan and OligoG surface pre-treatments on bacterial attachment to titanium and polymethylmethacrylate was also studied.

Results

OligoG potentiated the antimicrobial effect of triclosan, particularly when used in combination at 0.3% against S. mutans grown in artificial saliva. OligoG was less effective against established P. gingivalis biofilms. However, attachment of P. gingivalis, to titanium in particular, was significantly reduced after surface pre-treatment with OligoG and triclosan at 0.01% when compared to controls. Light microscopy and AFM showed that OligoG was biocidal to P. gingivalis, but not S. mutans.

Conclusions

OligoG and triclosan when used in combination produced an enhanced antimicrobial effect against two important oral pathogens and reduced bacterial attachment to dental materials such as titanium, even at reduced triclosan concentrations. Whilst the use of triclosan against oral bacteria has been widely documented, its synergistic use with OligoG described here, has not previously been reported. The use of lower concentrations of triclosan, if used in combination therapy with OligoG, could have environmental benefits.

Clinical importance

The potentiation of antimicrobial agents by naturally occurring oligomers such as OligoG may represent a novel, safe adjunct to conventional oral hygiene and periodontal therapy. The ability of OligoG to inhibit the growth and impair bacterial adherence highlights its potential in the management of peri-implantitis.     

Adjunctive application of chlorhexidine and ethanol-wet bonding on durability of bonds to sound and caries-affected dentine

Objectives

To examine the effect of adjunctive application of chlorhexidine (CHX) and ethanol-wet bonding (EWB) on bond durability and nanoleakage of hydrophobic adhesive to sound (SD) and caries-affected dentine (CAD).

Methods

Dentine surfaces of molars were etched after caries removal and randomly allocated to four groups (n = 12). In Groups 1 and 2, dentine surfaces were saturated with either 2 ml of 100% ethanol or 2 ml of ethanol with 2% CHX for 60 s. In Groups 3 and 4, dentine surfaces were saturated with either 15 μL of distilled water or 15 μL of distilled water with 2% CHX for 60 s. Two coats of primer, followed by neat resin were applied and light-cured for 40 s. Resin composite build-ups were placed and bonded specimens were sectioned for bond strength testing after 24 h and 12 months’ storage in artificial saliva. Bond strength data were analyzed using 3-way ANOVA and SNK tests. Interfacial nanoleakage was evaluated after 24 h and 12 months using a field-emission scanning electron microscopy and data were analyzed using Kruskal–Wallis test.

Results

Significant differences were observed for the three factors: “substrate” (p < 0.001), “rewetting agents” (p < 0.001) and “time” (p < 0.001) on bond strength. Incorporation of 2% CHX to EWB preserved bond strength to SD and CAD and reduced interfacial nanoleakage after 12 months. Incorporation of 2% CHX to WWB also preserved bond strength to SD after ageing.

Conclusions

Incorporation of chlorhexidine to ethanol-wet bonding has an interaction effect on preservation of bond durability to sound and caries-affected dentine.

Clinical significance

Incorporation of chlorhexidine to ethanol-wet bonding with hydrophobic adhesive enhances the success rate of aesthetic bonded restorations.     

Effect of cariogenic biofilm challenge on the surface hardness of direct restorative materials in situ

Objectives

The presence of cariogenic biofilm could result in surface degradation of composite and ionomeric restorative materials. Thus, this study evaluated in situ the alterations in the surface microhardness of these materials under biofilm accumulation and cariogenic challenge.

Methods

In a split-mouth, double-blind, cross-over study, 10 volunteers wore palatal intra-oral devices containing bovine enamel slabs restored with composite resin (CR – Z250) or resin-modified glass ionomer (RMGI – Vitremer). Two phases of 14 days were carried out, one for each restorative material. In one side of the device, biofilm was allowed to accumulate under a plastic mesh, whereas in the opposing side, regular brushing was carried out 3 times/day with a dentifrice containing 1100 μg F/g as NaF. A 20% sucrose solution was applied extra-orally 10×/day on each restored dental slab. Knoop microhardness was used to calculate the percentage of surface hardness loss (%SHL).

Results

All materials showed a decrease in surface hardness after the in situ period. The restorative materials presented the following average for %SHL: RMGI without biofilm accumulation = 8.9 and with biofilm accumulation = 25.6, CR without biofilm accumulation = 14.7 and with biofilm accumulation = 17.0.

Conclusion

Biofilm accumulation and the presence of cariogenic challenge promoted faster degradation of ionomeric materials, but this was not observed for composite resin.

Clinical significance

The oral environment affects the surface hardness of aesthetic restorative materials. Biofilm accumulation and cariogenic challenge promote surface degradation for ionomeric materials, but not for composite resin.     

Antibacterial Efficacy of a Human β-Defensin-3 Peptide on Multispecies Biofilms

Introduction

The aggregation of mixed bacterial flora into sessile biofilms on root canal surfaces can be one of the causes of persistent apical periodontitis. The aim of this study was to evaluate the antibacterial efficacy of human β-defensin-3 (HBD3) peptide on multispecies biofilms by using confocal laser scanning microscopy.

Methods

Actinomyces naeslundii, Lactobacillus salivarius, Streptococcus mutans, and Enterococcus faecalis were cultured in a peptone-yeast-glucose broth, and their culture suspensions were combined in equal proportions. The mixed bacteria were inoculated on sterile coverslips placed into the wells of tissue culture plates to permit the formation of mixed species biofilm. After incubation for 3 weeks, the samples were treated for 24 hours with saline (control), saturated calcium hydroxide solution (CH), 2% chlorhexidine solution (CHX), and 50 μg/mL HBD3 solution. A commercial biofilm/viability assay kit was used to assess cell viability and analyze the 3-dimensional architecture of biofilms. The percentage of dead cells was determined from the ratio of biovolumes for the red subpopulation and the total biofilm.

Results

Three medication groups showed a significant reduction of biovolume within the biofilms compared with the control group (P < .001). The HBD3-treated biofilms had a higher percentage of dead cells than the other medication groups (P < .05). The CH and CHX groups showed higher levels of bactericidal activity than saline (P < .05), and there was no significant difference between the 2 groups (P > .05).

Conclusions

HBD3 peptide exhibited more antibacterial activity against mature multispecies biofilms in vitro than either CH or CHX.     

Absorption and release of protein from hydroxyapatite-polylactic acid (HA-PLA) membranes

Objectives

The aim of this study was to investigate the kinetics of protein interactions with a novel hydroxyapatite-polylactide (HA-PLA) composite membrane material.

Methods

Trilayer PLA and HA-PLA composite membranes reinforced with PLA fibres were used to absorb and release protein which was measured by a BioRad assay. The proteins used were fetal calf serum and bovine serum albumin. PLA and HA-PLA composite films were manufactured to test permeability.

Results

Maximal protein absorption was seen within 5 min of treating materials; a nearly 8-fold increase in total absorption was seen with HA-containing composites compared to those without HA. These also exhibited a more gradual and sustained release of protein for periods of up to 96 h, for example at 24 h protein concentrations released were 2.20 ± 2.80 and 0.49 ± 5.38 μg/ml for membranes with and without HA respectively. In addition low pressure and temperature used during production of membranes also allowed greater and more sustained protein release. HA-PLA composite films also showed marked increased permeability compared to plain PLA films, for example after 24 h PLA only films 3.64 ± 1.01 μg/ml, PLA film with 25% HA: 44.99 ± 35.61 μg/ml, PLA film with 75% HA: 153.12 ± 65.57 μg/ml.

Conclusions

The results demonstrate that these composite membranes rapidly absorb protein and that the absorbed protein is released slowly for periods of up to 96 h, dependent on constituents of the material and the manufacturing conditions. Incorporation of HA into these membranes was the key factor for improved protein kinetics and membrane permeability.     

Corrosion behaviour of titanium in the presence of Streptococcus mutans

Objective

The main aim of this in vitro study was to evaluate the influence of Streptococcus mutans on the corrosion of titanium.

Methods

S. mutans biofilms were formed on commercially pure titanium (CP-Ti) square samples (10 mm × 10 mm × 1 mm) using a culture medium enriched with sucrose. Open circuit potential (OCP) and electrochemical impedance spectroscopy (EIS) measurements were used to evaluate the corrosion behaviour of CP-Ti in the presence of S. mutans in Fusayama's artificial saliva. The corrosion of biofilm-free CP-Ti samples was also evaluated in artificial saliva. Biofilms biomass was measured by spectrophotometry, using crystal violet staining, after 1, 2 and 7 days.

Results

The OCP values recorded on CP-Ti in the presence of S. mutans (−0.3 ± 0.02 V vs. SCE) was lower than those on biofilm-free CP-Ti (−0.1 ± 0.01 V vs. SCE) after 2 h of immersion in artificial saliva (p < 0.05). That reveals a high reactivity of titanium in presence of S. mutans. Impedance spectra revealed the formation of a compact passive film on titanium in artificial saliva or in the presence of a 2 days old S. mutans biofilm even though the corrosion resistance of CP-Ti has decreased in presence of a S. mutans biofilm.

Conclusion

The presence of bacterial colonies, such as S. mutans, negatively affected the corrosion resistance of the titanium.     

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.     

Novel antibacterial orthodontic cement containing quaternary ammonium monomer dimethylaminododecyl methacrylate

Objectives

Demineralized lesions in tooth enamel around orthodontic brackets are caused by acids from cariogenic biofilm. This study aimed to develop a novel antibacterial orthodontic cement by incorporating a quaternary ammonium monomer dimethylaminododecyl methacrylate (DMADDM) into a commercial orthodontic cement, and to investigate the effects on microcosm biofilm response and enamel bond strength.

Methods

DMADDM, a recently-synthetized antibacterial monomer, was incorporated into orthodontic cement at 0%, 1.5%, 3% and 5% mass fractions. Bond strength of brackets to enamel was measured. A microcosm biofilm model was used to measure metabolic activity, lactic acid production, and colony-forming units (CFU) on orthodontic cements.

Results

Shear bond strength was not reduced at 3% DAMDDM (p > 0.1), but was slightly reduced at 5% DMADDM, compared to 0% DMADDM. Biofilm viability was substantially inhibited when in contact with orthodontic cement containing 3% DMADDM. Biofilm metabolic activity, lactic acid production, and CFU were much lower on orthodontic cement containing DMADDM than control cement (p < 0.05).

Conclusions

Therefore, the novel antibacterial orthodontic cement containing 3% DMADDM inhibited oral biofilms without compromising the enamel bond strength, and is promising to reduce or eliminate demineralization in enamel around orthodontic brackets.     

Mechanisms of action of fluoridated acidic liquid dentifrices against dental caries

Objective

This study attempted to clarify the mechanisms of action of fluoridated acidic liquid dentifrices against dental caries.

Design

In the in vitro leg, enamel specimens were submitted to a pH-cycling model, treated with distinct dentifrices (0, 550 μgF/g pH 4.5 and pH 7.0, 1100 or 5000 μgF/g pH 7.0) and analyzed using hardness. Alkali-soluble fluoride (F) deposition was quantified on pre-demineralized specimens treated with the dentifrices. In the clinical leg, 2-to-4-year-old children who had been using liquid dentifrices for 6 months (550 μgF/g pH 4.5 or pH 7.0 or 1100 μgF/g pH 7.0) had their plaque samples collected 5 and 60 min after the last brushing. Fluoride uptake in whole plaque was evaluated.

Results

The reduction of the pH had a partial preventive effect on subsurface hardness loss only. [F] had a significant influence on the deposition of fluoride, surface and subsurface hardness loss. In vivo, the reduction of the pH was able to significantly increase plaque F uptake, leading to similar levels as those found for the neutral dentifrice containing twice [F].

Conclusion

The results obtained from in vitro studies whose design does not include the presence of dental plaque should be interpreted with caution.     

Effect of chlorhexidine incorporation into a self-etching primer on dentine bond strength of a luting cement

Objectives

This study investigated the effect of incorporating chlorhexidine in a self-etching primer on the bond strength of an adhesive resin cement to dentine and on antibacterial activity.

Methods

Experimental self-etching primers were prepared by adding chlorhexidine (CHX) diacetate to ED primer 2.0 (Kuraray Medical Inc.) to obtain chlorhexidine concentrations of 1.0 wt% and 2.0 wt%. Human occlusal dentine surfaces were conditioned for 30 s using the experimental primers or pure ED primer. Composite blocks were luted using Panavia F 2.0 (Kuraray Medical Inc.). After storage in water for 24 h, the bonded teeth were sectioned into 0.9 mm × 0.9 mm beams and stressed to failure in tension to examine microtensile bond strength (μTBS). Fractured surfaces were examined with a field-emission scanning electron microscopy (FE-SEM). The morphology of dentine surfaces that were conditioned with each primer was also observed under FE-SEM. An agar diffusion test was performed to examine the antibacterial effect of each primer against Streptococcus mutans and Enterococcus faecalis.

Results

Addition of chlorhexidine to the primer had significant effects (p < 0.001) on μTBS and antibacterial effect. No significant difference in μTBS was found between the groups that contained 0% and 1% chlorhexidine in the primer (p = 0.095). Conversely, the group that contained 2% chlorhexidine showed significantly lower μTBS and pronounced cohesive failures within the thin layer of the primer.

Conclusions

Incorporation of chlorhexidine into ED primer 2.0 showed significant antibacterial activities. Conditioning with the 1% chlorhexidine-containing primer had no adverse effect on μTBS.     

Time-kill behaviour against eight bacterial species and cytotoxicity of antibacterial monomers

Objectives

The objectives of this study were to investigate: (1) the antibacterial activity of two antibacterial monomers, dimethylaminododecyl methacrylate (DMADDM) and dimethylammoniumethyl dimethacrylate (DMAEDM), against eight different species of oral pathogens for the first time; (2) the cytotoxicity of DMAEDM and DMADDM.

Methods

DMAEDM and DMADDM were synthesized by reacting a tertiary amine group with an organo-halide. Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) against eight species of bacteria were tested. Time-kill determinations were performed to examine the bactericidal kinetics. Cytotoxicity of monomers on human gingival fibroblasts (HGF) was assessed using a methyl thiazolyltetrazolium assay and live/dead viability assay.

Results

DMADDM showed strong bactericidal activity against all bacteria, with MIC of 1.2–9.8 μg/mL. DMAEDM had MIC of 20–80 mg/mL. Time-kill determinations indicated that DMADDM and DMAEDM had rapid killing effects against eight species of bacteria, and eliminated all bacteria in 30 min at the concentration of 4-fold MBC. Median lethal concentration for DMADDM and DMAEDM was between 20 and 40 μg/mL, which was 20-fold higher than 1–2 μg/mL for BisGMA control.

Conclusions

DMAEDM and DMADDM were tested in time-kill assay against eight species of oral bacteria for the first time. Both were effective in bacteria-inhibition, but DMADDM had a higher potency than DMAEDM. Different killing efficacy was found against different bacteria species. DMAEDM and DMADDM had much lower cytotoxicity than BisGMA. Therefore, DMADDM and DMAEDM are promising for use in bonding agents and other restorative/preventive materials to combat a variety of oral pathogens.     

Influence of 2% chlorhexidine digluconate on bond strength of a glass-fibre post luted with resin or glass-ionomer based cement

Objectives

This study evaluated the influence of 2% chlorhexidine digluconate (CHX) on the bond strength (BS) of a glass-fibre post to the root canal, regarding the cements (dual-cured resin or resin-modified glass-ionomer cement), the root thirds and the time of storage.

Method

Eighty bovine roots were selected and endodontically treated, before being randomly assigned to the following groups according to the luting protocol: ARC (RelyX ARC); ARC + CHX; RL (RelyX Luting 2); and RL + CHX. After 24 h of luting, the roots were sliced to obtain 1 mm-thick slices. Half of each group was submitted to either 7-day or 6-month storage in artificial saliva (n = 10). The specimens were subjected to push-out tests with a crosshead speed of 0.5 mm/min. The data were analysed with four-way ANOVA and Tukey's test (P ≤ 0.05). The failure modes were analysed with a digital microscope (50× and 200×).

Results

ARC yielded a significantly higher BS compared to RL (P < 0.001). Despite CHX exerted a significant effect; it depends on the interaction with the luting cement and time (P < 0.001). Thus, CHX decreased the values of BS to those of ARC after 6 months (P < 0.001). On the 7th day of storage, the ARC + CHX presented higher BS to the cervical and middle thirds compared to RL + CHX (P = 0.012). Time solely was not a significant factor (P = 0.081). Adhesive cement-dentine type and mixed failures were predominant modes for the ARC groups. For the RL groups, the main failures were adhesive cement-post and mixed modes.

Conclusions

Glass-fibre posts luted with RelyX ARC dual-cure resin cement exhibited higher BS than those luted with RelyX Luting 2 resin-modified glass-ionomer cement. Furthermore, CHX was not effective to improve the BS and negatively affected the BS of RelyX ARC after 6 months of storage.

Clinical significance

The use of chlorhexidine solution seems not to improve the bond strength of fibre posts to root canals, disregarding the composition of the luting cement.     

Synergistic activity of lysozyme and antifungal agents against Candida albicans biofilms on denture acrylic surfaces

Denture related oral candidiasis is a recalcitrant fungal infection not easily resolved by topical antifungals. The antimycotic protein lysozyme, in saliva is an important host defense mechanism although its activity against Candida biofilms on denture acrylic has not been evaluated.

Objectives

(i) To establish a clinically relevant denture acrylic assay model to develop standardized Candida albicans biofilms, and (ii) assess the inhibitory effects of lysozyme alone and, the latter combined with antifungals (nystatin, amphotericin B, ketoconazole and 5-fluorocytosine) on sessile Candida cells and, finally (iii) to visualize the accompanying ultrastructural changes.

Design

The rotating-disc biofilm reactor was used to develop standardized 48 h Candida biofilms on acrylic discs in YNB/100 mM glucose medium and the biofilm metabolic activity was monitored using a tetrazolium reduction assay.

Results

The biofilm metabolic activity was similar in 18 identical denture acrylic discs (p < 0.05) thus validating the rotating-disc biofilm model. Very low concentrations of lysozyme (6.25 μg/ml) significantly (p < 0.01) inhibited Candida biofilm formation indicating that lysozyme may likely regulate intra-oral Candida biofilm development. Although 100 μg/ml lysozyme killed 45% of sessile Candida cells, further increasing its concentration (up to 240 μg/ml) had no such effect. Nystatin, amphotericin B, and ketoconazole in association with 100 μg/ml lysozyme exhibited effective synergistic killing of biofilm Candida in comparison to drug-free controls. Scanning electron and confocal scanning laser microscopy analysis confirmed the latter trends.

Conclusion

Our results indicate that agents found in biological fluids such as lysozyme could be a safe adjunct to antifungals in future treatment strategies for recalcitrant candidal infections.     

Mineral distribution and CLSM analysis of secondary caries inhibition by fluoride/MDPB-containing adhesive system after cariogenic challenges

Objective

To evaluate the inhibition zone formation (IZ) and mineral distribution along the interface of adhesive systems either containing fluoride and antibacterial primer or not, after chemical and biological artificial caries challenges.

Methods

Forty-eight third molars were used. Artificial caries was developed with S. mutans in a 4 mm × 4 mm area of occlusal dentin surface. Carious dentin was removed and cavities were restored with Adper Scotchbond Multi-Purpose (SBM) and Clearfil Protect Bond (CPB) (n = 24). Samples were submitted to secondary caries development by chemical (C) (acidic gel) or biological (B) (S. mutans culture) methods for 5 days. Four groups were tested (n = 12): (1) SC (SBM + C); (2) SB (SBM + B); (3) CC (CPB + C); (4) CB (CPB + B). The IZ and outer lesion (OL) formations were analyzed by confocal laser scanning microscopy (CLSM). The distribution of calcium (Ca) and phosphorus (P) content along the interface was analyzed by micro X-ray fluorescence spectrometer by energy-dispersive (μEDX).

Results

The frequency of IZ formation and mean values of IZ thickness differed among the groups. The CC group presented the lowest OL depth. μEDX analysis showed that CPB had the highest mineral loss by the biological method, but the lowest mineral loss by the chemical method. SC and SB groups showed intermediate values of mineral loss.

Conclusion

The mineral loss along the dentin/restoration interface was affected by the artificial caries method, and hybrid layer formation by adhesive systems used. The adhesive system containing fluoride and antibacterial primer did not prevent secondary caries formation.     

Antibacterial and physical properties of EGCG-containing glass ionomer cements

Objectives

To evaluate the effect of the addition of epigallocatechin-3-gallate (EGCG) on the antibacterial and physical properties of glass ionomer cement (GIC).

Methods

A conventional GIC, Fuji IX, was used as a control. EGCG was incorporated into GIC at 0.1% (w/w) and used as the experimental group. Chlorhexidine (CHX) was added into GIC at 1% (w/w) as a positive control. The anti-biofilm effect of the materials was assessed by a colorimetric technique (MTT assay) and scanning electron microscopy (SEM). The leaching antibacterial activity of the materials on Streptococcus mutans was evaluated by an agar-diffusion test. The flexural strength of the materials was evaluated using a universal testing machine and the surface microhardness was measured using a microhardness tester. The fluoride-releasing property of the materials was tested by ion chromatography.

Results

The optical density (OD) values of the GIC-EGCG group were significantly decreased at 4 h compared with the GIC group, but only a slightly decreased tendency was observed at 24 h (P > 0.05). No inhibition zones were detected in the GIC group during the study period. Significant differences were found between each group (P < 0.05). Compared with the control group, there was a significant increase in the flexural strength and surface microhardness for the GIC-EGCG group (P < 0.05). The fluoride ion release was not influenced by EGCG-incorporation (P > 0.05).

Conclusions

These findings suggested that GIC-containing 0.1% (w/w) EGCG is a promising restorative material with improved mechanical properties and a tendency towards preferable antibacterial properties.

Clinical significance

Modification of the glass ionomer cements with EGCG to improve the antibacterial and physical properties showed some encouraging results. This suggested that the modification of GIC with EGCG might be an effective strategy to be used in the dental clinic. However, this was only an in vitro study and clinical trials would need to verify true outcomes.