In vitro evaluation of composite containing DMAHDM and calcium phosphate nanoparticles on recurrent caries inhibition at bovine enamel-restoration margins

ObjectiveRecurrent caries is a primary reason for restoration failure caused by biofilm acids. The objectives of this study were to: (1) develop a novel multifunctional composite with antibacterial function and calcium (Ca) and phosphate (P) ion release, and (2) investigate the effects on enamel demineralization and hardness at the margins under biofilms.MethodsDimethylaminohexadecyl methacrylate (DMAHDM) and nanoparticles of amorphous calcium phosphate (NACP) were incorporated into composite. Four groups were tested: (1) Commercial control (Heliomolar), (2) Experimental control (0% DMAHDM + 0% NACP), (3) antibacterial group (3% DMAHDM + 0% NACP), (D) antibacterial and remineralizing group (3% DMAHDM + 30% NACP). Mechanical properties and Ca and P ion release were measured. Colony-forming units (CFU), lactic acid and polysaccharide of Streptococcus mutans (S. mutans) biofilms were evaluated. Demineralization of bovine enamel with restorations was induced via S. mutans, and enamel hardness was measured. Data were analyzed via one-way and two-way analyses of variance and Tukey’s multiple comparison tests.ResultsAdding DMAHDM and NACP into composite did not compromise the mechanical properties (P > 0.05). Ca and P ion release of 3% DMAHDM + 30% NACP was increased at cariogenic low pH. Biofilm lactic acid and polysaccharides were greatly decreased via DMAHDM, and CFU was reduced by 4 logs (P < 0.05). Under biofilm acids, enamel hardness at the margins was decreased to about 0.5 GPa for control; it was about 1 GPa for antibacterial group, and 1.3 GPa for antibacterial and remineralizing group (P < 0.05).ConclusionsThe novel 3% DMAHDM + 30% NACP composite had strong antibacterial effects. It substantially reduced enamel demineralization adjacent to restorations under biofilm acid attacks, yielding enamel hardness that was 2-fold greater than that of control composites. The novel multifunctional composite is promising to inhibit recurrent caries.     

Dual-functional adhesive containing amorphous calcium phosphate nanoparticles and dimethylaminohexadecyl methacrylate promoted enamel remineralization in a biofilm-challenged environment

ObjectiveThe cariogenic biofilm on enamel, restoration, and bonding interface is closely related to dental caries and composite restoration failure. Enamel remineralization at adhesive interface is conducive to protecting bonding interface and inhibiting secondary caries. This study intended to assess the remineralization efficiency of adhesive with dimethylaminohexadecyl methacrylate (DMAHDM) and nanoparticles of amorphous calcium phosphate (NACP) on initial caries lesion of biofilm-coated enamel.MethodsArtificial initial carious lesion was created via 72-hour immersion in demineralization solution and cariogenic biofilm was formed after 24-hour culture of Streptococcus mutans (S. mutans). Specimens were then divided into 4 groups: enamel control, enamel treated with NACP, DMAHDM and NACP+DMAHDM respectively. Samples next underwent 7-day cycling, 4 h in BHIS (brain heart infusion broth containing 1 % sucrose) and 20 h in AS (artificial saliva) per day. The pH of BHIS was tested daily. So did the concentration of calcium and phosphate in BHIS and AS. Live/dead staining, colony-forming unit (CFU) count, and lactic acid production of biofilms were measured 7 days later. The enamel remineralization efficiency was evaluated by microhardness testing and transverse microradiography (TMR) quantitatively.ResultsEnamel of NACP+DMAHDM group demonstrated excellent remineralization effectiveness. And the NACP+DMAHDM adhesive released a great number of Ca²⁺ and PO₄^3- ions, increased pH to 5.81 via acid neutralization, decreased production of lactic acid, and reduced CFU count of S. mutans (P < 0.05).SignificanceThe NACP+DMAHDM adhesive would be applicable to preventing secondary caries, strengthening enamel-adhesive interface, and extending the lifespan of composite restoration.     

Antibacterial and mechanical properties of arginine-containing glass ionomer cements

ObjectiveThe study investigated the effect of incorporating l-arginine (Arg) in a glass ionomer cement (GIC) on its mechanical properties and antibacterial potential.MethodsPre-determined proportions (1%, 2%, and 4% by wt.) of Arg were incorporated in GIC powder; while GIC without Arg served as control. The flexural strength, nanohardness, surface roughness, elemental analysis using SEM-EDX (n = 6) and F/Arg/Ca/Al/Si release in deionized water for 21 days were assessed. The antibacterial potential was evaluated in a multi-species biofilm model with Streptococcus mutans, Streptococcus sanguinis, Streptococcus gordonii, and Lactobacillus acidophilus for 72 h. Real-time qPCR was used to analyse biofilm bacterial concentrations. Propidium monoazide modification of real-time qPCR was performed to quantify viable/dead bacteria. The pH, lactic acid, ADS activity, and H₂O₂ metabolism were measured. Confocal microscopy was used to investigate the biofilm bacterial live/dead cells, density, and thickness.ResultsThere was no difference in flexural strength among the different groups (p > 0.05). No significant difference in nanohardness and surface roughness was observed between 4% Arg + GIC and control (p > 0.05). The 4% Arg + GIC showed significantly higher F/Arg/Al/Si release than the other groups (p < 0.05), reduced total bacterial concentration and growth inhibition of viable S. mutans and S. sanguinis (p < 0.05). Lactic acid formation for 4% Arg + GIC was significantly higher than 1% Arg + GIC (p < 0.05). The spent media pH of 4% Arg + GIC was higher than the other groups (p < 0.05), with proportionately lower ammonia and higher H₂O₂ released (p < 0.05).SignificanceAddition of 4% l-arginine in GIC enhanced its antibacterial activity via a biofilm modulatory effect for microbial homeostasis, with no detrimental effect on its mechanical properties.     

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.     

Fluoride-releasing sealant and caries-like enamel lesion formation in vitro

OBJECTIVE: The aim of this laboratory study was to evaluate the effects of a fluoride-releasing pit and fissure sealant (FS) and a conventional non-fluoride-containing pit and fissure sealant (CS) on caries-like lesion formation in enamel. DESIGN: Twelve extracted, macroscopically caries-free human molar teeth underwent fluoride-free prophylaxis and were sectioned into toothquarters. Cavity preparations were placed in buccal and lingual surfaces of the toothquarters, acid-etched and filled with sealant material (per the manufacturer's instructions) as follows: 1) mesiobuccal and mesiolingual quarters--FS (Fissurit-F, VocoChemie); and 2) distobuccal and distolingual quarters--CS (Fissurit, VocoChemie). Prior to caries formation in an acidified gel, the toothquarters were thermocycled (500 cycles) in artificial saliva. Sections were taken following caries initiation (6 week gel exposure) and caries progression (9 week gel exposure), and examined by polarized light microscopy to determine mean primary surface lesion depth and presence or absence of wall lesions. RESULTS: Mean surface lesion depths were significantly different between FS and CS groups following caries initiation (FS = 119 microns; CS = 157 microns, p < 0.05, paired t-test) and progression (FS = 169 microns; CS = 221 microns, p < 0.05, paired t-test). Wall lesions were reduced in FS (14% initiation; 25% progression) when compared with CS (19% initiation; 35% progression); however, this was not significantly different (p > 0.05, paired t-test). CONCLUSION: A pit and fissure sealant containing releasable fluoride provided a caries inhibiting effect with a significant reduction in lesion depth in the surface enamel adjacent to the fluoride-releasing sealant and a tendency toward reduction in the frequency of wall lesions. CLINICAL SIGNIFICANCE: Fluoride release by pit and fissure sealants may provide additional protection against caries formation in cuspal incline enamel and smooth surfaces adjacent to sealed pits and fissures, and act as a fluoride reservoir with long-term release of fluoride into the immediately adjacent oral environment.     

Intelligent pH-responsive dental sealants to prevent long-term microleakage

ObjectivesMicroleakage is a determinant factor of failures in sealant application. In this study, DMAEM (dodecylmethylaminoethyl methacrylate), a pH-responsive antibacterial agent, was incorporated into resin-based sealant for the first time. The objectives of this study were to: (1) investigate the long-term performance of DMAEM-modified sealants against oral microbial-aging; and (2) investigate the long-term preventive effect of DMAEM-modified sealants on microleakage.MethodsDepth-of-cure and cytotoxicity of DMAEM-modified sealants were measured. Then, an aging model using biofilm derived from the saliva of high caries experience children was conducted. After aging, microhardness and surface roughness were measured. Biofilm activity, lactic acid production and exopolysaccharide (EPS) production were measured. 16S rRNA gene sequencing were also performed. The effects of DMAEM on microleakage were tested using an in vitro microleakage assessment.ResultsThe addition of DMAEM with a mass fraction of 2.5–10% did not affect depth-of-cure values and cytotoxicity of sealants. Adding 2.5–10% DMAEM did not affect the surface roughness and microhardness after aging. Compared to control, adding 2.5–10% DMAEM reduced biofilm metabolic activity by more than 80%. The lactic acid production and EPS production were reduced by 50% in DMAEM groups. DMAEM-modified sealants maintained the microbial diversity of biofilm after aging, they also inhibited the growth of lactobacillus. The 5% and 10% DMAEM groups exhibited a significant reduction in microleakage compared to control.SignificanceThe long-term antibacterial activities against oral microbial-aging and the long-term microecosystem-regulating capabilities enabled DMAEM-modified sealant to prevent microleakage in sealant application and thus prevent dental caries.     

Hydroalcoholic extracts of Myracrodruon urundeuva All. and Qualea grandiflora Mart. leaves on Streptococcus mutans biofilm and tooth demineralization

ObjectivesThis study evaluated the effect of the hydroalcoholic extracts of Myracrodruon urundeuva All. and Qualea grandiflora Mart. leaves (alone or combined) on the viability of Streptococcus mutans biofilm and on the prevention of enamel demineralization.MethodsStrain of S. mutans (ATCC 21175) was reactivated in BHI broth. Minimum inhibitory concentration, minimum bactericidal concentration, minimum inhibition biofilm concentration and minimum eradication biofilm concentration were determined in order to choose the concentrations to be tested under biofilm model. S. mutans biofilm (5 × 10⁵ CFU/ml) was produced on bovine enamel, using McBain saliva under 0.2% sucrose exposure, for 3 days. The biofilm was daily treated with the extracts for 1 min. The biofilm viability was tested by fluorescence and the enamel demineralization was measured using TMR.ResultsMyracrodruon urundeuva All. (Isolated or combined) at the concentrationsc ≥0.625 mg/ml was able to reduce bacteria viability, while Qualea Grandflora Mart. alone had antimicrobial effect at 5 mg/ml only (p < 0.05). On the other hand, none of the extracts were able to reduce enamel demineralization.ConclusionsThe hydroalcoholic extracts of Myracrodruon urundeuva All. and Qualea grandiflora Mart. leaves (isolated or combined) have antimicrobial action; however, they do not prevent enamel caries under S. mutans biofilm model.     

Minimally-invasive dentistry via dual-function novel bioactive low-shrinkage-stress flowable nanocomposites

The objectives of this in vitro study were to develop a novel low-shrinkage-stress flowable nanocomposite with antibacterial properties through the incorporation of dimethylaminohexadecyl methacrylate (DMAHDM) and nanoparticles of amorphous calcium phosphate (NACP), and investigate the mechanical and oral biofilm properties, to be used in minimally-invasive techniques.MethodsThe light-cured low-shrinkage-stress flowable resin was formulated by mixing urethane dimethacrylate (UDMA) and triethylene glycol divinylbenzyl ether (TEG-DVBE) at a 1:1 mass ratio. Different mass fractions of glass, and either 5% DMAHDM or 20%NACP or both were incorporated. Paste flowability, ultimate micro tensile strength and surface roughness were evaluated. The antibacterial response of DMAHDM resin was assessed by using biofilms of human saliva-derived microcosm model. Virtuoso flowable composite was used as a control.Results(45% resin+5% DMAHDM+20% NACP+30% glass) formula yielded the needed outcomes. It had flow rate within the range of ISO requirement. The micro tensile strength was (39.1 ± 4.3) MPa, similar to (40.1 ± 4.0) MPa for commercial control (p > 0.05). The surface roughness values of the novel composite (0.079 ± 0.01) µm similar to commercial composite (0.09 ± 0.02) µm (p > 0.05). Salivary microcosm biofilm colony forming unit values were reduced by 5–6 logs (p < 0.05). Biofilm metabolic activity was also substantially reduced, compared to control composite (p < 0.05).SignificanceThe novel bioactive flowable nanocomposite achieved strong antibacterial activities without compromising the mechanical properties. It is promising to be used as pit and fissure sealants, and as fillings in conservative cavities to inhibit recurrent caries and increase restoration longevity.     

Effects of single species versus multispecies periodontal biofilms on the antibacterial efficacy of a novel bioactive Class-V nanocomposite

ObjectiveThe objectives of this studywere to: (1) develop a novel bioactive nanocomposite for Class V restorations with subgingival margins to inhibit periodontal pathogens; and (2) investigate if the bioactive nanocomposite could inhibit multi-species periodontal biofilms with a potency as strong as that against single species biofilms.MethodsNanocomposite was fabricated using dimethylaminohexadecyl methacrylate (DMAHDM), 2-methacryloyloxyethyl phosphorylcholine (MPC) and nanoparticles of amorphous calcium phosphate (NACP). Biofilms with 1, 3, 6 and 9 species of periodontal pathogens were grown on the composites and tested for live/dead staining, colony-forming units (CFU), metabolic activity, and biofilm matrix polysaccharide production.ResultsThe bioactive composite reduced protein adsorption by an order of magnitude (p < 0.05) and greatly reduced biofilm viability. It decreased the biofilm CFU by more than 3 orders of magnitude for all four types of periodontal biofilms, compared to control composite. With increasing the biofilm species from 1 to 9, the antibacterial efficacy of DMAHDM composite decreased; the CFU reduction folds decreased from 947 folds to 44 folds. In contrast, the MPC + DMAHDM composite maintained a CFU reduction folds of greater than 3000, showing a similar antibacterial potency from 1 to 9 species in the biofilms (p > 0.1).ConclusionDual agents MPC + DMAHDM achieved the greatest inhibition in biofilm, without decreasing its antibacterial potency when the biofilm species was increased from 1 to 9. A single agent became less effective when the biofilm species was increased from 1 to 9.SignificanceThe multifunctional MPC + DMAHDM composite is promising for root caries treatment and Class V restorations with subgingival margins to effectively inhibit multispecies periodontal biofilms, combat periodontitis and protect the periodontium.     

The effect of an antibacterial monomer on the antibacterial activity and mechanical properties of a pit-and-fissure sealant

BackgroundThe authors incorporated an antibacterial monomer, methacryloxylethyl cetyl dimethyl ammonium chloride (DMAE-CB), into a commercial pit-and-fissure sealant to evaluate the monomer's antibacterial activity and mechanical properties.MethodsThe authors incorporated DMAE-CB at 1 percent by weight into a commercially available sealant (Helioseal, Ivoclar Vivadent, Schaan, Lichtenstein). The same sealant, Helioseal, without DMAE-CB served as a negative control. The authors used Helioseal F (Ivoclar Vivadent), containing a fluoride-releasing resin, as a positive control. They determined effects of the cured sealants and their eluents on the growth of Streptococcus mutans by means of a film contact test and absorbance measurement, respectively. They measured and compared surface contact angle, degree of conversion, microhardness and microleakage.ResultsThe authors found that in comparison with the negative control sealant, the cured sealant containing DMAE-CB had an inhibitory effect on the growth of S. mutans (P N/A.05), whereas the eluents of neither of these sealants showed any detectable antibacterial activity. The positive control also showed no detectable bactericidal activity. Moreover, there were no significant differences in the contact angle, degree of conversion, microhardness and microleakage between the DMAE-CB group and the negative control group (P >.05).ConclusionsThe incorporation of DMAE-CB can provide sealant with contact antibacterial activity after polymerization by influencing the growth of S. mutans. The incorporation of small amounts of the monomer DMAE-CB showed no influence on the contact angle, degree of conversion, hardness and microleakage of the parental material.Clinical ImplicationsThe results provided inference concerning the potential clinical performance of the sealant with antibacterial activity.     

Organo-selenium containing dental sealant inhibits biofilm formation by oral bacteria

ObjectiveDental plaque is a complex structure (called a biofilm) that is produced by a community of oral bacteria. As microorganisms accumulate in the oral cavity, bacteria can assemble into biofilms that protect them from antibiotics and disinfectants, which contribute to dental cavities and oral infections that acts as the seed for further infections throughout the body. Therefore, there is great interest in developing dental sealants that can effectively eliminate biofilms formed from an assortment of oral bacteria species.MethodsIn previous papers, it was shown that both in vivo and in vitro use of organo-selenium dental sealants have the potential to be an effective method for preventing dental caries and plaque formation. However, our previous in vitro study only examined the effect of the organo-selenium sealants on Streptococcus mutans and salivarius. Since that time, this organo-selenium sealant has been changed to improve its curing time.ResultsWe showed a selenium containing sealant (SeLECT-DefenseTM) can completely eliminate biofilm formation on the sealant at selenium concentrations of 0.25% and higher, by S. salivarius, S. sanguinis, or S. mutans, individually or in combination. This selenium containing sealant can also completely inhibit the same bacteria from growing under the sealant, while control sealant cannot. The selenium containing sealant was tested for stability and it was found to still kill these same bacteria after soaking for the equivalent of one year in PBS (pH 7.4). It was also found that the combination of the three bacteria were also killed by the selenium sealant, thus ruling out potential synergism of the bacteria in forming resistance.SignificanceThe following study showed that this modified selenium dental sealant effectively eliminates species of bacteria both on and under the dental sealant.     

Verification of antibacterial activity to enamel surfaces of new type of surface coating

ObjectivePrevention is difficult to decrease dental caries only via the partial application of fluoride. The GC Co. has developed a coating material adhesive containing fluoride and zinc. It is thought that this zinc has an effect which prevents dental caries. The aim of this study was to evaluate the influence of the Caredyne Shield® (CS) on biofilm generation by S. mutans, as compared to an acidulated phosphate fluoride (APF) gel.Materials and methodsWe performed the comparative study of the biofilm inhibitory effect which used the enamel of bovine teeth. Specimens were separated into a control group, an APF gel group, and a CS group. Biofilms were generated by adherent S. mutans. We observed the antibacterial weigh by the creation state of Biofilms.ResultsSignificant difference was observed in the number of bacterial colonies formed after 24 h, the number of bacterial colonies formed from detached S. mutans from the CS-treated experimental group were fewer in number than in the other group (p < 0.01). The biofilm formed by S. mutans 72 h after dissemination on the enamel surface was visible by fluorescence microscopy (Live/Dead staining method) and under the scanning electron microscope, in the CS-treated group, no plastic structures were observed, as the models were free of the biofilm and only scattered S. mutans cells were observed.ConclusionWe showed in this study the efficacy of CS in controlling the formation of biofilm. From such a result, we conclude that CS is a novel anticaries agent.     

Clinical comparison of a flowable composite and fissure sealant: A 24-month split-mouth,randomized, and controlled study

Objectives

The aim of this clinical study was to evaluate the retention rate and caries-prevention effect of a flowable composite compared to a conventional resin-based sealant in a young population over a 24-month period.

Methods

Thirty-four patients, ranging in age from 16 to 22 years, diagnosed with at least 2 non-cavitated pit-and-fissure caries in the first and second molars were selected for this randomized split-mouth design trial. A total of 220 sealants, were placed in 117 upper molars and 103 lower molars. The teeth were sealed with a flowable resin composite (Tetric Evo Flow) or a sealant material (Helioseal F). Each restoration was independently evaluated in terms of retention and the presence of caries at baseline and at 1, 6, 12, and 24 months. Data were analyzed using non-parametric Mann–Whitney U and Friedman 1-way ANOVA tests at p < 0.05.

Results

Tetric Evo Flow showed complete retention with 100%, 95.5%, 93.8%, and 88.5% at 1, 6, 12, and 24-month evaluations, respectively, while Helioseal F retention rates were 98.1%, 95.5%, 94.8%, and 85.4%, respectively, for the same evaluation periods. At the 24-month recall, 4 (4.2%) total losses were observed in subjects treated with Tetric Evo Flow and 2 total losses (2.1%) for Helioseal F, respectively. No significant differences were observed between the materials in retention rates or caries incidence for each evaluation period (p > 0.05).

Conclusion

Placement of flowable composite as fissure sealants in the younger population seems to be as effective as conventional fluoride containing fissure sealants for the prevention of fissure caries.

Clinical relevance

The use of a flowable composite as a fissure sealant material, in conjunction with a total-etch, single bottle adhesive, yielded better retention than did the conventional fluoride containing resin-based fissure sealant over a 24-month period in young patients.     

Efficacy of red propolis hydro-alcoholic extract in controlling Streptococcus mutans biofilm build-up and dental enamel demineralization

ObjectiveThe efficacy of a red propolis hydro-alcoholic extract (RP) in controlling Streptococcus mutans biofilm colonization was evaluated. The effect of RP on dental demineralization was also investigated.MethodsChemical composition was determined by High Performance Liquid Chromatography (HPLC). Minimum Inhibitory and Bactericidal Concentration (MIC and MBC, respectively) were investigated against Streptococcus mutans (ATCC 25175). The cytotoxic potential of 3% RP in oral fibroblasts was observed after 1 and 3 min. Bovine dental enamel blocks (N = 24) were used for S. mutans biofilm formation (48 h), simulating ‘feast or famine’ episodes. Blocks/biofilms were exposed 2×/day, for 3 days, to a cariogenic challenge with sucrose 10% (5 min) and treated (1 min) with: 0.85% saline solution (negative control), 0.12% Chlorhexidine (CHX, positive control for biofilm colonization), 0.05% Sodium Fluoride (NaF, positive control to avoid demineralization) and 3% RP. Biofilms were assessed for viability (CFU/mL), and to observe the concentration of soluble and insoluble extracellular polysaccharides (SEPS and IEPS). Dental demineralization was assessed by the percentage of surface hardness loss (%SHL) and through polarized light microscopy (PLM).ResultsThe RP presented 4.0 pH and ºBrix = 4.8. The p-coumaric acid (17.2 μg/mL) and luteolin (15.23 μg/mL) were the largest contents of phenolic acids and flavonoids, respectively. MIC and MBC of RP were 293 μg/mL and 1172 μg/mL, respectively. The 3% RP showed 43% of viably cells after 1 min. Lower number (p < 0.05) of viable bacteria (CFU/mL) was observed after CHX (1.8 × 10⁵) followed by RP (1.8 × 10⁷) treatments. The lowest concentration (μg/CFU) of SEPS (12.6) and IEPS (25.9) was observed in CHX (p < 0.05) followed by RP (17.1 and 54.3), and both differed from the negative control (34.4 and 63.9) (p < 0.05). Considering the %SHL, all groups differed statistically (p < 0.05) from the negative control (46.6%); but NaF (13.9%), CHX (20.1%) and RP (20.7%) did not differ among them (p > 0.05). After all treatments, suggestive areas of caries lesions were observed by PLM, which were lower for CHX and NaF.ConclusionThe 3% RP reduced S. mutans colonization, decreased concentration of extracellular polysaccharides and reduced dental enamel demineralization.     

Antibacterial surface properties of fluoride-containing resin-based sealants

Objectives

The aim of the present study was to determine the antibacterial properties of three resin-based pit and fissure sealant products: Clinpro (3M ESPE), Embrace (Pulpdent), and UltraSeal XT plus (Ultradent).

Methods

The antibacterial effects of the sealants were tested in both an agar diffusion assay and a planktonic growth inhibition assay using Streptococcus mutans and Lactobacillus acidophilus. The materials were applied to paper and enamel disks in the former and on the side walls of 96-well microtiter plates on the latter.

Results

All materials showed either diffusible or contact antibacterial effects in the agar diffusion assays. The effect was diminished when enamel disks were used as substrate. In the planktonic growth inhibition assay, Clinpro had its effect reduced, but retained activity against both bacteria over time. L. acidophilus was more sensitive than S. mutans to UltraSeal. Embrace retained antibacterial activity against both bacteria over time.

Conclusions

Within the limitations of this in vitro study it can be concluded that all materials are capable of contact inhibition of L. acidophilus and S. mutans growth. Embrace has the longer lasting antibacterial activity when in solution, especially against S. mutans.     

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.     

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

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

Self-healing adhesive with antibacterial activity in water-aging for 12 months

ObjectiveSecondary caries and micro-cracks are the main limiting factors for dentin bond durability. The objectives of this study were to develop a self-healing adhesive containing dimethylaminohexadecyl methacrylate (DMAHDM) and nanoparticles of amorphous calcium phosphate (NACP), and investigate the effects of water-aging for 12 months on self-healing, dentin bonding, and antibacterial properties for the first time.MethodsMicrocapsules were synthesized with poly (urea-formaldehyde) (PUF) shells containing triethylene glycol dimethacrylate (TEGDMA) and N,N-dihydroxyethyl-p-toluidine (DHEPT). The adhesive contained 7.5% microcapsules, 10% DMAHDM, and 20% NACP (all mass). Specimens were water-aged at 37 °C for 1 day to 12 months. Dentin bond strength was measured using extracted human teeth. A single-edge-V-notched-beam (SEVNB) method was used to measure fracture toughness K_IC and self-healing efficiency. A dental plaque microcosm biofilm model was used with human saliva as inoculum.ResultsThe microcapsules + DMAHDM + NACP group showed no decline in dentin bond strength after water-aging for 12 months, which was significantly higher than that of other groups without DMAHDM (p < 0.05). A self-healing efficiency of 67% recovery in K_IC was obtained even after 12 months of water immersion, indicating that the self-healing ability was not lost in water-aging (p > 0.1). The bacteria-killing ability of this adhesive did not decline from 1 day to 12 months (p > 0.1), with biofilm CFU reduction by 3–4 orders of magnitude after the resin was water-aged for 12 months, compared to control resin.SignificanceThis novel adhesive with triple merits of self-healing, antibacterial and remineralization functions showed an excellent long-term durability in water-aging for 12 months. This multifunctional adhesive has the potential for dental applications to heal cracks, inhibit bacteria, provide ions for remineralization, and increase the restoration longevity.     

Antibacterial,remineralising and matrix metalloproteinase inhibiting scandium-doped phosphate glasses for treatment of dental caries

ObjectivesAntibiotic resistance is increasingly a growing global threat. This study aimed to investigate the potential use of newly developed scandium-doped phosphate-based glasses (Sc-PBGs) as an antibacterial and anticariogenic agent through controlled release of Sc³⁺ ions.MethodsSc-PBGs with various calcium and sodium oxide contents were produced and characterised using thermal and spectroscopic analysis. Degradation behaviour, ion release, antibacterial action against Streptococcus mutans, anti-matrix metalloproteinase-2 (MMP-2) activity, remineralisation potential and in vivo biocompatibility were also investigated.ResultsThe developed glass system showed linear Sc³⁺ ions release over time. The released Sc³⁺ shows statistically significant inhibition of S. mutans biofilm (1.2 log₁₀ CFU reduction at 6 h) and matrix metalloproteinase-2 (MMP-2) activity, compared with Sc-free glass and positive control. When Sc-PBGs were mounted alongside enamel sections, subjected to acidic challenges, alternating hyper- and hypomineralisation layers consistent with periods of re- and demineralisation were observed demonstrating their potential remineralising action. Furthermore, Sc-PBGs produced a non-toxic response when implanted subcutaneously for 2 weeks in Sprague Dawley rats.SignificanceSince Sc³⁺ ions might act on various enzymes essential to the biological mechanisms underlying caries, Sc-PBGs could be a promising therapeutic agent against cariogenic bacteria.     

Concentration dependence of quaternary ammonium monomer on the design of high-performance bioactive composite for root caries restorations

ObjectivesDental plaque build-up on the cervical area adjacent to gingival margins is a trigger factor for secondary caries around restored root caries lesions. Dimethylaminohexadecyl methacrylate (DMAHDM) and amorphous calcium phosphate nanoparticles (NACP) impart anti-caries effect by reducing the bacterial growth and releasing high concentrations of calcium and phosphate ions, respectively. The present study explored the optimization and formulation of dental composite with increased concentration of DMAHDM combined with NACP and its effect on mechanical behavior and antibacterial response.MethodsDMAHDM was incorporated into dental composite formulation at 3% and 5% with 20% NACP fillers. Mechanical properties were assessed by flexural strength and elastic modulus. The cationic charge density of the samples was determined using fluorescein staining assay. A human saliva-derived microcosm biofilm model was used to assess antibacterial response via colony-forming units, metabolic activities, lactic acid production, and live/dead assay. Surface roughness was measured after 48 h-biofilm formation.ResultsThe viability of human saliva microcosm biofilms was DMAHDM concentration-dependent, where all the microbiological assays were substantially reduced in the presence of 5%DMAHDM. The increased DMAHDM concentration mirrors an increased surface charge density of composites by 8–12 folds and reduced the growth of cariogenic species by 2–5 log (p ≤ 0.05). Metabolic activity and lactic acid were reduced by 70–90% and 48–99%, respectively. Increasing DMAHDM concentration up to 5% and its association with NACP fillers did not adversely affect the mechanical properties.SignificanceA highly potent antibiofilm bioactive composite for root caries restorations having DMAHDM-NACP could be flexibly tailored during formulation without detrimental outcome for mechanical function. The enhanced antibacterial performance of the novel bioactive composite has great potential to suppress the dental plaque build-up that triggers secondary caries around the restored root caries lesions.