Lidiany Karla Azevedo Rodrigues

Evaluation of the antimicrobial effect of photodynamic antimicrobial therapy in an in situ model of dentine caries

Photodynamic antimicrobial therapy (PACT) promotes bacterial death as a result of the photosensitization of microbial components. This study evaluated the effect of PACT on dentine caries produced in situ. Over the course of 14 d, 20 volunteers wore intra-oral devices containing human dentine slabs that were treated 10 times daily with a 40% sucrose solution. Afterwards, the antimicrobial effect of toluidine blue O, associated with 47 or 94 J cm(-2) of a light-emitting diode, was evaluated. Before and after the treatments, dentine samples were analysed with regard to the total number of microorganisms, total streptococci, mutans streptococci, and lactobacilli. Significant reductions in the bacterial count were observed for PACT with both energy densities tested, with the following values observed for 47 and 94 J cm(-2) of irradiation: for total streptococci, 3.45 and 5.18; for mutans streptococci, 3.08 and 4.16; for lactobacilli, 3.24 and 4.66; and for total microorganisms, 4.29 and 5.43, respectively. The control, treated with 94 J cm(-2) of irradiation alone, was also effective against all bacteria. To conclude, PACT was effective in killing oral microorganisms present in dentine caries produced in situ and may be a useful technique for eliminating bacteria from dentine carious lesions before restoration.

The antimicrobial activity of photodynamic therapy against Streptococcus mutans using different photosensitizers.

UNLABELLED Several photosensitizers have been used against oral bacteria without standardization. Singlet oxygen ((1)O(2)) is an aggressive chemical species that can kill cells through apoptosis or necrosis. OBJECTIVE to compare the antimicrobial activity of photodynamic therapy (PDT) with different photosensitizers at the same concentration against Streptococcus mutans. In addition, the (1)O(2) production of each photosensitizer was determined. The photosensitizers (163.5 μM) methylene blue (MB), toluidine blue ortho (TBO) and malachite green (MG) were activated with a light-emitting diode (LED; λ=636 nm), while eosin (EOS), erythrosine (ERI) and rose bengal (RB) were irradiated with a curing light (λ=570 nm). Light sources were operated at 24 J cm(-2). For each photosensitizer, 40 randomized assays (n=10 per condition) were performed under one of the following experimental conditions: no light irradiation or photosensitizer, irradiation only, photosensitizer only or irradiation in the presence of a photosensitizer. After treatment, serial dilutions of S. mutans were seeded onto brain heart infusion agar to determine viability in colony-forming units per milliliter (CFU mL(-1)). Generation of (1)O(2) was analyzed by tryptophan photooxidation, and the decay constant was estimated. Results were analyzed by one-way ANOVA and the Tukey-Kramer test (p<0.05). PDT with irradiation in the presence of the photosensitizers TBO and MG was effective in reducing S. mutans counts by 3 and 1.4 logs, respectively (p<0.01), compared to their respective untreated controls. MB generated 1.3 times more (1)O(2) than TBO, and both produced significantly higher concentrations of singlet oxygen than the other photosensitizers. Since in vitro bulk (1)O(2) production does not indicate that (1)O(2) was generated in the bacterial activity site, the bactericidal action against S. mutans cannot be related to in vitro singlet O(2) generation rate. In vitroS. mutans-experiments demonstrated TBO as the only photosensitizer that effectively reduced 99.9% of these microorganisms.

Novel dental adhesives containing nanoparticles of silver and amorphous calcium phosphate.

OBJECTIVES Secondary caries is the main reason for restoration failure, and replacement of the failed restorations accounts for 50-70% of all restorations. Antibacterial adhesives could inhibit residual bacteria in tooth cavity and invading bacteria along the margins. Calcium (Ca) and phosphate (P) ion release could remineralize the lesions. The objectives of this study were to incorporate nanoparticles of silver (NAg) and nanoparticles of amorphous calcium phosphate (NACP) into adhesive for the first time, and to investigate the effects on dentin bond strength and plaque microcosm biofilms. METHODS Scotchbond multi-purpose adhesive was used as control. NAg were added into primer and adhesive at 0.1% by mass. NACP were mixed into adhesive at 10%, 20%, 30% and 40%. Microcosm biofilms were grown on disks with primer covering the adhesive on a composite. Biofilm metabolic activity, colony-forming units (CFU) and lactic acid were measured. RESULTS Human dentin shear bond strengths (n=10) ranged from 26 to 34 MPa; adding NAg and NACP into adhesive did not decrease the bond strength (p>0.1). SEM examination revealed resin tags from well-filled dentinal tubules. Numerous NACP infiltrated into the dentinal tubules. While NACP had little antibacterial effect, NAg in bonding agents greatly reduced the biofilm viability and metabolic activity, compared to the control (p<0.05). CFU for total microorganisms, total streptococci, and mutans streptococci on bonding agents with NAg were an order of magnitude less than those of the control. Lactic acid production by biofilms for groups containing NAg was 1/4 of that of the control. SIGNIFICANCE Dental plaque microcosm biofilm viability and acid production were greatly reduced on bonding agents containing NAg and NACP, without compromising dentin bond strength. The novel method of incorporating dual agents (remineralizing agent NACP and antibacterial agent NAg) may have wide applicability to other dental bonding systems.

Novel calcium phosphate nanocomposite with caries-inhibition in a human in situ model.

OBJECTIVES Secondary caries at the restoration margins remains the main reason for failure. Although calcium phosphate (CaP) composites are promising for caries inhibition, there has been no report of CaP composite to inhibit caries in situ. The objectives of this study were to investigate the caries-inhibition effect of nanocomposite containing nanoparticles of amorphous calcium phosphate (NACP) in a human in situ model for the first time, and to determine colony-forming units (CFU) and Ca and P ion concentrations of biofilms on the composite restorations. METHODS NACP with a mean particle size of 116 nm were synthesized via a spray-drying technique. Two composites were fabricated: NACP nanocomposite, and control composite filled with glass particles. Twenty-five volunteers wore palatal devices containing bovine enamel slabs with cavities restored with NACP or control composite. After 14 days, the adherent biofilms were collected for analyses. Transverse microradiography determined the enamel mineral profiles at the margins, and the enamel mineral loss ΔZ was measured. RESULTS NACP nanocomposite released Ca and P ions and the release significantly increased at cariogenic low pH (p<0.05). Biofilms on NACP nanocomposite contained higher Ca (p=0.007) and P ions (p=0.005) than those of control (n=25). There was no significant difference in biofilm CFU between the two composites (p>0.1). Microradiographs showed typical subsurface lesions in enamel next to control composite, but much less lesion around NACP nanocomposite. Enamel mineral loss ΔZ (mean±sd; n=25) around NACP nanocomposite was 13.8±9.3 μm, much less than 33.5±19.0 μm of the control (p=0.001). SIGNIFICANCE Novel NACP nanocomposite substantially reduced caries formation in a human in situ model for the first time. Enamel mineral loss at the margins around NACP nanocomposite was less than half of the mineral loss around control composite. Therefore, the Ca and P ion-releasing NACP nanocomposite is promising for caries-inhibiting restorations.

Nanotechnology-based restorative materials for dental caries management

Nanotechnology has been applied to dental materials as an innovative concept for the development of materials with better properties and anticaries potential. In this review we discuss the current progress and future applications of functional nanoparticles incorporated in dental restorative materials as useful strategies to dental caries management. We also overview proposed antimicrobial and remineralizing mechanisms. Nanomaterials have great potential to decrease biofilm accumulation, inhibit the demineralization process, to be used for remineralizing tooth structure, and to combat caries-related bacteria. These results are encouraging and open the doors to future clinical studies that will allow the therapeutic value of nanotechnology-based restorative materials to be established.

Novel dental adhesive containing antibacterial agents and calcium phosphate nanoparticles

Secondary caries remains the main reason for dental restoration failure. Replacement of failed restorations accounts for 50%-70% of all restorations performed. Antibacterial adhesives could inhibit biofilm acids at tooth-restoration margins, and calcium phosphate (CaP) ions could remineralize tooth lesions. The objectives of this study were to: (1) incorporate nanoparticles of silver (NAg), quaternary ammonium dimethacrylate (QADM), and nanoparticles of amorphous calcium phosphate (NACP) into bonding agent; and (2) investigate their effects on dentin bonding and microcosm biofilms. An experimental primer was made with pyromellitic glycerol dimethacrylate (PMGDM) and 2-hydroxyethyl methacrylate (HEMA). An adhesive was made with bisphenol-A-glycerolate dimethacrylate (BisGMA) and triethylene glycol dimethacrylate (TEGDMA). NAg was incorporated into primer at 0.1 wt %. The adhesive contained 0.1% NAg and 10% QADM, and 0%-40% NACP. Incorporating NAg into primer and NAg-QADM-NACP into adhesive did not adversely affect dentin bond strength (p > 0.1). Scanning electron microscopy showed numerous resin tags, and transmission electron microscopy revealed NAg and NACP in dentinal tubules. Viability of human saliva microcosm biofilms on primer/adhesive/composite disks was substantially reduced via NAg and QADM. Metabolic activity, lactic acid, and colony-forming units of biofilms were much lower on the new bonding agents than control (p < 0.05). In conclusion, novel dental bonding agents containing NAg, QADM, and NACP were developed with the potential to kill residual bacteria in the tooth cavity and inhibit the invading bacteria along tooth-restoration margins, with NACP to remineralize tooth lesions. The novel method of combining antibacterial agents (NAg and QADM) with remineralizing agent (NACP) may have wide applicability to other adhesives for caries inhibition.

Relationship among microbiological composition and presence of dental plaque, sugar exposure, social factors and different stages of early childhood caries

OBJECTIVE This cross-sectional study aimed to investigate the relationship among microbiological composition of dental plaque, sugar exposure and social factors, as well as the presence of visible plaque in preschoolers with different stages of early childhood caries. DESIGN A total of 169 children were clinically examined according to the World Health Organisation criteria + early caries lesions and were divided into three groups: caries-free (n=53), early caries lesions-ECL (n=56), and cavitated caries lesions-CCL (n=60). The presence of clinically visible dental plaque on maxillary incisors was recorded. After that, dental plaque from all buccal and lingual smooth surfaces was collected and the number of colony-forming units of mutans streptococci and total microorganism, as well as the presence of lactobacilli was determined. Daily frequency of meals containing sugar was assessed by a diet chart whereas social factors were evaluated by a questionnaire. The data were analysed by chi-square test followed by multiple logistic regressions (alpha=0.05, confidence interval=95%). RESULTS High levels of mutans streptococci (OR=2.28), high total sugar exposure (OR=5.45) and presence of dental plaque (OR=2.60) showed significant association with ECL (p<0.05). High total microorganism count (OR=5.18), high solid sugar exposure (OR=2.50) and the presence of lactobacilli (OR=24.99) revealed significant association with CCL (p<0.05). CONCLUSIONS These results support the conclusion that dietary sugar experience influenced the microbiological composition of dental plaque. Moreover, the early stage of caries is highly affected by mutans streptococci and visible dental plaque on maxillary incisors whereas cavities are strongly related to lactobacilli and total microorganism.

In situ effects of restorative materials on dental biofilm and enamel demineralisation

OBJECTIVES Since secondary caries is one of the main reasons for replacing restorations, this study assessed the effects of different restorative materials on the microbiological composition of dental biofilm and on enamel demineralisation around the restoration. METHODS A randomized, double-blind, split-mouth in situ design was conducted in one phase of 14 days, during which, 20 volunteers wore palatal devices containing five human dental enamel slabs. Each slab was randomly restored with one of the following materials: Filtek-Z-250/Single Bond, control group (composite resin), Permite (amalgam), Fuji II (encapsulated resin-modified glass ionomer), Vitremer (resin-modified glass ionomer) and Ketac Molar (conventional glass ionomer). The volunteers used fluoride dentifrice, 3x/day and a 20% sucrose solution was dripped onto the slabs 8x/day. The biofilm formed on the slabs was analyzed to determine the counts of total streptococci, mutans streptococci and lactobacilli. Enamel demineralisation was determined by cross-sectional microhardness (CSMH) at 20 and 70 microm from the margin of the restoration. Kruskal-Wallis and analysis of variance, followed by least mean squares (LMS) test, were used to evaluate microbiota and CSMH among the groups. The significance level used was 5%. RESULTS No statistically significant differences were found in the cariogenic microbiota grown on the slabs. At a 20-mum distance, only Fuji II statistically differed from the other groups, showing the lowest demineralisation. At 70 microm, Fuji II significantly inhibited demineralisation when compared to Permite, Filtek-Z-250 and Ketac Molar. CONCLUSIONS In the context of fluoride dentifrice and under the cariogenic exposure conditions of this study, only the encapsulated resin-modified glass ionomer material provided additional protection against secondary caries.

In situ Mineral Loss Inhibition by CO2 Laser and Fluoride

Laser and fluoride treatments have been shown to inhibit enamel demineralization in the laboratory. However, the intra-oral effects of this association have not been tested. This study assessed in situ the effect of a Transversely Excited Atmospheric CO2 laser (λ = 9.6 μm) and the use of pressure fluoridated dentifrice on enamel demineralization. During two 14-day phases, 17 volunteers wore palatal appliances containing human enamel slabs assigned to treatment groups, as follows: (1) non-fluoride dentifrice, (2) CO2 laser irradiation plus non-fluoride dentifrice, (3) fluoride dentifrice, and (4) CO2 laser irradiation plus fluoride dentifrice. A 20% sucrose solution was dripped onto the slabs 8 times per day. The specimens treated with laser and/or fluoridated dentifrice presented a significantly lower mineral loss when compared with those from the non-fluoride dentifrice group. The results suggested that CO2 laser treatment of enamel inhibits demineralization in the human mouth, being more effective when associated with fluoride.

Combined Effects of Carbon Dioxide Laser and Fluoride on Demineralized Primary Enamel: An in vitro Study

This in vitro study aimed at evaluating the efficacy of a CO2 laser (10.6 µm) alone or combined with acidulated phosphate fluoride (APF) on the inhibition of lesion progression in primary enamel. The specimens were treated with/without CO2 laser and/or APF and submitted to pH cycling. Microhardness analysis was performed and the enamel mineral loss values were obtained. The groups treated with laser and/or APF presented lower mineral loss when compared to the control group (p < 0.05). Laser irradiation alone or combined with APF decreased lesion progression in primary enamel. However, the combined treatment did not show any significant additional effect.