Effect of bioactive compounds on the regulation of quorum sensing network-associated genes and virulence in Streptococcus mutans-A systematic review.

OBJECTIVE: The aim of this study was to systematically review the literature on the effect of bioactive compounds and their regulation of quorum sensing (QS)-related and/or -regulated-virulence genes expression in Streptococcus mutans. DESIGN: The search strategy was conducted through the electronic databases Pubmed, Scopus, and Web of Science for studies reporting the effects of natural and synthetic bioactive compounds on the regulation of QS-associated and/or -regulated genes of S. mutans. RESULTS: After full-text reading, 19 studies met the inclusion criteria, in most of them, QS-inhibitors from synthetic origin were evaluated, 16 articles described the effect of the compounds on biofilm formation cultivated in vitro and five studies described these effects on adhesion of biofilm-producing cells. Only 2 studies analyzed the potential target-component of the QS. CONCLUSIONS: Mostly, the bioactive compounds affected the expression of QS-associated and/or -regulated genes and virulence traits (e.g. adhesion, biofilm formation, acid stress tolerance) of S. mutans. Further studies are necessary to elucidate the target-specific QS-system constituent used by bioactive compounds to achieve QS inhibition as well as validate the use of these compounds in controlling dental caries.

Saliva proteomics from children with caries at different severity stages.

OBJECTIVE: To perform a comparative analysis of saliva protein profile of patients with early childhood caries at different levels of severity and caries-free individuals. MATERIALS AND METHODS: Stimulated saliva samples were collected from 126 children (2-6 years old), classified according to the ICDAS II, and divided into 3 groups (n = 42): caries-free (CF), enamel caries (EC), and dentine caries (DC). Samples were digested and analyzed by nanoUPLC coupled with a mass spectrometry. Data analyses were conducted with Progenesis QI for Proteomics Software v2.0. Gene Ontology (GO) terms and protein-protein interaction analysis were obtained. RESULTS: A total of 306 proteins (≈6 peptides) were identified. Among them, 122 were differentially expressed in comparisons among children with different caries status. Out of the 122 proteins, the proteins E2AK4 and SH3L2 were exclusively present in groups CF and EC, respectively, and 8 proteins (HAUS4, CAH1, IL36A, IL36G, AIMP1, KLHL8, KLH13, and SAA1) were considered caries-related proteins when compared to caries-free children; they were up-regulated proteins in the caries groups (EC and DC). CONCLUSION: The identification of exclusive proteins for caries-free or carious-related conditions may help in understanding the mechanisms of caries and predicting risk as well as advancing in caries control or anti-caries approaches.

Human In Situ Study of the effect of Bis(2-Methacryloyloxyethyl) Dimethylammonium Bromide Immobilized in Dental Composite on Controlling Mature Cariogenic Biofilm.

Cariogenic oral biofilms cause recurrent dental caries around composite restorations, resulting in unprosperous oral health and expensive restorative treatment. Quaternary ammonium monomers that can be copolymerized with dental resin systems have been explored for the modulation of dental plaque biofilm growth over dental composite surfaces. Here, for the first time, we investigated the effect of bis(2-methacryloyloxyethyl) dimethylammonium bromide (QADM) on human overlying mature oral biofilms grown intra-orally in human participants for 7⁻14 days. Seventeen volunteers wore palatal devices containing composite specimens containing 10% by mass of QADM or a control composite without QADM. After 7 and 14 days, the adherent biofilms were collected to determine bacterial counts via colony-forming unit (CFU) counts. Biofilm viability, chronological changes, and percentage coverage were also determined through live/dead staining. QADM composites caused a significant inhibition of Streptococcus mutans biofilm formation for up to seven days. No difference in the CFU values were found for the 14-day period. Our findings suggest that: (1) QADM composites were successful in inhibiting 1⁻3-day biofilms in the oral environment in vivo; (2) QADM significantly reduced the portion of the S. mutans group; and (3) stronger antibiofilm activity is required for the control of mature long-term cariogenic biofilms. Contact-killing strategies using dental materials aimed at preventing or at least reducing high numbers of cariogenic bacteria seem to be a promising approach in patients at high risk of the recurrence of dental caries around composites.

Insights into the Virulence Traits of Streptococcus mutans in Dentine Carious Lesions of Children with Early Childhood Caries.

Streptococcus mutans is an oral bacterium considered to play a major role in the development of dental caries. This study aimed to investigate the prevalence of S. mutans in active and arrested dentine carious lesions of children with early childhood caries and to examine the expression profile of selected S. mutans genes associated with survival and virulence, within the same carious lesions. Dentine samples were collected from 29 active and 16 arrested carious lesions that were diagnosed in preschool children aged 2-5 years. Total RNA was extracted from the dentine samples, and reverse transcription quantitative real-time PCR analyses were performed for the quantification of S. mutans and for analyses of the expression of S. mutans genes associated with bacterial survival (atpD, nox, pdhA) and virulence (fabM and aguD). There was no statistically significant difference in the prevalence of S. mutans between active and arrested carious lesions. Expression of the tested genes was detected in both types of carious dentine. The pdhA (p = 0.04) and aguD (p = 0.05) genes were expressed at higher levels in arrested as compared to active lesions. Our findings revealed that S. mutans is part of the viable microbial community in active and arrested dentine carious lesions. The increase in expression of the pdhA and aguD genes in arrested lesions is likely due to the unfavourable environmental conditions for microbial growth, inherent to this type of lesions.

CO2 laser irradiation enhances CaF2 formation and inhibits lesion progression on demineralized dental enamel-in vitro study.

This study evaluated if Carbon dioxide (CO2) (λ 10.6 µm) laser irradiation combined with acidulated phosphate fluoride gel application (APF gel) enhances "CaF2" uptake by demineralized enamel specimens (DES) and inhibits enamel lesion progression. Thus, two studies were conducted and DES were subjected to APF gel combined or not with CO2 laser irradiation (11.3 or 20.0 J/cm(2), 0.4 or 0.7 W) performed before, during, or after APF gel application. In study 1, 165 DES were allocated to 11 groups. Fluoride as "CaF2 like material" formed on enamel was determined in 100 DES (n = 10/group), and the surface morphologies of 50 specimens were evaluated by scanning electron microscopy (SEM) before and after "CaF2" extraction. In study 2, 165 DES (11 groups, n = 15), subjected to the same treatments as in study 1, were further subjected to a pH-cycling model to simulate a high cariogenic challenge. The progression of demineralization in DES was evaluated by cross-sectional microhardness and polarized light microscopy analyses. Laser at 11.3 J/cm(2) applied during APF gel application increased "CaF2" uptake on enamel surface. Laser irradiation and APF gel alone arrested the lesion progression compared with the control (p < 0.05). Areas of melting, fusion, and cracks were observed. CO2 laser irradiation, combined with a single APF application enhanced "CaF2" uptake on enamel surface and a synergistic effect was found. However, regarding the inhibition of caries lesion progression, no synergistic effect could be demonstrated. In conclusion, the results have shown that irradiation with specific laser parameters significantly enhanced CaF2 uptake by demineralized enamel and inhibited lesion progression.

Erosive potential of processed and fresh orange juice on human enamel.

PURPOSE: The purpose of this study was to analyze the erosive potential of processed and fresh orange juice on human enamel. METHODS: Sixty enamel slabs, incubated in human saliva to develop acquired pellicle, were exposed to processed and freshly squeezed juice from oranges and lime oranges. Daily erosive cycles were performed by immersing the slabs in the juices for 20 minutes over five days. During the intervals, the samples were immersed in artificial saliva. The pH, titratable acidity (TA), and buffer capacity (ß) of the juices were determined. Analysis of variance and Tukey's tests were used to compare the percentage of surface hardness loss and the amount of wear between groups. RESULTS: The ranges of pH, TA, and ß were 3.66 to 3.75, 3.33 to 110 mmol/L, and 2.98 to 40.97 mmol/L × pH, respectively. A similar erosive potential was found for all groups, except for the lime orange juices (P<.05). CONCLUSION: Both processed and freshly squeezed orange juices were erosive to enamel. However, the lime orange juice was acidless and, therefore, not able to produce any significant changes in enamel.

The influence of dentin demineralization on morphological features of cavities using Er:YAG laser.

OBJECTIVE: The purpose of this study was to evaluate the influence of erbium-doped: yttrium-aluminum-garnet (Er:YAG) laser parameters and different degrees of demineralization on morphological features, diameter, and depth of prepared cavities. BACKGROUND DATA: Minimally invasive dentin caries removal has been recommended. Ablation of deep caries lesions using Er:YAG laser should preserve remaining demineralized dentin; however, the influence of the degree of mineralization of this substrate had not been entirely described. MATERIALS AND METHODS: A randomized, factorial design was used to study the effects of two factors. Laser parameter was tested at two levels (250 mJ/4 Hz vs. 200 mJ/2 Hz) and degree of demineralization was tested at four levels (control, two-four-eight cycles). Twelve slabs of human dentin were divided into four groups according to the number of cycles induced by pH-cycling: G1, zero cycles; G2, two cycles, G3, four cycles, and G4, eight cycles. An Er:YAG laser was used at an output energy of 250 mJ/4 Hz and 200 mJ/2 Hz for all groups, for 10 sec at 12 mm distance focus/object. Circumference and depth of the cavities were measured on scanning electron microscopy (SEM) images using image analysis software. The mean values were subjected to two way analysis of variance (ANOVA) and Tukey tests. RESULTS: When using 250 mJ/4 Hz, the mean values of circumferential area increased significantly in relation to control (503.54 µm(2)) with increasing demineralization level (eight cycles) (555.45 µm(2)). Regardless of the demineralization level, there was also significant statistical difference in the studied measurements of the cavities when 250 mJ/4 Hz and 200 mJ/2 Hz were used. SEM also showed that laser cavity preparations left no smear layer, and the dentinal tubules were clear. CONCLUSIONS: The circumferential area and depth measurements were affected by laser parameter and demineralization level (eight cycles). Energy level output represents a relevant factor for increased circumferential area and depth measurements. High demineralized artificially caries-affected dentin may also imply higher ablation. Appropriated parameter of laser pulse frequency/power density for demineralized dentin should be used for effective less-invasive caries treatment.

Acid etching concentration as a strategy to improve the adhesive performance on Er:YAG laser and bur-prepared demineralized enamel.

OBJECTIVE: This study investigated the effect of phosphoric acid concentration on microshear bond strength (µSBS) of an etch-and-rinse adhesive system to demineralized irradiated enamel. BACKGROUND DATA: Er:YAG laser irradiation may increase acid resistance of enamel; however, its use is associated with reduced bonding effectiveness. MATERIALS AND METHODS: Three experimental etching agents and one bonding agent were fabricated. Enamel pretreatment was tested in two levels (bur and laser) and phosphoric acid was tested in four levels (control and at 35, 45, or 55%). Commercially available phosphoric acid and a bonding agent were used as control. Enamel samples were submitted to a pH-cycling model to induce demineralization. Half of the exposed demineralized enamel area was prepared with diamond bur and half was prepared with an Er:YAG laser (λ=2.94 µm, 300 mJ, 2 Hz). Specimens were randomly allocated to receive different acid etching concentrations and either the experimental or the commercial adhesive system. Microshear bond strength and the adhesive remnant index (ARI) scores were determined. µSBS's data were evaluated by two way analysis of variance (ANOVA) and Tukey's test. Kruskal-Wallis followed by Dunn's method and Mann-Whitney test were used for ARI comparisons (α=5%). RESULTS: For µSBS, no significant difference among the groups was found, either for phosphoric acid concentration or surface preparation. For ARI scores, statistically higher resin retention was found for lased groups (p<0.001). CONCLUSIONS: The increase of phosphoric acid concentration did not impair bond strength regardless of surface preparation; however, increased retention was found when demineralized enamel was prepared with laser.

In Situ Response of Nanostructured Hybrid Fluoridated Restorative Composites on Enamel Demineralization, Surface Roughness and Ion Release.

Recurrent caries at the tooth-restoration margins is the main reason for composite failure. Fluoride-releasing nanohybrid composite resin may reduce the recurrent caries rates. A fluoride-releasing resin (FCR) and non-fluoride-releasing resin (CR) were tested using an in situ model. Demineralization (ΔS), ion release and surface roughness of composite specimens were determined. The F concentration in the group FCR was higher than the CR group. ΔS (Mean ± SD) was 2579 ± 1582 and 1705 ± 1292, respectively, for FCR and CR. Surfaces roughness was altered by biofilm accumulation. The hybrid fluorated restorative composites containing nanoparticles have a slight anticaries action without alteration of surface smoothness of the material.

Fluoride releasing and enamel demineralization around orthodontic brackets by fluoride-releasing composite containing nanoparticles.

INTRODUCTION: Fluoride-containing materials have been suggested to control enamel demineralization around orthodontic brackets during the treatment with fixed appliances. The improvement of their properties has been made through innovations, such as the application of nanotechnology by incorporation of nanofillers. OBJECTIVE: This in vitro study evaluated the capacity of fluoride releasing and enamel demineralization inhibition of fluoride-releasing nanofilled cement around orthodontic brackets using an artificial caries biofilm model. MATERIALS AND METHODS: Forty bovine enamel discs were selected by evaluating surface microhardness and randomized into four groups (n = 10): non-fluoride-releasing microfilled composite, fluoride-releasing microfilled composite, resin-modified glass ionomer cement (RMGI), and fluoride-releasing nanofilled composite (FN). After brackets bonding in each disc, the specimens were subjected to a cariogenic challenge through a Streptococcus mutans biofilm model. After the experimental period, the biofilm formed around the brackets was collected for fluoride analysis and the mineral loss around the brackets was determined by integrated demineralization via cross-sectional microhardness measurement at 20 and 70 µm from the bracket margin. Additionally, samples of each group were subjected to energy-dispersive X-ray spectroscopy (EDX) analysis examined under a scanning electron microscopy (SEM). ANOVA followed by Tukey test were applied for fluoride concentration and mineral loss data, respectively. RESULTS: At both distances, only RMGI statistically differed from the other groups presenting the lowest demineralization, although there was a trend to a lower demineralization of enamel around brackets in FN group. Similar condition was found to fluoride concentration and EDX/SEM analysis. CONCLUSIONS: Under the cariogenic exposure condition of this study, the fluoride-releasing nanofilled material had similar performance to fluoride-releasing microfilled materials. CLINICAL RELEVANCE: The presence of nanofillers in the fluoride-releasing materials studied did not promote further benefits against caries lesion development around brackets and presented inferior demineralization inhibition than the resin-modified glass ionomer material.

A Comparative Study of the Photosensitizer Penetration into Artificial Caries Lesions in Dentin Measured by the Confocal Raman Microscopy.

This study utilized the confocal Raman microspectroscopy (CRM) technique for the first time to investigate the degree of the penetration of toluidine blue-orto (TBO) in artificial caries lesions produced by two distinct caries-inducing models. The dentin specimens (n = 10) were divided into three groups: control, in vitro and in situ. Thereafter, the lesion depth and the demineralization level were evaluated by cross-sectional microhardness (CSMH). CRM mapping across the dentin surface was assessed after the dye application. The CSMH and CRM data were analyzed by t-test and ANOVA, respectively (P < 0.05). The values of the lesion depth and the demineralization areas were higher for in situ samples (P < 0.05). The TBO penetration values (µm) for the control, in vitro and in situ groups were 44.8 ± 5.6, 46.1 ± 4.5 and 51.2 ± 8.5, respectively. There were no statistically significant differences among the groups (P > 0.05). The rate of TBO penetration was detected up to about <50 µm and the demineralization level did not influence the results. These results have showed promising parameters to develop new protocols for deep caries lesions management using photodynamic antimicrobial chemotherapy.

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.

Dentin erosion by whitening mouthwash associated to toothbrushing abrasion: a focus variation 3D scanning microscopy study.

The aim of this study was to determine the erosive potential of hydrogen peroxide (HP) containing mouthwash on dentin assessed by Focus variation three-dimensional (3D) microscopy. Twenty dentin slabs were selected and randomly allocated into two groups (n = 10): DW--Distilled water (pH = 7.27) and HP-1.5% (pH = 3.78). Each specimen was cyclically demineralized (4 × 60 s/day, 10 days) with HP or DW and brushed 3×/day (200 g, 150 strokes--toothpaste with 1,450 ppmF as NaF). Between the challenges, the specimens were exposed to artificial saliva. Afterward, dentin loss was analyzed using focus variation 3D microscopy, and the data were submitted to unpaired t-test (α = 0.05). Statistically significant difference was found between the mean wear rate (µm, ±SD) of HP (1.98 ± 0.51) and DW (1.45 ± 0.39). The results suggest that the use of HP-containing mouthwash associated to brushing may increase the risk of tissue loss and focus variation 3D microscopy may be used as a technique for quantifying dental wear.

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.

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.

Comparison of methods for quantifying dental wear caused by erosion and abrasion.

Various methods have been applied to evaluate the effect of erosion and abrasion. So, the aim of this study was to check the applicability of stylus profilometry (SP), surface hardness (SH) and focus-variation 3D microscopy (FVM) to the analysis of human enamel and dentin subjected to erosion/abrasion. The samples were randomly allocated into four groups (n = 10): G1-enamel/erosion, G2-enamel/erosion plus abrasion, G3-dentin/erosion, and G4-dentin/erosion plus abrasion. The specimens were selected by their surface hardness, and they were subjected to cycles of demineralization (Coca-Cola®-60 s) and remineralization (artificial saliva-60 min). For groups G2 and G4, the remineralization procedures were followed by toothbrushing (150 strokes). The above cycle was repeated 3×/day during 5 days. The samples were assessed using SH, SP, and FVM. For each substrate, the groups were compared using an unpaired t-test, and Pearson correlation coefficients were calculated (α = 5%). For enamel, both profilometry technique showed greater surface loss when the erosion and abrasion processes were combined (P <0.05). The correlation analysis did not reveal any relationships among SH, SP, and FVM to G2 and G4. There were significant correlation coefficients (-0.70 and -0.67) for the comparisons between the FVM and SH methods in enamel and dentin, respectively, in G1 and G3. Choosing the ideal technique for the analysis of erosion depends on the type of dental substrate. SP was not sufficiently sensitive to measure the effects on dentin of erosion or erosion/abrasion. However, SP, FVM and SH were adequate for the detection of tissue loss and demineralization in enamel.

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.

Assessment of cavitated and active non-cavitated caries lesions in 3- to 4-year-old preschool children: a field study.

BACKGROUND: The prevalence of early childhood caries (ECC) is high in developing countries; thus, sensitive methods for the early diagnosis of ECC are of prime importance to implement the appropriate preventive measures. AIM: To investigate the effects of the addition of early caries lesions (ECL) into WHO threshold caries detection methods on the prevalence of caries in primary teeth and the epidemiological profile of the studied population. DESIGN: In total, 351 3- to 4-year-old preschoolers participated in this cross-sectional study. Clinical exams were conducted by one calibrated examiner using WHO and WHO + ECL criteria. During the exams, a mirror, a ball-ended probe, gauze, and an artificial light were used. The data were analysed by Wilcoxon and Mc-Nemar's tests (α=0.05). RESULTS: Good intra-examiner Kappa values at tooth/surface levels were obtained for WHO and WHO+ECL criteria (0.93/0.87 and 0.75/0.78, respectively). The dmfs scores were significantly higher (P<0.05) when WHO+ECL criteria were used. ECLs were the predominant caries lesions in the majority of teeth. CONCLUSIONS: The results strongly suggest that the WHO+ECL diagnosis method could be used to identify ECL in young children under field conditions, increasing the prevalence and classification of caries activity and providing valuable information for the early establishment of preventive measures.

The effect of diode laser irradiation on dentin as a preventive measure against dental erosion: an in vitro study.

Increasing rates of non-carious cervical lesions due to dental erosion, exposure of dentinal tubules, and hypersensitivity to environmental stimuli have led to the development of new prevention strategies. This study evaluated the effects of a low-intensity diode laser (λ = 808 nm) on the dentinal chemical composition and prevention of demineralization. In addition, the study monitored temperature changes during the course of irradiation. Forty dentin specimens were randomly allocated into four groups (n = 10): G1 - No treatment (control), G2 - irradiated with 15 J/cm(2), G3 - irradiated with 30 J/cm(2), and G4 - irradiated with 60 J/cm(2). Each specimen was partially covered with nail varnish, treated according to the group irradiation levels, and exposed to an erosive challenge (1.0 M hydrochloric acid) for 5 min. Afterwards, dentin loss was profilometrically analyzed and examined by scanning electron microscopy (SEM) combined with energy dispersive X-ray (EDX). Intrapulpal temperatures were measured during the dentin irradiation. One-way ANOVA and Tukey tests (p < 0.05) were performed to assess differences. For all irradiated groups, intrapulpal temperature changes were less than 3°C. The G2 group showed statistically significant differences when compared to the other groups, representing the lowest temperature increase. A quantitative element analysis via EDX did not significantly differ (p < 0.05) for Ca, P, F, O, or C between the four groups when measured after irradiation/erosion. The mean wear rates (± SD, µm) were 35.66 ± 7.28; 40.70 ± 5.03; 38.17 ± 10.81 and 25.25 ± 6.87 for G1-G4, respectively. The G4 group statistically differed from all other groups representing the lowest wear rate. These results suggest that dentin irradiation, using a diode laser with levels set at 60 J/cm(2), may induce inhibitory effects on root dentin demineralization without causing any harmful thermal effects. However, the exact mechanism of the action of the laser remains unclear.