The effect of laser therapy for the treatment of dentin hypersensitivity on surface roughness and bacterial adhesion.

The aim of the study was to measure the degree of dentine surface roughness caused by five distinct lasers used to treat dentine hypersensitivity, as well as to evaluate the subsequent bacterial colonization on these irradiated surfaces. Sixty human maxillary premolar teeth without caries or restoration which were extracted for periodontal reasons were used in this study. Five different types of lasers were applied to the root dentin surface. Tested samples were divided into six groups of 10 samples each; control, diode (810 nm), diode (980 nm), Nd: YAG, Er: YAG, and Er, Cr: YSGG laser groups. The arithmetic mean of the surface roughness values (Ra) and the average roughness over a measurement area (Sa) were measured pre- and post-application using any of the laser types. Swab samples were then collected from the dentin surface. Following a 24-hour incubation period at 37 °C, the colony forming units were counted using a stereoscope. The results demonstrated a statistically significant difference in the surface roughness values pre- and post-application (Ra and Sa, respectively) in the Er, Cr: YSGG laser group (p = 0.037,p = 0.007). No significant difference was observed in the other groups (p > 0.05). There was no statistically significant difference in the number of bacterial colonies observed between the test and control groups. Diode and Nd: YAG lasers showed either a decrease or no change in surface roughness; however, the hard tissue lasers (Er: YAG, Er, Cr: YSGG) showed an increase. The Er: YAG and Nd: YAG laser groups exhibited decreased bacterial adhesion compared to the other groups.

In vitro effect of TiF4/NaF solution on the development of radiation-induced dentin caries.

OBJECTIVE: To evaluate the protective effect of an experimental solution containing TiF4/NaF on the development of radiation-induced dentin caries lesions. METHODOLOGY: bovine root samples were irradiated (70Gy) and distributed as following (n=12/group): Commercial Saliva (BioXtra), NaF (500 ppm F-), TiF4 (500 ppm F), TiF4/NaF (TiF4: 300 ppm F-, NaF: 190 ppm F-), and Phosphate buffer solution (PBS, negative control). Biofilm was produced using biofilm from irradiated patients and McBain saliva (0.2% of sucrose, at 37oC and 5% CO2) for five days. The treatments were applied 1x/day. Colony-forming units (CFU) were counted and demineralization was quantified by transversal microradiography. The ANOVA/Tukey test was applied for all parameters. RESULTS: All treatments reduced CFU for total microorganisms. TiF4 reduced Lactobacillus sp. (7.04±0.26 log10 CFU/mL) and mutans streptococci (7.18±0.28) CFU the most, when compared to PBS (7.58±0.21 and 7.75±0.17) and followed by NaF (7.12±0.31 and 7.34±0.22) and TiF4/NaF (7.16±0.35 and 7.29± 0.29). TiF4 and Commercial saliva showed the lowest integrated mineral loss (ΔZ-vol%.mm) (1977±150 and 2062±243, respectively) when compared to PBS (4540±335), followed by NaF (2403±235) and TiF4/NaF (2340±200). Commercial saliva was the only to significantly reduce mineral loss (LD-µm) (111±25) compared to PBS (153±24).Mean mineral loss (R-vol%) decreased by 35.2% for TiF4 (18.2±3.3) when compared to PBS (28.1±2.9) Conclusion: TiF4/NaF has a comparable anti-cariogenic effect to TiF4 and Commercial saliva under the model in this study.

Assessment of multispecies biofilm growth on root canal dentin under different radiation therapy regimens.

OBJECTIVES: To assess the growth of a multispecies biofilm on root canal dentin under different radiotherapy regimens. MATERIALS AND METHODS: Sixty-three human root dentin cylinders were distributed into six groups. In three groups, no biofilm was formed (n = 3): NoRT) non-irradiated dentin; RT55) 55 Gy; and RT70) 70 Gy. In the other three groups (n = 18), a 21-day multispecies biofilm (Enterococcus faecalis, Streptococcus mutans, and Candida albicans) was formed in the canal: NoRT + Bio) non-irradiated + biofilm; RT55 + Bio) 55 Gy + biofilm; and RT70 + Bio) 70 Gy + biofilm. The biofilm was quantified (CFUs/mL). Biofilm microstructure was assessed under SEM. Microbial penetration into dentinal tubules was assessed under CLSM. For the biofilm biomass and dentin microhardness pre- and after biofilm growth assessments, 45 bovine dentin specimens were distributed into three groups (n = 15): NoRT) non-irradiated + biofilm; RT55 + Bio) 55 Gy + biofilm; and RT70 + Bio) 70 Gy + biofilm. RESULTS: Irradiated specimens (70 Gy) had higher quantity of microorganisms than non-irradiated (p = .010). There was gradual increase in biofilm biomass from non-irradiated to 55 Gy and 70 Gy (p < .001). Irradiated specimens had greater reduction in microhardness after biofilm growth. Irradiated dentin led to the growth of a more complex and irregular biofilm. There was microbial penetration into the dentinal tubules, regardless of the radiation regimen. CONCLUSION: Radiotherapy increased the number of microorganisms and biofilm biomass and reduced dentin microhardness. Microbial penetration into dentinal tubules was noticeable. CLINICAL RELEVANCE: Cumulative and potentially irreversible side effects of radiotherapy affect biofilm growth on root dentin. These changes could compromise the success of endodontic treatment in oncological patients undergoing head and neck radiotherapy.

Aluminum zirconate nanoparticles in etch and rinse adhesive to caries affected dentine: An in-vitro scanning electron microscopy, elemental distribution, antibacterial, degree of conversion and micro-tensile bond strength assessment.

To incorporate different concentrations of Al2O9Zr3 (1%, 5%, and 10%) nanoparticles (NP) into the ER adhesive and subsequently assess the impact of this addition on the degree of conversion, µTBS, and antimicrobial efficacy. The current research involved a wide-ranging examination that merged various investigative techniques, including the application of scanning electron microscopy (SEM) for surface characterization of NP coupled with energy-dispersive x-ray spectroscopy (EDX), Fourier-transform infrared (FTIR) spectroscopy, µTBS testing, and microbial analysis. Teeth were divided into four groups based on the application of modified and unmodified three-step ER adhesive primer. Group 1 (0% Al2O9Zr3 NPs) Control, Group 2 (1% Al2O9Zr3 NPs), Group 3 (5% Al2O9Zr3 NPs), and Group 4 (10% Al2O9Zr3 NPs). EDX analysis of Al2O9Zr3 NPs was performed showing elemental distribution in synthesized NPs. Zirconium (Zr), Aluminum (Al), and Oxides (O2). After primer application, an assessment of the survival rate of Streptococcus mutans was completed. The FTIR spectra were analyzed to observe the characteristic peaks indicating the conversion of double bonds, both before and after the curing process, for the adhesive Etch and rinse containing 1,5,10 wt% Al2O9Zr3 NPs. µTBS and failure mode assessment were performed using a Universal Testing Machine (UTM) and stereomicroscope respectively. The µTBS and S.mutans survival rates comparison among different groups was performed using one-way ANOVA and Tukey post hoc (p = .05). Group 4 (10 wt% Al2O9Zr3 NPs + ER adhesive) specimens exhibited the minimum survival of S.mutans (0.11 ± 0.02 CFU/mL). Nonetheless, Group 1 (0 wt% Al2O9Zr3 NPs + ER adhesive) displayed the maximum surviving S.mutans (0.52 ± 0.08 CFU/mL). Moreover, Group 2 (1 wt% Al2O9Zr3 NPs + ER adhesive) (21.22 ± 0.73 MPa) samples displayed highest µTBS. However, the bond strength was weakest in Group 1 (0 wt% Al2O9Zr3 NPs + ER adhesive) (14.13 ± 0.32 MPa) study samples. The etch-and-rinse adhesive exhibited enhanced antibacterial activity and micro-tensile bond strength (µTBS) when 1% Al2O9Zr3 NPs was incorporated, as opposed to the control group. Nevertheless, the incorporation of Al2O9Zr3 NPs led to a decrease in DC. RESEARCH HIGHLIGHTS: 10 wt% Al2O9Zr3 NPs + ER adhesive specimens exhibited the minimum survival of S.mutans. 1 wt% Al2O9Zr3 NPs + ER adhesive samples displayed the most strong composite/CAD bond. The highest DC was observed in Group 1: 0 wt% Al2O9Zr3 NPs + ER adhesive.

Overexpression of proinflammatory cytokines in dental pulp tissue and distinct bacterial microbiota in carious teeth of Mexican Individuals.

The prevalence of dental caries in the Mexican adult population aged 20 to 85 years is around 93.3%, and 50% in Mexican children and adolescents. Worldwide, it is the most common non-communicable disease. One of the main etiological factors for dental caries is the oral microbiome and changes in its structure and function, with an expansion of pathogenic bacteria like Streptococcus mutans. The exposed dental pulp tissue triggers an innate immune response to counteract this bacterial invasion. The relation between oral dysbiosis and innate immune responses remains unclear. We aimed to understand the relationship between innate immune response and the oral microbiota by quantifying the expression of Toll-like receptors (TLRs) and proinflammatory markers (cytokines and a chemokine) in dental pulp tissue, either exposed or not to carious dentin, and to correlate this information with the oral microbiome found in healthy teeth and those with moderate caries. RNA was purified from pulp tissue, subjected to RT-qPCR and analysed with the ΔΔCt method. Supragingival dental plaque of non-carious teeth and dentin of carious teeth were subjected to 16S targeted sequencing. Principal coordinate analysis, permutational multivariate ANOVA, and linear discriminant analysis were used to assess differences between non-carious and carious teeth. Correlations were assessed with Spearman´s test and corrected for multiple comparisons using the FDR method. The relative abundance (RA) of Lactobacillus, Actinomyces, Prevotella, and Mitsuokella was increased in carious teeth; while the RA of Haemophilus and Porphyromonas decreased. Olsenella and Parascardovia were only detected in carious teeth. Significant overexpression of interleukin 1 beta (IL1 ß), IL6, and CXCL8 was detected in pulp tissue exposed to carious dentin. IL1ß correlated positively with TLR2 and Actinomyces; yet negatively with Porphyromonas. These findings suggest that immune response of pulp tissue chronically exposed to cariogenic microbiome is triggered by proinflammatory cytokines IL1ß and IL6 and the chemokine CXCL8.

Modified photoactivated methylene blue-incorporated quartz particles for dentin disinfection: A scanning electron microscope and spectroscopic analysis.

The aim of this study was to synthesize methylene blue-incorporated quartz particles (MB@QP) and to investigate its anti-bactericidal properties. Methylene blue was incorporated inside QP and characterized for morphology and chemical structure using scanning electron microscope (SEM) and Fourier transformed infrared spectroscopy (FTIR). Specimens were randomly divided into experimental and control groups (n = 9/groups). The dentin specimen infected with Enterococcus faecalis was treated using different treatment modalities: Control groups: treatment with 5.25% of sodium hypochlorite (NaOCl) for 60 s; MB: treatment with 1 ml MB solution and incubated for 60 s; MB-PDT (photodynamic therapy): treatment with 1 ml MB solution followed by irradiation using diode laser for 60 s; MB-QP-PDT: specimens treated with MB@QP and irradiated by the diode laser for 60s, and Er,Cr:YSGG laser alone. MB@PDT therapy showed the highest efficacy in reducing the survival rate of E. faecalis (0.49%) in comparison to control NaOCl (0.78%) and Er,Cr:YSGG laser treatment (2.17%). Encapsulating MB into QP followed by the PDT significantly improved the bactericidal capacity and significantly reduced the bacterial survival rate to 0.11% (p < .05) compared to other groups. The combination of MB incorporated into QP and PDT could be an alternative treatment modality to conventional disinfection method for eliminating bacteria from the tooth dentin. RESEARCH HIGHLIGHTS: Quartz particles are potent in delivering the photosensitizer. Photoactivated MB@QP has a higher efficacy in eliminating bacteria from tooth dentin.

Influence of EDTA and its Association with Benzalkonium Chloride on Enterococcus faecalis Adhesion to Dentin / Influencia del EDTA y su Asociación con Cloruro de Benzalconio en la Adhesión de Enterococcus faecalis a la Dentina

ABSTRACT: The aim of this in vitro study was to investigate the influence of ethylenediaminetetraacetic acid (EDTA) associated with the benzalkonium chloride (BAK) on the adhesion and formation of Enterococcus faecalis biofilms attached to coated dentin. Discs standard bovine dentin blocks were treated with the coating materials evaluated: Saline solution (control), 17 % EDTA, 17 % EDTA associated with 1 % BAK for 5 minutes and subsequently washed with saline solution. Afterwards, biofilms of E. faecalis (ATCC 29212) were grown on the surface of coated dentin blocks for time intervals of 1 hour and 7 days (n = 20) and were subsequently washed with phosphate-buffered saline (PBS). Bacterial viability and total biovolume were analyzed by confocal laser scanning microscopy (CLSM) using the Live/Dead technique. Nonparametric Kruskal-Wallis followed by Dunn tests were used to determine statistical differences (a = 5 %). The 17 % EDTA + 1 % BAK group showed significantly lower biovolume and bacterial viability values at the end of 1 hour (p < 0.05). After 7 days of contamination, the 17 % EDTA and 17 % EDTA + 1 % BAK groups showed similar results that differed statistically from those of the control group (p < 0.05). The saline solution group showed higher values. The use of BAK associated with EDTA on dentin blocks surfaces before exposure to contamination was able to interfere in the adhesion of E. faecalis to dentin. Also, dentin treatment by BAK associated with a chelating agent influences the secondary biofilm formation, which could have important effects on the long-term success of root canal treatment.

RESUMEN: El objetivo del estudio consistió en investigar in vitro, la influencia del ácido etilendiamino-tetraacético (EDTA) con cloruro de benzalconio (BAK) en la adhesión y formación de biopelículas de Enterococcus faecalis a la dentina. Discos de dentina bovina fueron tratadas con solución salina (control), 17 % de EDTA, 17% de EDTA asociado con 1 % de BAK durante 5 minutos y lavadas con solución salina. Las biopelículas de E. faecalis (ATCC 29212) se cultivaron sobre los discos de dentina durante intervalos de tiempo de 1 hora y 7 días (n = 20), lavados con solución salina tamponada con fosfato (PBS). La viabilidad bacteriana y el biovolumen total se analizaron mediante microscopía de barrido por láser (CLSM) utilizando la técnica Live / Dead. Se realizó prueba no paramétrica de Kruskal-Wallis, seguida por Dunn con una diferencia estadística (a = 5 %). El grupo de 17 % EDTA + 1 % BAK mostró valores significativamente menores de biovolumen y viabilidad bacteriana al final de 1 hora (p < 0,05). Después de 7 días de contaminación, los grupos de 17 % EDTA y 17 % EDTA + 1 % BAK mostraron resultados similares que diferían estadísticamente del grupo control (p < 0,05). La solución salina mostró valores más altos. La asociación de BAK con EDTA antes de la contaminación interfirió en la adhesión de E. faecalis. Además, el tratamiento de la dentina por BAK asociado con EDTA influye en la formación de biopelículas secundarias, lo que podría tener efectos importantes sobre el éxito a largo plazo del tratamiento del conducto radicular.

Antibacterial effect of diode laser on different cariogenic bacteria: An In-vitro study.

AIMS: The authors have used an in vitro model to appraise the antimicrobial efficacy of diode lasers with two different power outputs on Streptococcus mutans (SM), Lactobacillus casei (LC), and Actinomyces naeslundii (AN). METHODS AND MATERIALS: The coronal dentin of thirty human mandibular third molars was prepared with four cylindrical cavities left in contact with SM, LC, and AN for 72 h to facilitate bacterial penetration. Diode laser (810 nm for 30 s in two cycles) with 1.5 W (group I), 1 W (group II), and 2% chlorhexidine gluconate solution for 60 s (group III) was applied on three cavities and the fourth cavity was not subjected to any treatment (control). Similar amounts of dentin debris were collected from the cavity into sterile tubes. The bacterial count was determined by serial dilution and plate count method. Percentage of killing was calculated for comparative analysis. RESULTS: The percentage of SM killed after exposure was 73.68 ± 23.37, 51.75 ± 25.45, and 26.78 ± 21.8 in three groups, respectively, (P = 0.002; Kruskal-Wallis) with no significant difference between group I and group II (P = 0.089; Mann-Whitney). The percentage of AN killed after exposure was 37.77 ± 49.52, 22 ± 19.48, and 56.86 ± 23.93 in three groups, respectively, (P = 0.013; Kruskal-Wallis) with significant difference between group II and group III (P = 0.002; Mann-Whitney). The percentage of LC killed after exposure was 51.32 ± 39.07, 36.65 ± 38.48, and 75.41 ± 22.6 in three groups, respectively (P = 0.091; Kruskal-Wallis). CONCLUSIONS: Diode lasers exerted antibacterial effect of varying levels against all the three cariogenic bacteria. Although they are recommended as a supplementary antibacterial surface pretreatment technique for efficient removal of cariogenic bacteria, further clinical studies are required to confirm the in vitro findings.

A quaternary ammonium silane antimicrobial triggers bacterial membrane and biofilm destruction.

To study the antimicrobial effects of quaternary ammonium silane (QAS) exposure on Streptococcus mutans and Lactobacillus acidophilus bacterial biofilms at different concentrations. Streptococcus mutans and Lactobacillus acidophilus biofilms were cultured on dentine disks, and incubated for bacterial adhesion for 3-days. Disks were treated with disinfectant (experimental QAS or control) and returned to culture for four days. Small-molecule drug discovery-suite was used to analyze QAS/Sortase-A active site. Cleavage of a synthetic fluorescent peptide substrate, was used to analyze inhibition of Sortase-A. Raman spectroscopy was performed and biofilms stained for confocal laser scanning microscopy (CLSM). Dentine disks that contained treated dual-species biofilms were examined using scanning electron microscopy (SEM). Analysis of DAPI within biofilms was performed using CLSM. Fatty acids in bacterial membranes were assessed with succinic-dehydrogenase assay along with time-kill assay. Sortase-A protein underwent conformational change due to QAS molecule during simulation, showing fluctuating alpha and beta strands. Spectroscopy revealed low carbohydrate intensities in 1% and 2% QAS. SEM images demonstrated absence of bacterial colonies after treatment. DAPI staining decreased with 1% QAS (p < 0.05). Fatty acid compositions of dual specie biofilm increased in both 1% and 2% QAS specimens (p < 0.05). Quaternary ammonium silane demonstrated to be a potent antibacterial cavity disinfectant and a plaque inhibitor and can be of potential significance in eliminating caries-forming bacteria.

Influencia de la dentina en el efecto antibacteriano de 2 concentraciones de hipoclorito de sodio en el crecimiento de Enterococcus faecalis ATCC 29212 / Influence of the dentin on the antibacterial effect of 2 concentrations of sodium hypochlorite in the growth of Enterococcus faecalis ATCC 29212

Este estudio in vitro evaluó la influencia de la dentina sobre el efecto antibacteriano contra Enterococcus faecalis ATCC 29212 de dos concentraciones de Hipoclorito de Sodio (NaOCl) 2,5 % y 5 %. Se empleó polvo de dentina a partir de dientes humanos (84 µg/ml) y la supervivencia de la bacteria se evaluó realizando recuento de unidades formadoras de colonias (UFC) a los 10, 30 y 60 segundos. Los datos se analizaron con la prueba estadística ANOVA factorial no encontrándose diferencias estadísticamente significativas entre los grupos con dentina y sin dentina. En conclusión, la dentina en este estudio no influyó en el efecto antibacteriano del Hipoclorito de Sodio en ninguna concentración, ni en los tiempos.

This in vitro study evaluated the influence of dentin on the antibacterial effect against Enterococcus faecalis ATCC 29212 of two concentrations of Sodium Hypochlorite NaOCl 2.5 % and 5 %. Dentin powder was used from human teeth (84 mg/ml) and the survival of the bacteria was evaluated by counting colony forming units (CFU) at 10, 30 and 60 seconds. The data were analyzed with the statistical ANOVA factorial test, finding no statistically significant differences between the groups with and without dentin. In conclusion, the dentin in this study had no inhibitory effect on antibacterial activity of Sodium Hypochlorite and any concentration, nor over time.

Amplicon-based microbiome study highlights the loss of diversity and the establishment of a set of species in patients with dentin caries.

OBJECTIVES: To elicit patterns in pathogenic biofilm composition we characterized the oral microbiome present in patients with dentin caries in comparison to healthy subjects. METHODS: 16S amplicon sequencing was used to analyse a total of 56 patients; 19 samples of carious dentin (pooled from at least three teeth) and 37 supragingival samples (pooled from three healthy tooth surfaces). Oral and periodontal status and socio-demographic parameters were recorded. Group assignment, smoking and further socio-demographic parameters were used as explanatory variables in the microbiome composition analysis. RESULTS: Overall, a total of 4,110,020 DNA high-quality sequences were yielded. Using a threshold of similarity >97% for assigning operational taxonomic units (OTU), a total of 1,537 OTUs were identified. PERMANOVA showed significant differences in microbiome composition between the groups caries/healthy (p = 0.001), smoking/non-smoking (p = 0.007) and fluoride intake during childhood yes/no (tablets p = 0.003, salt p = 0.023). The healthy microbiome had a significantly higher diversity (alpha diversity, p<0.001) and a lower dominance (Berger-Parker index, p<0.001). It was dominated by Fusobacteria. A linear discriminant analysis effect size (LEfSe) yielded a set of 39 OTUs being more abundant in carious dentin samples, including Atopobium spp. (14.9 log2FoldChange), Lactobacillus casei (11.6), Acinetobacter spp. (10.8), Lactobacillus gasseri (10.6), Parascardovia denticolens (10.5), Olsenella profusa (10.4), and others. Also Propionibacterium acidifaciens (7.2) and Streptococcus mutans (5.2) were overabundant in caries lesions. CONCLUSIONS: The healthy microbiome was highly diverse. The advanced caries microbiome was dominated by a set of carious associated bacteria where S. mutans played only a minor role. Smoking and fluoride intake during childhood influenced the microbiome composition significantly. CLINICAL SIGNIFICANCE: The presented investigation adds knowledge to the still not fully comprehended patterns of oral microbiomes in caries compared with oral health. By analysing the genetics of biofilm samples from oral health and severe tooth decay we found distinct discriminating species which could be targets for future therapeutic approaches.

Effects of Erbium:Yttrium-Aluminum-Garnet Laser Irradiation on Bovine Dentin Contaminated by Cariogenic Bacteria.

Objective: This study was performed to determine the bactericidal effects of erbium:yttrium-aluminum-garnet (Er:YAG) laser irradiation and the morphological and chemical composition changes in bovine dentin. Methods: Dentin slabs were prepared from bovine incisors, and then cultured with Streptococcus mutans to produce bacteria-infected dentin samples. The samples were randomly divided into five groups with Er:YAG laser irradiation energy densities of 0, 6.37, 12.73, 19.11, and 25.47 J/cm2. After irradiation, samples were stained and observed by confocal laser scanning microscopy. The bactericidal abilities were measured using live/dead staining. The morphology and chemical components were investigated by scanning electron microscopy and energy-dispersive spectrometry. Results: After irradiation, the elimination of bacteria and the smear layer were significantly better in the high energy density groups (19.11, 25.47 J/cm2) than in the low energy density groups (6.37, 12.73 J/cm2; p < 0.001). On morphological examination, the group with minimum energy density (6.37 J/cm2) showed superficial melting. In the high energy density groups (12.73, 19.11, and 25.47 J/cm2), laser-irradiated dentin showed a clean surface with open orifices. Significant increases were observed in the weight percentages of calcium (from 19.75 ± 0.69 to 34.47 ± 2.91, p < 0.001) and phosphate (from 8.58 ± 0.43 to 15.10 ± 1.81, p < 0.001), whereas significant decreases were observed for oxygen (from 49.84 ± 0.69 to 36.39 ± 2.86, p < 0.001) and carbon (from 26.06 ± 3.58 to 12.80 ± 2.26, p < 0.01) with increasing energy density. Conclusions: This study confirmed that Er:YAG laser irradiation has bactericidal and dentin conditioning effects.

Oil droplet formation on pellicle covered tooth surfaces studied with environmental scanning electron microscopy.

Lipophilic components are known to modulate the process of bioadhesion on the tooth surface. However, the presence of lipid droplets at the acquired pellicle under oral conditions has not been demonstrated, yet. The purpose of the present study was to establish a method for direct visualisation of lipids on the surface of hydrated, pellicle covered tooth samples by environmental scanning electron microscopy (ESEM), and to use this technique for studying the effects of rinsing with edible oils on the acquired pellicle under in vivo conditions. In situ pellicle formation was performed by 3 min exposure of enamel and dentin specimens in the oral cavity of volunteers. Subsequently, the volunteers rinsed in vivo with safflower oil or linseed oil for 30 s, and the specimens were further carried intraorally for periods from 0 min up to several hours. After intraoral exposure the specimens were treated by osmium tetroxide vapour, and were subsequently analysed by ESEM. This technique was capable to directly visualise the presence of lipid droplets at the pellicle's surface under hydrated conditions. ESEM analyses revealed that surface bound nano- and micro-sized lipid droplets were present at the acquired pellicle's surface even several hours after rinsing with edible oils indicating that these droplets had tightly adhered to the pellicle surface. Pellicle modification by edible oil rinsing as demonstrated in the present study might have the potential to be beneficial as an adjunct in dental prophylaxis.

Efectividad bactericida del diamino fluoruro de plata a diferente concentración sobre estreptococos cariogénicos en muestras de saliva y dentina de escolares: un estudio in vitro / Bactericidal effectveness of silver diamine fluoride in different concentrations on cariogenic Streptococcus in salivary and dentinal samples of schoolchildren: an in vitro study

La OMS y la FDI han publicado que entre el 60 y 90% de los escolares padecen caries. En nuestro país, el Sistema de Vigilancia Epidemiológica de Patologías Orales (SIVEPAB) 2012, reporta un 85% de caries a nivel nacional en población pediátrica. Los agentes anticariogénicos como el diamino y el fluoruro de plata son un tratamiento alentador, este agente puede actuar como bactericida o bacteriostático en función de su concentración y su capacidad para inhibir el crecimiento de estreptococos del grupo viridans, y por ende, de la caries. Problema: ¿Cuál es la efectividad bactericida del diamino fluoruro de plata (Saforide®) a diferente concentración sobre la microbiota cariogénica de escolares? Objetivo: Determinar la eficacia bactericida del diamino fluoruro de plata (DFP) a diferentes concentraciones en el crecimiento bacteriano de Streptococcus mitis, S. mutans y S. salivarius en muestras de saliva y dentina en escolares. Material y métodos: Se llevó a cabo un estudio experimental con una variable independiente, el efecto bactericida del diamino fluoruro de plata y se tomó el halo de inhibición como la dependiente. Se utilizaron medidas descriptivas como prueba de comparación y análisis de varianza usando post-hoc Tukey≠ con una confianza del 95%, y análisis de datos exploratorios. Resultados: Se analizaron 100 muestras, de las cuales 48.3% correspondió a S. mutans, 41.4% a S. salivarius y 10.3% a S. mitis, se obtuvo una mayor zona de inhibición para las tres bacterias al 38% mostrando una diferencia estadísticamente significativa 12% (p < 0.05). También se observó un efecto bacteriostático al 12%, no así para el 38%, donde se encontró un efecto bactericida Conclusión: Nuestros resultados sugieren que al 38% de la concentración hay un claro efecto bactericida en el grupo de estreptococos viridans y el 12% no se recomienda para la detención de caries debido al efecto bacteriostático (AU)

WHO and FDI have ruled that 60-90% of schoolchildren are affected by caries. In our country, the System of Epidemiological Surveillance of Oral Pathologies (SIVEPAB) (SIVEPAB) 2012. Report a rate of 85% of caries nationally in pediatric population. Anticariogenic diamino agents such as silver fluoride are an encouraging decrease in treatment for these high rates of tooth decay in our country, this agent can act as bactericidal or bacteriostatic based on their concentration and their ability to inhibit endogenous metalloproteinase (MMP-2, 8, 9). Problem: What will be the bactericidal effectiveness of silver diamine fluoride different concentration on cariogenic Streptococci saliva samples taken from school and dentin? Objective: Determine the bactericidal effectiveness Silver diamine fluoride (SDF) to different concentration on bacterial growth of Streptococcus mitis, S. mutans, and S. salivarius in saliva samples and dentin in school. Material and methods: An experimental study was conducted as an independent variable the bactericidal effect of silver diamine fluoride was taken as dependent inhibition halo. Descriptive measures were used as a comparison test and analysis of variance using Post-hoc Tukey with 95% confidence, and exploratory data analysis. Results: One hundred samples, of which 48.3% corresponded to S. mutans, 41.4% to S. salivarius and 10.3% to S. mitis, were analyzed, we obtained a larger zone of inhibition for all three organisms at 38% showing a statistically significant difference from 12% (p < 0.05). It was also observed that the 12% sample bacteriostatic effect, not to the concentration of 38% was found a bactericidal effect. Conclusion: Our results suggest that 38% concentration has a bactericidal effect on Streptococcus viridans group and 12% showed not recommended for the arrest or detention of dentine caries bacteriostatic effect (AU)

Effect of Sweeteners on Root Dentine Demineralization Using a Microcosm Biofilm Model / Efecto de los Edulcorantes en la Desmineralización de la Dentina Radicular Utilizando un Modelo de Biofilm Microcosmo

ABSTRACT: The aim of the present study was to evaluate the effect of commercial sweeteners on root dentin demineralization using a microcosm biofilm model. Bovine dentin specimens with pre-determined surface hardness were randomized into six groups according to the studied sweeteners: sucralose, stevia, saccharin, aspartame. Sucrose was used as a positive control and an untreated group as a negative control. The specimens were submitted to biofilm development from one saliva donor and the cariogenic challenge occurred on subsequent five days, twice a day. At the end, the percentage of surface hardness loss (%SHL) and biomass was determined and submitted to ANOVA followed by Tukey's test. Sucrose presented the highest rate of demineralization, however, all sweeteners tested lead to a statistically higher root demineralization compared to the negative control (p <0.05). Sucrose caused greater demineralization in root dentin, however, the sweeteners were also able to induce it under this biofilm model.

RESUMEN: El objetivo del presente estudio fue evaluar el efecto de los edulcorantes comerciales en la desmineralización de la dentina radicular utilizando un modelo de biofilm microcosmo. Se asignaron al azar muestras de dentina bovina con una dureza de la superficie predeterminada de acuerdo con los edulcorantes estudiados: sucralosa, estevia, sacarina, aspartame. La sacarosa se utilizó como control positivo y un grupo no tratado como control negativo. Las muestras se enviaron al desarrollo de biopelículas de un donante de saliva y el desafío cariogénico se produjo en los siguientes cinco días, dos veces al día. Al final, se determinó el porcentaje de pérdida de dureza de la superficie (% PDS) y biomasa y se aplicó un estudio estadístico de ANOVA seguido de la prueba de Tukey. La sacarosa presentó la mayor tasa de desmineralización; sin embargo, todos los endulzantes probados condujeron a una desmineralización de la raíz estadísticamente mayor en comparación con el control negativo (p<0,05). La sacarosa causó una mayor desmineralización en la dentina de raíz, sin embargo, los edulcorantes también fueron capaces de inducirla bajo este modelo de biofilm.

Lack of evidence for the necessity of root canal obturation.

OBJECTIVE: Root canal obturation still is a relevant research topic and patients spend substantial amounts of financial resources for this step of endodontic treatment. Three experiments were conducted challenging the necessity of root canal obturation. METHOD AND MATERIALS: Applying micro computed tomography, the volume of dentin tubules that cannot be instrumented during root canal therapy was determined. Using a simple biofilm model of human tooth segments, the effect of root canal obturation on the persistency of bacteria was evaluated and freshly extracted root canal treated teeth were examined for bacteria remaining in dentin. RESULTS: The volume of dentinal canals was found to be at least three times greater than the volume of the root canal itself. Bacterial growth was observed both in specimens with and without root canal obturation implying that the treatment rendered was ineffective in removing bacterial biofilm and the obturation material was incapable of hindering bacterial regrowth. CONCLUSION: Despite showing adequate root canal obturation radiographically, persistent bacteria could be identified in all teeth extracted. While perfect disinfection of root canals is mandatory, root canal obturation seems questionable as current materials have no antibacterial activity, do not stabilize the tooth, and cannot seal the canal system if a coronal restoration is missing.

Comparison between static and semi-dynamic models for microcosm biofilm formation on dentin

Abstract Microcosm biofilm has been applied to induce carious lesions in dentin. However, no study has been done to compare the impact of the type of model for providing nutrients to microcosm biofilm formation on dentin. Objective This study compared the performance of two kinds of models (static and semi-dynamic) on the biofilm formation and the development of dentin carious lesions. Material and Methods In both models, biofilm was produced using inoculum from pooled human saliva mixed with McBain saliva for the first 8 h (5% CO2 and 37°C). Afterwards, for the static model, the samples were placed in 24-wells microplate containing McBain saliva with 0.2% sucrose, which was replaced at 24 h. In the semi-dynamic model, the samples were submitted to artificial mouth system with continuous flow of McBain saliva with 0.2% sucrose (0.15 ml/min, 37°C) for 10 h a day (for the other 14 h, no flow was applied, similarly to the static model). After 5 days, biofilm viability was measured by fluorescence and dentin demineralization by transverse microradiography. Results Biofilm viability was significantly lower for the static compared with semi-dynamic model, while dentin demineralization was significantly higher for the first one (p<0.05). The static model was able to produce a higher number of typical subsurface lesions compared with the semi-dynamic model (p<0.05). Conclusions The type of model (static and semi-dynamic) applied in the microcosm biofilm may have influence on it's viability and the severity/profile of dentin carious lesions.

Antimicrobial Potential of Laser Diode in Infected Dentin.

AIM: To evaluate the antibacterial effect of diode laser, associated or not with 2.5% sodium hypochlorite (NaOCl), against Enterococcus faecalis. MATERIALS AND METHODS: Eighty dentin blocks were obtained from single-rooted human teeth and sterilized. Seventy were inoculated with 0.01 mL of fresh bacterial inoculum (within 24 hours of preparation from pure culture) standardized to 1 McFarland turbidity. Contaminated blocks were incubated for 7 days at 37°C in humid conditions. Ten uncontaminated samples were incubated at 37°C during the contamination period to serve as a negative control group, while 10 of the infected specimens served as a positive control group. The dentin blocks were randomly divided into eight experimental groups (n = 10 each) according to the method of decontamination: 2.5% NaOCl alone; 2.5% NaOCl + photodynamic therapy (PDT) with methylene blue/660 nm laser at 18 J for 180 seconds; 2.5% NaOCl + PDT with methylene blue/660 nm laser at 8 J for 80 seconds; methylene blue alone; PDT alone with methylene blue/660 nm laser at 18 J for 180 seconds; PDT alone with methylene blue/660 nm laser at 8 J laser for 80 seconds; positive control group; and negative control group. Microbial growth was evaluated by culture medium turbidity and microbial concentration was analyzed by UV spectrophotometry (adjusted to read at wavelength l = 600 nM). RESULTS: Root canals treated with laser alone at 18 J for 180 seconds had higher bacterial contamination compared with groups in which NaOCl was used, with or without laser irradiation at 18 J for 180 seconds (p < 0.05). CONCLUSION: Photodynamic therapy with a 660 nm diode laser effectively reduced E. faecalis contamination. These findings can guide development of further studies in search of better alternatives for endodontic treatment. CLINICAL RELEVANCE: Chemical and mechanical root canal preparation plays an essential role in reducing microbial burden. However, microorganisms present in areas not mechanically reachable by endodontic instruments. As an alternative to fix this problem, the laser can be applied.

Antimicrobial Photodynamic Therapy as an Adjunct for Clinical Partial Removal of Deciduous Carious Tissue: A Minimally Invasive Approach.

This study aimed to evaluate the use of antimicrobial photodynamic therapy (aPDT) as an adjunct for minimally invasive treatment (partial removal of carious tissue-PRCT) of deciduous carious tissue evaluating its efficacy in reducing microorganisms. For that, a clinical study was design including children with deciduous molars with active deep caries lesions (DCL). PRCT was performed and remaining dentin was treated with 100 µg mL-1 methylene blue solution (5 min) and than irradiated with a low power laser emitting red light (InGaAIP-indium gallium aluminum phosphide; λ = 660 nm; 100 mW; 300 J cm-2 ; 90 s; 9 J). The colony forming units (CFU) count after PRCT and after PRCT + aPDT/mg of dentin were compared for total microorganisms, including Candida spp., the mutans streptococci group, Streptococcus spp. and Lactobacillus spp. The dentin was classified (color, consistency and humidity). The microbial reduction varied from 69.88% to 86.29% and was significantly observed for total microorganisms, mutans streptococci, Streptococcus spp. and Lactobacillus spp (P < 0.001). The dentin type did not influence reduction of microorganisms (P > 0.05). The aPDT presents a promising future for clinical use as an adjunct for the reduction of microorganisms in PRCT of DCL in all kinds of dentin.

Bactericidal Effect of Various Laser Irradiation Systems on Enterococcus faecalis Biofilms in Dentinal Tubules: A Confocal Laser Scanning Microscopy Study.

OBJECTIVE: The aim of this study was to compare the bactericidal effect of various laser irradiation systems on Enterococcus faecalis biofilms in dentinal tubules by using a novel dentin infection model and confocal laser scanning microscopy (CLSM). BACKGROUND DATA: Laser-activated irrigations have been proposed as an adjuvant to conventional protocols of root canal treatment to enhance the smear layer removal, which is a promising protocol for root canal disinfection. MATERIALS AND METHODS: E. faecalis were centrifuged into the dentinal tubules, cultured for 3 weeks, and then received 1- and 3-min treatments as follows: (A) 5.25% sodium hypochlorite (NaOCl) irrigation, (B) Nd:YAG laser irradiation, (C) diode laser irradiation, (D) Nd:YAP laser irradiation, (E) Er,Cr:YSGG laser-activated NaOCl irrigation, and (F) Er:YAG laser-activated NaOCl irrigation. Bacterial reductions were assessed by CLSM using a LIVE/DEAD® bacterial viability stain method. RESULTS: For each group, the bacterial reduction increased as the treatment time increased (p < 0.05). The Er,Cr:YSGG and Er:YAG laser significantly enhanced the bactericidal effect of NaOCl (p < 0.05). Under the conditions of the same treatment time, bacterial reductions were presented in the descending order of Er:YAG + NaOCl, Er,Cr:YSGG + NaOCl > Nd:YAP > Nd:YAG, diode > NaOCl. CONCLUSIONS: Under the conditions of present study, treatments of Er:YAG + NaOCl and Er,Cr:YSGG + NaOCl presented the strongest bactericidal effect among the tested protocols and are potential protocols for root canal disinfection.