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Effects of silver diamine fluoride preparations (SDFs) on cariogenic biofilm formation on root dentin (RD) were investigated. Streptococcus mutans (S. mutans) biofilms were formed on bovine RD blocks coated with one of three the SDFs (38%-SDF, 3.8%-SDF and 35%-SDF+potassium-iodide; SDF+KI) and a non-coated Control which were quantified (spectrometric-measurement) and thickness measured (optical coherence tomography) after 20 h. Bacterial viability test (BacLight) and biofilm-morphometry (SEM) of 2 h biofilms were also performed. The amounts of biofilms (bacteria and water insoluble glucan) and the thickness of biofilm were minimum on 38%-SDF specimen; 3.8%-SDF and SDF+KI had significantly more than that, but had significantly less than Control (p

This study investigated the ability of a surface prereacted glass–ionomer (S-PRG) coating material to inhibit the biofilm formation and demineralization of dentin. Dentin specimens were randomly divided into three groups: (1) no coating (control), (2) S-PRG filler-containing coat, and (3) a nonS-PRG filler–containing coat. Streptococcus mutans biofilms were grown on the dentin surfaces in a microcosm for 20 h. Then, the quantity of bacteria and water-insoluble glucan in the retained biofilm on the dentin surface were measured. Regarding demineralization inhibition test, specimens were demineralized for 5 days then sectioned into halves and observed under confocal laser scanning microscope (CLSM). One-way ANOVA and Tukey’s HSD were used for statistical analysis. The estimated mean surface roughness for specimens in the S-PRG group was statistically significantly higher than the estimates for both the nonS-PRG and the control group specimens. The quantity of bacteria and water-insoluble glucan/mm2 revealed that the S-PRG group prevented biofilm formation and bacterial adhesion to the dentin surface compared with the control and nonS-PRG groups. The S-PRG group recorded the highest acid-resistance ability with no surface loss. Application of S-PRG barrier coat on dentin surfaces can inhibit biofilm formation as well as protecting the dentin surface against demineralization. Coating material containing S-PRG fillers might be used for caries prevention, through inhibiting biofilm formation, enhancing mineralization, and reducing acidic attack by cariogenic bacteria.

Objective This study investigated the effect of silver diammine fluoride (SDF), nano-silver (AgNPs) and nano-silver/potassium fluoride (AgNPs/KF) on the morphology of dentin collagen and color staining on the dentin. Also, we investigated the anti-caries properties in terms of anti-bacterial effects and potentiality of dentin remineralization at collagen level. Methods Dentin specimens obtained from bovine tooth roots were finished with #2000 SiC and sonicated for 5 min. Specimens were divided into two main groups: sound and demineralized dentin. Each group was subdivided into five subgroups according to treatment solution: control (no treatment), SDF, nano-silver (AgNPs), AgNPs/KF and KF. Specimens were analyzed using transmission electron microscope (TEM), spectrophotometer, scanning electron microscope (SEM) and antibacterial test. Repeated Measures ANOVA was used for statistical analysis of color change while one-way ANOVA was used for CFU analysis. Results Regarding color change, SDF showed the highest values while the other groups exhibited insignificant changes. SDF showed alteration in the collagen fibrils while the other groups showed normal intact fibrils. Regarding antibacterial activity, SDF and AgNPs/KF showed the greatest antibacterial effect. Significance SDF altered the dentin collagen morphology and inferior ethetics, but still providing intrafibrillar remineralization. Nano-silver fluoride preparations (AgNPs/KF) can be an alternative material for SDF as it can preserve the dentin collagen, showing no color change overtime, and provide a high anti-caries property including intrafibrillar remineralization with a significant anti-bacterial effect.

This study compared the effect of a novel zinc containing, Caredyne Shield (CS), and a fluoroaluminocalciumsilicate-based, Nanoseal (NS) desensitizers on dentin tubule occlusion, inhibition of Streptococcus mutans (S. mutans) biofilm growth, and resistance to bacterial demineralization. Desensitizers were applied to simulated hypersensitive bovine dentin, with distilled water used as a control. S. mutans biofilms were grown on the surface of each specimen in an oral biofilm simulator. CS showed the least bacterial count and water insoluble glucan amount followed by NS. Transverse micro radiography revealed that both CS and NS showed significant reduction in mineral loss and lesion depth of the associated lesion. Scanning electron micrographs showed that the two desensitizers formed obvious depositions on the dentin surfaces, occlusion of tubules and mineral tag formation.

This study was designed to evaluate the cariogenic potential of Lactobacillus gasseri YIT 12321.L. gasseri YIT 12321 was isolated from the human oral cavity as a probiotic candidate strain. Three types of experimental biofilm formations on bovine enamel specimens were assessed using Streptococcus sobrinus ATCC 33478, L. gasseri YIT 12321, and a co-culture of S. sobrinus and L. gasseri YIT 12321 in vitro. L. gasseri YIT 12321 was analyzed for its ability to utilize seven carbohydrates. L. gasseri YIT 12321 was cultured in tryptic soy broth without dextrose and containing a test carbohydrate at 37 °C for 16 h.The decrease in pH in the region under the biofilm produced by L. gasseri YIT 12321 was unusually slow, and the pH was maintained above 5.5 for 16 h. The amount of biofilm and the reduction in enamel hardness were minimal in the L. gasseri YIT 12321 group among the three experimental groups. The amount of co-culture-derived biofilms was less than that of S. sobrinus biofilms. When S. sobrinus was grown using sucrose, the pH decreased to 3.98. In contrast, L. gasseri YIT 12321 cultures maintained the pH above the critical pH for dentin demineralization.L. gasseri YIT 12321 is suggested to have a low cariogenic potential. The application of this strain as a probiotic may contribute to oral safety.

The purpose of this study was to compare optical properties of root caries under two observing conditions using swept-source optical coherence tomography (SS-OCT). In vitro and natural root caries were observed by SS-OCT under wet and dry conditions, followed by confocal laser scanning microscope (CLSM) and transverse microradiography (TMR). Signal intensity (SI), distance between SI peaks (SI-distance) and optical lesion depth were obtained from OCT. Lesion depth was measured from CLSM; lesion depth (LDTMR) and mineral loss (ML) were obtained from TMR. In vitro root caries under wet and dry conditions showed different OCT images and SI patterns. Lesion depth of OCT and that of CLSM, SI-distance and LDTMR, LDTMR and ML significantly correlated. Under dry conditions, half natural root caries showed similar OCT images and SI patterns as in vitro root caries. The base of demineralized dentin could be detected more clearly under dry conditions than under wet conditions.

The aim of this study was to evaluate the bactericidal effect of ultraviolet (UV) light at 265-nm (UVC) and 310-nm (UVB) wavelengths from a newly developed UV light-emitting diode (LED) device against cariogenic bacteria in vitro. Suspensions of Streptococcus mutans (S. mutans) and Streptococcus sobrinus were directly irradiated by UVB or UVC for 2.5 min or 5 min. Numbers of colonies were counted and calculated as colony forming units (CFU) per milliliter. Fluorescence microscopy (FM) and optical density measurements at 490 nm (OD490) were also taken after irradiation. In addition, the bactericidal effects of irradiation against S. mutans under 0.5 mm-thick dentin were compared using culture tests and OD490 measurements. Direct UV-LED irradiation with both UVB and UVC showed strong bactericidal effects. UVB showed superior bactericidal effect through 0.5-mm-thick dentin than did UVC, especially after demineralization. These results suggest that UVB irradiation could be utilized for the prevention and management of dental caries.

Effects of silver diamine fluoride preparations (SDFs) on cariogenic biofilm formation on root dentin (RD) were investigated. Streptococcus mutans (S. mutans) biofilms were formed on bovine RD blocks coated with one of three the SDFs (38%-SDF, 3.8%-SDF and 35%-SDF+potassium-iodide; SDF+KI) and a non-coated Control which were quantified (spectrometric-measurement) and thickness measured (optical coherence tomography) after 20 h. Bacterial viability test (BacLight) and biofilm-morphometry (SEM) of 2 h biofilms were also performed. The amounts of biofilms (bacteria and water insoluble glucan) and the thickness of biofilm were minimum on 38%-SDF specimen; 3.8%-SDF and SDF+KI had significantly more than that, but had significantly less than Control (p0.05). Most S. mutans cells found dead and morphology damaged by 38%-SDF. Some dead bacteria and remarkably damaged biofilms were observed in case of 3.8%-SDF and SDF+KI. Inhibition potential of 3.8%-SDF and SDF+KI on S. mutans biofilm formation is almost similar, although not equivalent to 38%-SDF.

Although a significant association between periodontal disease and atherosclerotic cardiovascular disease has been reported, their cause-to-effect relationship remains controversial. This randomized controlled clinical trial aimed to investigate the effect of advanced self-care on atherosclerotic cardiovascular disease-related vascular function markers flow-mediated brachial artery dilatation (FMD) and serum asymmetric dimethylarginine (ADMA) level in patients with early-stage periodontal disease. The study was designed as a parallel group, 3-month follow-up, open-label, randomized controlled trial. The control group received standard care for periodontal diseases, whereas the test group additionally applied disinfectant using a custom-fabricated prescription tray for advanced self-care twice a day. Overall, 110 patients provided data for FMD and serum ADMA level. No significant improvements in FMD were observed in the control (mean increase, −0.1%; 95% confidence interval [CI], −1.0–0.8; P = 0.805) or test (mean increase, −0.3%; 95% CI, −1.1–0.4; P = 0.398) group. No significant changes in serum ADMA levels were observed (mean reduction, 0.01 μmol/L; 95% CI, −0.00–0.02; P = 0.366 and mean reduction, 0.00 μmol/L; 95% CI, −0.01–0.01; P = 0.349, respectively). No significant between-group differences were found in FMD (mean difference, −0.2%; 95% CI, −1.4–0.9; p = 0.708) or serum ADMA levels (mean difference, 0.01 nmol/L; 95% CI, −0.00–0.03; p = 0.122). Significant improvements in the average probing pocket depth were observed in the control and test groups. The bleeding on probing score in the test group was significantly reduced, while that in the control group was reduced, although not significantly. Periodontal care for a 3-month duration did not provide better endothelial function although improvements of periodontal status in patients with early-stage periodontal diseases. This trial is registered in UMIN Clinical Trials Registry (www.umin.ac.jp/ctr/; ID: UMIN000023395).

This study evaluated the effect of mechanical abrasion on the surface integrity, color change (ΔE) and antibacterial properties of demineralized and sound dentin surfaces treated with silver-diammine-fluoride (SDF). The dentin specimens were divided into two groups: sound and demineralized dentin, then divided into three sub-groups, control (no-treatment), SDF, and SDF + potassium-iodide (KI). Each sub-group was further divided into two groups, one exposed to mechanical brushing and the other without brushing. Specimens were analyzed for the ΔE, surface roughness/surface loss and antibacterial properties (CFU, optical density and fluorescent microscope). Repeated Measures ANOVA was used for statistical analysis of color change while one-way ANOVA was used for CFU analysis. SDF and SDI + KI groups showed significant reduction in ΔE with brushing in the sound dentin group unlike the demineralized group. The surface roughness values were higher for both SDF and SDF + KI groups but roughness values significantly decreased after brushing. Both SDF and SDF + KI groups revealed significantly less surface loss than control. The SDF group showed high anti-bacterial effect after brushing, unlike SDF + KI group. So, we concluded that mechanical brushing improved the esthetic outcome. While, SDF and SDF + KI could protect the dentin surface integrity. SDF-treated dentin possesses an antibacterial property even after mechanical brushing.

Objective We aimed to evaluate morphological, mechanical and chemical characteristics at resin–dentin interfaces using multiple combinations of adhesives and resins after a short-term biofilm-induced degradation. Methods Cervical cavities were prepared in bovine incisors, treated by Clearfil SE Bond 2 (SE) or FL-Bond II (FL), restored by Clearfil Majesty ES Flow (ES) or Beautifil Flow Plus (BFP) and grouped into SE-ES, SE-BFP, FL-ES and FL-BFP. After biofilm challenge, interfacial gaps and dentin wall lesions were examined by optical coherence tomography (OCT). Gap depth (GD), gap pattern scale (GPS) and dentin wall lesion depth (WLD) were evaluated from confocal laser scanning microscope. Microhardness of dentin lesions was measured with a Vickers microhardness tester. Chemical elements in resins and dentin wall lesions were analyzed by scanning electron microscopy and energy dispersive X-ray spectrometry (SEM/EDS). Morphological structures of interfacial gaps were observed by SEM. Results OCT could detect adhesive-dentin-bonded and adhesive-dentin-debonded gaps. SE-containing groups showed significantly lower GPS than FL-containing groups. FL-BFP showed significantly lower WLD than FL-ES. Microhardness of dentin wall lesions was higher than that of outer lesions and they showed significant differences in FL-BFP. SE-BFP showed a lower GPS curve and higher intensities of Ca and P in the upper half of dentin wall lesions than other groups. From SEM, microgaps between filler and matrix, break and loss of matrix, separation of adhesive matrix with hybrid layer occurred at interfacial gaps. Significance The morphological, mechanical and chemical characteristics of resin–dentin interfacial degradation depend on the component and chemistry of restorative materials.

This study investigated the ability of a surface prereacted glass-ionomer (S-PRG) coating material to inhibit the biofilm formation and demineralization of dentin.Dentin specimens were randomly divided into three groups: (1) no coating (control), (2) S-PRG filler-containing coat, and (3) a nonS-PRG filler-containing coat. Streptococcus mutans biofilms were grown on the dentin surfaces in a microcosm for 20 h. Then, the quantity of bacteria and water-insoluble glucan in the retained biofilm on the dentin surface were measured. Regarding demineralization inhibition test, specimens were demineralized for 5 days then sectioned into halves and observed under confocal laser scanning microscope (CLSM). One-way ANOVA and Tukey's HSD were used for statistical analysis.The estimated mean surface roughness for specimens in the S-PRG group was statistically significantly higher than the estimates for both the nonS-PRG and the control group specimens. The quantity of bacteria and water-insoluble glucan/mmApplication of S-PRG barrier coat on dentin surfaces can inhibit biofilm formation as well as protecting the dentin surface against demineralization.Coating material containing S-PRG fillers might be used for caries prevention, through inhibiting biofilm formation, enhancing mineralization, and reducing acidic attack by cariogenic bacteria.

Oral biofilm reactor (OBR) and pH cycling (pHC) artificial caries model were employed to evaluate the anti-demineralization effects of four composite filling systems on enamel–root dentin junction. Sixty-four enamel–root dentin blocks (6 mm × 6 mm × 2 mm) each with a cylindrical cavity were randomly assigned to the pHC and OBR group, then four subgroups (n = 8) and filled with either the Beautifil II (BEF, SPRG-filler-containing) or Estelite (EST) composite after the adhesive (either Single Bond Universal (SBU) or FL Bond II (FL, SPRG-filler-containing)). The demineralization lesions of filling interface were examined by micro-computerized tomography (μCT) and swept-source-optical coherence tomography (SS-OCT). According to the degree of interface damage, the caries lesions were sorted into four types: Type A and B (no attachment loss), Type C and D (attachment loss). EST/SBU showed the worst demineralization lesion and attachment loss (100% Type D), while BEF/FL exhibited the shallowest lesion depth (p &lt, 0.05, 145 ± 45 μm on enamel, 275 ± 35 μm on root dentin) and no attachment loss (75% Type A and 25% Type B). Using FL adhesive alone does not effectively reduce enamel demineralization. BEF plays a leading role in acid resistance. The combination of BEF and FL showed a cumulative synergistic effect on anti-demineralization.

The aim of this study was to evaluate the effect of resin infiltration on root caries induced by Streptococcus mutans biofilms. Human premolar specimens were divided to 5 groups: negative control (NC), Clearfil SE Bond (SEB), Icon-etch120s+Icon-infiltrant (HA120), Icon-etch10s+Icon-infiltrant (HA10) and K-etchant10s+Icon-infiltrant (PA10). The resin penetration was observed by fluorescent microscope. Biofilm-induced demineralization was conducted again and observed by swept-source optical coherence tomography and confocal laser scanning microscope. The maximum resin penetration depth (PDmax), lesion depth increase (∆LD), frequency of cervical enamel loss and dentinoenamel junction separation length were measured and statistically analyzed. HA120 showed 138.00±49.25 µm PDmax that was significantly larger than PA10 and SEB (p

Objective To evaluate early degradation at resin-dentin interface using non-invasive swept-source optical coherence tomography (SS-OCT) and confocal laser scanning microscope (CLSM). Methods Self-etch adhesives and resin-composites containing bisphenol-glycidyl-dimethacrylate (Bis-GMA), which is one of the most widely used monomers in restorative materials, were investigated in this study. Forty cervical cavities were prepared in bovine incisors and applied by the adhesive with/without Bis-GMA (AdhesiveBG/Adhesive), filled by the resin with/without Bis-GMA (ResinBG/Resin) and then challenged by cariogenic biofilm (37 °C, 24 h). Gap Formation and dentin demineralization around resin-composites were observed by SS-OCT and CLSM. Results Three types of resin-dentin interfacial degradation could be detected from SS-OCT. Type I—dentin demineralization around resin without gap, showing feather-shaped dark zones without bright scattered lines at resin-dentin interfaces. Type II—dentin demineralization around resin with adhesive-dentin bonded gaps, showing feather-shaped dark zones with bright scattered lines at resin-dentin interfaces. Type III—dentin demineralization around resin with adhesive-dentin debonded gaps, showing edge-shaped dark zones with bright scattered lines at resin-dentin interface. From CLSM, the groups were compared in gap scale (GS), gap depth (GD), gap width (GW) and dentin wall lesion depth (WLD). Bis-GMA-containing adhesive groups showed significantly lower GS than Bis-GMA-free adhesive groups. Bis-GMA-containing resin groups showed significantly lower WLD than Bis-GMA-free resin groups. However, they did not show significant differences in GD and GW. Conclusion Three types of early degradation at resin-dentin interface can be noninvasively detected by SS-OCT. Bis-GMA-containing and Bis-GMA-free restorative materials show differences in gap scale and dentin wall lesion depth. Clinical significance SS-OCT can nondestructively detect early resin-dentin interfacial degradation. Gap scale can be used as a parameter to evaluate the risk factor of gaps.

To observe the bacterial demineralization of the enamel and dentin around composite restorations bonded with one-step and two-step self-etch adhesive systems using swept-source optical coherence tomography (SS-OCT).Forty class V cavities (2.5-mm surface diameter, 2.0-mm maximum depth) were prepared on cervical areas of 20 human molars. The specimens were either treated with one-step adhesive (Clearfil Tri-S Bond ND Quick; TS) or two-step adhesive (Clearfil SE Bond; SE), restored with a flowable resin composite (Estelite Flow Quick). Specimens in the demineralized group were incubated for 2 weeks after Streptococcus mutans biofilm formation, while specimens in the control group were incubated for 2 weeks without biofilms. After SS-OCT observation, specimens were cut and examined under confocal laser scanning microscope (CLSM). The lesion depth (LD), interfacial gap depth (GD) and gap length (GL) obtained from SS-OCT and CLSM were analyzed by Pearson's correlation, LD by Independent-samples t-test, GD and GL by Welch t-test, the frequency of specimens with or without gap by Fisher's exact test at the 95% significant level.Specimens with interfacial gaps in the demineralized group showed significantly higher frequency than that in the control group (p0.05). There was a significant correlation between SS-OCT and CLSM values of LD, GD and GL (p0.05). SE showed significantly larger LD of dentin, but less GD and GL of enamel than TS (p0.05).SS-OCT nondestructively detected demineralization around composite restorations and interfacial gaps created by S. mutans biofilm in this in vitro model.

Typical methods for the elucidation of the function of a particular gene involve comparative phenotypic analyses of the wild-type strain and a strain in which the gene of interest has been disrupted. A gene-disruption DNA construct containing a suitable antibiotic resistance marker gene is useful for the generation of gene-disrupted strains in bacteria. However, conventional construction methods, which require gene cloning steps, involve complex and time-consuming protocols. Here, a relatively facile, rapid, and cost-effective method for targeted gene disruption in Streptococcus mutans is described. The method utilizes a 2-step fusion polymerase chain reaction (PCR) to generate the disruption construct and electroporation for genetic transformation. This method does not require an enzymatic reaction, other than PCR, and additionally offers greater flexibility in terms of the design of the disruption construct. Employment of electroporation facilitates the preparation of competent cells and improves the transformation efficiency. The present method may be adapted for the generation of gene-disrupted strains of various species.

The aim of this study was to evaluate the signal intensity and signal attenuation of swept source optical coherence tomography (SS-OCT) for dental caries in relation to the variation of mineral density. SS-OCT observation was performed on the enamel and dentin artificial demineralization and on natural caries. The artificial caries model on enamel and dentin surfaces was created using Streptococcus mutans biofilms incubated in an oral biofilm reactor. The lesions were centrally cross sectioned and SS-OCT scans were obtained in two directions to construct a three-dimensional data set, from the lesion surface (sagittal scan) and parallel to the lesion surface (horizontal scan). The integrated signal up to [Formula: see text] in depth (IS200) and the attenuation coefficient ([Formula: see text]) of the enamel and dentin lesions were calculated from the SS-OCT signal in horizontal scans at five locations of lesion depth. The values were compared with the mineral density obtained from transverse microradiography. Both enamel and dentin demineralization showed significantly higher IS200 and [Formula: see text] than the sound tooth substrate from the sagittal scan. Enamel demineralization showed significantly higher IS200 than sound enamel, even with low levels of demineralization. In demineralized dentin, the [Formula: see text] from the horizontal scan consistently trended downward compared to the sound dentin.

Objectives:The aim of this study was to compare the effects of certain commercially available mouthwashes on cariogenic bacteria and biofilms, following the acquisition of inhibition potentials of caries.Materials and Methods:Mouthwashes containing I) chlorhexidine gluconate (CHG; 0.0005% w/v), II) benzethonium chloride (BTC; 0.01% w/v), III) an essential oil (Listerine), and IV) povidone-iodine (PVP-I; 0.035% w/v) were tested on planktonic cariogenic bacteria, biofilms, and an ex vivo caries model. Bacterial aliquots were inoculated with each solution separately and vortexed for 10 seconds at room temperature. Bacterial viability was subsequently investigated by fluorescence microscopy (FM) after staining with a BacLight viability kit and the number of colony-forming units (CFUs) was counted. Similarly, mouthwash solutions were applied to artificial cariogenic biofilms, and bacterial viability of the biofilms was investigated as stated above. Inhibition potentials of two selected mouthwashes of carious lesions were investigated using biofilm-induced caries and a secondary caries model. In all steps, a phosphate-buffered saline (PBS) solution was included as a control.Results:Planktonic cariogenic bacteria and bacteria embedded in biofilms were killed in remarkably large numbers with Listerine and PVP-I treatment compared to PBS and other gargles. CFU counts also showed significant reduction after treatment with Listerine and PVP-I compared to other solutions (PConclusion:Bactericidal potencies of the mouthwashes varied significantly, suggesting that mouthwashes like Listerine can be useful for the prevention of caries and secondary caries.

Background: The aim of this study was to establish a new in vitro model for biofilm induced secondary caries studies using an oral biofilm reactor. Methods: An approximately 2 × 3 × 2 mm3 sized dentino-enamel Class I cavity was prepared in the middle of a square-shaped specimen from the mid-labial portion of bovine incisors. The cavities were partially filled with either Clearfil AP-X with SE-Bond or Clearfil AP-X without any bond. Artificial biofilms were then formed on the resin composite filled surfaces using three species of oral bacteria in an oral biofilm reactor for 20 hours followed by 7- or 30-day incubation periods. Results: The lesions were clearly visible on fluorescence microscopy and by scanning electron microscopy in the enamel at the interface of resin restorations in all samples. The data from image analysis showed that the lesion size was largest in the No-bond samples with statistically significant differences (p

Protein-resistant coatings have been studied for inhibiting biofilm formation on implant devices. In this study, titanium (Ti) surfaces were biofunctionalized with poly(ethylene glycol) (PEG) by electrodeposition and were evaluated as biofilm substrates under an oral simulated environment. Streptococcus gordonii, an early colonizer of oral biofilms, was inoculated on Ti and PEG-electrodeposited Ti (PEG-Ti) surfaces and was analyzed quantitatively and topographically. Streptococcus mutans supplemented with sucrose, a late colonizer mainly found in dental plaque, was also used to form biofilms on the surfaces of Ti and PEG-Ti for 20 h followed by sonication as a means of detaching the biofilms. The results indicated that the attachment of S. gordonii on PEG-Ti surfaces was inhibited compared with Ti, and the S. mutans biofilm was easier to be detached from the surface of PEG-Ti than that of Ti. Moreover, the presence of PEG electrodeposited on Ti surface inhibited salivary protein adsorption. The degree of detachment of biofilms from PEG-Ti was associated with the inhibition of the salivary protein adsorption, suggesting weak basal attachment of the biofilms to the electrodeposited surfaces. Therefore, controlling protein adsorption at the initial stage of biofilm formation may be an effective strategy to protect metal surfaces from bacterial contamination not only in dental manipulations but also in orthopedic applications.

This study was designed to evaluate the efficacy of resin coating on the regional microtensile bond strength (MTBS) of a resin cement to the dentin walls of Class II cavities. Twenty mesio-occlusal cavities were prepared in human molars. In 10 cavities, a resin coating consisting of a self-etching primer bonding system, Clearfil SE Bond, and a low-viscosity microfilled resin, Protect Liner F, was applied. The other 10 teeth served as a non-coating group. After impression taking and temporization, they were kept in water for one day. Composite inlays were then cemented with a dual-cure resin cement, Panavia F 2.0, and stored in water for one day. Thereafter, MTBSs were measured. Two-way ANOVA (p=0.05) revealed that the MTBS of resin cement to dentin was influenced by resin coating, but not by regional difference. In conclusion, application of a resin coating to the dentin surface significantly improved the MTBS in indirect restorations.

The surface properties of three different resin composite materials which influence Streptococcus mutans biofilm formation were evaluated using an artificial mouth system (AMS). Specimens were prepared from Clearfil AP-X, Grandio, and Reactmer Paste, and each material was divided into two groups: (1) surface was ground with 800-grit silicon paper (SiC#800); or (2) surface was polished with up to 1- microm diamond paste (DP1 microm). Biofilms were grown on the surface of each specimen for 20 hours, and then subjected to vortex agitation followed by measurement of retained biofilms. Surfaces with retained biofilms were also inspected by SEM. Significant differences were detected in surface roughness (Ra) between the two polishing conditions for all materials. The quantity of retained biofilm was significantly less (p < 0.05) on Clearfil AP-X DP 1 microm than on Clearfil AP-X SiC#800. With Reactmer Paste, their surfaces registered the lowest amount of retained biofilm--but there were no significant differences between the two polishing conditions. In conclusion, polishing did not render all resin composites equally resistant to biofilm formation.

E2f1 is an important regulator of T cell proliferation, differentiation, and apoptosis that controls the transcription of a group of genes that are normally regulated at the G1 to S phase transition in the cell cycle. Insulin-dependent diabetes mellitus (IDDM) and Sjögren’s syndrome (SS) are highly regulated autoimmune diseases that develop spontaneously in NOD mice. The aim of the present in vivo study was to explore the functional importance of the E2f1 molecule in IDDM and SS, in the context of whole animal physiology and pathophysiology, using E2f1-deficient NOD mice. For the experiment, we produced NOD mice homozygous for a nonfunctional E2f1 allele onto a NOD background. E2f1-deficient NOD mice developed an early and increased onset of diabetes as compared with their littermates. These mice also exhibited a defect in T lymphocyte development, leading to excessive numbers of mature T cells (CD4+ and CD8+), due to a maturation stage-specific defect in the apoptosis of thymocytes and peripheral T cells. We also found that they also exhibited a more rapid and increased entry into the S phase following antigenic stimulation of spleen cells and thymocytes in vitro. Furthermore, E2f1-deficient mice showed a profound decrease of immunoregulatory CD4+CD25+ T cells, while the spleen cells of NOD mice lacking E2f1 showed a significant increase of the proinflammatory cytokine IFN-γ following antigenic stimulation in vitro. Consistent with these observations, E2f1 homozygous mutant NOD mice were highly predisposed to the development of IDDM and SS.

The use of peptides for various aspects of medical science has been a significant advance. Peptide-based vaccines are promising, but weak immunogenic potency is impeding the clinical application. We have remarkably enhanced the immunogenicity of peptide antigens by addition of motifs that bind to cell attachment proteins, such as arginine-glysine-aspartate (RGD), to the amino acid sequence. The modified peptides induced antigen-specific serum antibodies by intranasal immunization without adjuvants. RGD, an integrin-binding motif was the strongest, among several molecules tested in this experiment, giving an average of 10 times enhancement of antibody titers when incorporated into several peptide antigens. The peptides also acted as an efficient adjuvant following the intranasal immunization with protein antigens. Our data support the feasibility of developing peptide vaccines and peptide adjuvants for intranasal vaccination.

Objective: Aspects of an Artificial Mouth System (AMS) to model the caries process in bovine enamel biofilm induced white spot enamel lesions (WSEL) were analyzed. Methods: Square shaped and polished bovine enamel slabs were covered with modeling wax and 2.5 mm diameter window was prepared to expose enamel surface at the center of each slab. Artificial biofilms were grown inside an AMS on the slabs using freshly cultured S. mutans (MT8148) in suspended in phosphate buffered saline (PBS), heart infusion (HI) with 1% sucrose and PBS for 20, 30 and 40 hrs at 37°C. The severity of demineralization was quantified by Quantitative Light-induced Fluorescence (QLF, Inspektor, Netherlands). All experiments were repeated three times (n=5 for each group). Results: A circular WSEL could clearly be detected at the center of each slab by naked eyes, which was prominent after dehydration by mild air drying. QLF data showed that severity of demineralization occurred with the increase of time interval, after 40 hours both ΔF (23+/-2.9) and ΔQ (148.75+/- 26.78) were significantly more compared to ΔF (10.14+/-2.3) and ΔQ (47.63+/-30.67) of 20 hours. Conclusion: An advanced in vitro model of biolfilm induced WSEL was developed that showed promises to be useful in studying enamel demineralization. DOI: http://dx.doi.org/10.3329/cdcj.v9i1.11829 City Dental College Journal Vol.9(1) 2012 4-6

NOD/LtSz-prkdc(scid)/prkdc(scid) (non-obese diabetic-severe combine immunodeficiency; NOD-scid) mice grafted with human peripheral blood lymphoid cells have been used as an in vivo humanized mouse model in various studies. However, cytotoxic human T cells are induced in this model during immune responses, which gives misleading results. To assist in grafting of human lymphocytes without the induction of cytotoxic human T cells, we investigated the effects of T helper type 1 (Th1) and Th2 cytokines on human lymphocyte grafting and migration, as well as the production of immunoglobulin deposited in glomeruli and human immunodeficiency virus-1 (HIV-1) infection using NOD-scid mice. Administration of interleukin-18 (IL-18) and IL-12 enhanced the grafting of human CD4+ and CD8+ T cells in the mice, whereas co-administration prevented grafting due to interferon-gamma-dependent apoptosis. Immunoglobulin A (IgA) deposits were observed in mice treated with IL-18 alone, but not in those given phosphate-buffered saline, IL-12 alone, or IL-18 + IL-12. A high rate of HIV infection was also observed in the IL-18-treated group. Together, these results indicate that IL-18 may be effective for the grafting and migration of CD4+ and CD8+ T cells, except for the induction of apoptosis and regulation of class-switching IgA. IL-18-administered NOD-scid mice provide a useful small humanized model for the study of HIV infection and IgA nephropathy.

SUMMARY The chemokine stromal-cell derived factor-1 (SDF-1) controls maturation, trafficking, and homing of certain subsets, lymphoid cells including immunogenic B and T cells, as a ligand of the CXCR4 chemokine receptor. Insulin-dependent diabetes mellitus (IDDM) and Sjogren's syndrome (SS), both highly regulated autoimmune diseases, develop spontaneously in non-obese diabetic (NOD) mice. To investigate the role of SDF-1 in the development of autoimmune diseases, we injected groups of NOD female mice with antibodies to SDF-1 (anti-SDF-1), which resulted in a 30% reduction of diabetes up to 30 weeks of age, delayed average diabetes onset by 10 weeks, and suppressed insulitis. Autoimmune sialoadenitis was evident in anti-SDF-1-injected mice (SDF-1-Ig group) at the same level as in all groups of mice, whether injected with non-specific antibodies or not. In addition, in the SDF-1-Ig group, a greater number of immunoglobulin M (IgM) x IgD x B220 low CD38 + CD43 + CD23 x progenitor B cells and IgM + IgD + B220 high CD43 x CD38 + CD24 + CD23 + mature B cells remained in the bone marrow, whereas infiltration of mature IgM + B cells was less extensive in peripheral tissues. Our results suggested that anti-SDF-1 antibodies injection was effective in inhibiting diabetes and insulitis without affecting autoimmune sialoadenitis or SS in NOD mice. SDF-1 may be an essential chemokine for trafficking and migration of autoreactive B cells in the development of diabetes.

Titanium and stainless steel wires used for retainers in orthodontic procedures were coated with Ag ions and the effects of the coating on common oral pathogens and their pathogenicity were investigated. Two species of cariogenic and three species of periodontopathic bacteria were assessed. Biofilms of Streptococcus sobrinus and two VSC gases produced by P. gingivalis were also examined. Ag ion-coated wires showed marked antibacterial activities compared with uncoated wires; in most cases, the differences were statistically significant (p

A surface protein antigen (PAc) of Streptococcus mutans, particularly the A-region of the molecule, has been reported to interact with salivary components on the tooth surface. It might be a candidate antigen inducing the production of antibodies against the adherence of S. mutans to the tooth surface. We investigated the effects of monoclonal antibodies (MoAbs) obtained by immunization of synthetic PAc peptides that completely correspond to the amino acid sequence of part of the A-region. These MoAbs recognize several core B-cell epitopes in the sequence. Two (KH5 and SH2) of these antibodies reacted with both S. mutans and Streptococcus sobrinus, but not with Streptococcus sanguis, Streptococcus salivarius, Porphyromonas gingivalis or Lactobacillus casei. They clearly inhibited the real-time adherence of S. mutans to salivary components in a biosensor. KH5, which showed a real-time inhibition (71%), also significantly prevented the recolonization of S. mutans on the tooth surface in rats. These results suggested that the core B-cell epitope (-Y---L--Y----) recognized by KH5 was the essential sequence in the antigenic epitopes of PAc protein recognized specifically by the inhibitory antibody. Therefore, the amino acid residues were found to be important in the initial attachment of S. mutans to the tooth surface. These results provide for the mechanism of PAc molecule in the initial attachment of S. mutans on the tooth surface and more effective designs for the removal of S. mutans and S. sobrinus from the oral cavity.

感染した接合上皮と結合組織を無菌化することを目的として, 歯周ポケットにCO2レーザーを照射した際の歯肉組織の変化について検討した。実験には, デフォーカス型のCO2レーザーと6頭の雑種成犬を用いた。実験に先立ち, イヌには毎日15～20分のブラッシングを1週間続けて行った。この時点での歯周ポケットは, 最高で3mmであった。実験的な歯肉炎は, 軟食を与えてブラッシングも中止することにより引き起こし, 4週後には平均で1～2mm歯周ポケットが増加した。小臼歯と大臼歯の頬側歯肉45部位に, 0.3から2.5Jの範囲で出力を変化させて照射した。他の15部位については, 照射せずにコントロールとした。照射後7日と14日に, 臨床的および組織学的に歯肉組織の反応を観察した。照射直後には, 軽度の歯肉炎が観察された。組織の破壊程度は出力に関係しており, すなわち最大の組織破壊 (到達深度が1.4mmで創傷領域が0.33mm2) は最高出力 (2.5J) の場合に起こった。7日後には, 照射された歯肉の再上皮化がほぼ完全になされており, 結合組織にも新生したコラーゲン線維が形成されていた。特に0.5～1.5Jの照射出力の時は, コントロールよりも健康的な歯肉が臨床的にも組織学的にも認められた。この時期には, 炎症性の細胞は顕著に減少していた。14日後になると, レーザーを照射した歯肉は完全に治癒し, 傷害を受けた痕跡は全く観察されなかった。今回の実験から, 周囲組織を損傷せずに感染した接合上皮を無菌化することができるCO2レーザーの出力範囲を考察することができた。

Exposed root surfaces due to gingival recession are subject to biofilm stagnation that can result in caries formation. Cervical enamel and dentin demineralization induced by a cariogenic biofilm was evaluated using swept-source optical coherence tomography (SS-OCT). The cementoenamel junction (CEJ) sections of extracted human teeth were subjected to demineralization for 1, 2, or 3 weeks. A suspension of Streptococcus mutans was applied to form a cariogenic biofilm using an oral biofilm reactor. After incubation, demineralization was observed by SS-OCT. For the analysis of SS-OCT signal, the value of the area under the curve (AUC) of the signal profile was measured. Statistical analyses were performed with 95% level of confidence. Cervical demineralization was displayed as a bright zone in SS-OCT. The demineralization depth of dentin was significantly deeper than that of enamel ([Formula: see text]). Enamel near the CEJ demonstrated a significant increase of AUC over the other enamel region after the demineralization. The gaps along the dentinoenamel junction were additionally observed in SS-OCT. SS-OCT was capable of monitoring the cervical demineralization induced by a cariogenic biofilm and is considered to be a promising modality for the diagnosis of cervical demineralization.

Objectives A low pH environment is created by cariogenic bacteria. This study was aimed to measure pH of carious lesions intraorally using a micro-pH sensor, and assess predominant acid-producing cariogens by qPCR to differentiate caries activities. Methods 103 dentine lesions classified as active or arrested caries based on the clinical and radiological examinations were collected from patients after intraoral measurement of the lesion surface pH using a micro-pH sensor. Quantitative detection of genomic DNA copies of target cariogenic bacteria (mutans streptococci and Lactobacillus spp.) in each lesion was performed using real-time PCR. Correlation between the pH ranges and the number of bacterial species was examined by Spearman test. Results 50 samples were diagnosed as active and 53 as arrested lesions. Statistically significant difference was observed on average surface pH value between active and arrested lesions ( p Lactobacillus spp. was higher in active lesions than in arrested lesions (76% vs. 58% of samples, respectively). When the carious lesions were categorised into four different pH ranges (up to 5.5, from 5.6 to 5.8, from 5.9 to 6.1 and 6.2 or above), increased prevalence of Lactobacillus spp. was observed with decrease of pH levels. A significant negative relationship was found between pH value and number of Lactobacillus spp. ( r = −0.209, p Conclusions Intraoral pH measurement might be clinically useful to determine acidity of the local environment of carious lesions as one aspect of the caries activity assessment. Clinical significance The population of certain bacteria may indicate activity of carious lesions. Intraoral pH measurement of the carious lesions using a micro-pH sensor may be a clinically feasible method for assessment of lesion acidity.

Objective The aim of the study was to elucidate any alterations in nanomechanical properties and structural integrity of enamel surfaces after preservation in different liquid media. Methods The nanohardness and modulus of elasticity of bovine enamel blocks were measured before (control) and after storage in all of the following liquid media: human pooled saliva, physiological saline (NaCl), Deionized water (Milli-Q water) and phosphate buffered saline (PBS) for 1, 7, 14 and 30 days. Control and the 30 days preserved samples were examined by scanning electron microscopy (SEM). Data were analysed by one-way ANOVA and Dunnett T 3 test. Results Hardness and modulus of elasticity of enamel in saliva up to 14 days showed no significant decrease compared to controls. In the case of PBS, both hardness and modulus did not change up to 30 days. The hardness values of Milli-Q water (12.06%) and NaCl (14.19%) samples were reduced significantly after just for 1 day and after 30 days were reduced to 55.59% and 61.18%, respectively. The Young's modulus of NaCl samples showed significant reduction (8.35%) after 1 day, but not for Milli-Q water samples. From 7 to 30 days, significant reductions for both samples were detected. SEM photomicrographs of samples in saliva showed widening of the inter-prismal space, while in PBS samples the structural damage was less. In the Milli-Q water group, prominent structural damage was observed and in case of NaCl, the enamel prisms were eroded. Conclusions Human saliva and PBS are better for short term preservation of enamel samples compared to NaCl and Deionized water.

The purpose of this study was to evaluate the inhibition of biofilm formation on newly developed coating materials with self-cleaning properties. A series of experimental coating materials containing fluoroalkylated acrylic acid oligomer (FAAO) were applied to resin composite substrates. The surfaces of the coating materials were analyzed by X-ray photoelectron spectroscopy (XPS) and contact angle measurement. Biofilm formation on the surface was assessed using Streptococcus mutans biofilms inside an oral simulator in vitro. The results indicated that an increase in the concentration of FAAO in the coating materials enhanced surface hydrophilicity and oil-repellency. Biofilm assays demonstrated that the amount of biofilm retained on the coating materials gradually decreased when the concentration of FAAO increased in the materials. It was concluded that the coating materials incorporated with FAAO possessed self-cleaning properties and displayed signs of inhibiting biofilm formation on their surfaces.

The purpose of this study was to evaluate the adherence of biofilms to the surfaces of two indirect resin composites, Estenia C&B and Gradia. Slabs were prepared from the materials, and then either ground with 800-grit silicon carbide paper or polished with diamond pastes up to 1 microm. Artificial biofilms of Streptococcus mutans were grown on the composite slabs in an artificial mouth system for 20 hours. Thereafter, the amounts of retained biofilm on the surfaces were measured after sonication. Surface characteristics of the resins--such as surface roughness, amount of residual monomers, and distribution of filler particles--were examined. Two-way ANOVA revealed that the amount of retained biofilm varied (p

Sealing of exposed root dentinal surfaces with adhesive materials is expected to be a promising approach for preventing root dentin caries. The aim of this study was to evaluate the effects of surface coating with all-in-one adhesives on inhibiting Streptococcus mutans biofilm attachment.Bovine root dentin was cut and ground with #600-grit SiC paper. Each of the three all-in-one adhesives, Hybrid Bond (HB), Reactmer Bond (RB) or Shake One (SO) was single-coated on the dentin surfaces according to the manufacturers' instructions. The dentin surface without coating served as the control. The surface roughness of the coated and non-coated dentin surfaces was recorded by means of laser microscope measurements. S. mutans artificial biofilms were then grown on the surface of each specimen in a microcosm for 20h. The amounts of bacteria and water insoluble glucan in the retained biofilm on the surface of the specimens were measured. All numerical data were statistically analyzed using one-way ANOVATukey's HSD (p0.05).All of the coated groups showed significantly lower susceptibility to biofilm attachment compared with the non-coated root dentin (p0.05). Among the coated groups, SO showed the lowest susceptibility for biofilm formation.Three all-in-one adhesive materials could be effective for root surface coating. A fluoride-releasing adhesive, SO demonstrated the strongest potentiality in resisting biofilm formation.

Objectives: Dental caries prevention programs using chlorhexidine (CHX) have been proposed, but CHX's effect in reducing levels of mutans streptococci (S. mutans and S. sobrinus) appears to last for only a few months. The aim of this study was to attempt to eradicate mutans streptococci from the oral cavity using intensive professional mechanical tooth cleaning (PMTC) and topical application of CHX in custom-made trays. Methods: Seven adult dentate subjects participated in this study (mean age 53.7 +/- 5.6, age range 46 to 62, mean DMFT, 9.1 +/- 4.2). For each subject, PMTC was carried out eight times within ten days. After each PMTC, 1% CHX was applied twice to the tooth surface using custom-made trays. In addition, as home treatment, subjects were required to carry out tooth brushing three times a day, and apply 0.2% CHX in custom trays after brushing in the morning and evening. In addition, subjects rinsed with 0.2% CHX solution after lunch. Salivary levels of mutans streptococci were evaluated using Dentocult- SM at baseline and on days 9, 20, 70, 120. Results: Mutans streptococci were eradicated by day 120 from 4 of the 7 seven subjects participating in this study. Those 3 subjects still harboring mutans streptococci exhibited deep periodontal pocketing. Conclusions: Eradication of mutans streptococci from the oral cavity is feasible using a combination of CHX application in custom-made trays and intensive PMTC. (J. Oral Sci. 46, 179-183, 2004)

An oral biofilm is a community of surface-attached microorganisms that coats the oral cavity, including the teeth, and provides a protective reservoir for oral microbial pathogens, which are the primary cause of persistent and chronic infectious diseases in patients with dry mouth or Sjögren's syndrome (SS). The purpose of this study was to establish an animal model for studying the initial adhesion of oral streptococci that cause biofilm formation in patients with dry mouth and SS in an attempt to decrease the influence of cariogenic organisms and their substrates. In nonobese diabetogenic (NOD) mice that spontaneously develop insulin-dependent diabetes mellitus (IDDM) and SS, we replaced major histocompatibility complex (MHC) class II (Ag7Eg7) and class IDbwith MHC class II (AdEd) and class IDdfrom nondiabetic B10.D2 mice to produce an animal model that inhibited IDDM without affecting SS. The adhesion of oral streptococci, includingStreptococcus mutans, onto tooth surfaces was then investigated and quantified in homologous recombinant N5 (NOD.B10.D2) and N9 (NOD.B10.D2) mice. We found that a higher number of oral streptococci adhered to the tooth surfaces of N5 (NOD.B10.D2) and N9 (NOD.B10.D2) mice than to those of the control C57BL/6 and B10.D2 mice. On the basis of our observation, we concluded that these mouse models might be useful as animal models of dry mouth and SS for in vivo biological studies of oral biofilm formation on the tooth surfaces.

Oral-genital contact is one of the risk factors for the transmission of human immunodeficiency virus (HIV) in adults. In recent reports, oral exposure to simian immunodeficiency virus (SIV) was found to have important implications for the achievement of mucosal transmission of HIV in a rhesus monkey animal model. In the present study, we aimed first to establish a small animal model which did not develop tonsils suitable for HIV oral mucosa transmission, using non-obese diabetic/severe combined immunodeficiency (NOD/SCID) mice and NOD/SCID B2m(null) mice grafted with human peripheral blood leucocytes (hu-PBL) and stimulated with interleukin (IL)-4, and second to investigate whether oral exposure to cell-free R5 and X4 HIV-1 could lead to oral transmission of HIV through intact or traumatized mucosal tissues in humanized mice. Both low and high concentrations of cell-free R5 and X4 viruses failed to cause oral transmission with or without trauma in hu-PBL-NOD/SCID and NOD/SCID Beta2m(null) mice, which presented a number of CD4+ cells in gingival tissues and oral cavities with or without tissue injury. The present results show that IL-4-administrated NOD/SCID B2m(null) mice are useful as a small-humanized model for the study of HIV oral infection, which may help to define the window of opportunity for oral transmission by the HIV virus in animal model experiments.

The chemokine stromal-cell derived factor-1 (SDF-1) controls maturation, trafficking, and homing of certain subsets, lymphoid cells including immunogenic B and T cells, as a ligand of the CXCR4 chemokine receptor. Insulin-dependent diabetes mellitus (IDDM) and Sjögren's syndrome (SS), both highly regulated autoimmune diseases, develop spontaneously in non-obese diabetic (NOD) mice. To investigate the role of SDF-1 in the development of autoimmune diseases, we injected groups of NOD female mice with antibodies to SDF-1 (anti-SDF-1), which resulted in a 30% reduction of diabetes up to 30 weeks of age, delayed average diabetes onset by 10 weeks, and suppressed insulitis. Autoimmune sialoadenitis was evident in anti-SDF-1-injected mice (SDF-1-Ig group) at the same level as in all groups of mice, whether injected with non-specific antibodies or not. In addition, in the SDF-1-Ig group, a greater number of immunoglobulin M (IgM)- IgD- B220(low) CD38+ CD43+ CD23- progenitor B cells and IgM+ IgD+ B220(high) CD43- CD38+ CD24+ CD23+ mature B cells remained in the bone marrow, whereas infiltration of mature IgM+ B cells was less extensive in peripheral tissues. Our results suggested that anti-SDF-1 antibodies injection was effective in inhibiting diabetes and insulitis without affecting autoimmune sialoadenitis or SS in NOD mice. SDF-1 may be an essential chemokine for trafficking and migration of autoreactive B cells in the development of diabetes.