Objective: The banding pattern of Streptococcus mutans strains generated using repetitive extragenic palindromic PCR (rep-PCR) with enterobacterial repetitive intergenic consensus (ERIC) primers has been shown previously. In the present study, the sequencing and analysis of gene sequences of the 6 most intense and commonly occurring amplicons from S. mounts serotype c was investigated.Methods: The sequences of the amplicons from S. mutans serotype c strain by using rep-PCR with ERIC1R and ERIC2 primers were compared to the whole genome sequence of S. mutans UA159 and NN2025 strains.Results: The amplicons were found to contain partial gene sequences coding for proteins such as the hypothetical protein, glucan binding protein A, putative methylated-DNA-protein-cysLeine 5-methylLransferase, putative D-3-phosphoglyceraLe dehydrogenase, putative excinuclease ABC (subunit A) and putative GTE-binding protein. The locations of 5 of the 6 amplicons were found to be assembled downstream in the UA159 genome. The coding direction of the amplicons in the NN2025 genome was in reverse orientation relative to that in the UA159 strain, except in the 2170-bp amplicon.Conclusions: The repeating sequence elements of ERIC in S. mutans serotype c are located on one side of the genome and have less frequency and similarity compared to those in enterobacteria. (C) 2013 Japanese Association for Oral Biology. Published by Elsevier B.V. All rights reserved.

We designed three experimental 10-methacryloyloxydecyl dihydrogen phosphate (MDP)-based one-step (EX) adhesives consisting of MDP, urethane dimethacrylate, and triethylene glycol dimethacrylate adhesives with different water contents (98.4, 196.8, and 294.4 mg/g), and 4-methacryloyloxyethyl trimellitic anhydride (4-META) or 2-hydroxyethyl methacrylate (HEMA)-containing onestep adhesive. The effect of the amount of MDP-calcium (MDP-Ca) salt produced through demineralization of enamel and dentin on the bonding performance was examined. The efficacy of 4-META and HEMA was then discussed. When the amount of water in EX adhesive was increased, the production amount of MDP-Ca salt of enamel increased, but not the dentin. The enamel bond strength slightly increased with increasing the production amount of MDP-Ca salt, in contrast to the dentin. However, addition of 4-META in the EX adhesive (water content=98.4 mg/g) increased both bond strengths, although the production amounts of MDP-Ca salt significantly decreased. The 4-META enhances both bond strengths more effectively than the HEMA.

Background Streptococcus mutans and Streptococcus sobrinus are considered to be important bacterial species in the initiation of human dental caries. Therefore, the establishment of a reliable genotyping method to distinguish S. mutans from S. sobrinus is of central importance. Objective We assessed the usefulness of repetitive extragenic palindromic polymerase chain reaction (rep-PCR) using ERIC primer banding patterns in differentiating S. mutans and S. sobrinus. Design Five S. mutans and two S. sobrinus prototype strains and 50 clinical isolates (38 S. mutans serotype c, 4 S. sobrinus serotype d, and 8 S. sobrinus serotype g) were examined. The banding patterns of amplicons generated were compared among the prototype strains and clinical isolates, to find common bands that distinguish S. mutans and S. sobrinus. Results Multiple banding patterns were seen with all strains tested. The representative strains of S. mutans tested revealed six unique, strong bands at 2,000 bp, 1,700 bp, 1,400 bp, 1,100 bp, 850 bp, and 250 bp, whereas S. sobrinus had seven strong bands at 2,000 bp, 1,800 bp, 1,100 bp, 900 bp, 800 bp, 600 bp, and 550 bp. The band at 1,100 bp was the only band that was observed in both S. mutans and S. sobrinus. Furthermore, most clinical S. mutans isolates revealed identical banding patterns. All S. mutans had amplicons at 1,700 bp, 850 bp, and 250 bp, whereas those of S. sobrinus were at 1,100 bp, 900 bp, and 800 bp. Conclusions These results indicate that using rep-PCR with the ERIC primers can distinguish between S. mutans and S. sobrinus.

OBJECTIVES:The effect of visible light passing through a resin composite with different particle sizes of spherical silica filler on the curing depth and Knoop hardness of the resin composite was examined. The null hypotheses tested were that the particle size of silica filler has no effect on (1) the transmitted amount of visible light passing through the resin composite, (2) the curing depth of the resin composite, and (3) the ratio of the Knoop hardness of the hardened resin composite. METHODS:A series of different experimental resin composite pastes with different particle sizes of silica filler were prepared. The amount of visible light passing through each of the experimental resin pastes during the hardening process was determined. The curing depth and Knoop hardness of the resin composites were measured. Further, to characterize the polymerization conversion, the ratio of the Knoop hardness of the bottom surface against the irradiated surface of the hardened resin composites was determined. RESULTS:Increases in the particle size of the silica filler resulted in decreases in the transmitted amount of visible light, curing depth, and ratio of the Knoop hardness of the resin composite. The rate of polymerization conversion of the resin composite occurring deep within the cavity was found to be less than the rate of polymerization conversion occurring at the upper surface. SIGNIFICANCE:To increase the rate of polymerization conversion deep within the resin composite, it is important to reduce the particle size of the silica filler.

We determined the number of reacted and unreacted 10-methacryloyloxydecyl dihydrogen phosphate (10-MDP) molecules with calcium during the demineralization process of hydroxyapatite or dentin by 10-MDP-based one-step (Clearfil Tri-S Bond, TS) or two-step self-etch adhesive (Clearfil SE Bond Primer, SE). We then examined the effects of the number of reacted and/or unreacted 10-MDP molecules on the initial bond strength and bond durability of the resultant adhesive layer. The null hypotheses were that (1) the etching efficacy of tooth apatite by 10-MDP used in TS was the same as that in SE, and (2) the unreacted 10-MDP polymer included within the adhesive layer does not affect bond durability. Addition of hydroxyapatite or dentin to the TS and SE resulted in decreases in the NMR peak intensities for 10-MDP. The peak intensity for 10-MDP showed a greater reduction in SE than in TS, consistent with the observation that SE provided significantly higher initial mean bond strengths than TS. Further, the unreacted 10-MDP polymer within the adhesive layer did not decrease the mean bond strength, despite the application of 20,000x thermo-cycling.

To compare wear performance and resistance to crack propagation (K1C) of commercial restorative materials and their flowable variations. A potential correlation between three-body wear and fracture toughness, modulus of elasticity, fracture work, Vickers hardness, and filler content was investigated.Seven restoratives (five composites, one ormocer, and one compomer) and their corresponding flowable materials were used to determine and compare the three-body wear with a bolus of millet-seed shells and rice food (Willytec). The wear characteristics were measured by profilometry after 50,000, 100,000, 150,000, and 200,000 loading cycles. The fracture toughness value, K1C (MPam1/2), for each single-edged notched specimen was measured in a three-point bending test (universal testing machine 1446, Zwick). Fracture work and modulus of elasticity were calculated from the load curves. Vickers hardness was measured (HV hardness tester, Zwick) according to DIN 50133. The veneering composite Sinfony (3M ESPE) was used as a reference material.Heavily filled composites experienced less wear than their flowable variations. The nanofiller composites revealed better wear results than hybrid composites, compomers, and ormocers. After 200,000 load cycles, the lowest wear rates were detected for Grandio (14 microm; Voco), and the highest mean values were found for Dyract AP (104 microm; Dentsply DeTrey). The values for fracture toughness (K1C) ranged from 0.82 to 3.64 MPam1/2. Highest K1C data was exhibited by the nanocomposite Nanopaq (Schutz Dental). All tested restorative materials exhibited higher fracture toughness than their low-viscosity variations.The wear resistance of the newer generation composites with incorporated nanofiller or microfiller particles increased to a high extent. Flowables show less resistance against wear and crack propagation because of their lower filler content. The reduced mechanical properties limit their use as a restorative to small noncontact, low-stress clinical situations.

OBJECTIVE:Hydroxyapatite (HA) is used as a construction material for artificial supplementation of enamel tooth surfaces to improve oral hygiene. This study examined in vitro HA interactions with mutans streptococci (MS) and bacterial adherence to small (nanosize) crystal form of HA beads having a protean hexagonal structure. The adsorption and physical effects of HA employed in vivo is also described.METHOD AND MATERIALS:[3H-thymidine]-labeled streptococci were incubated with HA noncoated or coated with salivary components or salivary agglutinin peptide (SRCRP2), a receptor for streptococcal surface proteins. Bacterial adhesion activities on HA were measured by uptake of [3H-thymidine]. Application of HA paste in an individual tray was tried on the tooth surface, and its effects on the colony ratio of MS/total streptococci (TS) in saliva were analyzed by culture technique.RESULTS:The adhesion assay showed that the binding of streptococci to HA was inhibited by coating with salivary components, whereas coating with SRCRP2 had nearly no influence on binding with or without Ca+. Further, treatment with HA decreased the adherence of Streptococci mutans to roughened enamel surfaces by one-third. In vivo application of a HA dentifrice to individual teeth demonstrated that the colony number ratio of MS/TS slowly decreased.CONCLUSION:MS adhesion to HA was restricted by both salivary components, except for SRCRP2, and the physical effects of HA; in addition, the material itself has a unique effect for removing MS from the oral cavity.

We investigated the effects of lasers irradiation on the exposed dentinal tubule. Human tooth specimens with exposed dentinal tubule orifices were used. Three types of lasers (CO(2) laser, Er:YAG laser and Ga-Al-As laser) were employed. The parameters were 1.0 W in continuous-wave mode with an irradiation time of 30 s for the CO(2) laser, 30 mJ in continuous-wave mode with an irradiation time of 60 s for the Er:YAG laser, and 1.0 W in continuous-wave mode with an irradiation time of 60 s for the Ga-Al-As laser. A non-irradiated group was used as a control. After laser irradiation, the dentinal surface of each sample was observed using SEM. Afterwards, all samples were immersed in methylene blue dye solution in order to evaluate the penetration of the dye solution and observe the change in dentinal permeability after laser irradiation. SEM observation showed that the control group had numerous exposed dentinal tubule orifices, whereas these orifices were closed in the laser-irradiated groups. There was consistent dye penetration into the pulp chamber in the control group, whereas no dye penetration was evident in the laser-irradiated groups. Therefore, laser appears to be a promising treatment for reducing permeation through exposed dentinal tubules.

This study sought to investigate the degradation mechanism of 4-methacryloyloxy ethyl trimellitic acid, 4-MET, which is commonly used as an acidic monomer in solvated self-etching primers or one-step bonding agents. To this end, we examined the effects of solvent type used—such as ethanol, methanol, and acetone—on the degradation mechanism of 4-MET by using the 13C NMR technique.The degradation mechanism of 4-MET was strongly dependent on the type of solvent used. When an alcohol-based solution was used for 4-MET, the esterification of 1- or 2-carboxylic acid in 4-MET occurred. However, when an acetone solvent was used for 4-MET, the esterification reaction did not occur. Increases in the aging period of 4-MET solvated solutions resulted in the hydrolysis of the benzoyl ester portion in 4-MET. The 2-hydroxyethyl methacrylate, produced as a subproduct, also became hydrolyzed. In addition, methacrylic acid, non-esterified and esterified trimellitic acid, as well as ethylene glycol were produced as subproducts. In particular, the production of trimellitic acid and ethylene glycol affected the bonding efficacy and durability of the resin to the tooth created by self-etching primers or one-step bonding agents that contained the altered 4-MET.

The aim of this study was to enhance the bond strength of one-step bonding agents to dentin. In particular, the focus was on using Catabrush the applicator system of AQ Bond Plus. Catabrush was supplemented with N-phenylglycine and aromatic sulfinate as polymerization accelerators, as N-phenylglycine was reportedly beneficial in improving the bond strength to dentin. The results indicated that the bond strength to dentin was significantly augmented and the photo-polymerization as well as the chemical polymerization were both improved even in the moistened dentin when 1.0 wt% N-phenylglycine was added to AQ Bond Plus agent, hence implying significantly higher bond strength to dentin. It was therefore concluded that N-phenylglycine is useful as a polymerization accelerator to be adopted in the applicator system for one-step bonding agents.

本研究では，レーザーの波長の違いが象牙質に与える影響について調べることを目的として，象牙質の赤外線の吸収波長特性から得られた吸収波長を選択し，照射エネルギー密度を一定にした各々の赤外自由電子レーザーを健全象牙質あるいは脱灰象牙質に照射して，その削除深さと各々の波長の違いが象牙質の構造変化に与える影響について調べた．その結果，以下の結論が得られた．健全象牙質では波長9.4μｍは6.4μｍの2倍以上の削除深さが得られたが，脱灰象牙質では9.4と6.4μｍの波長で比較的近似した削除深さが得られた．レーザー照射後の象牙質表面のSEM所見に関しては，波長6.4μｍでは一部において，脱灰象牙質の表層が除去され，トームス線維様の構造物が顕在化する像が示された．波長9.4μｍでは健全象牙質において脱灰象牙質よりも象牙質表面の凹凸が顕著に認められた．波長10.6μｍでは健全，脱灰象牙質の両者において照射による構造変化は認められなかった．

It is well understood that the application of a self-etching primer enhances the bonding at the resin-teeth interface. In this study, we designed a self-etching primer consisting of N-methacryloyl glycine (NMGly) and N-methacryloyl-2-aminoethyl phosphonic acid (NMEP). The demineralization effects on the hydroxyapatite or dentin by the carboxylic acid in the NMGly and by the phosphonic acid in the NMEP and their effects on the bond strength of the resin to the tooth were examined. The application of the NMGly–NMEP solution to the enamel resulted in an increase in the bond strength when additional amounts of NMEP were added to the NMGly aqueous solution. This increase was due to the phosphonic acid in the NMEP demineralizing the enamel. Conversely, the addition of the NMEP to the NMGly solution resulted in a decrease in the bond strength to the dentin. The optimal concentration of the NMEP in the NMGly–NMEP solution resulted in bond strengths of over 20MPa for both the enamel and dentin.

In this study, we examined the effect of the transmitted amount of visible light through a resin composite on the curing depth and polymerization conversion. Transmitted amount of visible light was strongly dependent on the magnitude of refractive index difference that existed between the resin and silica filler. More specifically, the differences arose from the type of base monomer used. The transmitted amount of visible light exhibited a good correlation with the curing depth and Knoop hardness ratio of the bottom surface against the top surface of the resin composite. To improve the polymerization conversion of the cavity floor, it is important to reduce the refractive index difference that exists between the base resin and silica filler.

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.

Porphyromonas gingivalis is a Gram-negative anaerobic bacterial species implicated as an important pathogen in the development of adult periodontitis. We previously cloned a gene encoding dipeptydilaminopeptidase IV (DAPIV) from P. gingivalis. In the present study, for immunological diagnosis and development of passive immunization, we produced a mouse monoclonal antibody (MAb) capable of inhibiting the DAPIV activity of P. gingivalis using highly purified recombinant DAPIV as an immunogen. The constructed MAb, designated as MAb-Pg-DAP-1, significantly inhibited DAPIV activity in P. gingivalis, as well as slightly inhibited that in other gram-negative bacteria such as Porphyromonas endodontalis and Prevotella loesheii, whereas no inhibition was seen in the gram-positive bacteria Streprococcus mutans and Actinomyces viscosus. Furthermore, the MAb did not inhibit DAPIV enzyme activity in human serum. This novel MAb may be useful for the development of immunological diagnosis capability and in passive immunization.