Yusuke Takahashi

Osaka City University, Ōsaka, Ōsaka, Japan

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Publications (13)48.31 Total impact

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    ABSTRACT: The protective effects of N-acetyl cysteine (NAC) against cytotoxicity induced by conventional dental resin monomers have been widely documented. However, its effectiveness to detoxify cationic antibacterial monomers has not yet been elucidated. The aim of the present study was to investigate the possible protective effects of NAC against the cytotoxicity of 12-methacryloyloxydodecylpyridiniumbromide (MDPB) and explore the role of adduct formation in NAC-directed detoxification. The influences of NAC on the cytotoxicity of MDPB were studied in mouse osteoblast-like MC3T3-E1 cells using the MTT assay. Ultra-performance liquid chromatography (UPLC) and liquid chromatography-mass spectrometry (LC-MS) analysis were performed to investigate the possible chemical reaction between NAC and MDPB. While only slight reduction in the cytotoxicity of MDPB by NAC was observed immediately after mixing with MDPB, remarkable protection against MDPB-induced cell death was detected when the mixture was tested after 24h of pre-incubation. UPLC and LC-MS analysis revealed that chemical binding of MDPB and NAC occurred under neutral conditions after 24h of pre-incubation. Our findings suggest that NAC reduces the toxicity of the cationic antibacterial monomer MDPB, and adduct formation is partially responsible for the detoxification ability of NAC against MDPB-induced cell damage.
    Dental materials: official publication of the Academy of Dental Materials 10/2013; · 2.88 Impact Factor
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    ABSTRACT: Tertiary dentin is deposited inside teeth after various stimuli and serves as a major defensive wall to preserve pulp cells. However, the molecular mechanisms of the activation of quiescent odontoblasts, immature pulp cells and tertiary dentin formation are still unclear. Therefore, we performed a comprehensive gene expression analysis of pulp cells after cavity preparation of 9-week-old rat molars to clarify the critical molecules in tertiary dentinogenesis. As a result, mRNA expression of various molecules was up- or down-regulated. Notably, several members of the matrix metalloprotease family and their endogenous inhibitors were up-regulated after cavity preparation. In situ hybridization showed that tissue inhibitor of metalloprotease 1 (Timp1) was widely and continuously distributed in the pulp beneath the cavity in vivo. We also observed accumulation of β-catenin in the pulp cells beneath the cavity by fluorescence immunohistochemistry. Furthermore, Timp1 transcription was repressed by a dominant-negative TCF4 in immature undifferentiated mesenchymal cells, but not altered in mature odontoblast-like cells. These results indicate that cavity preparation may activate the Wnt/β-catenin pathway, and the Wnt/β-catenin pathway and Timp1 may be correlatively involved in pulp repair. Timp1 might play crucial roles in reactivation of immature pulp cells for tertiary dentinogenesis.
    Journal of Biochemistry 10/2012; · 3.07 Impact Factor
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    ABSTRACT: The antibacterial monomer 12-methacryloyloxydodecylpyridinium bromide (MDPB) is a strong bactericide when unpolymerized and has the potential to be utilized in various resinous biomaterials. To analyze the antibacterial characteristics of this monomer in detail, the ability of high concentrations of unpolymerized MDPB to kill Streptococcus mutans in planktonic or biofilm forms within a short time-period of contact, and the inhibitory effects of low concentrations of MDPB on the metabolic function of S. mutans, were examined. High concentrations of MDPB showed effective killing of planktonic and biofilm S. mutans cells within 60 s, and complete killing was obtained by contact with 1,000 μg ml(-1) of MDPB for 60 s. At a concentration of 4-8 μg ml(-1) , MDPB demonstrated growth inhibition, inducing elongation of the lag phase and of the doubling time, when the bacterial number was low. Inhibition of the production of acid from S. mutans by 8 μg ml(-1) of MDPB may have been caused by the inhibition of lactate dehydrogenase activity. At high concentrations, MDPB is lethal to both planktonic and biofilm forms of S. mutans in a short time-period, and at low concentrations, MDPB inhibits metabolic enzymatic activity.
    European Journal Of Oral Sciences 04/2011; 119(2):175-81. · 1.42 Impact Factor
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    ABSTRACT: Polyphasic calcium phosphates (Poly-CaP), a complex of hydroxyapatite (HAp) and soluble calcium phosphates including alpha-tricalcium phosphate and tetracalcium phosphate, demonstrate promoting effects on hard tissue formation by osteoblasts. We hypothesized that a Poly-CaP block with a soluble calcium phosphates phase on one side and an insoluble HAp phase on the other side is useful for vital pulp therapy as it may promote dentin regeneration and provide the surface effective to achieve sealing. The purpose of this study was to investigate the efficacy of Poly-CaP as a direct pulp capping material by examining the Ca-release profile, the in vivo ability to induce reparative dentinogenesis, and the bonding of HAp surface with adhesive systems. Poly-CaP prepared by annealing crude HAp disc was immersed in buffer solution at pH 7.4 or 4.0, and the concentration of Ca released was measured until 15 days. The pulp of 9-week-old Wister rat molar was exposed and capped with Poly-CaP or HAp block, and dentin bridge formation and pulpal inflammation was evaluated histopathologically after 2 or 4 weeks. Etch & rinse or self-etching adhesive was bonded to HAp surface, and the interface was observed using SEM. Poly-CaP exhibited continuous release of Ca with significantly greater amount than HAp at both pH conditions (P<0.05, Student's t-test). Animal tests demonstrated formation of complete dentin bridge at higher rate for Poly-CaP compared with HAp after 4 weeks (P<0.05, Steel-Dwass test). Impregnation of resin into etched HAp surface, with production of intimate contact at the bonding interface, was seen for all adhesives. Poly-CaP is a potentially useful material for direct pulp capping with the advantages to promote dentin bridge formation and to provide tight sealing by adhesives.
    Journal of dentistry 10/2010; 38(10):828-37. · 3.20 Impact Factor
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    ABSTRACT: Dental tissue disease and trauma provides an excellent model for the interaction between tissue defence and regenerative processes and has application to many of the body's other tissues. Following dental tissue infection, characterised by caries, the molecular and cellular mediators of the immune/inflammatory processes clearly impact on the dental tissues' natural regenerative responses. This review of the literature was performed to better understand how these two processes interact and identify whether cross-talk may provide novel areas for future research and subsequent translation into clinical application. A review of the literature was performed using the PubMed database resource and this was followed by extensive hand searching using reference lists from relevant articles. Frequently, the dental tissue inflammatory and regenerative processes are seen as both distinct and antagonistic and subsequently have often been studied in isolation; however, both direct and indirect data are now emerging which indicate significant inter-relationship. Whilst the ensuing inflammatory process will result in dental tissue breakdown and molecular signalling which may impede regeneration, low grade inflammation, potentially induced by mechanical trauma and tissue necrosis, may promote regenerative mechanisms, including angiogenic and stem cell processes. Notably, the locally derived growth factors, neuropeptides, cytokines and chemokines, released from the host dentine matrix and by resident pulpal cells, immune cells, neurons and/or dying cells, will modulate defence and repair processes within the tissue.
    Journal of dentistry 09/2010; 38(9):687-97. · 3.20 Impact Factor
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    ABSTRACT: A dentin primer incorporating an antibacterial monomer 12-methacryloyloxydodecylpyridinium bromide (MDPB) shows strong antibacterial effects, and may provide better prognosis for direct capping of infected pulp exposed by caries removal compared with conventional adhesives. However, influences of MDPB on healing of the pulp have not yet been fully elucidated. The purpose of this study was to compare the influences of unpolymerized MDPB on proliferation, differentiation and mineralization of odontoblast-like MDPC-23 cells with those of other resin monomers, Bis-GMA, MDP, TEGDMA and HEMA. The inhibitory effects of MDPB on the proliferation of MDPC-23 were lower than those of Bis-GMA. While MDPB strongly affected the differentiation compared with the other monomers, it was less inhibitory than Bis-GMA and MDP on the mineralization ability of odontoblast-like cells. These findings indicate that MDPB has superior biocompatibility than Bis-GMA in terms of hard tissue formation by odontoblastic cells, suggesting its possible less negative influences on dentinogenesis.
    Biomaterials 12/2009; 31(7):1518-32. · 8.31 Impact Factor
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    ABSTRACT: Glass-ionomer cements (GICs) are known to have inhibitory effects on bacterial growth, but the biochemical mechanism of this property has not been fully understood. This study aimed to evaluate inhibitory effects of GIC on the acid production of caries-related oral streptococci, and to identify the components responsible for the inhibition. An eluate was prepared by immersing set GIC in phosphate-buffered saline at 37 degrees C for 24h. Fluoride and other elements in the eluate were quantified by fluoride ion electrode and atomic absorption photometry, respectively. Streptococcus mutans NCTC 10449 and Streptococcus sanguinis NCTC 10556 were used to evaluate the pH fall and the rate of acid production after the addition of glucose in the presence or absence of the eluate. Acidic end products from glucose were also assayed by carboxylic acid analyzer. The eluate contained silicon (1.24+/-0.26 mM), fluoride (0.49+/-0.02 mM) and aluminum (0.06+/-0.00 mM), and inhibited the pH fall and the acid production rate of both streptococci at acidic pH, with a concomitant decrease in lactic acid production. These effects were comparable to those of a potassium fluoride solution containing the same concentration of fluoride as the eluate. These results indicate that the GIC eluate inhibits the acid production of caries-related oral streptococci at acidic pH and that the effect is due to fluoride derived from the GIC. Thus, adjacent to GIC fillings, bacterial acid production and the subsequent bacterial growth may decrease, establishing a cariostatic environment.
    Dental materials: official publication of the Academy of Dental Materials 02/2009; 25(6):703-8. · 2.88 Impact Factor
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    ABSTRACT: This study examined the in vivo bonding ability to sound dentin of antibacterial adhesive systems incorporating an antibacterial monomer MDPB based on morphological evaluation of the resin-dentin interface. Class V cavities were prepared on the buccal surfaces of the teeth of a beagle dog and a composite filling performed using (1) commercial self-etching system Liner Bond 2 (LB primer+LB bond), (2) experimental primer containing 5% MDPB and LB bond, (3) LB primer and experimental bonding-resin containing 2.5% MDPB, or (4) combination of experimental primer and bonding-resin. After 7 days, the tooth crown was cut and fixed in half-Karnovsky's solution, and the sectioned surface observed under scanning electron microscopy (SEM) after treatment with phosphoric acid and NaOCl. The ultrastructure of the bonding interface was also examined by transmission electron microscopy (TEM). Microtensile bond strengths (microTBS) of each group were measured using extracted teeth. SEM demonstrated that all groups produced a 1-2microm thick hybrid layer with funnel shaped resin tags, although the length of tags was shorter for the group in which MDPB-containing bonding-resin was used. TEM examination supported good adhesion of the comprehensive adhesive system employing MDPB-containing primer/bonding-resin, showing integrity between resin and dentin. There were no significant differences in microTBS among the four groups tested (p>0.05, ANOVA). This study confirmed that the experimental antibacterial adhesive systems employing MDPB-containing primer or/and bonding-resin could produce an effective bond under in vivo conditions.
    Dental Materials 02/2007; 23(2):170-6. · 3.77 Impact Factor
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    ABSTRACT: Since atraumatic restorative treatment (ART) involves removal of carious lesions with manual instruments, improvement of filling materials to guarantee greater success should be considered. This study aimed to evaluate antibacterial, physical, and bonding properties of glass-ionomer cements (GIC) containing chlorhexidine (CHX), and to determine optimal concentrations for incorporation of agents to obtain antibacterial GICs for use with the ART approach. CHX diacetate combined with CHX dihydrochloride was added to control GIC powder to obtain concentration ratios of 1/0, 2/0, 3/0, 1/1, or 2/2% w/w. Antibacterial activity of each cement against Streptococcus mutans, Lactobacillus casei or Actinomyces naeslundii was examined using agar-diffusion methods, and release of CHX was analyzed by HPLC. Compressive strength, bond strength to dentin, and setting time were measured, and compared with those of control samples. All experimental GICs exhibited inhibition of three bacteria, but sizes of inhibition zones and concentrations of CHX released were not dependent upon CHX content. Incorporation of CHX diacetate at 2% or greater, significantly decreased compressive strength, and bond strength to dentin was adversely affected by addition of CHX diacetate at 2% or more (p<0.05, ANOVA, Fisher's PLSD test), although setting time was extended a little by addition of any concentrations of CHX. The present results demonstrate that experimental GICs containing CHX are effective in inhibiting bacteria associated with caries, and incorporation of 1% CHX diacetate is optimal to give appropriate physical and bonding properties.
    Dental Materials 08/2006; 22(7):647-52. · 3.77 Impact Factor
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    ABSTRACT: This study aimed to investigate the antibacterial effects of the dentin primer of a commercially available self-etching adhesive system, Clearfil Protect Bond, which contains antibacterial monomer 12-methacryloyloxydodecylpyridinium bromide (MDPB). Inhibitory effects against Streptococcus mutans, Lactobacillus casei, or Actinomyces naeslundii were examined by an agar-disc diffusion method using the Clearfil Protect Bond primer containing 5% MDPB and an acidic adhesion-promoting monomer MDP, the primer only with MDP, and the primer with 1% cetylpyridinium chloride. The minimum inhibitory/bactericidal concentrations (MIC/MBC) of each primer for the three bacterial species were determined by serial microdilution assays. For testing the bactericidal effects seen in dentin, the primer was applied to demineralized dentin blocks in which S. mutans had been impregnated, and numbers of viable bacteria were counted. For all three bacteria, the sizes of the inhibition zones produced by Clearfil Protect Bond primer were significantly greater than for the other primers (p<0.05, ANOVA and Scheffe's F-test). The MIC/MBC values of Clearfil Protect Bond primer were less than those of the primer without MDPB, and comparable to those of the primer containing cetylpyridinium chloride. No bacterial recovery was obtained after application of Clearfil Protect Bond primer to the bacteria-impregnated dentin, although the primer without MDPB showed some bactericidal effect. Clearfil Protect Bond primer has strong antibacterial activity based upon MDPB against S. mutans, L. casei and A. naeslundii, and the capability to disinfect cavities containing residual bacteria.
    Dental Materials 07/2006; 22(6):527-32. · 3.77 Impact Factor
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    ABSTRACT: This study examined the hypothesis that experimental primer containing the antibacterial monomer 12-methacryloyloxydodecylpyridinium bromide (MDPB), which was previously reported to show bactericidal effects in vitro, inhibits bacteria in cavities under in vivo conditions. The number of bacteria resulting from applying primer solution to cavities in dog teeth infected with Streptococcus mutans was determined. The infected cavities were also restored using primer and the pulp response was histopathologically examined after 7, 30 and 75 days. No bacteria were recovered after applying the experimental primer, although the bactericidal effects of the proprietary primer were insignificant. Restoration with the experimental primer resulted in little or no pulpal inflammation for all periods; whereas, mild to moderate inflammatory response was observed when using proprietary primer. These results indicate that the experimental primer containing MDPB could exhibit in vivo antibacterial effects, suggesting its possible clinical benefit.
    Operative Dentistry 01/2004; 29(4):369-75. · 1.31 Impact Factor
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    ABSTRACT: Porphyromonas gingivalis is found in subgingival biofilm and is associated with periodontal disease. Bacteria in biofilms are able to resist higher antimicrobial concentrations than in suspension. Little is known about the susceptibility of P. gingivalis in biofilms to antimicrobial agents. The effects of chlorhexidine gluconate, minocycline hydrochloride, and metronidazole on P. gingivalis biofilms were examined in vitro. P. gingivalis strain 381 biofilms were prepared on 32 hydroxyapatite disks. At 0, 24, 72, and 144 hours after perfusion of the three antimicrobial agents, two disks from each device were used to assess the antimicrobial effects by adenosine triphosphate (ATP) bioluminescence, and for morphological investigation by scanning electron microscopy (SEM). Close relationships were found between the results of the ATP analyses and the SEM observations in all groups examined. A significant decrease (P < 0.001) in ATP content was found between the chlorhexidine-treated and control groups. The extracellular matrix structure and P. gingivalis cells were altered in the presence of chlorhexidine. Minocycline hydrochloride also caused a decrease (P < 0.05) in the ATP content and morphological change on P. gingivalis biofilms. Metronidazole showed no significant efficacy against P. gingivalis biofilms. Chlorhexidine gluconate was effective at reducing the viability of P. gingivalis 381 cells in biofilms, while minocycline hydrochloride and metronidazole exhibited weaker effects.
    Journal of Periodontology 11/2003; 74(11):1647-51. · 2.40 Impact Factor
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    ABSTRACT: This study examined the antibacterial activity of prepolymerized resin filler, in which the bactericide quaternary ammonium was immobilized. The experimental filler was prepared by grinding prepolymerized resin blocks of methacrylate monomers, silica particles, and the antibacterial monomer methacryloyloxydodecylpyridinium bromide (MDPB). The number of Streptococcus mutans after incubation for 18h in contact with the experimental filler with or without protein adsorption by saliva treatment was determined, and adherence of bacteria to the filler surface was evaluated by scanning electron microscopy. Elution of unpolymerized MDPB from the filler and its influence on bacterial growth were also investigated. The growth of S. mutans was completely inhibited by contact with the experimental filler without saliva treatment. Although the effects were attenuated, the saliva-treated filler still exhibited growth inhibition at >99.9%. Less bacteria attached to the experimental filler than the control filler without MDPB, indicating that the reduction in bacterial number after contact with the experimental filler was not due to bacterial adherence to the particles. Unpolymerized MDPB at 1 microg/ml was eluted from the filler particles but was confirmed to have little effect on bacterial growth. The results indicate that the bactericide-immobilized filler containing MDPB shows significant bacteriostatic effects without releasing antibacterial components, and is useful for incorporation into various resin-based restoratives.
    Biomaterials 09/2003; 24(20):3605-9. · 8.31 Impact Factor