Bactericidal efficacy of metronidazole against bacteria of human carious dentin in vivo

The bactericidal efficacy of metronidazole (MN) against bacteria of carious dentin was estimated by measuring bacterial recovery with or without application of MN to dentinal lesions in vivo. More than 10(3) colony-forming units of bacteria per milligram of sample were recovered from the samples of carious dentin. However, none were recovered from the samples of dentinal lesions which had been covered by alpha-tricalcium phosphate (TCP) cement containing MN in situ for 1 day, 1 month, 1 year and/or 2 years in vivo. Placebo experiments in which carious dentin was covered by TCP cement without MN showed that TCP itself was not bactericidal. Bactericidal efficacy of MN in situ is clearly demonstrated, indicating that MN is a valuable material in caries treatment to disinfect carious dentin.     

Bactericidal efficacy of metronidazole against bacteria of human carious dentin in vitro

The bactericidal efficacy of metronidazole against bacteria in carious dentin was clarified by measuring (1) the difference between bacterial recovery from suspensions of carious dentin on metronidazole-containing BHI-Blood agar plates (10 micrograms/ml) and control plates and (2) the difference between bacterial recovery from carious lesions of freshly extracted teeth, covered by alpha-tricalcium phosphate (TCP) cement containing metronidazole (5%) for 1-3 days and that covered by TCP only. More than 10(3) bacteria per milligram sample were recovered from carious dentin. More than 99% of the bacteria were, however, not recovered when samples were inoculated on metronidazole-containing BHI-Blood agar plates or when the lesions were covered by TCP cement containing metronidazole, indicating that metronidazole effectively disinfected the carious dentin.     

In vivo antibacterial effects of dentin primer incorporating MDPB

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.     

Effects of the antibacterial monomer 12-methacryloyloxydodecylpyridinium bromide (MDPB) on bacterial viability and metabolism

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.     

Water sorption and colour stability of composites containing the antibacterial monomer MDPB

OBJECTIVES: Previously, we have reported that composites incorporating the newly developed antibacterial monomer methacryloyloxydodecylpyridinium bromide (MDPB) inhibited bacterial growth and plaque accumulation on their surfaces without releasing antibacterial components, and that the antibacterial effect was not lost even after ageing in a wet environment for a long period. The purpose of this study was to investigate the influence of the incorporation of MDPB on the water sorption characteristic and colour stability of Bis-GMA based composites. METHODS: Cured disc-shaped specimens of composites containing 0.4% or 0.5% MDPB were fabricated, and water uptake, the diffusion coefficient and the surface contact angle for water were determined. Discoloration of MDPB-containing composites was evaluated after immersion in water at 37 degrees C and 60 degrees C for 4 weeks and compared with that of the control composite without MDPB. RESULTS: There were no significant differences in water uptake, diffusion coefficient and contact angle between the control and MDPB-containing composites. Although composites incorporating MDPB showed greater discoloration than the control when maintained at 37 degrees C, no difference was found after 4 weeks of immersion in water at 60 degrees C. CONCLUSIONS: Incorporation of small amounts of the antibacterial monomer MDPB showed no influence on the water sorption characteristic and affected a little the colour stability of the Bis-GMA based composite, causing rapid discoloration.     

Antibacterial activity and bonding characteristics of an adhesive resin containing antibacterial monomer MDPB.

OBJECTIVES: The aims of this study were to investigate the antibacterial activity of an adhesive resin incorporating the antibacterial monomer 12-methacryloyloxydodecylpyridinium bromide (MDPB) as well as its bonding characteristics in terms of bond strength into dentin and curing ability. METHODS: An experimental adhesive resin was prepared by incorporation of 2.5% MDPB into proprietary adhesive (LB Bond), and the inhibitory effect of cured specimen against the growth of Streptococcus mutans on its surface was determined. Bond strength to human dentin and degree of conversion of the experimental adhesive in combination with 1, 2, or 5% MDPB-containing primers, which were previously reported to possess bactericidal effect, were evaluated by conventional tensile bond strength test and Fourier transformation infrared spectroscopy. RESULTS: The cured experimental adhesive exhibited an inhibitory effect on S. mutans growth, reducing the number of bacteria to approximately 3% of control adhesive without MDPB. Tensile bond strength of experimental adhesive was not significantly different from that of the control (p>0.05), and use in combination with MDPB-containing primer did not show any adverse influence on bond strength. The degree of conversion of the bonding system was not different (p>0.05) between control and experimental adhesive irrespective of addition of MDPB into primer. SIGNIFICANCE: These results indicate that an adhesive resin with antibacterial activity after curing could be produced by incorporation of MDPB without influencing bond strength or curing performance, suggesting that the comprehensive bonding system including MDPB-containing primer and adhesive should be highly effective in its antibacterial effect before and after curing.     

Mechanism of detoxification of the cationic antibacterial monomer 12-methacryloyloxydodecylpyridiniumbromide (MDPB) by N-acetyl cysteine

Objectives

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.

Methods

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.

Results

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 24 h of pre-incubation. UPLC and LC–MS analysis revealed that chemical binding of MDPB and NAC occurred under neutral conditions after 24 h of pre-incubation.

Significance

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.     

Micro-tensile bond strength of self-etching primer adhesive systems to human coronal carious dentin

The aim of this study was to evaluate the micro-tensile bond strengths of three self-etching primer adhesive systems to normal dentin (ND), caries-affected dentin (CAD) and caries-infected dentin (CID). Human extracted molars with caries were used, and flat dentin surfaces ground by 600-grit SiC paper were prepared. The surfaces were dyed using Caries-Detector solution, treated with Clearfil SE Bond, Mac-Bond II and UniFil Bond, and then covered with resin composites according to manufacturer's instructions. After immersion in 37 degrees C water for 24 h, the teeth were serially sectioned into multiple slices. Each slice was distinguished into ND, CAD and CID groups by the degree of staining, and the bond strength was measured in a universal testing machine. Scanning electron microscopic (SEM) observation was also performed. For statistical analysis, anova and Scheffe's test were used (P < 0.05). The bond strengths of the three adhesive systems to CAD and CID were significantly lower than those to ND. There was significant difference in the bond strength to ND between Clearfil SE Bond and UniFil Bond, but no significant differences to CAD and CID among the three adhesive systems. On SEM, the hybrid layers in CAD and CID showed more porous structures compared with ND. The results indicated that the bond strengths to CAD and CID were not affected by a variety of self-etching primer adhesive systems because of the porous hybrid layer formation in carious dentin.     

The effect of antibacterial monomer MDPB on the growth of organisms associated with root caries

MDPB, 12-methacryloyloxydodecylpyridinium bromide, was tested for its ability to inhibit the growth of organisms associated with active root caries lesions and to modify the growth characteristics of these organisms at sub-MICs. MICs and MBCs of MDPB for independent isolates (n=5) of the following taxa: Streptococcus mutans, Streptococcus oralis, Streptococcus salivarius, Actinomyces naeslundii, Actinomyces israelii, Actinomyces gerensceriae, Actinomyces odontolyticus, Lactobacillus spp., and Candida albicans were determined, and the effects at sub-MIC on microbial growth kinetics were assessed. All isolates were sensitive to inhibition by MDPB. The median MICs and MBCs of MDPB for these organisms were in the range of 3.13 to 25.0 microg/ml and 6.25 to 50.0 microg/ml, respectively. As for the influence of pH, inhibition was sensitive to acidic pH. Even at sub-MICs, the growth of all strains, measured as cell yield and doubling time, was significantly reduced. Based on the results of this study, MDPB exhibited the potential to inhibit the growth of microbiota associated with active root caries lesions.     

Identification of bacteria in association with immune components in human carious dentin

Actinomyces viscoxus, A. naselundii. Streptococcus mutans serotype "c" and S. mutans serotype "d/g" were identified in human carious dentin using histological and immunofluorescent techniques. A. viscosus was most frequently found in association with patient's immnoglobulins and complement, followed by S. mutans serotype "d/g", S. mutans serotype "c", and A. naeslundii .     

Identification of bacteria in association with immune components in human carious dentin

Actinomyces viscosus, A. naselundii, Streptococcus mutans serotype "c" and S. mutans serotype "d/g" were identified in human carious dentin using histological and immunofluorescent techniques. A. viscosus was most frequently found in association with patient's immunoglobulins and complement, followed by S. mutans serotype "d/g", S. mutans serotype "c", and A. naeslundii.     

纳米银对牙本质表面粪肠球菌生物膜的杀菌作用

目的 研究纳米银对牙本质表面粪肠球菌生物膜的杀菌作用,为探寻新型有效的根管消毒药物提供实验依据.方法 制备牙本质片309个,随机分成3组,在牙本质表面形成粪肠球菌生物膜后,分别用0.1%纳米银溶液、1.313%次氯酸钠溶液和0.9%氯化钠溶液处理.3组分别于处理0(即处理前)、1、6、12、24 h时取出样本,应用活菌...     

Diffusion of resin monomers through human carious dentin in vitro

Abstract The diffusion of 2-hydroxyethylmediacrylate (HEMA) and triethylene glycol dimethacrylate (TEGDMA) from light cured bonding resin-composite resin restorations through human carious dentin was investigated. Extracted human molar teeth with different degrees of caries were obtained from consenting donors. Teeth were classified into three groups according to caries severity (mild, moderate and severe) using subjective criteria. The outer carious lesions were then removed guided by a proprietary caries detector dye. Teeth with exposure of the pulp space after caries removal were excluded from the study. A polypropylene chamber was attached to the cemento-enamel junction of each tooth to contain 1 ml distilled water. Each cavity was restored with a HEMA containing bonding resin then a TEGDMA-containing resin composite. Water samples were retrieved over a lime course and analyzed by high performance liquid chromatography. There was great variation between teeth in HEMA and TEGDMA permeability. The cumulative amounts released were of similar magnitude to those observed in non-carious teeth for the mild and moderately-severe groups. However, the cumulative amounts released were markedly greater in severely carious teeth than in those with moderate or mild caries.     

Column-like structures of human dentin in carious and artificial lesions

Abstract— Serial sections of 36 human teeth with minor to medium-sized carious lesions, adequate sections from previous studies of artificial lesions adjacent to fillings of amalgam (AM), silicate cement (SI) and glass ionomer cement (GI) were studied by ordinary light microscopy and microradiography. Six artificial primary root lesions were included. In carious lesions, dentin columns were visible in the demineralization zone and partly also in the translucent/radiopaque zone. A typical feature of the artificial lesions adjacent to SI fillings was highly radiopaque columns alternating with radiolucent ones. In the GI group such columns were seen at the cavity wall and the advancing front of the outer lesion. In the AM group dark columns were unveiled as pegs in the latter localization. The findings led to the assumption that the dark columns of intact dentin are preferred pathways of acid diffusion in artificial and, possibly, carious lesions. Increased radiopacity of columns in the SI and GI groups is explained by continued exposure of the tooth surface to fluoride dissolved from these fluoride-rich filling materials.     

Effects of dual antibacterial agents MDPB and nano-silver in primer on microcosm biofilm,cytotoxicity and dentine bond properties

ObjectivesThe objective of this study was to investigate the effects of dentine primer containing dual antibacterial agents, namely, 12-methacryloyloxydodecylpyridinium bromide (MDPB) and nanoparticles of silver (NAg), on dentine bond strength, dental plaque microcosm biofilm response, and fibroblast cytotoxicity for the first time.MethodsScotchbond Multi-Purpose (SBMP) was used as the parent bonding agent. Four primers were tested: SBMP primer control (referred to as “P”), P + 5% MDPB, P + 0.05% NAg, and P + 5% MDPB + 0.05% NAg. Dentine shear bond strengths were measured using extracted human teeth. Biofilms from the mixed saliva of 10 donors were cultured to investigate metabolic activity, colony-forming units (CFU), and lactic acid production. Human fibroblast cytotoxicity of the four primers was tested in vitro.ResultsIncorporating MDPB and NAg into primer did not reduce dentine bond strength compared to control (p > 0.1). SEM revealed well-bonded adhesive–dentine interfaces with numerous resin tags. MDPB or NAg each greatly reduced biofilm viability and acid production, compared to control. Dual agents MDPB + NAg had a much stronger effect than either agent alone (p < 0.05), increasing inhibition zone size and reducing metabolic activity, CFU and lactic acid by an order of magnitude, compared to control. There was no difference in cytotoxicity between commercial control and antibacterial primers (p > 0.1).ConclusionsThe method of using dual agents MDPB + NAg in the primer yielded potent antibacterial properties. Hence, this method may be promising to combat residual bacteria in tooth cavity and invading bacteria at the margins. The dual agents MDPB + NAg may have wide applicability to other adhesives, composites, sealants and cements to inhibit biofilms and caries.