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.     

Bactericidal effect of dentin primer containing antibacterial monomer methacryloyloxydodecylpyridinium bromide (MDPB) against bacteria in human carious dentin

The aim of this study was to investigate the bactericidal effect of a dentin primer incorporating the antibacterial monomer 12-methacryloyloxydodecylpyridinium bromide (MDPB) against bacteria in human dentinal carious lesions. To evaluate the antibacterial activity of MDPB against anaerobes, the minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) against obligate anaerobes and facultative anaerobic strains of lactobacilli were determined. Bacteria were recovered from carious dentin samples obtained from the teeth of patients, and the bactericidal activities of the experimental primer containing 5% MDPB and three commercially available primers were compared by counting the number of viable cells after contact with diluted solutions of each primer for 30 s. MDPB showed strong antibacterial activity against anaerobes, the MIC and MBC values ranging from 3.9 to 31.3 micrograms/ mL-1 and 15.6-125 micrograms/ mL-1, respectively. Experimental primer containing MDPB was the most bactericidal among the materials tested (ANOVA, Fisher's PLSD test, P < 0.05) and was able to kill the bacteria completely even when diluted 40 times, while the three commercial products exhibited little activity at 40 times dilution. These results indicate that incorporation of MDPB into dentin primer could be beneficial for eliminating the residual bacteria in cavities.     

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.     

Cysteine cathepsins in human carious dentin

Matrix metalloproteinases (MMPs) are important in dentinal caries, and analysis of recent data demonstrates the presence of other collagen-degrading enzymes, cysteine cathepsins, in human dentin. This study aimed to examine the presence, source, and activity of cysteine cathepsins in human caries. Cathepsin B was detected with immunostaining. Saliva and dentin cysteine cathepsin and MMP activities on caries lesions were analyzed spectrofluorometrically. Immunostaining demonstrated stronger cathepsins B in carious than in healthy dentin. In carious dentin, cysteine cathepsin activity increased with increasing depth and age in chronic lesions, but decreased with age in active lesions. MMP activity decreased with age in both active and chronic lesions. Salivary MMP activities were higher in patients with active than chronic lesions and with increasing lesion depth, while cysteine cathepsin activities showed no differences. The results indicate that, along with MMPs, cysteine cathepsins are important, especially in active and deep caries.     

Predominant obligate anaerobes in human carious dentin

With the adoption of an anaerobic glove box system, we isolated and identified the predominant micro-organisms in lesions of carious dentin from four permanent third molars. The overwhelming majority of the micro-organisms isolated from both shallow and deep layers of carious dentin were obligate anaerobes, suggesting that conditions in the dentin were strictly anaerobic. The predominant obligate anaerobes were Gram-positive rods which were identified as members of the following genera: Propionibacterium, Eubacterium, Arachnia, Lactobacillus, Bifidobacterium, and Actinomyces. Clostridia, Gram-negative rods (Bacteroides and Fusobacterium) and Gram-positive cocci (Peptococcus, Peptostreptococcus, and Streptococcus) were also isolated, although they constituted a minor part of the flora.     

龋蚀检知液对细菌的影响

目的：了解龋蚀检知液对牙本质细菌的影响。方法：试管稀释法药敏试验及常规牙本质细菌分离鉴定。结果：经χ２检验，染色前后牙本质细菌的几何均数和检出率无显著差异。结论：检知液染色龋坏牙本质对其中的细菌无影响。     

Observations on endopeptidases in human carious dentin

abstract – The enzymatic hydrolysis of hemoglobin (Hb) and N-α-benzoyl-DL-arginine-2-naphthylamine (BANA) was observed both in sound- and in carious human dentin. The enzyme activity expressed per mg protein in the sample and per min was at least ten times higher in carious than in sound dentin, as was also the DNA content in the samples. The hydrolysis of Hb and BANA was high over a relatively wide pH range, the optimum pH being 5.5 with Hb, and 7.0 with BANA, as substrates. EDTA, cetylpyridinium chloride and Hg²⁺ ions inhibited strongly the hydrolysis of BANA at pH 7.0 but only slightly at pH 10.4. The origin of these endopeptidase-like enzymes was thought to be mainly microbial. These enzymes were thought to hydrolaze proteins of the organic stroma of dentin during the carious process.     

Nanomechanical properties of hydrated carious human dentin.

Most restorative materials are bonded to caries-affected dentin that has altered structure. We tested the hypothesis that hydrated dentin of the transparent zone did not have increased hardness or elastic modulus. Nanoindentation by modified AFM was used to determine site-specific elastic modulus and hardness for components of hydrated dentin from 8 carious and non-carious human teeth. Indentations in intertubular dentin were made at intervals from pulp through the affected layers (subtransparent, transparent, and discolored zones). The values of intertubular dentin increased slightly from near the pulp into the transparent zone, then remained constant or decreased slightly through transparent dentin (E, 18.3 GPa; H, 0.8 GPa; confirming the hypothesis), and decreased markedly through the discolored region. Peritubular dentin values were unaltered in transparent dentin, and intratubular mineral had values between those of normal peritubular and intertubular dentin. Superficial areas contained distorted tubules without peritubular dentin or intratubular mineral.     

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.     

Acid profile in carious dentin

Organic acids in carious dentin from 69 permanent teeth were analyzed by gas chromatography. Lactate, acetate, propionate, and butyrate were detected in most samples, and limited amounts of isobutyrate, valerate, isovalerate, caproate, and isocaproate were occasionally detected. Lactate, acetate, and propionate were major acids and altogether accounted for about 90% of total acid in most samples of carious dentin. However, the proportion of these three acids varied among the samples. Some samples contained over 85% lactate, while others contained mainly acetate and propionate. A high percentage of acetate was usually accompanied by an appreciable amount of propionate. All seven samples in carious dentin under fillings or restorations had little lactate, but a high percentage of acetate plus propionate. The differences in acid profiles of carious dentin may reflect differences in the microbial ecology of carious dentin, and a stage of progress of dentin caries or a type of dentin caries.     

Serum proteins and secretory component in human carious dentin.

By the use of the peroxidase-labeled antibody method, significant localization of IgG, IgA, albumin and transferrin was demonstrated in the deep lesion of 20 carious teeth, where the secretory component was absent. These serum proteins formed a distinct zone, surrounding the overlying, shallow lesion infected with bacteria.     

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

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.