Biomedical subjects

S Ebisu

Publications and source records attributed to S Ebisu.

At least 37 records · Page 2Linked to original sources

PubMed2001

The localization of periodontal-disease-associated bacteria in human periodontal pockets.

Some Gram-negative anaerobes are associated with the incidence and progression of periodontal disease. In periodontal pockets, however, the localization of those bacteria is unknown. We investigated the localization of 5 bacterial species in human periodontal pockets. Fifteen teeth with a part of periodontal pockets from 10 adult periodontitis patients were obtained, and the localization of bacteria was examined immunohistochemically. Positive reactions with anti-Prevotella nigrescens antibody were located at the epithelium-associated plaque area in the middle pocket zones. In the middle and deep pocket zones, Fusobacterium nucleatum and Treponema denticola were especially localized in the unattached plaque area, but Eikenella corrodens was observed in the tooth-attached plaque area. Actinobacillus actinomycetemcomitans, detected in 2 of 15 samples examined, was found in the unattached plaque area, in the middle pocket zone. The present findings indicated that the 5 bacterial species examined localized at distinct regions in human periodontal pockets.

Adult↗

PubMed2000

Microstructure and mechanical property of synthesized hydroxyapatite prepared by colloidal process.

Synthesized hydroxyapatite (HAp) was prepared by the colloidal process. Mixed slurry composed of 25 vol% HAp powder and 75 vol% aqueous solution containing a small amount of polycarboxylic acid ammonium as dispersant was produced. Viscosity of the HAp slurry, microstructure and mechanical property of the synthesized HAp prepared by this process depends on the quantity of dispersant. A mainly uniform microstructure was obtained in both green and sintered HAp with 2 wt% dispersant by preventing the agglomeration of powder in the slurry. A homogeneous microstructure was maintained even after annealing at temperatures between 1200 and 1400 degrees C. Maximum bending strength of 88.5 MPa was also obtained at an optimum dispersant concentration.

Colloids↗

PubMed2000

Cytotoxic effects of composite restorations employing self-etching primers or experimental antibacterial primers.

OBJECTIVE: The aim of this study was to investigate the cytotoxic effects of composites which employ proprietary self-etching primers or experimental primers containing an antibacterial monomer 12-methacryloyloxydodecylpyridinium bromide (MDPB) on human pulpal cells by in vitro dentine barrier tests. METHODS: Experimental primers were prepared by the addition of MDPB to each of two control proprietary primers at 1, 2 and 5%. Direct and indirect composite specimens were placed using each primer on one side of a dentine disk assembled in a simple pulp chamber device. Human pulp cells were incubated on the other side of the disk. After 48 h of incubation, the uptake of [3H]-thymidine by the cells was compared with that for negative controls using wax. Tests were repeated four times for each material. The diffusion of monomers from each specimen was determined using the same device. RESULTS: The specimens with control primers showed 26-35% reduction in cell activity. There were no significant differences in cytotoxicity between the control and experimental primers-specimens (Kruskal-Wallis test, p > 0.05). 2-Hydroxyethyl methacrylate at more than 1 mg ml-1 diffused from all specimens and was considered to be the cause of cytotoxic effects. The concentrations of MDPB diffused from the experimental primers-specimens were less than the toxic level, even for 5% MDPB-containing specimens. CONCLUSIONS: The results demonstrate that composites employing proprietary self-etching primers produced cytotoxic effects on human pulpal cells in vitro, although the toxicity was not severe. Incorporation of MDPB into a proprietary primer of up to 5% had no significant influence on the cytotoxicity observed.

Acid Etching, Dental↗

PubMed2000

A new electrical method for detecting marginal leakage of in vitro resin restorations.

OBJECTIVES: Ingress of bacteria at sites of marginal leakage has been suggested to cause pulpal inflammation. The purpose of this in vitro study was to evaluate the validity of a new electrical method to detect marginal leakage of restoratives by comparing the results obtained with those of a dye penetration test. METHODS: After cavities were prepared on the buccal coronal surfaces and root surfaces of 16 extracted non-carious human molar teeth, eight specimens were treated with a dentin bonding system (bonding group) and the other eight specimens were not treated (non-bonding group). Resin composites were filled in the cavities of all specimens, and physiological saline was applied to the margin of the restorative. Any excess saline was wiped off, leaving only the electrolyte, which had penetrated into the marginal gap. The change in conductance was measured continuously across the margin of each specimen during this process. The marginal leakage of specimens was confirmed using the dye penetration test, and the results were evaluated by the microleakage score. RESULTS: In both coronal and root surface cavities, the changes in conductance in the non-bonding group after filling were significantly larger than those of the bonding group (p<0.05). The change in conductance of each specimen correlated with the microleakage score (p<0.05). CONCLUSIONS: It was concluded that the relative electrical method could detect marginal leakage in both coronal and root surface cavities.

Composite Resins↗

PubMed2000

Polymerization contraction stress of resin composite restorations in a model Class I cavity configuration using photoelastic analysis.

PURPOSE: An important factor that contributes to deterioration of resin composite restorations is contraction stress that occurs during polymerization. The purpose of this article is to familiarize the clinician with the characteristics of contraction stress by visualizing the stresses associated with this invisible and complex phenomenon. MATERIALS AND METHODS: Internal residual stresses generated during polymerization of resin composite restorations were determined using micro-photoelastic analysis. Butt-joint preparations simulating Class I restorations (2.0 mm x 5.0 mm, 2.0 mm in depth) were prepared in three types of substrates (bovine teeth, posterior composite resin, and transparent composite resin) and were used to examine contraction stress in and around the preparations. Three types of composite materials (a posterior composite, a self-cured transparent composite, and a light-cured transparent composite) were used as the restorative materials. The self-cured composite is an experimental material, and the others are commercial products. After treatment of the preparation walls with a bonding system, the preparations were bulk-filled with composite. Specimens for photoelastic analysis were prepared by cutting sections perpendicular to the long axis of the preparation. Fringe patterns for directions and magnitudes of stresses were obtained using transmitted and reflected polarized light with polarizing microscopes. Then, the photoelastic analysis was performed to examine stresses in and around the preparations. RESULTS: When cavity preparations in bovine teeth were filled with light-cured composite, a gap was formed between the dentinal wall and the composite restorative material, resulting in very low stress within the restoration. When cavity preparations in the posterior composite models were filled with either self-cured or light-cured composite, the stress distribution in the two composites was similar, but the magnitude of the stress was greater in the light-cured material. When preparations in the transparent composite models were filled with posterior composite and light-cured transparent composite material, significant stress was generated in the preparation models simulating tooth structure, owing to the contraction of both restorative materials. CLINICAL SIGNIFICANCE: Polymerization contraction stress is an undesirable and inevitable characteristic of adhesive restorations encountered in clinical dentistry that may compromise restoration success. Clinicians must understand the concept of polymerization contraction stress and realize that the quality of composite resin restorations depends on successful management of these stresses.

Animals↗

PubMed2000

Antibacterial activities and release kinetics of a newly developed recoverable controlled agent-release system.

We attempted to develop a resin with a recoverable antibacterial activity based on the desorption/adsorption of a cationic bactericide by the ion-exchange mechanism. The aims of this study were to investigate the release kinetics of the agent and the antibacterial activity of this newly designed resin system. An experimental resin was prepared by the addition of methacrylic acid as a cation-exchanger and a cationic antibacterial agent, cetylpyridinium chloride (CPC), to triethyleneglycol dimethacrylate. The amount of CPC desorbed from the experimental resin into buffer solutions at pH 4-8 was measured. The adsorption of CPC to control resin and re-adsorption of CPC to the experimental resin, which had once desorbed the agent, were also determined. The antibacterial activity of experimental resin against Streptococcus mutans was evaluated, and the relationship between bacterial acid production and antibacterial effect was assessed. The experimental resin desorbed CPC at pH < or = 6, and the amount of agent desorbed increased with increasing acidity. The control resin adsorbed CPC when immersed in CPC aqueous solution at a rate determined by the concentration of the agent and immersion time. The experimental resin, once desorbed CPC, could re-adsorb the bactericide by being exposed to a solution of the agent. Less plaque formed on the experimental resin, and the growth and survival of S. mutans was inhibited in the condition in which acid was produced. These results demonstrate that the resin system proposed was able to desorb and re-adsorb the cationic bactericide by an ion-exchange mechanism and could show an inhibitory effect on S. mutans growth and plaque formation.

Adsorption↗

PubMed2000

Estrogenicity of fissure sealants and adhesive resins determined by reporter gene assay.

It is controversial whether the dental resinous materials containing 2,2-bis[4-(2-hydroxy-3-methacryloyloxypropoxy)phenyl]propane (Bis-GMA), which is synthesized from the estrogenic compound bisphenol A (BPA), include unreacted BPA and/or can mimic the effects of natural steroid hormones. In the present study, the estrogenic activities of 3 fissure sealants and 5 adhesive resins, which were all unpolymerized, were determined by means of a reporter gene assay, and the relevance of the components to the estrogenicity was investigated. Two commercially available sealants were confirmed to have estrogenic activity, although none of the tested materials contained BPA. In contrast, hydrophobic monomer bisphenol A dimethacrylate (BPA-DMA), which is also estrogenic, was found to be included in these estrogenic sealants in an amount greater than the minimum concentration to show estrogenicity. This suggests that the estrogenicity of the two proprietary sealants was associated with BPA-DMA rather than with BPA.

Analysis of Variance↗

PubMed2000

Use of Bacillus brevis for synthesis and secretion of Des-B30 single-chain human insulin precursor.

A synthetic gene encoding a single chain human insulin precursor [B-chain (1-29)-A-chain] linked to the C-terminal lysine of human epidermal growth factor (1-28) (EGF-SCI) was constructed. This gene was expressed using Bacillus brevis. EGF-SCI was isolated from the supernatant of the culture broth. Treatment of EGF-SCI with lysyl endopeptidase resulted in the formation of des-B30 human insulin. The identification of the formed des-B30 human insulin was made by the measurement of molecular weight and amino acid analysis. The binding coefficient to anti-human insulin antibody was comparable to that of human insulin.

Amino Acid Sequence↗

PubMed2000

Identification of periodontal disease-associated bacteria in the "plaque-free zone".

BACKGROUND: Subgingival plaque bacteria live within a biofilm covered with glycocalyx, and little is known of the bacterial species associated with biofilm formation at the bottom of human periodontal pockets, the so-called "plaque-free zone"(PFZ). METHODS: Seventy-seven extracted teeth from 56 patients with severe advanced adult periodontitis were examined. Porphyromonas gingivalis, Campylobacter rectus, Fusobacterium nucleatum, Actinobacillus actinomycetemcomitans, Treponema denticola, Prevotella nigrescens, and Actinomyces viscosus were examined by scanning immunoelectron microscopic techniques, using both secondary and back-scattered imaging, with rabbit antibodies specific for each bacteria. RESULTS: Secondary electron images showed that rods, filaments, and spirochete-shaped bacteria formed small aggregates in the PFZ. Some of the bacteria were covered with an amorphous film-like structure. By back-scattered electron imaging, positive reactions with anti-P. gingivalis were found in 8 of 13 samples examined, and film-like structures coated several cells of 6 positive samples examined. Labeled cells with anti-C. rectus, anti-T. denticola and anti-P. nigrescens were detected in 3 of 11, 5 of 10, and 1 of 8 samples examined. A. viscosus were found in 6 of 11 of the samples. A. viscosus tended to overlay the amorphous capsula and aggregate. F. nucleatum and A. actinomycetemcomitans were not detected in any samples examined. CONCLUSIONS: These findings indicated that P. gingivalis, C. rectus, T. denticola, P. nigrescens, and A. viscosus were present in the PFZ, and that some specified bacteria were possibly related to plaque-biofilm formation of subgingival plaque.

Actinomyces viscosus↗

PubMed2000

Effect of surface roughness of porcelain on adhesion of bacteria and their synthesizing glucans.

STATEMENT OF PROBLEM: In some instances of porcelain restoration, refinishing is inevitable. In terms of plaque accumulation on porcelain, refinishing could be a substitute method for glazing. PURPOSE: This study compared the amount of adhesion of plaque components (bacterial cells and glucans) on porcelain disks with various degrees of surface roughness to assess the effects of surface roughness on the amount of plaque accumulation. MATERIAL AND METHODS: Radiolabeled cell suspensions were incubated with porcelain disks for 3, 8, and 24 hours at 37 degrees C, and the amounts of adhered cells and glucans were measured by using a liquid scintillation method. RESULTS: The amount of cells and glucans adhered on porcelain increased with incubation time. The surface roughness value and the amount of plaque adhesion decreased with the increase in polishing level. However, the greatest amount of plaque was adhered on glazed surfaces, although their surfaces were smoother than the surfaces polished with 120- or 600-grit abrasive papers. CONCLUSION: With the exception of glazed surfaces, a positive correlation between surface roughness and the amount of plaque accumulation was observed. Repolishing with a diamond paste would not induce problems of plaque accumulation, compared with an intact glazed surface.

Analysis of Variance↗

PubMed1999

Post-irradiation polymerization of composites containing bis-GMA and TEGDMA.

OBJECTIVE: This study was conducted to evaluate the effect of the amount of TEGDMA on post-irradiation polymerization of bis-GMA based composites. METHODS: Four experimental composites containing bis-GMA and TEGDMA at the ratios (w/w) of 75/25, 70/30, 67/33, and 50/50 were prepared. The degree of conversion of each composite was measured by Fourier transformation infrared spectroscopy immediately after being cured and after 24 h of storage, and the post-irradiation polymerization was determined. In addition, the change in respective amounts of residual bis-GMA and TEGDMA in cured specimens over 24 h was determined by high performance liquid chromatography. Data were analyzed by means of ANOVA, followed by a Fisher's PLSD test at a significance level of 0.05. RESULTS: At both stages of as-cured and after 24 h-sotrage, the composites containing greater amounts of TEGDMA showed a greater degree of conversion. The amount of post-irradiation polymerization decreased as the content of the incorporated TEGDMA increased. The reduction in the amount of TEGDMA remaining unreacted during post-irradiation polymerization also decreased as the content of TEGDMA increased, while the amount of unreacted bis-GMA showed almost constant values. SIGNIFICANCE: An increase in the amount of TEGDMA-incorporation in bis-GMA based composites resulted in less post-irradiation polymerization. As the change in the amount of residual unreacted TEGDMA in cured specimens showed the same trend as the results of the post-irradiation polymerization. TEGDMA is considered to be the main contributor to post-irradiation polymerization of bis-GMA based composites.

Analysis of Variance↗

PubMed1999

Bactericidal activity and cytotoxicity of antibacterial monomer MDPB.

The aim of this study was to investigate bactericidal characteristics and cytotoxicity of the newly developed antibacterial monomer 12-methacryloyloxydodecylpyridinium bromide (MDPB). To evaluate the bactericidal activity of MDPB against oral streptococci, the minimum bactericidal concentration (MBC) for seven species and time-kill kinetics against Streptococcus mutans were determined. The cytotoxic effects of MDPB on human pulpal cells were assessed by [3H]-thymidine uptake after contact with MDPB solutions at various concentrations. MDPB showed strong bactericidal activity against seven streptococci, the MBC value ranging from 31.1 to 62.5 micrograms ml-1. Time-kill determination indicated a rapid killing effect of MDPB at 250 micrograms ml-1 or over, and all cells were killed within 1 min by MDPB at 500 micrograms ml-1 or over. No cytotoxic effect was observed on contact with MDPB at concentrations of 10 micrograms ml-1 or less, and the toxicity of MDPB was considered to be similar to those of other monomers used for dental materials. These results suggest that MDPB can be effectively incorporated in dental resin-based materials to provide bactericidal activity against oral bacteria.

Anti-Infective Agents, Local↗

PubMed1999

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.

Absorption↗

PubMed1999

Comparison of polymerization contraction stresses between self- and light-curing composites.

OBJECTIVES: The objective of this study was to examine the distributions and the magnitudes of the internal stresses in self- and a light-curing composite restorations resulting from polymerization shrinkage. METHODS: Butt-joint box-shaped cavities (5.0 x 2.0 mm2, 2.0 mm in depth) prepared in composite molds were filled with either a self- or light-curing transparent resin composite. The restorations were cross-sectioned perpendicular to the longitudinal axes of the cavities and observed using polarizing microscopes. The principal stresses in the restorations, normal and shear stresses at the cavity wall were evaluated by photoelastic analysis. RESULTS: The distributions of the principal stresses and the stresses generated at the cavity wall in both the self- and the light-curing composite restorations were similar. The maximum stress generated at the cavity wall in the light-curing composite restorations was twice as large as that seen in the self-curing restorations. CONCLUSIONS: The results of this study indicated that the difference in the magnitude of the internal stresses between self- and light-curing composites was not related to the distribution of the stresses. The velocity of polymerization appeared to be the most important factor contributing to the magnitude of the internal stresses generated in the composite restorations in this study.

Birefringence↗

PubMed1999

Evaluation of post-treatment solutions for clinical use with the calcium phosphate precipitation method.

OBJECTIVE: We have proposed a calcium phosphate precipitation (CPP) method that occludes dentine tubules with apatitic minerals for the treatment of dentine hypersensitivity. The current CPP method uses 1 mol/l NaOH as the post-treatment solution. However, its high pH is not desirable for clinical use. The aim of this study was to evaluate several solutions, especially buffer solutions, as post-treatment solutions for the CPP method. METHODS: The CPP solution was mixed with several post-treatment solutions at various mixing ratios in test tubes. The precipitates were collected, and freeze-dried in a vacuum. Their weights were measured to evaluate the feasibility of buffer solution for the post-treatment solutions. Among the solutions, we selected one buffer solution and have done further evaluation using human dentine disks. The degree of occlusion of dentine tubules was evaluated by scanning electron microscopic observation and measurement of dentine permeability. Also, the composition of the precipitate was analyzed using energy-dispersive X-ray microanalysis and powder X-ray diffraction. RESULTS: Among the solutions, 1 mol/l NaHCO3 gave sufficient amounts of precipitate at a relatively mild pH, and thus was considered to be a good candidate for a post-treatment solution. Scanning electron microscopic observation showed that dentine tubules were occluded with the precipitate to a depth of approximately 10 microns from the surface when the CPP treatment was done twice, and dentine permeability was reduced to 98.8%. The precipitate was dicalcium phosphate dihydrate (DCPD; CaHPO4-2H2O) when the post-treatment solution was free of NaF. In contrast, the precipitate was an apatitic mineral with CaF2 as a by-product when the post-treatment solution contained NaF. CONCLUSIONS: We concluded that 1 mol/l NaHCO3 containing 0.3 mol/l NaF would be suitable as the post-treatment solution for the CPP method as it has a relatively mild pH and occludes dentinal tubules well.

Adult↗

PubMed1999

Interleukin-6 (IL-6) and IL-8 are induced in human oral epithelial cells in response to exposure to periodontopathic Eikenella corrodens.

Periodontitis is the inflammatory response in periodontal tissues elicited by bacterial colonization in periodontal pockets. In this response, pocket epithelial cells are the first cells to come into contact with bacteria. To elucidate this mechanism, we determined the adherence of the periodontopathic bacterium Eikenella corrodens 1073, which has a GalNAc-sensitive lectin-like adhesin (EcLS), to a human oral epithelial carcinoma cell line (KB) and the induction of proinflammatory cytokine production in the cells following exposure to this bacterium in vitro. In the adherence assay, EcLS played a role as the adhesin of this bacterium in adherence to KB cells. In a reverse transcriptase PCR, significant interleukin-8 (IL-8) and IL-6 mRNA levels were induced in response to exposure to this bacterium. In an enzyme-linked immunosorbent assay after an 8-h bacterial exposure, the IL-8 and IL-6 protein levels were 13.5- and 8.3-fold higher than those in the nonexposed controls, respectively. These protein responses were time dependent. Interestingly, when E. corrodens was separated from KB cells by cell culture inserts, a slight stimulation of the IL-6 and IL-8 mRNA and secreted protein levels was seen. These results imply that the direct contact of E. corrodens 1073 with oral epithelial cells is not necessarily required for the stimulation of IL-6 and IL-8 secretion. We suggest that E. corrodens induces the epithelial cells to secrete proinflammatory cytokines which serve as an early signaling system to host immune and inflammatory cells in underlying connective tissues.

Adhesins, Bacterial↗

PubMed1999

Structural conversion from non-native to native form of recombinant human epidermal growth factor by Brevibacillus choshinensis.

Brevibacillus choshinensis (Bacillus brevis) HPD31 is a very efficient producer of recombinant human epidermal growth factor (EGF). The produced EGF is secreted into the medium with high efficiency. However part of the EGF that accumulates in the medium, exists as multimeric forms which are biologically inactive. We found the bacterium has the activity to structurally convert multimeric forms to the monomeric, native ones. Optimal temperature and pH for the conversion were 40 degrees C and pH 9, respectively. The reaction was promoted in the presence of reduced glutathione or cysteine. But the cells which had been sonicated or exposed to moderate heat treatment completely lost the activity. Thus, it was presumed that the activity might be due to the enzyme(s) that catalyze the protein disulfide exchanging reaction, and that they resides on the surface of viable cells.

Bacillus↗

PubMed1998

Overexpression of cation-dependent mannose 6-phosphate receptor prevents cell death induced by serum deprivation in PC12 cells.

PC12 cells express well cation-independent mannose 6-phosphate receptors (CI-MPR), but not cation-dependent (CD)-MPR as much. To examine CD-MPR dependency of transport of cathepsins B and D to lysosomes in PC12 cells, we prepared the cells overexpressing CD-MPR. Immunoreactivity for cathepsin B became more distinct and larger in size in the transfected cells than in wild-type cells. No difference in the distribution of cathepsin D was seen between these two cells. The viability of the cells following serum deprivation was significantly higher in the transfected cells than in wild-type cells. This increased viability of the transfected cells was blocked by CA074, a specific inhibitor of cathepsin B, while pepstatin A suppressed the action of CA074. The results suggest that CD-MPR preferentially transport cathepsin B in PC12 cells, and cathepsins B and D participate in the regulation of PC12 cell apoptosis.

Amino Acid Sequence↗