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Biomedical subjects

Yoshihiko Hayashi

Publications and source records attributed to Yoshihiko Hayashi.

11 recordsLinked to original sources

Chitosan monomer promotes tissue regeneration on dental pulp wounds.

The present study was undertaken to evaluate the applicability of chitosan monomer (D-glucosamine hydrochloride) as a pulp capping medicament. Both in vitro and in vivo experiments were carried out to study the cell metabolism and wound healing mechanisms following the application of chitomonosaccharide. After 3 days of osteoblast culture, alkaline phosphatase (ALP) activity significantly increased in the chitosan group. Reverse transcription polymerase chain reaction analysis revealed that chitosan induced an increase in the expression of ALP mRNA after 3 days and bone morphogenetic protein-2 mRNA after 7 days of osteoblast incubation. Inflammatory cytokine, interleukin (IL)-8, synthesis in fibroblasts was strongly suppressed in the medium supplemented with chitosan monomer. Histopathological effects were evaluated in rat experiments. After 1 day, inflammatory cell infiltrations were observed to be weak when compared with the application of chitosan polymer. After 3 days, a remarkable proliferation of fibroblasts was seen near the applied chitosan monomer. The inflammatory cell infiltration had almost completely disappeared. After 5 days, the fibroblastic proliferation progressed, and some odontoblastic cells appeared at the periphery of the proliferated fibroblasts. These findings indicate that the present study is the first report that chitosan monomer acts as a biocompatibility stable medicament even at the initial stage of wound healing in comparison with the application of chitosan polymer.

Alkaline Phosphatase↗

Effect of a caries-detecting solution on the tensile bond strength of four dentin adhesive systems.

This study investigated the effect of a caries-detecting solution on the tensile bond strength (TBS) to sound bovine dentin--which was either rinsed thoroughly of or contaminated with the caries-detecting solution. Caries Detector (1.0% acid red in propylene glycol) was applied on flat dentin surfaces for 10 seconds, rinsed, and dried with syringe air. In another group, Caries Detector was not rinsed but air-dried. Then, the surfaces were treated with one of the following adhesive systems: Clearfil Protect Bond, Clearfil SE Bond, One-Up Bond F, or Single Bond. Furthermore, an ingredient of Caries Detector, either 1.0% acid red aqueous solution or propylene glycol, was applied to evaluate the effect of each component. In the control groups, Caries Detector was not applied to the dentin surfaces. Finally, a resin composite was light-cured and the TBS measured. Fractured specimens and treated dentin surfaces were observed by SEM. Caries Detector did not reduce the tensile bond strength of any adhesive system (p>0.05) when rinsed thoroughly. On the other hand, when dentin surface was contaminated with Caries Detector, TBS decreased significantly with Clearfil SE Bond and Single Bond. As for the ingredients of Caries Detector, the effect of acid red on TBS was not significant, but that of propylene glycol was significant.

Adhesives↗

Five-year clinical evaluation of two adhesive systems in non-carious cervical lesions.

OBJECTIVE: This controlled clinical trial evaluated the 5-year clinical performance of a self-etching primer system including selective enamel-etching with phosphoric acid and a one-bottle adhesive system. METHODS: Seventy-two non-carious cervical lesions in 8 patients (4 male and 4 female) with a mean age of 61.3 years (range 45-78) participated in the study. An enamel bevel was placed and dentin lightly ground, and cavities restored with clearfil liner bond II (LB) or single bond (SB) in conjunction with a hybrid resin composite (Clearfil AP-X). In the case of 27 cavities for LB, the enamel was pretreated with 37% phosphoric acid for 10 s. Each patient received both types of restoration, which were distributed on a random basis. All restorations (37 restorations for LB and 35 restorations for SB) were placed by one dentist. The restorations were evaluated blind after 5 years using modified USPHS criteria. The data were statistically analyzed using the Fisher's exact test. RESULTS: All but one restoration (which was replaced by a crown after the 2-year recall) were evaluated after 5 years. 100% retention rates were recorded for both restorative groups. No caries was detected in association with any restorations. The only minor problem was marginal discoloration; superficial and localized marginal discoloration occurred around 18% of the restorations, and mainly at the dentin margin. There were no significant differences in the marginal integrity between the LB and SB groups. CONCLUSIONS: Restorative materials used in this study demonstrated a good clinical effectiveness in the restoration of non-carious cervical lesions for 5 years.

Aged↗

Characteristics of nociceptors in the periodontium--an in vitro study in rats.

Recent studies have indicated that nociceptors can be classified into various types according to their physiological properties. These studies have clarified that the frequency distribution of various nociceptor types is different among body sites and animal species. In the present study, we investigated the physiological properties of rat's periodontal nociceptors in an in vitro jaw-nerve preparation. Responses were recorded from functional single filaments in the inferior alveolar nerve. To determine the nociceptor type, calibrated von Frey filaments, heat, and bradykinin (BK) stimuli were used. We found five subtypes of nociceptors in the periodontal ligaments of the lower incisor: Adelta-high threshold mechanonociceptors (Adelta-HTM, n=28), Adelta-mechanoheat nociceptors (Adelta-MH, n=6), Adelta-polymodal nociceptors (Adelta-POLY, n=26), C-high threshold mechanonociceptors (C-HTM, n=3) and C-polymodal nociceptors (C-POLY, n=4). Most nociceptors were Adelta-innervated, while only a small number of C-innervated nociceptors were found. The present results suggest that periodontal nociceptors transmit mainly fast pain, and may thus play a role in rapid detection of injure-related stimuli during mastication.

Animals↗

Early gene expression analyzed by cDNA microarray and RT-PCR in osteoblasts cultured with water-soluble and low molecular chitooligosaccharide.

Chitosan has a variety of biological activities. However, little is known about how chitosan modulates the hard tissue forming cells. When we cultured an osteoblastic cell line in alpha-MEM supplemented with 10% FBS and 0.005% chitooligosaccharide for 3 days, alkaline phosphatase (ALP) activity was significantly high compared with the control culture group (p<0.05). This study was focused on gene expression in osteoblasts cultured with water-soluble chitooligosaccharide. cDNA probes were synthesized from isolated RNA and labeled with fluorescent dye. They were hybridized with Human 1.0((R)) cDNA microarray, and fluorescent signal was analyzed. cDNA microarray analysis revealed that 16 genes were expressed at >/=1.5-fold higher signal ratio levels in the experimental group compared with the control group after 3 days. RT-PCR analysis showed that chitosan oligomer induced an increase in the expression of two genes, CD56 antigen and tissue-type plasminogen activator. Furthermore, the expression of mRNAs for BMP-2 was almost identical in the experimental and control groups after 3 days of culture, but slightly increased after 7 days of culture with chitosan oligomer. These results suggest that a super-low concentration of chitooligosaccharide could modulate the activity of osteoblastic cells through mRNA levels and that the genes concerning cell proliferation and differentiation can be controlled by water-soluble chitosan.

Bone Morphogenetic Protein 2↗

The effect of light-curing modes on the microleakage of cervical resin composite restorations.

OBJECTIVE: This study investigated the effect of light-curing modes on the microleakage of cervical resin composite restorations. METHODS: Eighty wedge-shaped cervical cavities, C-factor approximately 1.3, were prepared on the labial surface of bovine incisors. The cavities were treated with a two-step experimental self-etching primer adhesive system containing an antibacterial monomer (ABF), according to the manufacturer's instructions, and filled with a single increment of a hybrid resin composite. The restorations were allocated randomly into four groups of 20: Group 1-the restorations were light-cured for 40 s with a conventional light-curing unit (New Light VL-II); Group 2-the restorations were cured with Candelux using a two-step mode; Group 3-the restorations were cured with Astralis 7 using a ramping mode; Group 4-the restorations were light-cured for 3 s with a plasma arc light-curing unit (Wave Light). Half of the specimens from each group were subjected to thermocycling (5-60 degrees C, 15 s dwell time, 5000 cycles) prior to immersion in 0.5% basic fuchsin. The teeth were sectioned and the degrees of dye penetration were scored. The data were analyzed with either the Kruskal-Wallis test or the Mann-Whitney U-test (p < 0.05) RESULTS: No significant differences were found in the microleakage scores among the light-curing modes used. The restorations tended to display more microleakage when they were subjected to thermocycling, but the differences were not statistically significant. CONCLUSIONS: These findings suggest that light-curing modes may have no effect on the microleakage of cervical cavities with a small C-factor, as is the case in this study.

Animals↗

Dimensional changes of ring-shaped pattern.

Ring shaped wax patterns, having the same outside diameter and different inside diameters, were invested with a gypsum-bonded cristobalite investment. The wax pattern was eliminated in an electric furnace at 120 degrees C. A fusible alloy with a melting point of 47 degrees C was cast at room temperature. The dimensional deviations between the fusible alloy casting and the wax pattern were calculated using the inside diameter, ring width and outside diameter. On the other series, a gold alloy casting of the same size was fabricated in the usual manner of the dental precise casting procedure, and the dimension was compared with that of the wax pattern. In the comparison of 2 types of patterns, dimensional change by setting expansion was different. Dimensional change of the small inside diameter specimen differed at 3 portions measured, but that of the large inside diameter specimen was comparable at 3 portions. Concerning the resultant gold alloy casting, dimensional change at the outside diameter differed from each other, but those at ring width and inside diameter were comparable to each other. The difference in the inside diameter influenced dimensional change by setting expansion as well as that of the resultant casting.

Analysis of Variance↗

Sandblasting of inlay margin--marginal abrasion and bond strength.

Specimens (such as metal inlays) with 30 degrees or 45 degrees marginal bevel were prepared by casting with a 12% Au-Pd-Ag alloy or a gold alloy. A form of the marginal bevel was traced on a profile projector before and after sandblasting, and the length of the abraded margin measured. All the blasting conditions abraded the marginal bevel, while the blasting at 20 mm for 2 seconds brought about the least abrasion of approximately 10 microm in the 45 degrees specimen cast with Au-Pd-Ag alloy. The gold alloy specimens were abraded more than the Au-Pd-Ag alloy ones; those with marginal bevel of 30 degrees were abraded more than those of 45 degrees. On the other hand, the effect of different blasting conditions on the bond strength of units bonded with resin cement was evaluated (under selected blasting conditions known to cause relatively less damage to the marginal level). Specimens treated by sandblaster exhibited a comparable tensile bond strength, while specimens without sandblasting but applied with only an alloy primer showed a statistically low value.

Air Abrasion, Dental↗

Microleakage of cervical cavities restored with flowable composites.

PURPOSE: To investigate the microleakage of cervical cavities restored with flowable composites in conjunction with an all-in-one adhesive system (AQ Bond) and an experimental two-step self-etching primer system (ABF) after thermocycling or flexural load cycling. METHODS: 180 wedge-shaped cervical cavities on the labial surfaces of bovine incisors were treated with AQ Bond or ABF according to the manufacturers' instructions. The treated cavities were filled with one of three flowable composites (AElite Flo, Metafil Flo or Protect Liner F). Ten of 30 specimens for each group were immersed in 0.5% basic fuchsin solution immediately after finishing of the restorations. Specimens to be subjected to thermocycling (5-60 degrees C, 15 seconds dwell time, 5000 cycles) or to flexural load cycling (approximately 0.5 mm labio-lingual displacement at the incisal edge, 10,000 cycles, 1 Hz) prior to immersion in the dye were also prepared. After 24-hour storage in the dye solution, the teeth were sectioned longitudinally through the center of the restorations, and the degrees of dye penetration scored. The microleakage scores of a hybrid composite (Clearfil AP-X) obtained from our previous study were used for comparison. The data were analyzed with the Kruskal-Wallis test and the Mann-Whitney U-test (P < 0.05). RESULTS: No significant differences in the microleakage patterns and scores were found among the flowable composites. On the other hand, significant differences in the microleakage patterns and scores were observed between the flowable and the hybrid composites. Thermocycling caused deterioration of marginal integrity, and the flowable composites showed more microleakage than the hybrid composite. In contrast, flexural load cycling did not cause deterioration of the marginal integrity when the cavities were filled with the flowable composites. Statistical analysis also revealed that the adhesive systems played a large role in the microleakage patterns.

Animals↗

Mineralization of matrix vesicles isolated from a human osteosarcoma cell line in culture with water-soluble chitosan-containing medium.

Chitosan is a natural bioactive material. Although it has been reported that chitosan promotes osteogenesis in bone lesions, little is known about how chitosan modulates this process. The present study was designed to investigate the effect of water-soluble chitosan relative to initiation of biologic mineralization, especially in the matrix-vesicles-(MVs) mediated process in vitro. A human osteoblastic cell line (NOS-1) was used. After 3 days of incubation, the number of cells and alkaline phosphatase (ALP) activity increased significantly in the chitosan group. RT-PCR analysis revealed that chitosan induced an increase in the expression of bone morphogenetic protein-2 mRNA after 7 days of incubation. MVs were isolated from NOS-1 cells using a collagenase digestion and ultracentrifugation method. ALP activity of MVs isolated from chitosan-supplemented cells was significantly higher than that of the control group. Furthermore, isolated MVs were incubated in medium supplemented with Na-beta-glycerophosphate without fetal bovine serum. Needle-like crystals were observed in association with MVs after 24 h of incubation. These needle-like crystals were densely accumulated in the chitosan group. The present findings suggest that water-soluble chitosan would promote osteoblast proliferation and differentiation and may be useful for the acceleration of initial biologic mineralization.

Alkaline Phosphatase↗

Effect of low-viscosity resin-based composite on the microleakage of cervical restorations.

PURPOSE: To investigate the effect of a low-viscosity resin-based composite (LVC) liner on the microleakage of cervical resin-based composites bonded with new adhesive systems. METHODS: 180 wedge-shaped cervical cavities on bovine incisors were treated with AQ Bond (all-in-one), ABF (experimental two-step self-etching primer containing antibacterial monomer) or Clearfil SE Bond (two-step self-etching primer) systems according to the manufacturers' instructions. Half of the treated cavities in each adhesive system were lined with an LVC (Protect Liner F) and filled with a hybrid resin-based composite (Clearfil AP-X). Ten specimens from each group were immersed in 0.5% basic fuchsin solution immediately after finishing the restorations. The specimens to be subjected to thermocycling (5-60 degrees C, 15 seconds dwell time, 5,000 cycles) or to flexural load cycling (approximately 0.5 mm labio-lingual displacement at the incisal edge, 10,000 cycles, 1 Hz) prior to immersion in the dye solution were also prepared. The teeth were sectioned through the center of the restorations and the degrees of dye penetration were scored. The data were analyzed with the Mann-Whitney U-test or the Kruskal-Wallis test (P < 0.05). RESULTS: The use of the LVC liner significantly improved marginal sealing of AQ Bond. However, the LVC liner could not prevent the deterioration in the marginal integrity caused by thermocycling or flexural load cycling. In the case of ABF, the use of the LVC liner showed a trend toward less microleakage. For Clearfil SE Bond, the LVC liner prevented the deterioration in the marginal seal caused by thermocycling.

Animals↗