Masanori Hashimoto

Collagen Degradation by Host-derived Enzymes during Aging

Incompletely infiltrated collagen fibrils in acid-etched dentin are susceptible to degradation. We hypothesize that degradation can occur in the absence of bacteria. Partially demineralized collagen matrices (DCMs) prepared from human dentin were stored in artificial saliva. Control specimens were stored in artificial saliva containing proteolytic enzyme inhibitors, or pure mineral oil. We retrieved them at 24 hrs, 90 and 250 days to examine the extent of degradation of DCM. In the 24-hour experimental and 90- and 250-day control specimens, we observed 5- to 6-μm-thick layers of DCM containing banded collagen fibrils. DCMs were almost completely destroyed in the 250-day experimental specimens, but not when incubated with enzyme inhibitors or mineral oil. Functional enzyme analysis of dentin powder revealed low levels of collagenolytic activity that was inhibited by protease inhibitors or 0.2% chlorhexidine. We hypothesize that collagen degradation occurred over time, via host-derived matrix metalloproteinases that are released slowly over time.

In vivo Degradation of Resin-Dentin Bonds in Humans Over 1 to 3 Years

The longevity of resin restorations is currently an area of great interest in adhesive dentistry. However, no work has been conducted to investigate the durability of resin-dentin bond structures using human substrate in vivo. The purpose of this study was to investigate the degradation of the resin-dentin bond structures aged in an oral environment for 1, 2, or 3 years. Cavities were prepared in primary molars, and an adhesive resin system (Scotchbond Multi-Purpose) was applied to the cavity. After I to 3 years, following the eruption of the succedaneous permanent teeth, the resin-restored teeth were extracted. Immediately after extraction, those teeth were sectioned perpendicular to the adhesive interface and trimmed to produce an hourglass-shaped specimen. Then, a micro-tensile test was performed at a crosshead speed of 1.0 mm/min. The mean bond strengths were statistically compared with one-way ANOVA and Fishers PLSD test (p < 0.05). Further, all fractured surfaces were observed by SEM, and the area fraction of failure mode was calculated by means of a digital analyzer on SEM photomicrographs. There were significant differences in tensile-bond strength among all 3 groups (p < 0.05), with mean values ranging from 28.3 ± 11.3 MPa (control), to 15.2 ± 4.4 MPa (1 to 2 years), to 9.1 ± 5.1 MPa (2 to 3 years). Moreover, under fractographic analysis, the proportion of demineralized dentin at the fractured surface in specimens aged in an oral environment was greater than that in control specimens. Furthermore, degradation of resin composite and the depletion of collagen fibrils was observed among the specimens aged in an oral environment. Analysis of the results of this study indicated that the degradation of resin-dentin bond structures occurs after aging in the oral cavity.

In vitro degradation of resin–dentin bonds analyzed by microtensile bond test, scanning and transmission electron microscopy

Our knowledge of the mechanisms responsible for the degradation of resin-dentin bonds are poorly understood. This study investigated the degradation of resin-dentin bonds after 1 year immersion in water. Resin-dentin beams (adhesive area: 0.9mm(2)) were made by bonding using a resin adhesive, to extracted human teeth. The experimental beams were stored in water for 1 year. Beams that had been stored in water for 24h were used as controls. After water storage, the beams were subjected to microtensile bond testing. The dentin side of the fractured surface was observed using FE-SEM. Subsequently, these fractured beams were embedded in epoxy resin and examined by TEM. The bond strength of the control specimens (40.3+/-15.1MPa) decreased significantly (p<0.01) after 1 year of water exposure (13.3+/-5.6MPa). Loss of resin was observed within fractured hybrid layers in the 1 year specimens but not in the controls. Transmission electron microscopic examination revealed the presence of micromorphological alterations in the collagen fibrils after 1 year of water storage. These micromorphological changes (resin elution and alteration of the collagen fibrils) seem to be responsible for the bond degradation leading to bond strength reduction.

Aging Affects Two Modes of Nanoleakage Expression in Bonded Dentin

Water sorption into resin-dentin interfaces precedes hydrolytic degradation. We hypothesized that these processes are morphologically manifested by the uptake of ammoniacal silver nitrate, which is thought to trace hydrophilic domains and water-filled channels within matrices. Water sorption is thought to be nonuniform and can be traced by the use of silver nitrate. Human teeth bonded with an experimental filled-adhesive were aged in artificial saliva (experimental) or non-aqueous mineral oil (control). Specimens retrieved for up to a 12-month period were immersed in 50 wt% ammoniacal silver nitrate and examined by transmission electron microscopy for identification of the changes in their silver uptake. Reticular silver deposits initially identified within the bulk of hybrid layers in the experimental group were gradually reduced over time, but were subsequently replaced by similar deposits that were located along the hybrid layer-adhesive interface. Silver uptake in water-binding domains of the adhesive layers increased with aging, resulting in water tree formation. These water-filled channels may act as potential sites for hydrolytic degradation of resin-dentin bonds.

Fluid Movement across the Resin-Dentin Interface during and after Bonding

This study evaluated the extent of water penetration through resin-dentin interfaces before and after being sealed with adhesives. Four adhesive resin systems (2 total-etch adhesives and 2 self-etching primer adhesives) were used in this study. Dentin disks were placed in a split-chamber device, and in situ fluid movement across dentin was measured, with and without physiological pressure, during bonding procedures or 24 hrs after bonding. The fluid movement across dentin occurs via dentin tubules after acid-etching. Large outward or inward fluid shifts across dentin were observed during air-drying and light-curing for resin application. The amount of fluid movement across resin-bonded dentin when total-etch adhesives were used was significantly greater than that with self-etching adhesives. The milder acid-etching effects of self-etching primers may retain hybridized smear plugs within the tubules that reduce outward fluid flow, resulting in superior dentin sealing.

The effect of hybrid layer thickness on bond strength: demineralized dentin zone of the hybrid layer.

OBJECTIVES The purpose of this study was to evaluate the correlation between hybrid layer thickness and bond strength using specimens acid-conditioned for varying lengths of time. METHODS The dentin surfaces of human premolars, sectioned to remove the enamel from the labial surface, were conditioned with 35.0% phosphoric acid of an adhesive resin system (Scotchbond Multi-Purpose; 3M) for 15 (as directed by the manufacturer), 60, 120, or 180 s (experimental acid-conditioning times). The bonded specimens were then sectioned perpendicular to the adhesive interface to measure the hybrid layer thickness by SEM. The specimens for the micro-tensile test were sectioned perpendicular to the adhesive interface and trimmed to an hourglass-shape. Then, the micro-tensile test was performed at a crosshead speed of 1.0 mm/min. The bond strengths and hybrid layer thickness were statistically compared with Students t-test (p < 0.05). All fractured surfaces were also observed by SEM. RESULTS Significant differences between the groups exposed to acid for 15 and 60 s, and those exposed for 120 and 180 s were observed in hybrid layer thickness and bond strength (p < 0.05). SEM observation of the fractured surfaces revealed that a demineralized dentin zone without resin impregnation remained within the hybrid layer. SIGNIFICANCE A demineralized dentin zone was formed in the bond structures after prolonged acid-conditioning, resulting in low bond strength. The shrinkage of the hybrid layer due to desiccation during the SEM examination process provided evidence of the presence of the demineralized dentin zone within the hybrid layer.

The Extent to which Resin can Infiltrate Dentin by Acetone-based Adhesives

The combined methodologies of fractography and laser-Raman spectroscopic analysis were used for evaluation of the resin-dentin bonds made with wet and dry bonding. Resin-dentin-bonded beams were produced by means of 2 acetone-based adhesives (One-Step and Prime & Bond NT). The micro-tensile bond test was conducted, and the fractured surfaces of all specimens were examined by SEM and an image analyzer. The amount of resin infiltration within the hybrid layer was quantified by means of a laser-Raman spectroscope. In Raman analysis, the amount of resin impregnation within the hybrid layer of the dry bonding was found to be significantly lower (approximately 50%) than that in the wet one. Under fractographic analysis, a correlation was found between the bond strength and the failure mode. Based on those findings, it was suggested that the integrity between the bonding resin and the top of the hybrid layer played a major role in bond strength.

Over-etching effects on micro-tensile bond strength and failure patterns for two dentin bonding systems

OBJECTIVES The purpose of this study was to determine (1) the weakest zone of resin-dentin bonds and (2) the relation between bond strength and failure mode to clarify the effect of demineralized dentin. METHODS Human premolars were sectioned to expose the dentin surfaces, and the dentin surfaces were conditioned with phosphoric acid for 15, 60, 120, or 180s. Resin-dentin bonded specimens were produced using two adhesives: One-Step (Bisco) and OptiBond Solo (Kerr). Each sample was sectioned to produce a beam (adhesive area: 0.9mm(2)). Microtensile bond tests were then conducted, and the mean bond strengths (n=12 for each group) were statistically compared using two-way ANOVA and Duncans multiple-range test (p<0.05). The fractured surfaces of all specimens were examined using SEM, and the areas of failure were measured using an image analyzer. RESULTS For One-Step, the bond strength decreased with increase in acid-conditioning time (15s: 50.7+/-9.7, 60s: 40.8+/-11.0, 120s: 23.6+/-4.9 and 180s: 12.1+/-4.6MPa) (p<0.05). For OptiBond Solo, the bond strength in the case of 15s acid-conditioning time (42.6+/-7.9MPa) was significantly greater than that for the other times (60s: 31.9+/-10.3, 120s: 31.8+/-14.4 and 180s: 31.8+/-7.4MPa) (p<0.05). Fractography showed that the area percentage of the hybrid layer increased with increase in etching time for both systems. CONCLUSIONS The integrity of the hybrid layer, especially the top part, has an effect on bond strength.

Kinetic studies on the CO oxidation on a Rh(111) surface by means of angle-resolved thermal desorption

The kinetics of the reaction of adsorbed CO with oxygen adatoms was studied in the temperature range of 100–600 K with LEED and angle‐resolved thermal desorption. At small oxygen coverages the CO2 formation peaked from 500 to 400 K with increasing CO exposure. The activation energy decreased from 45 to 35 kcal/mol. When the oxygen coverage was large, a new CO2 formation peak appeared around 400 K. The activation energy was 30 kcal/mol. LEED observations revealed that the surface was covered by separate domains of CO and oxygen. The former CO2 is produced outside the domains or on the perimeters, whereas the latter is formed in the oxygen domains. The angular distribution of the desorption of CO2 in the former varied as cos4 θ, where θ is the desorption angle. The desorption of CO2 in the latter showed a sharper angular distribution of cos15 θ.

The Glass-ionomer Phase in Resin-based Restorative Materials

Glass-ionomer (GI) fillers are added to restorative materials, but it is unclear if they truly react with these materials. This TEM study evaluated the existence of the GI phase in a conventional GIC (ChemFlex), a resin-modified GIC (Fuji II LC), a giomer (Reactmer Paste), a compomer (Dyract AP), and a composite (Spectrum TPH), before and after water uptake. Wafers were stored at 100% RH for 24 hrs, or in water for 7 or 84 days. ChemFlex glass particles were surrounded by 300-nm-thick silica gel layers. In Fuji II LC, we found thinner hydrogel layers (100 nm) that became thicker upon water storage. No appreciable change occurred in Reactmer Paste. Only a very thin hydrogel layer occurred in Dyract AP, and none was seen in SpectrumTPH after water storage for 84 days. We conclude that the variable extent of the GI phase is determined by differences in the resin composition of the restoratives.