Inhibition of enamel demineralization by an ion-releasing tooth-coating material.

PURPOSE: To evaluate the inhibitory effect of a surface pre-reacted glass-ionomer (S-PRG) filler-containing tooth-coating material on enamel demineralization. The outer surface of the S-PRG filler is in a state in which ions are readily released. METHODS: Human enamel blocks were incubated in lactic acid solution (pH 4.0) with and without a disk (n=6) made of the cured tooth-coating material. Test solutions were changed every 24 hours and incubation was continued for 4 days. The pH and amount of fluoride released were measured with an electrode and ion meter, respectively. The concentrations of ions (aluminum, boron, calcium, phosphorus, silicon, sodium, and strontium) were measured by inductively coupled plasma atomic emission spectroscopy. The surface of the enamel block was observed with a scanning electron microscope. RESULTS: Enamel demineralization was not observed in an enamel block incubated with a disk of the tooth-coating material. Ions released from S-PRG filler had an acid buffering action in the low pH lactic acid solution. However, in the enamel block-only solution showing high levels of calcium ion release, the degree of demineralization was correlated with morphological changes of the enamel surface. CLINICAL SIGNIFICANCE: Due to the buffering effects of the pre-reacted glass-ionomer surface by ion release, the S-PRG filler-containing tooth-coating material inhibited enamel demineralization by neutralizing the acidic environment at an early time point.

Inhibition of enamel demineralization by buffering effect of S-PRG filler-containing dental sealant.

The buffering capacity and inhibitory effects on enamel demineralization of two commercially available dental sealants were evaluated in this study. The effects of filler particles were also examined. Disks of enamel and cured sealant materials of BeautiSealant (silica or S-PRG filler) or Teethmate F-1 were incubated in lactic acid solutions (pH 4.0) for 1-6 d. The pH changes and amounts of ions released in the solutions were assessed, and enamel surfaces were observed using a scanning electron microscope. The pH of the solution with BeautiSealant (S-PRG filler) was neutralized from pH 4.0 to pH 6.1 (after incubation for 1 d) and from pH 4.0 to pH 6.7 (after incubation for 6 d). In addition, no release of calcium ions was detected and the enamel surface was morphologically intact in scanning electron microscopy images. However, the pH of the solution with Teethmate F-1 remained below pH 4.0 during incubation from days 1 to 6. Calcium release was increased in solutions up to and after 6 d of incubation. Scanning electron microscopy images showed that the structures of hydroxyapatite rods were exposed at the specimen surfaces as a result of demineralization. Ions released from S-PRG filler-containing dental sealant rapidly buffered the lactic acid solution and inhibited enamel demineralization.

Bioactive Self-Polymerizing Resin with Surface Pre-Reacted Glass Ionomer Fillers for Suppressed Enamel Demineralization.

The treatment of damaged enamel surfaces involves modification of the enamel surface with artificial materials or the development of a pseudo-enamel, with research focusing on bioactive and biomimetic materials. In this study, a bioactive auto-polymerizing resin (APR) was developed by adding surface-pre-reacted glass ionomer (S-PRG) fillers of different quantities to APR. Its bioactive effects were evaluated via pH neutralization, ion release, and inhibition of enamel demineralization studies. The pH and fluoride ion release were measured using ion-specific electrodes, revealing that the APR disk with the S-PRG filler immediately neutralized the lactic acid solution (pH 4.0) through ion release. Inductively coupled plasma atomic emission spectrometry revealed that the Sr ion release peaked on the first day, with the other ions following the order F > B > Si > Al > Na, exhibiting a weekly decrease in the same order. Scanning electron microscopy was used to examine the enamel block morphology of the disks after 7 d of incubation, revealing enamel demineralization in disks without the S-PRG filler, whereas no demineralization occurred in disks with the S-PRG filler. APR containing the S-PRG filler demonstrated acid buffering suppressed enamel demineralization and bioactive properties.

Protective Effects of GIC and S-PRG Filler Restoratives on Demineralization of Bovine Enamel in Lactic Acid Solution.

This study was aimed at investigating the protective effects of glass ionomer cement (GIC) and surface pre-reacted glass ionomer (S-PRG) fillers used as dental restorative materials on demineralization of bovine enamel. GlasIonomer FX ULTRA (FXU), Fuji IX GP Extra (FIXE), CAREDYNE RESTORE (CDR) were used as GICs. PRG Barrier Coat (BC) was used as the S-PRG filler. They were incubated in a lactic acid solution (pH = 4.0) for six days at a temperature of 37 °C. The mineral was etched from the enamel surface, and a large number of Ca and P ions were detected in solution. The Al, F, Na, Sr, and Sr ions were released in GICs and S-RPG fillers. The Zn ion was released only in CDR and the B ion was released only in BC. The presence of apparent enamel prism peripheries was observed after six days of treatment for the group containing only enamel blocks. pH values for the FXU, FIXE, CDR, BC, and enamel block groups after six days were 6.5, 6.6, 6.7, 5.9, and 5.1, respectively. Therefore, the observed pH neutralization effect suppressed progression of caries due to the release of several ions from the restoratives.

Crystal growth by fluoridated adhesive resins.

OBJECTIVE: This investigation was carried out to evaluate the long-term effects of fluoride-releasing adhesive resins on structural changes in standardized fluid-filled gaps simulating microleakage between the materials and the tooth surface in vitro. METHODS: Three commercially available fluoride-releasing resin adhesives (One-Up Bond F, OptiBond Solo, and Reactmer Bond) were used in this study. Cured disks of resin adhesive were placed over flat human tooth surfaces (enamel and dentin), separated by a standardized 40microm interfacial gap and stored in distilled water for 24h (control group) or 1000 days (experimental group). After 1000 days of water storage, the resins were detached from the teeth and the opposing surfaces were examined by scanning electron microscopy (SEM). In addition, chemical structural analysis was performed by laser-Raman spectroscopy. RESULTS: The SEM microphotographs showed numerous crystal types on the enamel, dentin, and resin surfaces after 1000 days of water storage for OptiBond Solo and Reactmer Bond. However, there was no crystal formation in the control specimens and the aged specimens of One-Up Bond F. Raman analysis showed several peaks (463, 618, and 990cm(-1)) from the crystals of OptiBond Solo that were not identified in the enamel, dentin, or cured resin. SIGNIFICANCE: In conclusion, two of the three tested fluoride-release resin adhesives (OptiBond Solo and Reactmer Bond) have the ability to induce crystal growth within gaps between the adhesive and teeth in long-term water storage. These results suggest that the two adhesive resins have self-reparative ability with regard to bond leakage.

Effect of water on bonding of one-bottle self-etching adhesives.

This study evaluated the effect of water on dentin substrate bonding of one-bottle self-etching adhesives. Dentin substrates were divided into two groups: wet and dry dentin. Wet dentin is the normal substrate for bond testing whereas dry dentin was dehydrated in a desiccator for 24 hours. Bonded dehydrated teeth were then divided into two subgroups: stored in water or in desiccator for 24 hours. Microtensile bond strength of resin to dentin was measured using three one-bottle self-etching adhesives. In addition, nanoleakage evaluation was performed through the analyses of SEM and TEM micrographs. The bond strength of dry-dentin group was significantly greater than that of wet-dentin. Further, the amount of nanoleakage within the adhesive interface of dry-dentin group was less than that of wet-dentin. Results showed that bond strength and nanoleakage formation depended on the bonding substrate (wet versus dry dentin) before bonding. One-bottle self-etching adhesives might suck the water from dentinal tubules during bonding by osmosis, leading to nanoleakage formation and thus a decline in bond strength.

In vitro durability of one-bottle resin adhesives bonded to dentin.

Recently, one-bottle resins adhesives have been developed to reduce the number of clinical steps of resin application. They are now widely used in clinical dental practice. However, little is known regarding the detailed mechanism of bond degradation. Therefore, this study evaluated the durability of one-bottle resin adhesives using long-term water storage testing. Resin-dentin bonded specimens were prepared using five commercially available one-bottle resin adhesives. The specimens were sectioned perpendicular to the adhesive interface to produce beams and stored in distilled water for 24 hours (control), 100, 200, and 300 days. After the water storage, each beam was subjected to a microtensile bond test and then SEM fractography was performed on the fractured surface. Compared to the bond strength at 24 hours after bonding (control), the bond strength of all tested adhesives were significantly decreased after 100 or more days in water. SEM fractography revealed a typical type of deterioration in the adhesive-composite interface that might cause a decline in bond strength after aging.

Diffusion-induced water movement within resin-dentin bonds during bonding.

It is thought that water-filled channels and nanovoids in resin-dentin bonds represent potential sites for degradation of bonds or hydrolysis of collagen or both. How water gains access to bonded interfaces is not clear. This study evaluated the diffusion-induced water uptake through resin-dentin interfaces during bonding. Two light-cured total-etch adhesive systems (Excite and One-Step Plus) and a chemical-cured adhesive (Amalgambond Plus) were used in this study. Dentin disks were placed in a split-chamber device, and the fluid movement across dentin was measured, with and without a physiological pressure, during bonding procedures and 24 h after bonding. For light-cured adhesives in the experimental groups, a 6 min interval of dark storage was conducted prior to light-curing, to evaluate the diffusion of water through the uncured resin monomers, and to test the effect of prolonged infiltration time of adhesives on water permeability of bonds. Prolonged adhesive infiltration reduced the water permeability of resin-dentin bonds for light-cured adhesives. Water gradients produced by bonding systems contribute to water movement across the dentin-adhesive interfaces during bonding procedures. Differences in chemical formulations for adhesive systems may lead to differences in the extent of diffusion-induced water movement and the amount of water within the resin-dentin bonds.

The effects of common errors on sealing ability of total-etch adhesives.

OBJECTIVES: This study evaluated the effect of errors commonly made in using total-etch adhesives, on the resulting bond strength, fluid movement and nanoleakage of resin dentin bonds. METHODS: Two total-etch adhesives were used for bonding to dentin according to the manufacturers' recommendations, with meticulous solvent evaporation (control), or with the introduction of common bonding errors-wet bonding without solvent evaporation (no evaporation), and dry bonding. RESULTS: The 24-hour bond strength of the control was significantly higher than the other groups (p <0.05). For all groups, the higher initial permeability declined significantly after 24 h. The fluid movement across bonded dentin was similar in the control and dry bonding for both adhesives, whereas significantly higher permeability (p<0.05) was recorded for the no evaporation groups even after 24 h. Extensive silver impregnation within hybrid layers was seen by TEM in the no evaporation and dry bonding specimens after 24 h. Dry bonding caused collapse of the collagen matrix and interfered with resin infiltration. In contrast, inadequate solvent evaporation and/or residual water during dentin bonding results in dilution or incomplete polymerization of the resin, leading to severe nanoleakage formation. SIGNIFICANCE: Increased permeability associated with incomplete solvent evaporation in total-etch adhesives may lead to poor bond strength.

Permeability of adhesive resin films.

The purpose of this study was to quantify the permeability of adhesive resin films to water by measuring convective flow across thin films. Cured resin films were prepared with the use of five commercially available adhesive resins and an experimental resin. Two types of resin films were prepared from solvated comonomer blends within each product; resin films made before or after evaporation of solvent (10 s of drying with air-syringe). The permeability of the resin films was measured 30 min or 24 h after polymerization by placing the films in a split-chamber device. Fluid filtration rate through the resin films was measured with the use of 20 cm of water pressure. Osmotically induced water movement was measured by applying hypertonic aqueous solutions of CaCl2 or HEMA to the resin films. The results showed that evaporation of solvent before polymerization reduced the permeability of adhesive resin films, compared to permeabilities obtained without solvent evaporation. The cured adhesive resin films were all permeable to water, but to varying degrees.

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.

Secretion of macrophage migration inhibitory factor differs from interleukin-6 in hydrogen peroxide- and LPS-stimulated human fibroblasts.

To investigate the mechanism for secretion of macrophage migration inhibitory factor (MIF) in cultured human fibroblasts, we compared it with the secretion of interleukin-6 (IL-6) after stimulation with lipopolysaccharides (LPS) and H2O2. MIF content of the medium of 2.0 x 10(6) cells/20 ml after 20 h culture of nonstimulated fibroblasts was 0.30 +/- 0.06 ng/ml, whereas LPS-stimulation (10 microg/ml) only led to a 1.5-fold increase as compared with the nonstimulated cells. In contrast, a significant increase of IL-6 was induced by LPS-stimulation (6048 +/- 488 pg/ml in LPS-stimulated cells vs. 58 +/- 36 pg/ml in control cells). On the other hand, higher concentrations of H2O2 (0.6-1.2 mM) caused an increase of MIF secretion into the culture medium irrespective of LPS-stimulation; with 1.2 mM H2O2-stimulation for 20 h, it was increased to 40-fold as compared with the nonstimulated cells. However, lower concentrations (0.1-0.4 mM) did not cause this. Interestingly, H2O2-stimulation not only failed to increase IL-6 production from fibroblasts, but also repressed induction of IL-6 by LPS-stimulation in a dose-dependent manner. Genistein, a tyrosine kinase inhibitor, and H-7, a protein kinase C inhibitor, also inhibited IL-6 secretion but not MIF secretion in both LPS- and H2O2-stimulated fibroblasts. From analysis of trypan blue exclusion, formazan formation, morphological changes, and intracellular MIF content by Western blotting, we found that MIF secretion by H2O2 seemed to be mainly due to cell death and subsequent leakage of intracellular MIF. Taken together, these results suggest that MIF secretion differs from IL-6 via LPS-mediated signaling pathways.

Degradation patterns of different adhesives and bonding procedures.

Various types of resin adhesives and procedures are available in the clinical field, so comprehensive understanding of degradation is required for each material and bonding procedure. The objective of this study was to investigate the bond durability for different adhesives and bonding procedures. Resin-dentin bonded beams were prepared with the use of two adhesives (One-Up Bond F/self-etching primer system and One Bond/total-etch adhesive) and two experimental groups for the bonding procedure (wet and dry bonding of the total-etch adhesive). Those samples were soaked in water for 24 h(control), 6 and 12 months. After the water immersion, the bond strengths were measured by the microtensile bond test, and subsequently fractography was performed with the use of SEM. Statistically significant reduction of the bond strength (p < 0.05) was apparent after 12 months of water exposure in the range 22-48% of the control. The bonding resin was eluted from the hybrid layer of the self-etching and the total-etch adhesives for the wet bonding. Micromorphological alterations were found due to the hydrolysis of collagen fibrils with the total-etch adhesive for the dry bonding mode. These pathologic alterations were in accord with the bond strength.

Deproteinizing effects on resin-tooth bond structures.

This study investigated the effects of NaOCl on resin-tooth bonds to simulate the situations of long-term durability and caries invasion. Resin-tooth bonded specimens were produced with the use of two resin adhesives (Excite and One-Bond). Resin-tooth bonded beams (adhesive area; 0.9 mm2) were serially sectioned and the specimens were immersed in 10% NaOCl medium for 0 (control), 2, 4, and 6 h after being stored in water for 24 h. After immersion, microtensile bond tests were performed. SEM fractography was conducted to calculate each failure mode by image analysis. In addition, the adhesive interface was examined with the use of TEM. In the control specimens, enamel bond strengths had no difference between Excite (45.6 +/- 15.0) and One-Bond (56.9 +/- 12.9). On the other hand, dentin bond strengths had significant difference between Excite (80.6 +/- 21.2) and One-Bond (50.7 +/- 11.2). The bond strengths decreased with increased storage time for both systems with enamel and dentin bonds. The deteriorated mineralized dentin of beams resulted in bond-strength reduction for resin-enamel bonds. For dentin bonding, the adhesive interface was gradually dissolved from the outer to the center portion of the beam. The depletion of collagen fibrils within the demineralized dentin or hybrid layer deformation was found under SEM and TEM examinations. These morphological changes are responsible for bond strength reduction of resin-dentin bonds.

SEM and TEM analysis of water degradation of human dentinal collagen.

Recently several long-term studies have reported evidence of the hydrolytic degradation of collagen fibrils based on fractured surface observations after bond testing. Those studies suggested that one cause of the decline in the bond strength was the degradation of the collagen fibrils within the bonds. However, one concern has been raised that the dentinal collagen fibrils may be stable in water that does not contain oral bacteria or enzymes. Therefore, the present study aimed to clarify the micromorphological change in naked collagen fibrils after 500 days of water storage. To prepare exposed collagen fibrils, sectioned and polished human dentin surfaces were acid conditioned for 15 s with the use of two commercially available acid conditioners: All-Etch (10% phosphoric acid) and Uni-Etch (32% phosphoric acid) (Bisco, Inc.). Those specimens were stored in distilled water at 37 degrees C for 1 day (control) for 500 days. After the storage periods, the samples were examined with the use of SEM and TEM. Under SEM and TEM examination, micromorphological alterations (disarrangement of collagen web, widening the interfibrillar space, and the thinning diameter of collagen fibrils) were found in the specimens after 500 days in water.

Sublethal, 2-week exposures of dental material components alter TNF-alpha secretion of THP-1 monocytes.

OBJECTIVES: The aim of this study was to investigate the hypothesis that dental material components alter cytokine secretion from monocytes if applied for several weeks at sublethal doses. The current study significantly extended exposure times of monocytes to the components over times published in previous studies. These exposure times approached the estimated average life span of monocytes in the bloodstream. METHODS: Human THP-1 monocytes were exposed to 2-hydroxyethylmethacrylate (HEMA, 0-1.2mmol/l), triethyleneglycoldimethacrylate (TEGDMA, 0-0.75mmol/l), Hg(2+) (0-2 micromol/l), or Ni(2+) (0-20 micromol/l) for 2 weeks. The cells were then collected and additionally incubated for 24h, with or without bacterial lipopolysaccharide (LPS), a common component of dental plaque. TNF-alpha secretion from THP-1 was determined using by enzyme-linked immunosorbent assay. RESULTS: None of the dental material components induced TNF-alpha from THP-1 by themselves, but LPS alone strongly induced TNF-alpha secretion as expected. HEMA and TEGDMA significantly suppressed (40-70%) TNF-alpha secretion from cells stimulated with LPS. Hg(2+) at 2.0 micromol/l doubled TNF-alpha secretion from THP-1s stimulated with LPS over LPS alone. Ni(2+) did not significantly affect TNF-alpha secretion, with or without LPS exposure. Significance. The results in this study suggest that sublethal, 2-week exposures of some dental material components may alter TNF-alpha secretion from THP-1 monocytes when the cells are challenged. These alterations may influence the biological response of tissues to materials in an inflammatory intraoral environment.

Effects of multiple adhesive coatings on dentin bonding.

Simple changes to bonding techniques can improve resin-dentin bond strengths. This study evaluated the effect of multiple consecutive coatings of adhesive resin on dentin by measuring both microtensile bond strength and nano-leakage following exposure to ammoniacal silver nitrate. Resin-dentin bonded specimens were prepared using two total-etch adhesives (OptiBond Solo Plus/Kerr or Single Bond/3M ESPE). During bonding, resin application and air evaporation were done 1, 2, 3, 4, 6 or 8 times on acid-etched, moist dentin surfaces. Mean microtensile bond strengths were evaluated by two-way ANOVA and Fisher's PLSD test (p<0.05; n=16 for each group). Additionally, nanoleakage of silver nitrate was evaluated by transmission electron microscopy (TEM). The results indicated that bond strengths increased with each coating up to four coats. Nanoleakage decreased with each coat, becoming very small after four or more coats. This adhesive application method can be easily applied to clinical practice, thereby improving the quality of resin-dentin bonds.

In vitro testing of all-in-one adhesives as fissure sealants.

PURPOSE: To evaluate the clinical efficacy of all-in-one adhesives as fissure sealants. METHODS: Resin-enamel bonded specimens were fabricated between polished and ground enamel surfaces using two all-in-one adhesives (One-Up Bond F and Xeno CF Bond) and two resin sealants (Teethmate F-1 and Helioseal F). Specimen beams were prepared and microtensile bond tests were performed at a crosshead speed of 1 mm/minute. The bond strengths obtained were subjected to one-way ANOVA and Scheffe's F test (P < 0.05: n=12 for each group). To evaluate the etching effect of phosphoric acid and the all-in-one adhesives, the treated enamel surfaces (37% phosphoric acid, 40% phosphoric acid, or all-in-one adhesives) were examined by FE-SEM. RESULTS: The bond strengths of the two all-in-one adhesives tested to the polished enamel surface were significantly lower (P < 0.05) than to ground enamel. However, no significant differences (P > 0.05) were found between the polished and ground enamel surfaces for resin sealants using phosphoric acid. Under SEM examination, 37% or 40% phosphoric acid etched the enamel surface more than all-in-one adhesives.

Responses of RAW264.7 macrophages to water-dispersible gold and silver nanoparticles stabilized by metal-carbon σ-bonds.

Nanometals are currently receiving considerable attention for industrial and biomedical applications, but their potentially hazardous and toxic effects have not been extensively studied. This study evaluated the biological responses of novel water-dispersible gold (Au-NPs) and silver nanoparticles (Ag-NPs) stabilized by Au-C or Ag-C σ-bonds in cultured macrophages (RAW264.7), via analysis of the cell viability, the integrity of the plasma membrane, and the inflammatory and morphological properties. The cultured RAW264.7 was exposed to metal-NPs at various concentrations. The Ag-NPs showed cytotoxicity at high NP concentrations, but the cytotoxic effects of the Au-NPs were smaller than those of the Ag-NPs. For the microscopic analysis, both types of particles were internalized into cells, the morphological changes in the cells which manifested as an expansion of the vesicles' volume, were smaller for the Au-NPs compared with the Ag-NPs. For the Ag-NPs, the endocytosis abilities of the macrophages might have induced harmful effects, because of the expansion of the cell vesicles. Although an inflammatory response was observed for both the Au- and Ag-NPs, the harmful effects of the Au-NPs were smaller than those of the Ag-NPs, with minor morphological changes observed even after internalization of the NPs into the cells.

Micromorphological cellular responses of MC3T3-E1 and RAW264.7 after exposure to water-dispersible silver nanoparticles stabilized by metal-carbon σ-bonds.

With the continuous progress in nanomaterial development for biomedicine, the potential cytotoxicity of nanoparticles is drawing more attention and concern for clinical applications. The purpose of this study was to evaluate biological responses of new water-dispersible silver nanoparticles (Ag-NPs) stabilized by Ag-C σ-bonds in cultured murine macrophages (RAW264.7) and osteoblast-like cells (MC3T3-E1) using cell viability and morphological analyses. For RAW264.7, Ag-NPs seemed to induce cytotoxicity that was dependent on the Ag-NP concentration. However, no cytotoxic effects were observed in the MC3T3-E1 cell line. In microscopic analysis, Ag-NPs were taken up by MC3T3-E1 cells with only minor cell morphological changes, in contrast to RAW264.7 cells, in which particles aggregated in the cytoplasm and vesicles. The ability of endocytosis of macrophages may induce harmful effects due to expansion of cell vesicles compared to osteoblast-like cells with their lower uptake of Ag-NPs.