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.

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.

Immobilization induces acute nitric oxide production in the rat hypothalamus: a role of ionotropic glutamate receptors in the paraventricular nucleus.

Production of nitric oxide (NO) in the hypothalamic paraventricular nucleus (PVN) was examined by microdialysis in rats subjected to immobilization (IMO) stress. A dialysis probe was implanted in the posterior magnocellular subdivision of the PVN and nitrite (NO(2)(-)), an oxidized product of NO, was measured continuously. NO(2)(-) concentration in dialysate was enhanced to 156% after 30 min of IMO compared with the NO(2)(-) level before IMO. Intraperitoneal administration of N(G)-monomethyl-l-arginine (10 mg/kg), a NO synthase inhibitor, before IMO completely inhibited the increase of NO production that IMO was to induce. Depletion of catecholamines innervating the PVN by an injection of 6-hydroxydopamine into the lateral ventricle before the microdialysis had no suppressive effect on the increase of NO production by IMO. In contrast, NO(2)(-) levels in the PVN were lowered by continuous perfusion of the solution containing the ionotropic glutamate receptor antagonists 2-amino-5-phosphonovaleric acid (500 microm) and 6-cyano-7-nitroquinoxaline-2, 3 dione (50 microm) through the dialysis probe, and the IMO-induced increase of NO production was attenuated by the treatment. These results suggest that catecholaminergic drive to the hypothalamus is not necessary for the IMO-induced increase of NO production and that ionotropic glutamate receptors play a role in the basal and IMO-induced NO production.

Neuronal expression of catechol O-methyltransferase mRNA in neonatal rat suprachiasmatic nucleus.

We examined the expression profile of catechol O-methyltransferase (COMT) mRNA and its protein in the neonatal rat hypothalamus by in situ hybridization and immunohistochemistry to clarify the sites of dopamine degradation. Strong COMT mRNA expression was observed in the suprachiasmatic nucleus (SCN) throughout its rostrocaudal extent at postnatal day 1 (P1) and P2, and the mRNA levels decreased gradually until P16. COMT mRNA was predominantly localized to the ventral and medial parts of the SCN. Intense COMT immunoreactivity was demonstrated in the ventral SCN and was detected in neuronal perikarya and processes at P1. Ependymal and microglial cells also exhibited strong COMT immunoreactivity. These results indicate that COMT may directly be involved in dopaminergic signaling in the neonatal SCN.

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.

Changes in occlusal contact area and average bite pressure during treatment of anterior crossbite in primary dentition.

The purpose of this study was to examine changes in functional parameters of patients with anterior crossbite in primary dentition during orthodontic treatment. Occlusal contact area (OCA), average bite pressure (ABP) and integrated occlusal load (IOL) were measured. Data obtained before the start of treatment (period A), data obtained when crossbite had improved (period B) and data obtained when the appliance had been removed (period C) were compared. OCA showed the lowest value in period B, and then gradually increased. ABP increased until period B and then declined or became constant. OCA and IOL showed significant differences in periods A and B and periods B and C (p < 0.05). The results suggest that about 6 months is required for stability of the occlusion and acquisition of function in the new occlusion after improvement of crossbite.

Resin-enamel bonds made with self-etching primers on ground enamel.

Self-etching primer adhesives have recently been introduced to simplify bonding. However, the fundamental bonding mechanism of self-etching primers to enamel is not yet fully understood. This study aimed to investigate resin-enamel bonds of self-etching primer adhesives on ground enamel. Two self-etching primer adhesives (Clearfil Liner Bond 2 V and Clearfil SE Bond) were used in this study. A total-etch adhesive (One-Step) was used as a control. Resin-enamel beams were subjected to the microtensile bond test. Subsequently, the failure modes of all specimens were quantified using image analysis. Undemineralized and demineralized ultrathin sections of the resin-enamel bonded specimens were examined using transmission electron microscopy. The bond strengths of Clearfil SE Bond (39.8 +/- 11.9 MPa) and One-Step (46.2 +/- 12.7 MPa) were significantly greater than that of Clearfil Liner Bond 2V (30.4 +/- 6.2 MPa). Scanning electron microscopy of the fractured surfaces revealed the failure direction and weakest portion within each bond. Transmission electron microscopy showed a thin hybridized complex of resin in enamel produced by the self-etching primers without the usual micrometer-sized resin tags seen in resin-enamel bonds produced using the total-etch adhesive. The morphological features of the resin-enamel bonds produced by two self-etching primers tested were different from that created with the total-etch adhesive.

Histological study of deposited cementum in human deciduous teeth with pathological root resorption.

Physiological root resorption is a characteristic feature of human deciduous teeth. Pathological root resorption due to apical periodontitis, dental trauma or excessive orthodontic force is also observed in deciduous roots. The root resorption is not continuous, and has resting periods. In the resting period, cementum deposits in resorbed root surface. The deposited cementum in permanent teeth has been reported in detail. However, the deposited cementum in deciduous teeth is unclear. The present study examined apices of roots of human deciduous incisors with apical periodontitis and roots of sound deciduous incisors by light and transmission electron microscopy. Root dentin and original cementum had a severe irregular caved surface. Cementum was partially deposited on the caved root surfaces. The deposited cementum had made the caved root surface relatively flat. The cementum was lax and had some defects. The deposited cementum was belt-like in shape and had a stratified structure. Each layer had various structures consisting of abundant microfibrils and fine granular materials, microfibrils, granular materials, and collagen fibrils, a few fibrils and granular materials and a relatively homogeneous structure. The original cementum had many collagen fibrils, such as intrinsic and extrinsic fibers, and no granular materials or homogeneous structure. Therefore, structure of the deposited cementum was very different from that of original cementum in deciduous teeth and from that of deposited cementum in permanent teeth.

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.

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.

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.

Effects of tooth-conditioning agents on bond strength of a resin-modified glass-ionomer sealant to enamel.

OBJECTIVES: The purpose of this study was to evaluate the effects of tooth-conditioning agents on bond strength of resin-modified glass-ionomer sealant (Fuji III LC) to bovine enamel as well as on dissolution of calcium ions from the bovine enamel surfaces. METHODS: The enamel surfaces of bovine lower incisors were treated with 10 and 20% polyacrylic acid, 12% citric acid and 35% phosphoric acid for 20s. Fuji III LC was applied to the etched enamel surfaces, and the shear bond strength of each specimen was measured using an Instron Universal Testing Instrument. The amounts of calcium ions dissolved from the treated enamel surfaces were also measured using a polarized Zeeman atomic absorptiometer. RESULTS: In specimens pretreated with distilled water, 10% polyacrylic acid, 20% polyacrylic acid, 12% citric acid and 35% phosphoric acid, the mean values of shear bond strength were 5.5, 12.5, 15.2, 15.2 and 15.1MPa, respectively, and the amounts of Ca(2+) dissolved from the enamel surfaces were 5.6, 41.4, 88.5, 131.6 and 588.3 microg/cm(2), respectively. CONCLUSIONS: The adhesion of a resin-modified glass-ionomer sealant to bovine enamel was significantly improved by the use of tooth-conditioning agents. Especially, treatment of an enamel surface with 20% polyacrylic acid results in good shear both strength and relatively small degree of enamel erosion.

Expression of receptor activator of NF-kappa B ligand and osteoprotegerin in culture of human periodontal ligament cells.

The receptor activator of NF-kappa B ligand (RANKL) and its decoy receptor, osteoprotegerin (OPG), are the important proteins implicated in osteoclastogenesis. In this study, we investigated the expressions of RANKL and OPG in cultured human periodontal ligament (PDL) cells and their roles in osteoclastogenesis. Northern blotting revealed that the OPG mRNA was down-regulated remarkably by application of 10-8 m one-alpha, 25-dihydroxyvitamin D3[1,25-(OH)2D3] and 10-7 m dexamethasone (Dex). In contrast, RANKL mRNA was up-regulated by the same treatment. Western blotting demonstrated decrease of OPG by the application of 1,25-(OH)2D3 and Dex. Tartrate-resistant acid phosphatase-positive multinuclear cells were markedly induced when the PDL cells were cocultured with mouse bone marrow cells in the presence of an anti-OPG antibody together with 1,25-(OH)2D3 and Dex. These results indicate that PDL cells synthesize both RANKL and OPG and that inactivation of OPG may play a key role in the differentiation of osteoclasts.

Micromorphological changes in resin-dentin bonds after 1 year of water storage.

The purpose of this study was to evaluate the degradation of resin-dentin bonds after 1 year of water storage. Resin-dentin-bonded specimens were prepared with the use of an adhesive resin system (One-Step: Bisco). Half of the experimental specimens were sectioned perpendicular to the adhesive interface to produce a beam (adhesive area: 0.9 mm(2)) before being stored in distilled water at 37 degrees C for 1 year. The remaining half of the bonded specimens were sectioned into beams of similar dimensions after 1 year of water storage. Additional bonded specimens that had been stored in water for 24 h before sectioning into beams were used as controls. The beams in the two experimental groups and the control group were subjected to microtensile bond testing. Fractography was performed on all fractured beams with the use of FE-SEM. There were significant (p <.05) differences in bond strength among the control specimens (55.9 +/- 12.9 MPa), specimens that had been sectioned into beams after water storage (68.9 +/- 18.6 MPa), and specimens that had been sectioned into beams before water storage (28.1 +/- 9.3 MPa). Fractography revealed that the resin material was gradually extracted from the periphery to the center portion of the beam. This probably accounted for the decrease in bond strength after 1 year of water storage.

Early onset of NMDA receptor GluR epsilon 1 (NR2A) expression and its abundant postsynaptic localization in developing motoneurons of the mouse hypoglossal nucleus.

Oro-facial sensorimotor function conducted by the brainstem is vital to newborn mammals, and N-methyl-D-aspartate (NMDA) receptors play an important role in the regulation. Here we examined the expression of NMDA receptor subunits in the mouse hypoglossal nucleus from embryonic day 13 (E13) through postnatal day 21 (P21). Compared with other brainstem regions, early onset of GluRepsilon1 (NR2A) mRNA expression was conspicuous to the embryonic hypoglossal nucleus. The expression peaked at P1-P7, when other brainstem regions just started to express it. At P1, GluRepsilon1 subunit was localized to asymmetrical synapses on motoneuron dendrites, particularly, on the postsynaptic junction membrane. In developing motoneurons, expressions of GluRepsilon2 (NR2B), GluRepsilon4 (NR2D), and GluRzeta1 (NR1) mRNAs were accompanied. Until P21, however, all of these subunits were down-regulated with particular reduction for GluRepsilon2 and GluRepsilon4 mRNAs. Similar patterns of temporal expressions were observed in motoneurons of other brainstem motor nuclei. Taking that high levels of GluRepsilon1, GluRepsilon2, and GluRzeta1 subunits are also found in the adult hippocampus and cerebral cortex, it can be assumed that NMDA receptors in developing motoneurons are highly potent and potentially involved in structural and functional development of the brainstem motor system.