Development of a novel dental resin cement incorporating FGF-2-loaded polymer particles with the ability to promote tissue regeneration.

OBJECTIVE: Aiming to achieve bioactive dental resins that promote healing of surrounding tissues, we developed novel poly(2-hydroxyethyl methacrylate/trimethylolpropane trimethacrylate) (polyHEMA/TMPT) particles. These particles have been reported to be useful as a non-biodegradable carrier for fibroblast growth factor-2 (FGF-2) in vitro. The aim of this study was to evaluate the ability of an adhesive resin incorporating FGF-2-loaded polymer particles to promote tissue regeneration in vitro and in vivo. METHODS: Experimental adhesive resins were prepared by incorporating FGF-2-loaded polyHEMA/TMPT particles into a 4-META/MMA-based adhesive resin, and the release profiles of FGF-2 were evaluated. The proliferation of osteoblast-like cells in the eluate from cured experimental resin was assessed. When the experimental resin was implanted into rat calvaria defects, bone regeneration was evaluated by microcomputed tomography and histological observations. RESULTS: Sustained release of FGF-2 from the experimental resin was observed for 14 days. Eluate from the cured experimental resin significantly promoted the proliferation of osteoblast-like cells. Significantly greater bone regeneration was observed using the experimental resin compared with the control resin without FGF-2. SIGNIFICANCE: 4-META/MMA-based adhesive resin incorporating FGF-2-loaded polymer particles is useful to promote tissue regeneration, suggesting that its application would be beneficial for root-end filling or the repair of fractured roots in cases with severely damaged periodontal tissue.

Bonding effectiveness and multi-interfacial characterization of two direct buildup resin core systems bonded to post-space dentin.

OBJECTIVES: The aim of the present study was to evaluate the bonding effectiveness of two resin core buildup systems using conventional methods in the field of adhesive dentistry and a new non-destructive method. MATERIALS AND METHODS: Twenty-four single-rooted human teeth were built up with dual-cure one-step self-etch adhesive and composite systems (SY1: Clearfil DC bond and Clearfil DC core automix, SY2: Clearfil bond SE one and Clearfil DC core automix one). The prepared samples were sectioned into approximately 1 × 1-mm-thick beams and subjected to micro-tensile bond strength (µTBS) testing (n = 24). The fractured beams after µTBS testing were analyzed by SEM and energy-dispersive X-ray (EDX) spectrometry. The three teeth filled with each resin core system were sectioned and embedded in epoxy resin to observe the dentin-bonding interface under TEM (n = 6). Moreover, three of each resin core-filled teeth without any processing were examined using µCT (n = 6). RESULTS: Two-way ANOVA revealed that the two factors "root region" (p < 0.001, F = 15.22) and "system" (SY1 < SY2; p < 0.001, F = 22.52) had a significant influence. The µTBS gradually decreased from the coronal side to the apical side of the root canal. Morphological evaluation revealed that SY2 was superior in terms of resin curing at the apical side. µCT non-destructive evaluation clearly revealed gap formation in SY1. CONCLUSION: SY2, which included a new light-independent catalyst, showed better bonding effectiveness and adhesive interface to dentin compared to that of SY1. CLINICAL RELEVANCE: The new catalyst, which is activated by contact with adhesive and resin composite, can be used for resin core buildup restorations.

Immunohistochemical analysis of dentin matrix protein 1 (Dmp1) phosphorylation by Fam20C in bone: implications for the induction of biomineralization.

Dmp1 is an acidic phosphoprotein that is specifically expressed in osteocytes. During the secretory process, the full-length, precursor Dmp1 is cleaved into N- and C-terminal fragments. C-terminal Dmp1 is phosphorylated, becoming a highly negatively charged domain that may assist in bone mineralization by recruiting calcium ions and influencing subsequent mineral deposition. It has been recently reported that the Golgi-localized protein kinase Fam20C phosphorylates Dmp1 in vitro. To investigate this phosphorylation in situ, we determined the locations of phosphorylated Dmp1 and Fam20C in rat bones using immunohistochemistry. During osteocytogenesis, osteoblastic, osteoid, and young osteocytes (but not old osteocytes) express Dmp1 mRNA and contain Dmp1 protein in the Golgi apparatus. These Dmp1-producing cells were distributed across the surface layer of cortical bone. Using immunofluorescence, we found that N- and C-terminal Dmp1 fragments were predominantly distributed along the lacunar walls and canaliculi of mineralized bone, respectively, but were not present in the osteoid matrix. We also found that Fam20C and its substrate, C-terminal Dmp1, colocalized in the Golgi of osteoblastic, osteoid, and young osteocytes. Furthermore, phosphorylated C-terminal Dmp1 was present in the Golgi of young osteocytes. Double-labeling immunoelectron microscopy revealed that phosphorylated C-terminal Dmp1 localized to the canalicular wall in mineralized bone. These findings suggest that C-terminal Dmp1 is phosphorylated within osteocytes and then secreted into the pericanalicular matrix of mineralized bone. Phosphorylated, negatively charged C-terminal Dmp1 in the pericanalicular matrix may play an important role in bone mineralization by recruiting calcium ions.

Bonding effectiveness of self-adhesive and conventional-type adhesive resin cements to CAD/CAM resin blocks. Part 1: Effects of sandblasting and silanization.

The present study assessed the effect of sandblasting and silanization on resin cement bond strengths to CAD/CAM resin blocks. Twenty four blocks (KATANA AVENCIA BLOCK) were divided into two resin cement groups (PANAVIA V5 [PV5] and PANAVIA SA CEMENT HANDMIX [PSA]), and further divided into four subgroups representing different surface treatment methods: no treatment (Ctl), silanization (Si), sandblasting (Sb), and Sb+Si. After resin application, microtensile bond strengths (µTBSs) were measured immediately, 1, 3 and 6 months after water storage. In addition, surfaces resulting from each of the treatment methods were analyzed by scanning electron microscopy (SEM). Three-way analysis of variance revealed a statistically significant effect for the parameters 'surface treatment' (p<0.001, F=370), 'resin cement' (p<0.001, F=103, PSA

Bonding effectiveness of self-adhesive and conventional-type adhesive resin cements to CAD/CAM resin blocks. Part 2: Effect of ultrasonic and acid cleaning.

The present study assessed the effect of ultrasonic and acid cleaning on resin cement bonding to CAD/CAM resin blocks. One of two resin cements, PANAVIA V5 (PV5) or PANAVIA SA CEMENT HANDMIX (PSA), were bonded to one of 24 CAD/CAM blocks (KATANA AVENCIA BLOCK). Each cement group was divided into four subgroups: no cleaning (Ctl), ultrasonic cleaning (Uc), acid cleaning (Ac) and Uc+Ac. Micro-tensile bond strengths (µTBSs) were measured immediately and 1, 3, and 6 months after water storage. Block surfaces after each treatment were analyzed by scanning electron microscopy. Analysis of variance revealed a statistically significant effect for the parameters 'surface treatment' (p<0.001, F=40), 'resin cement' (p<0.001, F=696) and 'water aging' (p<0.001, F=71). The PV5 group exhibited higher µTBS values than the PSA group. Although cleaning after sandblasting was effective in removing residual alumina particles, it did not affect the long-term bonding durability with non-contaminated CAD/CAM resin blocks.

Decrease in fluorescence lifetime by glycation of collagen and its application in determining advanced glycation end-products in human dentin.

Advanced Glycation End-products (AGEs) are produced by the Maillard reaction, which causes cross-linking of collagen and results in changes in the mechanical properties of collagen tissues. Several types of AGE fluoresce, and measurement of this fluorescence is effective for determining the presence of AGEs. Because fluorescence intensity by steady-state fluorometry is affected by sample surface condition and light source, we focused on fluorescence lifetime measurement (FLM). We found that fluorescence lifetime of collagen gel decreased with glycation progress. In vivo application of FLM for determination of AGEs was confirmed in human dentin.

Accumulation of advanced glycation end-products in human dentine.

Cross-linking of collagen by Advanced Glycation End-products (AGEs) occurs by non-enzymatic glycation (Maillard reaction). The purpose of this study was to examine whether AGEs are formed in human dentinal collagen, and to consider any possible influence of AGEs on dentinal physiology. Mechanical characteristics, fluorescence spectra and immunohistochemical analyses of demineralized dentine sections from young subjects were compared with those of aged ones. The same investigations were performed with young dentine artificially glycated by incubation in 0.1M ribose solution. Indentation measurement indicated that the sections from aged dentine were mechanically harder than those from young dentine. The hardness of young dentine increased after incubation in ribose solution. Fluorescence peak wavelength of the young dentine was shorter than that of the aged one, but shifted towards the peak wavelength of the aged one after incubation in ribose solution. These changes were considered to be due to accumulation of AGEs. Existence of AGEs in dentinal collagen was confirmed by immunohistochemical analysis. The obtained results suggest that AGEs accumulation occurs in dentinal collagen and is affected by both human age and physiological conditions such as glucose level in blood because dentinal collagen receives nourishment via dental pulp and tubules.

Structural modifications of dentinal microcracks with human aging.

Several studies have reported the mechanism of crack propagation with aging. Although structural modifications of dentinal microcracks with aging have been evaluated by observing the cracked surface using scanning electron microscope (SEM), very few attempts have been made at sectional observation of the microcracks inside dentine using transmission electron microscopy (TEM). The objectives of this study were to: (1) examine the process of dentinal microcrack formation using TEM and (2) to morphologically evaluate the relation between dentinal microcrack propagation and human aging. Molars from 'young' (16-28 years) and 'aged' (62-76 years) subjects were evaluated. Dentine blocks were cracked with an indenter and sectioned using a diamond knife and ultramicrotome after embedding in epoxy resin. Microcracks were observed by TEM and ultra-high-voltage electron microscope tomography to determine the characteristics of crack propagation in the young and aged teeth. The results show that, in young teeth, crack propagation tended to pass through the dentinal tubules, while in aged teeth, it tended to deflect to the outer side of peritubular dentine (PTD), especially in coronal dentine. The advantage of this method is that it allows visualization and evaluation of the ultrastructural propagation of microcracks in dentine. The differences in crack propagation between young and aged dentine could be explained by differences in PTD thickness.

Detection of dentinal microcracks using infrared thermography.

INTRODUCTION: It is difficult to make a definite diagnosis of a cracked tooth solely based on an inspection within the root canal, especially in case of microcracks. At present, there seems to be no established method to detect dentinal microcracks in roots; therefore, the current detection techniques need to be improved. Vibrothermography (VibroIR) helps to detect microcracks by the friction heat generated from ultrasonic vibration. The purpose of this study was to establish a novel method using VibroIR to detect dentinal microcracks. METHODS: The root canals of 20 roots with cracks and control roots were prepared after removing the tooth crowns. A tapered indenter was inserted into the root canal and pressed until a microcrack was created under an optical microscope. Using VibroIR, the detection trials for dentinal microcracks were performed with an ultrasonic vibration power ranging from 0.43 to 1.48 W at an angle of 0°, 30°, 45°, 60°, and 90° between the ultrasonic vibration point and the microcrack line. After the detection test, the microcrack width was measured with an optical microscope. RESULTS: Frictional heat was detected in the microcracks with thermography at 0.89 to 1.48 W and at an ultrasonic vibration point angle less than 60° from the crack line for 10 seconds. Microcracks with a width of 4 to 35.5 µm were detected with this method. CONCLUSIONS: VibroIR may be an effective method for the diagnosis of root dentinal microcracks.

Mechanical and morphological evaluation of the bond-dentin interface in direct resin core build-up method.

OBJECTIVES: The purpose of this study was to evaluate the interfacial adhesion between resin and root canal dentin in the direct resin core build-up method in terms of microtensile bond strength (µTBS) and dentin micro morphology. METHODS: Single-rooted human teeth were decoronated at the cementoenamel junction and endodontically treated. Post spaces were prepared in the roots to a depth of 10mm. The spaces were then treated with a dual-cure bonding system, and filled with dual-cure resin composite. After 24-h storage in water at 37 °C, they were trimmed into approximately 1.0-mm(2) beams for µTBS. Bond strength was analyzed with one-way ANOVA and Tukey's test. The fractured surfaces were examined by scanning electron microscopy (SEM) and energy-dispersive X-ray spectrometry (EDX). Sectioned specimens were observed by ultra-high-voltage transmission electron microscopy. RESULTS: The bond strength of root dentin decreased gradually from the coronal to apical side, and the bond strength of the coronal section was significantly higher than that of the radicular section. Moreover, the failure modes in the coronal and apical sides of the specimens differed. The apical specimens fractured within the core material, while the coronal specimens fractured at the bonding layer. SEM and EDX analyses revealed that the core material penetrated into dentinal tubules in the apical region. SIGNIFICANCE: In the direct resin core build-up method, the interfacial adhesion of resin to root canal dentin may be insufficient in the apical region of the root canal due to poor polymerization.

Ultra-structural observation of human enamel and dentin by ultra-high-voltage electron tomography and the focus ion beam technique.

Human enamel and dentin are hard and brittle mineralized tissues. It is difficult to prepare thin specimens (under 200 nm) of these tissues for transmission electron microscope observation without demineralizing them. We present a novel method of creating three-dimensional ultra-structural images of human enamel and dentin, using the focus ion beam (FIB) method and ultra-high-voltage electron microscope tomography. Thin specimens (less than 2 µm) of enamel and dentin were prepared using the FIB method. This method is appropriate for nano-fabrication of thin specimens for brittle materials such as enamel and dentin. It allows penetration of an ultra-high-voltage electron beam of a 3000 kV maximal acceleration voltage into a specimen, enabling projections of the specimen to be taken. It facilitates tomography of the enamel rods, sheaths, dentinal tubules and collagen fibrils with a high resolution of 2 nm. The component percentages in ultra-structures of dentin can be expressed numerically by using three-dimensional reconstruction images of tomograms. The matrix of peritubular dentin differed from that of intertubular dentin by having relatively fewer collagen fibrils. The major advantage of this method is its ability to visualize ultra-structural tomograms of highly calcified specimens without demineralization.

Dental haptic simulator to train hand skill of student - calibration method to realize visual haptic environment.

In prior work, we have developed dental training simulator to train hand skill of student (HHDTS). In the present study, we performed calibration between haptic device coordinate system and half-mirror coordinate system for our system to realize real clinical situation. As a result, the user can overlay the CG (turbine) onto the stylus of haptic device and intract to the CG (tooth) directly.

Virtual reality simulation training for dental surgery.

In dentistry the exquisite hand skill is required for daily treatments, However, dental students have little chance to treat patients in the clinical training So, present study constructed a process simulation to train the hand skill for tooth extraction, especially impacted wisdom tooth that was an example of the difficult case by using virtual reality haptic device.

Relationship between the color of carious dentin with varying lesion activity, and bacterial detection.

OBJECTIVES: To investigate the relationship between the color of carious dentin with varying lesion activity, and bacterial detection in the lesions. METHODS: In 26 extracted human molars with coronal dentin caries and four extracted sound human molars, dentin was removed by a round bur every 150 microm from the dentin surface, in the direction of the pulp chamber. Before and after removal, images of nine-color samples and the dentin surface stained with a caries detector dye (1% acid red in propylene glycol) were taken simultaneously by a charge-coupled device (CCD), and dentinal tissue samples were taken with a new round bur. From the images, corrected L*, a* and b* values (CIE 1976 L*a*b* color system) of the dentin surfaces were calculated from the color changes of the nine-color samples. Bacterial DNA in the dentinal tissues was detected by polymerase chain reaction. RESULTS: Before removal of dentin, the L* of sound molars (L*>50) was significantly larger than that of carious molars (L*<50) (ANOVA, Scheffe's F-test, P<0.05). In addition, the carious molars were divided into type I (a*>20, characteristics of active caries) and type II (a*<20, characteristics of arrested caries), and there was a significant difference in the a* value (P<0.05). For both carious types, the area under the receiver operating characteristic curve of L* was significantly larger than that of a* or b* (univariate Z score test, P<0.05), and the rate of bacterial detection decreased as the L* of dentinal tissue increased, and bacterial DNA was not detected when L* was >60. CONCLUSIONS: Sound and types I and II carious dentin were discriminated by the combination of L* and a* values of dentinal tissue stained with the caries detector dye before removal of dentin. In carious lesions, the a* values of carious dentin stained with the dye were related to the carious lesion activity before removal of carious tissue, and the L* values were related to the degree of caries progression.

Evaluating the objectivity of caries removal with a caries detector dye using color evaluation and PCR.

OBJECTIVES: This laboratory study evaluated the objectivity of caries removal with a caries detector dye by color and bacterial evaluations. METHODS: In 41 cases of dentin caries (32 extracted human molars), carious tissues were removed using a caries detector dye. Images of dentin surfaces with color-matching stickers were acquired using a CCD camera, and dentinal tissue samples were collected with new round burs. Corrected L*, a* and b* values (CIE 1976 L*a*b* color system) of dentin surfaces were calculated from the sticker color changes. In addition, bacterial DNA in dentinal tissues was detected by the polymerase chain reaction. RESULTS: The intra-class correlation coefficient of the corrected L*, a* and b* values was 0.34, 0.30 and 0.49, respectively. There were significant inter-operator differences (P<0.05). Seventeen of 41 specimens contained bacterial DNA after caries removal. CONCLUSIONS: Results showed that objectivity of caries removal using the caries detector dye with visual inspection was low.

Fatigue of tooth-colored restoratives in aqueous environment.

The purpose of this study was to investigate the interaction between mechanical and chemical fatigue in resin composites and dental ceramics, and the effects thereof on fatigue resistance of tooth-colored restoratives. To this end, the fatigue fracture resistance of restoratives under dry and aqueous conditions were analyzed by a dynamic fatigue crack propagation test using beam-shaped specimens with a precrack. Fatigue crack propagation characteristics were expressed by the correlation between fatigue crack growth rate (da/dN) and stress intensity factor range (deltaK). In addition, fatigue crack growth threshold (deltaKth) was calculated. Following the fatigue test, a fractographic examination was performed using scanning electron microscopy. Fatigue crack initiation was retarded in resin composites under aqueous condition, but dental ceramics were susceptible to slow crack growth after crack initiation. SEM images of the fatigue facture surfaces reflected inorganic and organic filler particles of different sizes in composites and the bonding at crystal-glass interface in ceramics. It was concluded that water exerted different effects on the fatigue resistance of composites and ceramics.

The accuracy of electrical method for microleakage evaluation by a three-dimensional analysis.

OBJECTIVES: This in vitro study aimed to investigate the accuracy of an electrical method for the evaluation of microleakage by a three-dimensional analysis of dye penetration. METHODS: Coronal cavities were prepared on buccal, palatal or lingual surfaces in extracted human molars. The cavities were then filled with resin composites and were subjected to 10,000 load cycles (425g). Before cavity preparation and after load cycling, physiological saline was applied and wiped off, and the change in conductance was measured across the margin of the restoration in each specimen. After dye penetration, the specimens were reduced by 100 microm in a direction parallel to the cavity floor, from the surface of the restoration to the cavity floor. The sequence of reducing the sections by 100 microm and image taking was repeated. Three-dimensional images of dye penetration were made and the proportions of the interface showing penetration were calculated. RESULTS: Pearson's correlation coefficients between changes in conductance and the surface area of dye penetration, between these and the rate of dye penetration, and between these and the depth of dye penetration were 0.932, 0.920 and 0.732, respectively. The correlations were significant (p<0.05). CONCLUSIONS: The results of this electrical method for microleakage evaluation showed stronger correlations with the three-dimensional amount of marginal leakage (surface area of dye penetration and rate of dye penetration) than the two-dimensional amount (depth of dye penetration).

Relationship between colors of carious dentin and laser fluorescence evaluations in caries diagnosis.

This in vitro study investigated the relationship between assessments of dentin caries using a laser fluorescence device (DIAGNOdent) and a caries detector dye during caries removal. The dentin of eight extracted carious molars was removed at 300-microm interval points from the dentin surface toward the pulp chamber. Before and after each removal, images of the carious surfaces were taken in association with color-matching stickers (for color correction) and the surfaces were evaluated by DIAGNOdent based on fluorescence emission from the tooth surface. For the L* values (CIE 1976 L*a*b* color system), there was a strong negative correlation between DIAGNOdent results and the corrected L* values of the carious surfaces (Pearson's correlation coefficient: -0.853); additionally, there was a significant correlation between them (p<0.05). However, there were no significant correlations between the DIAGNOdent results and the corrected a* and b* values of the carious surfaces (Pearson's correlation coefficients: 0.108 and 0.018 respectively). In conclusion, DIAGNOdent was shown to be applicable for caries diagnosis during caries removal.

Three-dimensional evaluation of gap formation of cervical restorations.

OBJECTIVES: In some studies gap formation has been evaluated in just one section of the restorative. This in vitro study aimed to design a quantitative three-dimensional method for evaluation of the contraction gap in restoratives. METHODS: Cervical cavities were prepared on buccal, palatal or lingual surfaces in human extracted molars and were then filled with resin composites. Specimens were reduced every 100 microm in a direction parallel to the tooth axis, and perpendicular to the cavity floor from one proximal side to the other. The sequence of reducing the sections by 100 microm, image taking (250x) and observation of these images (maximal 2500x) were repeated. Three-dimensional images of the contraction gap were made using analytical software and the proportions of the interface with gap formation calculated. RESULTS: The mean proportions of the interface with gap formation of the self-etching system (Clearfil Liner Bond II Sigma) was 41.7+/-6.3% and that of the self-priming system (Single Bond) was 38.2+/-3.9%; there was no significant difference. CONCLUSIONS: Approximate three-dimensional images of the in vitro contraction gap could be drawn and the mean proportions of the interface with gap formation could be more precisely calculated than by previous methods.