Developmental roles of glomerular epithelial protein-1 in mice molar morphogenesis.

Glomerular epithelial protein-1 (Glepp1), a R3 subtype family of receptor-type protein tyrosine phosphatases, plays important role in the activation of Src family kinases and regulates cellular processes such as cell proliferation, differentiation, and apoptosis. In this study, we firstly examined the functional evaluation of Glepp1 in tooth development and morphogenesis. The precise expression level and developmental function of Glepp1 were examined by RT-qPCR, in situ hybridization, and loss and gain of functional study using a range of in vitro organ cultivation methods. Expression of Glepp1 was detected in the developing tooth germs in cap and bell stage of tooth development. Knocking down Glepp1 at E13 for 2 days showed the altered expression levels of tooth development-related signaling molecules, including Bmps, Dspp, Fgf4, Lef1, and Shh. Moreover, transient knock down of Glepp1 revealed alterations in cellular physiology, examined by the localization patterns of Ki67 and E-cadherin. Similarly, knocking down of Glepp1 showed disrupted enamel rod and interrod formation in 3-week renal transplanted teeth. In addition, due to attrition of odontoblastic layers, the expression signals of Dspp and the localization of NESTIN were almost not detected after knock down of Glepp1; however, their expressions were increased after Glepp1 overexpression. Thus, our results suggested that Glepp1 plays modulating roles during odontogenesis by regulating the expression levels of signaling molecules and cellular events to achieve the proper structural formation of hard tissue matrices in mice molar development.

The role of O-GlcNAcylation mediated by OGT during tooth development.

To understand the mechanisms underlying tooth morphogenesis, we examined the developmental roles of important posttranslational modification, O-GlcNAcylation, which regulates protein stability and activity by the addition and removal of a single sugar (O-GlcNAc) to the serine or threonine residue of the intracellular proteins. Tissue and developmental stage-specific immunostaining results against O-GlcNAc and O-GlcNAc transferase (OGT) in developing tooth germs would suggest that O-GlcNAcylation is involved in tooth morphogenesis, particularly in the cap and secretory stage. To evaluate the developmental function of OGT-mediated O-GlcNAcylation, we employed an in vitro tooth germ culture method at E14.5, cap stage before secretory stage, for 1 and 2 days, with or without OSMI-1, a small molecule OGT inhibitor. To examine the mineralization levels and morphological changes, we performed renal capsule transplantation for one and three weeks after 2 days of in vitro culture at E14.5 with OSMI-1 treatment. After OGT inhibition, morphological and molecular alterations were examined using histology, immunohistochemistry, real-time quantitative polymerase chain reaction, in situ hybridization, scanning electron microscopy, and ground sectioning. Overall, inhibition of OGT resulted in altered cellular physiology, including proliferation, apoptosis, and epithelial rearrangements, with significant changes in the expression patterns of ß-catenin, fibroblast growth factor 4 (fgf4), and sonic hedgehog (Shh). Moreover, renal capsule transplantation and immunolocalizations of Amelogenin and Nestin results revealed that OGT-inhibited tooth germs at cap stage exhibited with structural changes in cuspal morphogenesis, amelogenesis, and dentinogenesis of the mineralized tooth. Overall, we suggest that OGT-mediated O-GlcNAcylation regulates cell signaling and physiology in primary enamel knot during tooth development, thus playing an important role in mouse molar morphogenesis.

Comparison of microstructures and mechanical properties of 3 cobalt-chromium alloys fabricated with soft metal milling technology.

STATEMENT OF PROBLEM: Although several manufacturers market soft metal milling blanks and systems, comprehensive comparative studies of differences in properties across commercially available soft metal milling alloys are lacking. PURPOSE: The purpose of this in vitro study was to compare the microstructures and mechanical properties of 3 soft metal milling cobalt-chromium (Co-Cr) alloys (Ceramill Sintron, Soft Metal, and Sintermetall). MATERIAL AND METHODS: Disk-shaped specimens (for surface characterization and hardness test) and dumbbell-shaped specimens (for tensile test as per International Organization for Standardization (ISO) 22674) were prepared by following each soft metal milling manufacturer's instructions. The crystal structures and microstructures of the 3 alloys were evaluated with optical microscopy, X-ray diffractometry (XRD), and scanning electron microscopy with electron backscattered diffraction (EBSD). The mechanical properties were investigated with a tensile test and Vickers hardness test (n=6). The results of the mechanical (tensile and hardness) tests were analyzed with 1-way ANOVA and the post hoc Tukey multiple comparison test (α=.05). RESULTS: The Sintermetall specimen showed a finer microstructure and more porosity than the other 2 alloys. The XRD and EBSD analyses showed that the γ (face-centered cubic, fcc) matrix phase was predominant in the Ceramill Sintron alloy and the ε (hexagonal close-packed, hcp) matrix phase was predominant in the Soft Metal alloy. The Sintermetall alloy showed a slightly higher amount of ε phase than γ phase, with more chromium carbide formation than the other 2 alloys. The Ceramill Sintron alloy showed a significantly higher tensile strength than the other 2 alloys (P<.05), but a significantly lower 2% offset yield strength than the other 2 alloys (P<.05). The highest elongation was found in the Ceramill Sintron alloy, followed by the Sintermetall and Soft Metal alloys. The elastic modulus was the highest in the Sintermetall alloy, followed by the Soft Metal and Ceramill Sintron alloys. No significant differences in Vickers hardness values were detected among the 3 alloys (P=.263). CONCLUSIONS: The different commercially available soft metal milling blanks and systems produced dissimilar alloys in terms of crystal structures and microstructures and, as a result, different mechanical properties.

Developmental Roles of FUSE Binding Protein 1 (Fubp1) in Tooth Morphogenesis.

FUSE binding protein 1 (Fubp1), a regulator of the c-Myc transcription factor and a DNA/RNA-binding protein, plays important roles in the regulation of gene transcription and cellular physiology. In this study, to reveal the precise developmental function of Fubp1, we examined the detailed expression pattern and developmental function of Fubp1 during tooth morphogenesis by RT-qPCR, in situ hybridization, and knock-down study using in vitro organ cultivation methods. In embryogenesis, Fubp1 is obviously expressed in the enamel organ and condensed mesenchyme, known to be important for proper tooth formation. Knocking down Fubp1 at E14 for two days, showed the altered expression patterns of tooth development related signalling molecules, including Bmps and Fgf4. In addition, transient knock-down of Fubp1 at E14 revealed changes in the localization patterns of c-Myc and cell proliferation in epithelium and mesenchyme, related with altered tooth morphogenesis. These results also showed the decreased amelogenin and dentin sialophosphoprotein expressions and disrupted enamel rod and interrod formation in one- and three-week renal transplanted teeth respectively. Thus, our results suggested that Fubp1 plays a modulating role during dentinogenesis and amelogenesis by regulating the expression pattern of signalling molecules to achieve the proper structural formation of hard tissue matrices and crown morphogenesis in mice molar development.

An endoplasmic reticulum stress regulator, Tmbim6, modulates secretory stage of mice molar.

To understand the role of endoplasmic reticulum (ER)-stress in mice molar development, we studied Tmbim6 that antagonizes the unfolded protein response, using Tmbim6 knockout (KO) mice and in vitro organ cultivation with knocking down using small interfering RNA. During molar development, Tmbim6 is expressed in developing tooth at E14-E16, postnatal0 (PN0), and PN6. Mineral content in Tmbim6 KO enamel was reduced while dentin was slightly increased revealing ultrastructural changes in pattern formation of both enamel and dentin. Moreover, odontoblast differentiation was altered with increased Dspp expression at PN0 followed by altered AMELX localizations at PN5. These results were confirmed by in vitro organ cultivation and showed altered Bmp signaling, proliferation, and actin rearrangement in the presumptive ameloblast and odontoblasts that followed the altered expression of differentiation and ER stress-related signaling molecules at E16.5. Overall, ER stress modulated by Tmbim6 would play important roles in patterned dental hard tissue formation in mice molar within a limited period of development.

Effect of Post-Sintering Conditions on the Mechanical Properties of a New Co-Cr Alloy Produced by New Subtractive Manufacturing.

In this study, we investigated the effect of sintering temperature (1300, 1350, or 1400 °C) and holding time (1 or 2 h) on the mechanical properties of a cobalt-chromium (Co-Cr) alloy (Soft Metal) produced by milling/post-sintering, using a tensile test (n = 6). Prior to the test, the different nanostructures arising from the sintering conditions were also analyzed. The phase ratio of γ (face-centered cubic) phase to ɛ (hexagonal close-packed) phase increased mainly with increasing temperature. The formation of Cr23C6 carbide was greatest in the 1350 °C groups when compared to the other temperature groups. The 1400 °C groups had a substantially greater grain size than the 1300 °C and 1350 °C groups, together with a significant number of annealing twins inside the matrix phases. Overall, the 1350 °C groups showed the most superior properties. The 1400 °C groups showed a mean 0.2% yield strength under 500 MPa. The holding times did not significantly affect the mechanical properties (p > 0.05).

Influence of Oxidative Etching Solution Temperatures on the Surface Roughness and Wettability of a Titanium Alloy.

Recently, a simple surface modification treatment of titanium (Ti) was developed to produce nano-and micro-scale features on the surfaces via simple immersion in an oxidative aqueous solution (30% hydrogen peroxide/5% sodium bicarbonate). However, this treatment method of Ti surfaces requires a relatively long immersion time (4 h) in the oxidative solution. In this study, we investigated whether an increase in the temperature of the oxidative etching solution can shorten the immersion time of Ti effectively. Polished grade 5 dental Ti (Ti-6Al-4V) discs were immersed in the oxidative aqueous solution either for 30 or 60 min. The temperature of the etching solution was maintained at 25 (similar to room temperature), 35, or 45 °C during etching. The etched surfaces were studied in terms of micro- and nano-structures, surface roughness, and wettability (surface energy). The increase in the temperature of the solution accelerated the etching effect of Ti and created both micro- and nano-structures on the surfaces more effectively. In particular, immersion for 60 min at the solution temperature of 35 °C significantly increased the surface roughness and wettability, although the etching effect was enhanced further at the solution temperature of 45 °C.

Marginal fit of metal-ceramic crowns fabricated by using a casting and two selective laser melting processes before and after ceramic firing.

STATEMENT OF PROBLEM: Few studies have investigated changes in the marginal fit of metal-ceramic restorations fabricated by selective laser melting (SLM) techniques after the application of veneering ceramic. PURPOSE: The purpose of this in vitro study was to evaluate the marginal fit (silicone replica technique) and internal porosity (cross-section analysis) of cobalt-chromium (Co-Cr) alloy metal crowns prepared by using 2 SLM processes together with a casting technique before and after ceramic veneering. MATERIAL AND METHODS: Cast single Co-Cr crowns and SLM-processed crowns with large (SLML) or small (SLMS) porosity were prepared (n=20/group), and half were subjected to ceramic veneering. On a single Co-Cr master die, the marginal discrepancy (MD) and absolute marginal discrepancy (AMD) of the crowns were measured by using the silicone replica technique, in which each replica was cut into 4 sections before and after ceramic veneering (n=10 for each subgroup). After marginal fit measurements, each metal coping was cross-sectioned into 4 parts, and 5 rectangular optical microscope images were acquired on both outer corners of each quarter. The porosity was then calculated as the ratio of the black-to-white pixels on the binarized images. The data were analyzed by 2-way ANOVA and the post hoc test (Tukey or Student t test) (α=.05). RESULTS: Before ceramic veneering, the 2 SLM groups showed significantly larger MDs than the casting group (56.4 ±10.4 µm) (P<.05). A significant increase in MD after ceramic veneering was detected only in the SLML group (P<.001). The AMD values showed a similar trend with MD values. The 2 SLM groups (in particular, SLML) showed a significantly higher amount of porosity than the casting group before ceramic veneering (P<.001). Only the SLML group showed a significant decrease in the amount of porosity after ceramic veneering (P<.001). CONCLUSIONS: Within the limitations of this in vitro study, large internal porosity within the SLM-fabricated Co-Cr metal copings affected the marginal fit of the metal-ceramic crowns. However, all the MD values of the 3 groups were lower than the acceptable range even after the application of veneering ceramic.

Antifungal Effect of a Dental Tissue Conditioner Containing Nystatin-Loaded Alginate Microparticles.

In this in vitro study, nystatin-alginate microparticles were successfully fabricated to control the release of nystatin from a commercial dental tissue conditioner. These nystatin-alginate microparticles were spherical and had a slightly rough surface. The microparticles incorporated into the tissue conditioner were distributed homogeneously throughout the tissue conditioner matrix. The incorporation of the microparticles did not deteriorate the mechanical properties of the original material. The agar diffusion test results showed that the tissue conditioner containing the microparticles had a good antifungal effect against Candida albicans. The nystatin-alginate microparticles efficiently controlled the release of nystatin from the tissue conditioner matrix over the experimental period of 14 days. Moreover, the nystatin-alginate microparticles incorporated in the tissue conditioner showed effective antifungal function even at lower concentrations of nystatin. The current study suggests that the tissue conditioner containing the nystatin-alginate microparticle carrier system has potential as an effective antifungal material.

Surface Roughness of a 3D-Printed Ni-Cr Alloy Produced by Selective Laser Melting: Effect of Process Parameters.

The selective laser melting (SLM) process parameters, which directly determine the melting behavior of the metallic powders, greatly affect the nanostructure and surface roughness of the resulting 3D object. This study investigated the effect of various laser process parameters (laser power, scan rate, and scan line spacing) on the surface roughness of a nickel-chromium (Ni-Cr) alloy that was three-dimensionally (3D) constructed using SLM. Single-line formation tests were used to determine the optimal laser power of 200 W and scan rate of 98.8 mm/s, which resulted in beads with an optimal profile. In the subsequent multi-layer formation tests, the 3D object with the smoothest surface (Ra = 1.3 µm) was fabricated at a scan line spacing of 60 µm (overlap ratio = 73%). Narrow scan line spacing (and thus large overlap ratios) was preferred over wide scan line spacing to reduce the surface roughness of the 3D body. The findings of this study suggest that the laser power, scan rate, and scan line spacing are the key factors that control the surface quality of Ni-Cr alloys produced by SLM.

Resin Bonding to Type IV Gold Alloy Conditioned with a Novel Mercapto Silane System: Effect of Incorporation of a Phosphate Monomer.

Self-assembled monolayers of thiols have been used to link a range of materials to planar gold surfaces or gold nanoparticles in nanoscience and nanotechnology. Novel mercapto silane systems are a promising alternative to dental noble metal alloys for enhanced resin bonding durability Goldbased alloys for full-cast restorations contain various base metal elements, which may bond to acidic functional monomers chemically, in addition to noble metal elements. This study examined how the additional incorporation of a phosphate monomer (di-2-hydroxyethyl methacryl hydrogenphosphate, DHP) into novel mercapto silane primer systems affected the resin bond strength to a type IV gold alloy pretreated with the primers. One of three commercial primers (Alloy Primer and M. L. Primer) and three experimental primer systems ((1) blend of γ-mercaptopropyltrimethoxysilane (SPS) and γ-methacryloxypropyltrimethoxysilane (MPS) (both 1.0 wt%), (2) 1.0 wt% DHP-containing primer, and (3) blend of SPS, MPS, and DHP (each 1.0 wt%)) was applied to the alloy surfaces after sandblasting. Resin cylinders (diameter: 2.38 mm) were bonded to the surfaces and light-cured. All bonded specimens were stored in water at 37 °C for 24 h and then half of them additionally water immersed for 7 days (37 °C) and thermocycled 10,000 times before the shear bond strength test (n = 10). The mercapto silane systems (SPS + MPS) were found to show superior resin bonding durability to the commercial primers and the only DHP-containing primer, regardless of additional incorporation of the phosphate monomer.

A microcomputed tomography evaluation of the marginal fit of cobalt-chromium alloy copings fabricated by new manufacturing techniques and alloy systems.

STATEMENT OF PROBLEM: Although new digital manufacturing techniques are attracting interest in dentistry, few studies have comprehensively investigated the marginal fit of fixed dental prostheses fabricated with such techniques. PURPOSE: The purpose of this in vitro microcomputed tomography (µCT) study was to evaluate the marginal fit of cobalt-chromium (Co-Cr) alloy copings fabricated by casting and 3 different computer-aided design and computer-aided manufacturing (CAD-CAM)-based processing techniques and alloy systems. MATERIAL AND METHODS: Single Co-Cr metal crowns were fabricated using 4 different manufacturing techniques: casting (control), milling, selective laser melting, and milling/sintering. Two different commercial alloy systems were used for each fabrication technique (a total of 8 groups; n=10 for each group). The marginal discrepancy and absolute marginal discrepancy of the crowns were determined with µCT. For each specimen, the values were determined from 4 different regions (sagittal buccal, sagittal lingual, coronal mesial, and coronal distal) by using imaging software and recorded as the average of the 4 readings. For each parameter, the results were statistically compared with 2-way analysis of variance and appropriate post hoc analysis (using Tukey or Student t test) (α=.05). RESULTS: The milling and selective laser melting groups showed significantly larger marginal discrepancies than the control groups (70.4 ±12.0 and 65.3 ±10.1 µm, respectively; P<.001), whereas the milling/sintering groups exhibited significantly smaller values than the controls (P=.004). The milling groups showed significantly larger absolute marginal discrepancy than the control groups (137.4 ±29.0 and 139.2 ±18.9 µm, respectively; P<.05). In the selective laser melting and milling/sintering groups, the absolute marginal discrepancy values were material-specific (P<.05). Nonetheless, the milling/sintering groups yielded statistically comparable (P=.935) or smaller (P<.001) absolute marginal discrepancies to the control groups. CONCLUSIONS: The findings of this in vitro µCT study showed that the marginal fit values of the Co-Cr alloy greatly depended on the fabrication methods and, occasionally, the alloy systems. Fixed dental prostheses produced by using the milling/sintering technique can be considered clinically acceptable in terms of marginal fit.

Antimicrobial Activity of Glass lonomer Cement Incorporated with Chlorhexidine-Loaded Zeolite Nanoparticles.

A functional dental restorative system with antimicrobial properties was developed using zeolite (ZE) nanoparticles (NPs) as a drug delivery carrier. ZE NPs loaded with chlorhexidine (CHX) were prepared using the ionic immobilization method. The resulting CHX-loaded ZE NPs were then incorporated into commercial dental glass ionomer cement (GIC). The average size of the CHX-loaded ZE NPs was about 100 to 200 nm, and the NPs were dispersed homogeneously in the GIC. The in vitro release profile of encapsulated GIC containing CHX showed an early release burst of approximately 30% of the total CHX by day 7, whereas GIC containing CHX-loaded ZE NPs showed a sustained release of CHX without the early release burst in a 4-week immersion study. The agar diffusion test results showed that the GIC incorporated with CHX-loaded ZE NPs showed a larger growth inhibition zone of Streptococcus mutans than GIC alone, indicating that this innovative delivery platform potently imparted antimicrobial activity to the GIC. Moreover, these findings suggest that a range of antimicrobial drugs that inhibit the growth of oral bacteria can be incorporated efficiently into dental GIC using CHX-loaded ZE NPs.

Osteogenic Evaluation of Porous Calcium Phosphate Granules with Drug Delivery System Using Nanoparticle Carriers.

Novel porous biphasic calcium phosphate (BCP) granules incorporated with drug-releasing poly(lactic-co-glycolic acid) (PLGA) nanoparticles were developed as a drug delivery platform for bone regeneration. The charge interaction between the BCP and PLGA nanoparticle surfaces was manipulated to create this combination system. Spherical BCP granules with open micro-channels and PLGA nanoparticles loaded with dexamethasone (DEX) as a model drug were fabricated using a liquid nitrogen method and standard emulsion method, respectively. Polyethyleneimine was coated on the DEX-loaded PLGA nanoparticle surfaces, resulting in a net positively charged surface. Such modified nanoparticles were immobilized physically on the negatively charged BCP granule surfaces. An in vitro evaluation of MG 63 cells cultured for 1 and 2 weeks on the BCP granules containing DEX-loaded PLGA nanoparticles showed greater cell proliferation, differentiation, and a more extensively connected-tissue network than those cultured on the BCP granules alone. This innovative platform for bioactive molecule delivery more potently induced osteogenesis in vitro, which might be exploited in implantable bioceramic bone graft materials for stem cell therapy or improved in vivo performance.

Effect of Atmospheric Plasma Treatment to Titanium Surface on Initial Osteoblast-Like Cell Spreading. .

Plasma treatments are becoming a popular method for modifying the characteristics of a range of substrate surfaces. Atmospheric pressure plasma is cost-efficient, safe and simple compared to high-pressure plasma. This study examined the effects of atmospheric pressure plasma to a titanium (Ti) surface on osteoblast-like cell (osteoblast) spreading and cellular networks. The characteristics of the Ti surface before and after the atmospheric plasma treatment were analyzed by X-ray photoemission spectroscopy (XPS), scanning electron microscopy (SEM), contact angle measurements, and an optical 3D profiling system. The morphology of osteoblasts attached to the Ti surfaces was observed by SEM and confocal laser scanning microscopy. The atmospheric pressure plasma made the Ti surfaces more hydrophilic. The osteoblasts that adhered to the untreated surface were round and spherical, whereas the cells covered a larger surface area on the plasma-treated surface. The plasma-treated Ti surface showed enhanced cell spreading and migration with more developed cellular networks. In conclusion, an atmospheric plasma treatment is a potential surface modifying method that can enhance the initial the cell affinity at the early stages in vitro.

Simple Heat Treatment of Zirconia Ceramic Pre-Treated with Silane Primer to Improve Resin Bonding.

Establishing a strong resin bond to dental zirconia ceramic remains difficult. Previous studies have shown that the conventional application of silane does not work well with zirconia. This paper reports that a silane pre-treatment of dental zirconia ceramic combined with subsequent heat treatment has potential as an adhesive cementation protocol for improving zirconia-resin bonding. Among the various concentrations (0.1 to 16 vol%) of experimental γ-methacryloxypropyltrimethoxysilane (γ-MPTS) primers assessed, the 1% solution was found to be the most effective in terms of the shear bond strength of the resin cement to dental zirconia ceramic. A high shear bond strength (approx. 30 MPa) was obtained when zirconia specimens were pre-treated with this primer and then heat-treated in a furnace for 60 min at 150 degrees C. Heat treatment appeared to remove the hydrophilic constituents from the silane film formed on the zirconia ceramic surface and accelerate the condensation reactions between the silanol groups of the hydrolyzed silane molecules at the zirconia/resin interface, finally making a more desirable surface for bonding with resin. This estimation was supported by Fourier transform infrared spectroscopy of the silanes prepared in this study.

The Effect of Novel Mercapto Silane Systems on Resin Bond Strength to Dental Noble Metal Alloys.

Self-assembled monolayers of thiols (RSH), which are key elements in nanoscience and nanotechnology, have been used to link a range of materials to planar gold surfaces or gold nanoparticles. In this study, the adhesive performance of mercapto silane systems to dental noble metal alloys was evaluated in vitro and compared with that of commercial dental primers. Dental gold-palladium-platinum (Au-Pd-Pt), gold-palladium-silver (Au-Pd-Ag), and palladium-silver (Pd-Ag) alloys were used as the bonding substrates after air-abrasion (sandblasting). One of the following primers was applied to each alloy: (1) no primer treatment (control), (2) three commer- cial primers: V-Primer, Metal Primer II, and M.L. Primer, and (3) two experimental silane primer systems: 2-step application with 3-mercaptopropyltrimethoxysilane (SPS) (1.0 wt%) and then 3-methacryloxypropyltrimethoxysilane (MPS) (1.0 wt%), and a silane blend consisting of SPS and MPS (both 1.0 wt%). Composite resin cylinders with a diameter of 2.38 mm were bonded to the surfaces and irradiated for 40 sec using a curing light. After storage in water at 37 °C for 24 h, all the bonded specimens were thermocycled 5000 times before the shear bond strength test. Regardless of the alloy type, the mercapto silane systems (both the 2-step and blend systems) consistently showed superior bonding performance than the commercial primers. Contact angle analysis of the primed surfaces indicated that higher resin bond strengths were produced on more hydrophilic alloy surfaces. These novel mercapto silane systems are a promising alternative for improving resin bonding to dental noble metal alloys.

In Vitro Evaluation of Hydroxyapatite-Coated Titanium Implant with Atmospheric Plasma Treatment.

Plasma treatments are becoming a popular method for modifying the characteristics of a range of substrate surfaces. Atmospheric pressure plasma (APP) is cost-efficient, safe and simple compared to high-pressure plasma. This study examined the effects of a low-temperature APP treatment of a hydroxyapatite (HA)-coated Ti (HA-Ti) surface. The APP treatment made the HA-Ti surfaces more hydrophilic without changing surface morphologies. The APP-treated HA-Ti (APP-HA-Ti) surface showed enhanced cell spreading, cell proliferation, and alkaline phosphatase (ALP) levels with more developed cellular networks, and the formed extracellular matrix (ECM) was fused perfectly with the HA substrate than that on the HA-Ti surface. In conclusion, an APP treatment is a potential surface-modifying method that can enhance the cell affinity at the early stages in vitro.

Effect of adhesive resin flexibility on enamel fracture during metal bracket debonding: an ex vivo study.

OBJECTIVE: To test the null hypothesis that neither the flexural properties of orthodontic adhesive resins nor the enamel pre-treatment methods would affect metal bracket debonding behaviours, including enamel fracture. MATERIALS AND METHODS: A dimethacrylate-based resin (Transbond XT, TX) and two methyl methacrylate (MMA)-based resins (Super-Bond C&B, SB; an experimental light-cured resin, EXP) were tested. Flexural strength and flexural modulus for each resin were measured by a three-point-bending test. Metal brackets were bonded to human enamel pretreated with total-etch (TE) or self-etch adhesive using one of the three resins (a total of six groups, n = 15). After 24 hours of storage in water at 37°C, a shear bond strength (SBS) test was performed using the wire loop method. After debonding, remaining resin on the enamel surfaces and occurrence of enamel fracture were assessed. Statistical significance was set at P < 0.05. RESULTS: The two MMA resins exhibited substantially lower flexural strength and modulus values than the TX resin. The mean SBS values of all groups (10.15-11.09MPa) were statistically equivalent to one another (P > 0.05), except for the TE-TX group (13.51MPa, P < 0.05). The two EXP groups showed less resin remnant. Only in the two TX groups were enamel fractures observed (three cases for each group). LIMITATIONS: The results were drawn only from ex vivo experiments. CONCLUSIONS: The hypothesis is rejected. This study suggests that a more flexible MMA resin is favourable for avoiding enamel fracture during metal bracket debonding.

Developmental regulations of Perp in mice molar morphogenesis.

Teraspanin transmembrane protein, Perp (P53 apoptosis effector related to PMP22), which is found in the plasma membrane as a component of the desmosome, is reported to be involved in the morphogenesis of the epithelium and the enamel formation of the incisor. However, its expression pattern and signaling regulation during molar development have not been elucidated in detail. We have examined the precise expression patterns of Perp in developing lower molars and employed the knock-down of Perp by antisense oligodeoxynucleotide treatment during in vitro organ cultivation at embryonic day 13 to define the precise developmental function of Perp. Perp was expressed mainly in the dental lamina and stellate reticulum regions at the bud and cap stages. After Perp knock-down, the tooth germ showed disruption of the dental lamina and stellate reticulum with altered apoptosis and proliferation. The changed expression levels of related signaling molecules from the enamel knot and desmosome were evaluated by real-time quantitative polymerase chain reaction. A renal capsule transplantation method was employed to examine the effects of Perp knock-down on molar crown development. Ultrastructural observations revealed that enamel was deposited more densely in an irregular pattern in the cusp region, and that dentin was hypo-mineralized after Perp knock-down at the cap stage. Thus, Perp might play important roles in the formation and integration of stellate reticulum, dental lamina structure and enamel formation through signaling interactions with the enamel knot and desmosome-related signaling molecules at the cap stage of lower molar development.