Functional modulation of lysophosphatidic acid type 2 G-protein coupled receptor facilitates alveolar bone formation.

Lipid biosynthesis is recently studied its functions in a range of cellular physiology including differentiation and regeneration. However, it still remains to be elucidated in its precise function. To reveal this, we evaluated the roles of lysophosphatidic acid (LPA) signaling in alveolar bone formation using the LPA type 2 receptor (LPAR2) antagonist AMG-35 (Amgen Compound 35) using tooth loss without periodontal disease model which would be caused by trauma and usually requires a dental implant to restore masticatory function. In this study, in vitro cell culture experiments in osteoblasts and periodontal ligament fibroblasts revealed cell type-specific responses, with AMG-35 modulating osteogenic differentiation in osteoblasts in vitro. To confirm the in vivo results, we employed a mouse model of tooth loss without periodontal disease. Five to 10 days after tooth extraction, AMG-35 facilitated bone formation in the tooth root socket as measured by immunohistochemistry for differentiation markers KI67, Osteocalcin, Periostin, RUNX2, transforming growth factor beta 1 (TGF-ß1) and SMAD2/3. The increased expression and the localization of these proteins suggest that AMG-35 elicits osteoblast differentiation through TGF-ß1 and SMAD2/3 signaling. These results indicate that LPAR2/TGF-ß1/SMAD2/3 represents a new signaling pathway in alveolar bone formation and that local application of AMG-35 in traumatic tooth loss can be used to facilitate bone regeneration and healing for further clinical treatment.

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.

Resveratrol enhances bone formation by modulating inflammation in the mouse periodontitis model.

OBJECTIVE: To evaluate the effect of resveratrol on periodontal bone regeneration after local delivery and to determine its effect on inflammatory mediators. BACKGROUND: Resveratrol is considered an anti-inflammatory polyphenolic stilbene involved in the modulation of inflammation. MATERIALS AND METHODS: Periodontitis was induced in mouse molars using a 5-day ligature model followed by the left second molar extraction and 50 µM resveratrol treatment for 1 and 2 weeks. We then examined specimens treated for 1 week histologically and with immunostaining. Microfocus-computed tomography (micro-CT) was used to examine the bone volume formation. RESULTS: After 1 week of treatment, proinflammatory cytokine levels (TNF-alpha and IL6), cells exhibiting neutrophil and macrophage marker (MPO), cell proliferation marker (Ki67), and preosteoblastic marker (RUNX2) reactivity decreased in the resveratrol-treated specimens compared to the control group. In contrast, we observed a higher number of CD31-, F4/80-, and osteocalcin- (OCN-) positive cells in the resveratrol-treated specimens. After 2 weeks, micro-CT confirmed an increased bone mass in the region of the extraction socket in the resveratrol-treated group. CONCLUSION: After 1 week, the resveratrol-treated specimens revealed evidence of inflammation modulation compared to the control group. These data suggest that resveratrol not only affects inflammation control but also is useful for treating periodontitis-related tissue defects and bone regeneration.

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.

The Effects of Distribution of Image Matched Fiducial Markers on Accuracy of Computer-Guided Implant Surgery.

PURPOSE: Image registration of the optical intraoral scan to computed tomography image is essential for computer-guided implant surgery. The remaining teeth, which are considered to be congruent structures observed in the scan and radiographic images, are used to perform the image registration. The purpose of this study was to evaluate the effects of the distribution of matching fiducial points on the accuracy of the image registration. MATERIALS AND METHODS: A partially edentulous model with three anterior remaining teeth was prepared. Two mini dental implants were inserted in the posterior edentulous areas on both sides, and computed tomography and surface scan data were obtained. Three groups were set according to the distribution of the image matching points used: localized distribution, unilateral distribution, and bilateral distribution. Fifteen graduate students performed the registration process in each group using the same image matching method. The accuracy of image registration was evaluated by measuring the geometric discrepancies between the radiographic and registered scan images in the anterior, middle, and posterior regions. One-way and two-way analysis of variance with the Tukey HSD post hoc test were used for statistical analysis (α = 0.05) RESULTS: In general, the registration discrepancy was lowest in the bilateral distribution group, followed by the unilateral distribution and localized distribution groups (p< 0.001). In the regional analysis, the registration error tended to increase as the measurement region moved farther from the matching points. The distribution of the matching points and measurement regions had a statistical interaction in the accuracy of image registration. CONCLUSION: The accuracy of image registration of the surface scan to the computed tomography is affected by the matching point distribution that can be improved by placing artificial markers in the edentulous areas.

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.

Effects of oleanolic acid acetate on bone formation in an experimental periodontitis model in mice.

OBJECTIVE: We evaluated the role of oleanolic acid acetate (OAA), a triterpenoid commonly used in the treatment of liver disorders, inflammatory diseases, and metastasis, in bone formation after tooth loss by periodontitis. BACKGROUND: Periodontitis causes the sequential degradation of the alveolar bone and associated structures, resulting in tooth loss. Several studies have attempted to regenerate the bone for implantation following tooth loss. METHODS: Maxillary left second molar was extracted from 8-week-old male mice following induction of periodontitis by ligature for 5 days. The extraction socket was treated with 50 ng/µL OAA for 1, 2, and 3 weeks. Detailed morphological changes were examined using Masson's trichrome staining, and the precise localization patterns of various signaling molecules, including CD31, F4/80, interleukin (IL)-6, and osteocalcin, were observed. The volume of bone formation was examined by Micro-CT. Osteoclasts were enumerated using tartrate-resistant acid phosphatase (TRAP) staining. For molecular dissection of signaling molecules, we employed the hanging-drop in vitro cultivation method at E14 for 1 day and examined the expression pattern of transforming growth factor (TGF)-ß superfamily and Wnt signaling genes. RESULTS: Histomorphometrical examinations showed facilitated bone formation in the extraction socket following OAA treatment. In addition, OAA-treated specimens showed the altered localization patterns of inflammatory and bone formation-related signaling molecules including CD31, F4/80, IL-6, and osteocalcin. Also, embryonic tooth germ mesenchymal tissue cultivation with OAA treatment showed the significant altered expression patterns of signaling molecules such as transforming growth factor (TGF)-ß superfamily and Wnt signaling. CONCLUSIONS: Oleanolic acid acetate induces bone formation and remodeling through proper modulation of osteoblast, osteoclast, and inflammation with regulations of TGF-ß and Wnt signaling.

Effects of image and education on the precision of the measurement method for evaluating prosthesis misfit.

STATEMENT OF PROBLEM: Various methods for measuring prosthesis misfit have been suggested. Although the cross-sectional images between the crown and abutment are used to evaluate the misfit, the effects of the image and the observer's knowledge on the precision of measurement are unknown. PURPOSE: The purpose of this in vitro study was to investigate the effects of the image and of education on the precision of prosthesis misfit measurement methods using inter- and intraobserver variability analyses. MATERIAL AND METHODS: The cross-sectional images in the margin were obtained using the computer-aided replica technique (CART), silicone replica technique (RT), and sectioning technique (ST). Twenty-five observers measured the absolute marginal discrepancy values in the images obtained from each group 4 times at an interval of 2 weeks; the observers went through different education sessions regarding the selection of the measurement points. The precision of measurement was determined and compared using the likelihood-ratio test statistic (α=.05) and the intraclass correlation coefficient with the linear mixed-effects model. RESULTS: The CART group exhibited the smallest deviations in the measurement variations, followed by the ST and RT groups (P<.001). Additional education on misfit measurements generally decreased the deviation values in all the groups, but without any significant differences. CONCLUSIONS: The cross-sectional image obtained from the measurement methods and education on the evaluation method affected the precision of the prosthesis misfit measurement. Digital methods might be a useful tool to significantly enhance the precision of the measurements.

Effect of Heat Treatment on Mechanical Properties of Nickel-Titanium Instruments.

INTRODUCTION: The aim of this study was to evaluate the torsional resistance, cyclic fatigue resistance, and bending stiffness of nickel-titanium (NiTi) file systems with different heat treatments and cross-sectional designs. METHODS: WaveOne Primary treated with memory-triple (MT) heat treatment (WOMT) was compared with WaveOne Primary (WO) and WaveOne Gold Primary (WOG). Torsional resistance test was performed using a customized device, and the distortion angle, ultimate strength, and toughness were evaluated. For cyclic fatigue resistance test, the instruments were reciprocated with continuous 4 mm up-and-down movement until fracture in a customized device, and the time to fracture was compared. Fracture surfaces of each group were examined under the scanning electron microscope. Bending stiffness was measured using a custom-made device. The results were analyzed using one-way analysis of variance and the Tukey's post hoc comparison at a significance level of 95%. RESULTS: WOMT showed higher ultimate strength and toughness than the other systems (P < .05). WOMT also showed highest cyclic fatigue resistance among the tested groups (P < .05). WO had the highest bending stiffness than others, whereas WOMT had a larger residual angle than others (P < .05). CONCLUSIONS: This new MT heat treatment technique makes NiTi file more flexible and improves its mechanical properties. In addition, the effect of heat treatment on flexibility was found to be more significant than that of the cross-sectional area.

Effects of Side Flattening on Torsional and Cyclic Fracture Resistance of Nickel-Titanium File.

INTRODUCTION: The purpose of this study was to evaluate the effect of side flattening of cutting flutes on the cyclic resistance and torsional resistance of nickel-titanium files. METHODS: Both novel flattened Platinum V.EU (PL) and standard nonflattened CC Premium V.EU (CC) rotaries were tested. For cyclic fatigue tests, all the files were rotated in an artificial root canal with a curvature of 45° and a radius of 6.06 mm at 300 rpm (n = 15 in each group). The number of cycles to failure (NCF) was calculated. For torsional tests, the files were rotated at 2 rpm clockwise until fracture occurred. The maximum torque value at fracture was measured and the toughness and distortion angle were computed. Subsequently, 5 fragments were randomly selected in each experiment, the cross-section and longitudinal direction of the fragments were photographed using a scanning electron microscope. An unpaired t-test was performed at a significance level of 95%. RESULTS: There was a statistically significant difference in NCF between CC and PL (P < .05). CC showed higher NCF than PL. There was no statistically significant difference between CC and PL with regards to the parameters related to torsional resistance (distortion angle, ultimate strength, and toughness) (P > .05). CONCLUSION: Within the limitations of this study, side flattening of the file did not improve cyclic resistance or torsional resistance of the files. As side flattening may reduce a file's cyclic resistance, such files should be used with caution in clinical practice.

Effect of MT Technology of Heat Treatment on Reciproc: Comparison of Reciproc, Reciproc Blue, and Reciproc MT.

INTRODUCTION: This study aimed to evaluate the effects from the memory-triple (MT) heat treatment on the fatigue resistance of the Reciproc by comparison with the file systems of same geometry. METHODS: Reciproc files subjected to MT heat treatment technology were designated as Group RMT and were compared with the original Reciproc (Group REC) and Reciproc Blue (Group REB). Each NiTi file from 3 groups (n = 15) was operated reciprocally with a repetitive up-and-down movement in the curved canal with 4 mm of pecking distance inside of the simulated canal at body temperature. When each file fractured, the time until fracture was recorded. The length of the fractured fragment was measured. Fractured fragments were observed under scanning electron microscope (SEM) to evaluate the topographic features of the surface. Differential scanning calorimetry (DSC) analysis was performed to estimate phase transformation temperatures. One-way analysis of variance and Duncan post hoc comparison were applied to compare among the groups at a significance level of 95%. RESULTS: RMT showed significantly higher fracture resistance (P < .05), whereas there was no difference in fatigue resistance between REC and REB. SEM examination showed the files from the 3 groups had similar topographic features. RMT showed a peak of austenite peak (Ap) at a temperature (52°C) higher than body temperature, whereas REC and REB showed Ap at 37 and 32°C, respectively. CONCLUSIONS: Under the condition of this study, the new heat treatment technique of MT technology could enhance the fatigue fracture resistance of the reciprocating files made of M-wire and Blue-wire.

Prohibitin modulates periodontium differentiation in mice development.

Introduction: Prohibitin (PHB) is an essential scaffold protein that modulates signaling pathways controlling cell survival, metabolism, inflammation, and bone formation. However, its specific role in periodontium development remains less understood. This study aims to elucidate the expression pattern and function of PHB in periodontium development and its involvement in alveolar bone formation. Methods: Immunolocalization of PHB in the periodontium of postnatal (PN) mice were examined. Phb morpholino was micro-injected into the right-side mandible at PN5, corresponding to the position where the alveolar bone process forms in relation to the lower first molar. The micro-injection with a scramble control (PF-127) and the left-side mandibles were used as control groups. Five days post-micro-injection, immunohistochemical analysis and micro-CT evaluation were conducted to assess bone mass and morphological changes. Additionally, expression patterns of signaling molecules were examined following Phb downregulation using 24-h in vitro cultivation of developing dental mesenchyme at E14.5. Results: The immunostaining of PHB showed its localization in the periodontium at PN5, PN8, and PN10. The in vitro cultivation of dental mesenchyme resulted in alterations in Bmps, Runx2, and Wnt signalings after Phb knock-down. At 5 days post-micro-injection, Phb knocking down showed weak immunolocalizations of runt-related transcription factor (RUNX2) and osteocalcin (OCN). However, knocking down Phb led to histological alterations characterized by decreased bone mass and stronger localizations of Ki67 and PERIOSTIN in the periodontium compared 1 to control groups. The micro-CT evaluation showed decreased bone volume and increased PDL space in the Phb knock-down specimens, suggesting its regulatory role in bone formation. Discussion: The region-specific localization of PHB in the margin where alveolar bone forms suggests its involvement in alveolar bone formation and the differentiation of the periodontal ligament. Overall, our findings suggest that Phb plays a modulatory role in alveolar bone formation by harmoniously regulating bone-forming-related signaling molecules during periodontium development.

Effect of Periodic Changes in Rotation Speed on Torsional Stress and Screw-in Force by Alternative Rotation Technique.

INTRODUCTION: This study evaluated the effect of periodic changes in rotation speed on torsional stress and screw-in force using the dedicated alternative rotation technique (ART) motion of the EQ-M (Metabiomed, Cheongju, Korea) endodontic motor. METHODS: Two ART modes of the EQ-M motor in 2 alternative techniques (ART30 and ART50) and continuous rotation were compared using ProTaper Next X2 (Dentsply Sirona, Charlotte, NC) files and simulated resin blocks (n = 12 per group). ART30 and ART50 were operated by continuous rotation of 350 rpm for 360° and then rotated at 30% increased speed from the base speed for 180° and at 50% increased speed for 180°, respectively. Before the test, the simulated resin blocks were pre-enlarged using ProTaper Gold S1 and S2 (Dentsply Sirona) and fixed on a metal stage connected to the force- and torque-measuring unit. During shaping the simulated canal in an automatic up-and-down manner, the parameters of maximum torque, sum of torque, maximum screw-in force, and maximum apical force were measured. The data were statistically analyzed using 1-way analysis of variance and the Tukey post hoc comparison test at the 95% significance level. RESULTS: The ART30 and ART50 groups showed a lower maximum torque, sum of torques, screw-in force, and apical drive force than the continuous rotation group. There was no significant difference between the ART30 and ART50 groups (P > .05). When the screw-in force increased suddenly, the torque correspondently increased. CONCLUSIONS: Under the limitations of this study, the ART mode could reduce the torsional stress and apical forces of the screw-in during instrumentation in comparison with continuous rotation.

Comparison of the effects from coronal pre-flaring and glide-path preparation on torque generation during root canal shaping procedure.

This study aimed to evaluate the effect of glide-path preparation and coronal pre-flaring on torque generation of rotary nickel-titanium files. Sixty mesiobuccal root canals of 3D printed resin teeth (Dentalike) were randomly divided into 4 groups (n = 15) according to the preparation method; Group 1: OneCurve only; Group 2: OneG and OneCurve; Group 3: OneFlare and OneCurve; Group 4: OneFlare, OneG, and OneCurve. During the final canal preparation procedure, the torque generated was recorded. The sum of generated torque and maximum torque were calculated. Statistical analysis was performed with one-way ANOVA and Duncan's post hoc comparison. The summed torque was reduced by glide-path preparation and coronal pre-flaring (P < 0.05), whereas the maximum torque was not reduced by the glide-path preparation (P > 0.05). It will be advantageous to create a glide-path with coronal pre-flaring to reduce the stress and reactive forces to the NiTi file and root dentin.

Pilot Evaluation of Sealer-Based Root Canal Obturation Using Epoxy-Resin-Based and Calcium-Silicate-Based Sealers: A Randomized Clinical Trial.

The purpose of this randomized controlled clinical trial was to assess the short-term clinical effectiveness of sealer-based root canal obturation using calcium-silicate-based sealer (CSBS) and epoxy-resin-based sealer (ERBS). A total of eighty patients (eighty teeth) were enrolled and seventy-two patients (seventy-two teeth) were randomly assigned into four different sealer groups: AH Plus (AH, Dentsply Sirona), ADseal (AD, Meta Biomed), CeraSeal (CS, Meta Biomed), or EndoSeal TCS (ES, Maruchi). At the first visit, root canal treatment was performed using ProTaper. Next, the NiTi file system and passive ultrasonic irritation was applied with 2.5% NaOCl. At the second visit, the teeth were obturated with a sealer-based obturation technique using the assigned sealer. The quality of filling obturation was evaluated with postoperative radiographs in terms of the presence of voids and sealer extrusion. The patients were recalled at 1-week, 1-month, and 3-month intervals, and pain levels were measured using visual analog scales (VAS), and clinical examination performed with percussion and palpation. Radiographs were also taken. The results were statistically compared by using the χ2 test or Fisher exact test. Among the 80 teeth included, 72 teeth received allocated intervention and 71 teeth were included in the analysis (98.6% recall). There was no significant difference among the sealers in void and sealer extrusion (p > 0.05). Postoperative pain was not indicated prominently regardless of the type of sealer. For a 3-month follow-up period, all cases were maintained successfully except for one case from AD. According to the findings, there was no significant difference between CSBS and ERBS in terms of postoperative pain or the healing process, and the variation in filling quality appears to be linked to the properties of each product.

Facilitation of Reparative Dentin Using a Drug Repositioning Approach With 4-Phenylbutric Acid.

For hard tissue formation, cellular mechanisms, involved in protein folding, processing, and secretion play important roles in the endoplasmic reticulum (ER). In pathological and regeneration conditions, ER stress hinders proper formation and secretion of proteins, and tissue regeneration by unfolded protein synthesis. 4-Phenylbutyric acid (4PBA) is a chemical chaperone that alleviates ER stress through modulation in proteins folding and protein trafficking. However, previous studies about 4PBA only focused on the metabolic diseases rather than on hard tissue formation and regeneration. Herein, we evaluated the function of 4PBA in dentin regeneration using an exposed pulp animal model system via a local delivery method as a drug repositioning strategy. Our results showed altered morphological changes and cellular physiology with histology and immunohistochemistry. The 4PBA treatment modulated the inflammation reaction and resolved ER stress in the early stage of pulp exposure. In addition, 4PBA treatment activated blood vessel formation and TGF-ß1 expression in the dentin-pulp complex. Micro-computed tomography and histological examinations confirmed the facilitated formation of the dentin bridge in the 4PBA-treated specimens. These results suggest that proper modulation of ER stress would be an important factor for secretion and patterned formation in dentin regeneration.

Gene profiling in dorso-ventral patterning of mouse tongue development.

BACKGROUND: The tongue is a muscular fleshy organ in the oral cavity that is anatomically divided into the dorsal, ventral, anterior, and posterior part. The intricate tissue organisation and diverse origins of the tongue make it a complex organ of the oral cavity. OBJECTIVES: To reveal the signalling molecules involved in the formation of the dorsal and ventral parts of the tongue through microarray analysis. METHODS: Dorsal and ventral tongue tissues were isolated from embryonic day 14 mice by micro-dissection. RNA was extracted from the dorsal and ventral tongue tissues separately for microarray analysis. Microarray data were confirmed by quantitative reverse transcription polymerase chain reaction and whole-mount in situ hybridisation. RESULTS: Microarray analysis revealed expression of 33,793 genes. Of these, 931 genes were found to be equally expressed in both the dorsal and ventral parts of the tongue. On limiting the fold-change cut-off to over 1.5-fold, 725 genes were expressed over 1.5-fold in the ventral part and 1,672 in the dorsal part of the tongue. The qPCR and whole-mount in situ hybridisation revealed the expressions of angiopoietin 2 (Angpt2), fibroblast growth factor 18 (Fgf18), mesenchyme homeobox gene1 (Meox1), and SPARC-related modular calcium binding 2 (Smoc2) in the ventral part of the tongue. CONCLUSIONS: Numerous signalling molecules can be selected from our microarray results to examine their roles in tongue development and disease model systems. In the near future, the selection of candidate genes and their functional evaluations will be performed through loss- and gain-of-function mutation studies.

The effects of 4-Phenylbutyric acid on ER stress during mouse tooth development.

Introduction: During tooth development, proper protein folding and trafficking are significant processes as newly synthesized proteins proceed to form designated tissues. Endoplasmic reticulum (ER) stress occurs inevitably in tooth development as unfolded and misfolded proteins accumulate in ER. 4-Phenylbutyric acid (4PBA) is a FDA approved drug and known as a chemical chaperone which alleviates the ER stress. Recently, several studies showed that 4PBA performs therapeutic effects in some genetic diseases due to misfolding of proteins, metabolic related-diseases and apoptosis due to ER stress. However, the roles of 4PBA during odontogenesis are not elucidated. This study revealed the effects of 4PBA during molar development in mice. Methods: We employed in vitro organ cultivation and renal transplantation methods which would mimic the permanent tooth development in an infant period of human. The in vitro cultivated tooth germs and renal calcified teeth were examined by histology and immunohistochemical analysis. Results and Discussion: Our results revealed that treatment of 4PBA altered expression patterns of enamel knot related signaling molecules, and consequently affected cellular secretion and patterned formation of dental hard tissues including dentin and enamel during tooth morphogenesis. The alteration of ER stress by 4PBA treatment during organogenesis would suggest that proper ER stress is important for pattern formation during tooth development and morphogenesis, and 4PBA as a chemical chaperone would be one of the candidate molecules for dental and hard tissue regeneration.

Fiber-reinforced composite post removal using guided endodontics: a case report.

Although several techniques have been proposed to remove fiber-reinforced composite (FRC) post, no safe and efficient technique has been established. Recently, a guided endodontics technique has been introduced in cases of pulp canal obliteration. This study describes 2 cases of FRC post removal from maxillary anterior teeth using this guided endodontics technique with a dental operating microscope. Optically scanned data set from plaster cast model was superimposed with the data set of cone-beam computed tomography. By implant planning software, the path of a guide drill was selected. Based on them, a customized stent was fabricated and utilized to remove the FRC post. Employing guided endodontics, the FRC post was removed quickly and safely with minimizing the loss of the remaining tooth structure. The guided endodontics was a useful option for FRC post removal.