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.

Are There Differences in the Causes and Complications of Mandibular Third Molar Extraction in Older Patients Compared to Younger Patients?

BACKGROUND: Although most impacted third molars (ITMs) are extracted in the teens and early 20s, some undergo extractions after their 40s. It is unclear whether the reasons for extraction, the degree of impaction, and complications differ in patients in an older age group compared with a younger age group. PURPOSE: The purpose of this study was to measure the association between age and 1) reason for extraction and 2) postoperative complications. STUDY DESIGN, SETTING, SAMPLE: This was a retrospective cohort study of patients who had undergone surgical extraction of at least one mandibular ITM at a single institution. We excluded 1) age under 20 years, 2) follow-up period of less than 1 week, and 3) tooth extraction under general anesthesia. PREDICTOR VARIABLE: The primary predictor variable was age, classified into 3 groups (20s:20-29; 30s:30-39; over 40s: 40 and greater). MAIN OUTCOME VARIABLE(S): The primary outcome variables were the reason for extraction (prophylactic or symptomatic) and the presence of complications. The secondary outcome variable was type of complication (postoperative infection, dry socket, neurosensory disturbance, presenting pain over 1 month, retained root requiring secondary treatment). COVARIATES: The covariates were sex, laterality of ITM, and difficulty of extraction as measured by the difficulty index, a measure based on depth, orientation, and ramus relationship/space available, with a higher score indicating greater difficulty. ANALYSES: χ2 test was performed to analyze the association of categorical outcome variables and covariates. Level of statistical significance was set at P < .05. RESULTS: Of a total of 831 eligible subjects, there were 555 (66.8%), 159 (19.1%), and 117 (14.1%) in the 20s, 30s, and over 40s age groups, respectively. The percentage of symptomatic extraction of ITM was significantly higher in the over-40 age group compared with the 20s group (92.3 vs 69.4%, (P < .001). Complication rate also significantly differed between over 40s group and the 20s group (7.7 vs 1.8%, P < .001). Difficulty index and indications for ITM extraction were significantly different between groups (P < .001). CONCLUSION AND RELEVANCE: Symptoms, difficulty, and complications related to ITM increase at over 40 years of age. This should be taken into consideration during the joint clinical decision-making process with patients with ITM.

Microplastic contamination in urban aquatic environments: Occurrence characteristics in urban streams and stormwater runoff from urban surfaces.

This study investigates microplastics in urban environments, focusing on their abundance, types, and relationships with hydrological parameters. Microplastic analyses encompassed two steps: (1) examining urban streams including discharges from a wastewater treatment plant (WWTP) during non-rainy seasons, and (2) analyzing stormwater runoff from urban surfaces for microplastic content during rainy seasons. In urban streams, WWTP discharge exhibited higher microplastic concentrations compared to other streams, indicating WWTP discharge is a dominant source of microplastic pollution. The most prevalent microplastics detected were polypropylene, polyethylene, and their copolymer, although a variety of other types were also found. Concentrations of microplastics were notably influenced by specific urban land uses, as evidenced by a strong correlation (0.95) between microplastic concentrations and areas characterized by industrial and transportation activities. During rainy seasons, microplastics followed the pattern of stormwater runoff, but the highest concentrations, significantly exceeding those in urban streams, were observed before the peak runoff. These maximum concentrations and their timing of occurrence were linked to antecedent dry days, rain intensity, and runoff rate, showing significant statistical correlations. Regardless of their sizes, a diverse range of microplastic types was identified in these conditions, with no consistent pattern across different rain events. This highlights the complex nature of urban microplastic pollution. This study reveals that aquatic ecosystems are significantly affected by two primary factors: (1) the consistent contribution of microplastics from WWTP discharges, and (2) the short-term, but severe, impacts of microplastic pollution associated with stormwater runoff. Furthermore, it suggests the development of alternative strategies to mitigate microplastic pollution in aquatic ecosystems, informed by the findings on the characteristics of microplastics in urban environments. This research underscores the urgent need for integrated urban environmental management strategies, paving the way for future studies to further explore and address the multifaceted challenges posed by microplastic pollution in aquatic ecosystems.

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.

What Is the Perception of an Esthetic Lower Facial Third Profile in the Korean Layperson Population?

PURPOSE: The purpose of this study is to investigate whether there were differences in perception of facial profile esthetics at the upper lip, gnathic, and genial levels according to observer gender and age in the Korean layperson population. METHODS: Ten male (M1 to M10) and 10 female (F1 to F10) silhouettes with differing facial profiles at the level of the upper lip, mandibular, and chin were created from a male and female facial profile deemed well-balanced and orthognathic by reviewer consensus. These silhouettes were presented to 288 participants who were asked to rank the 5 most attractive male and female profiles. The participants were composed of 144 males and 144 females divided into 4 age groups: group I (under 20 years), group II (20 to 39 years), group III (40 to 59 years), and group IV (over 60 years). The most preferred profile and observer scores were investigated and calculated. Descriptive analysis, χ2 test, and analysis of variance were used for statistical analysis according to age and gender. RESULTS: The most preferred male and female silhouettes were orthognathic profiles. There was no significant difference according to gender when rating male (P = .281) and female (P = .442) silhouettes. No statistically significant difference was observed even when analyzed according to age groups. Although the difference in observer scores among each ranking showed a statistical difference according to age groups, the overall rankings showed a similar pattern both in male and female silhouettes. In the same age group, no difference in rank scores according to gender was observed. Severe concave profiles were found to have the lowest rank. CONCLUSIONS: An orthognathic profile was rated as the most desirable in the Korean layperson population, with few differences in perception of esthetic facial profile according to age and gender.

Follow-up evaluation of temporomandibular joints using magnetic resonance imaging after mandibular trauma: Case series analysis of young adult males.

BACKGROUND/AIMS: A relationship between mandibular fractures and traumatic temporomandibular joint (TMJ) conditions has been suggested in many studies. Although magnetic resonance imaging (MRI) is the best option for a TMJ evaluation, few studies have evaluated the TMJ condition after a mandibular fracture using MRI follow-up. The aim of this study was to evaluate the TMJ for post-traumatic conditions following a mandibular fracture using follow-up MRI. MATERIALS AND METHODS: Fourteen TMJs of seven young adult males (aged 19-21 years) with mandibular fractures were analyzed by MRI, and 12 TMJs of six patients were evaluated by follow-up MRI after the trauma. Regarding the intensity of MRI, the pathologic condition of TMJ was classified into acute joint inflammation, marrow edema, and joint space widening. RESULTS: Thirteen joints (92.9%) showed pathologic conditions, including 11 with acute joint inflammation (84.6%), 10 with joint space widening (76.9%), and six with marrow edema (46.2%). Five out of 12 evaluated joints were injected with dexamethasone. Among these, four joints healed within one week, and one healed within one month. Among the seven untreated TMJs, four and one joint healed within one week and one month, respectively, but two joints of one patient did not improve until one month. Although that patient received arthrocentesis, the right joint showed osteoarthritis six months after the trauma. CONCLUSIONS: Most TMJs were acutely damaged due to mandibular trauma and healed within one week to one month. A follow-up examination could be considered at one month after the injury to confirm the possibility of traumatic TMJ disorder, such as osteoarthritis.

Bobby sox homolog regulates tooth root formation through modulation of dentin sialophosphoprotein.

Tooth root development occurs through the interaction of multiple growth factors and transcription factors expressed in Hertwig's epithelial root sheath (HERS) and dental mesenchyme. Previously, we demonstrated that bobby sox homolog (Bbx) regulates odontoblast differentiation of human dental pulp stem cells. Here, we generated Bbx knockout (Bbx-/- ) mice to address the functional role of Bbx in tooth formation. During tooth development, Bbx was expressed in both dental epithelium and mesenchyme. However, molar and incisor morphology in Bbx-/- mice at postnatal Day 0 (P0) exhibited no prominent abnormalities compared with their wild-type (Bbx+/+ ) littermates. Until P28, the crown morphology in Bbx-/- mice was not distinctively different from Bbx+/+ littermates. Meanwhile, the length of the mandibular base in Bbx-/- mice was notably less at P28. Compared with Bbx+/+ mice, the mesial and distal root lengths of the first molar were reduced by 21.33% and 16.28% at P14 and 16.28% and 16.24% at P28, respectively, in Bbx-/- mice. The second molar of Bbx-/- mice also showed 10.16% and 6.4% reductions at P28 in the mesial and distal lengths, compared with Bbx+/+ mice, respectively. The gene expression analysis during early tooth root formation (P13) showed that the expression of dentin sialophosphoprotein (Dspp) was significantly decreased in Bbx-/- mice. Collectively, our data suggest that Bbx participates in tooth root formation and might be associated with the regulation of Dspp expression.

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.

Intraoral human herpes viruses detectable by PCR in majority of patients.

OBJECTIVES: To identify factors which influence the intraoral prevalence of human herpes viruses (HHVs) using mucosal swabs, saliva samples and qPCR analysis. METHODOLOGY: In this cross-sectional observational study, matched saliva and oral swabs were collected from a total of 115 subjects: 70 immunocompetent subjects with no mucosal abnormalities, 22 with mucosal abnormalities and 23 therapeutically immunocompromised individuals. Extracted DNA was analysed by multiplex qPCR for detection and quantification of HHVs 1-6. RESULTS: At least one human herpes virus was detected in 77.1% of immunocompetent individuals with no mucosal abnormalities, with EBV the most commonly detected at 61.4%. HHV-6 was detected in 17.1%, HSV-1 in 4.3% and CMV in 1.1%. Detection was higher in saliva than in oral swabs. There was no detection of HSV-2 or VZV. Neither presence of oral mucosal abnormality nor therapeutic immunocompromise was related to increased detection of human herpes virus. CONCLUSION: Commensal detection rates of EBV are high, and caution in clinical correlation of positive detection is warranted. Commensal CMV rates are low, and detection is likely to be clinically relevant. This study presents a comprehensive commensal detection rate of HHVs 1-6 by qPCR in saliva and swabs.

Craniofacial and dental injuries associated with stand-up electric scooters.

BACKGROUND/AIM: With the recent increased share of stand-up electric scooters (e-scooters), it is common to see people riding e-scooters on the roads in Korea. The aim of this study was to investigate traumatic injuries to the craniofacial region related to e-scooter accidents and to determine the role of dentists (especially oral and maxillofacial surgeons) in the evaluation of patients with trauma at the emergency department due to an e-scooter accident. MATERIALS AND METHODS: This retrospective study investigated the medical records of patients who visited the Gangnam Severance Hospital Emergency Care Center for trauma related to e-scooter use from January 1, 2017 to March 31, 2020. Medical records were reviewed to determine the injuries sustained to the craniofacial region related to e-scooter use, including location of the injury (eg, cranium, craniofacial bone, teeth, soft tissue) and the type of trauma (eg, fracture, laceration, abrasion, contusion, concussion). RESULT: A total of 256 patients' medical records were evaluated. Among them, 125 patients (48.8% of all patients) had sustained craniofacial trauma. Laceration (n = 56, 44.8%) was the most common type of craniofacial injury, followed by cerebral concussion (n = 49, 39.2%), dental injury (n = 27, 21.6%), and craniofacial bone fracture (n = 16, 12.8%). CONCLUSION: Dentists should always consider the possibility of brain trauma and perform a complete craniofacial and oral examination when assessing patients after e-scooter accidents as outlined by the International Association of Dental Traumatology guidelines. Additionally, it is necessary to educate e-scooter riders about the importance of wearing protective devices, such as helmets, to reduce the risk of injuries to the craniofacial region.

Three-dimensional assessment of the anterior and inferior loop of the inferior alveolar nerve using computed tomography images in patients with and without mandibular asymmetry.

BACKGROUND: The purpose of this study was to investigate the differences in configuration and dimensions of the anterior loop of the inferior alveolar nerve (ALIAN) in patients with and without mandibular asymmetry. METHOD: Preoperative computed tomography images of patients who had undergone orthognathic surgery from January 2016 to December 2018 at a single institution were analyzed. Subjects were classified into two groups as "Asymmetry group" and "Symmetry group". The distance from the most anterior and most inferior points of the ALIAN (IANant and IANinf) to the vertical and horizontal reference planes were measured (dAnt and dInf). The distance from IANant and IANinf to the mental foramen were also calculated (dAnt_MF and dInf_MF). The length of the mandibular body and symphysis area were measured. All measurements were analyzed using 3D analysis software. RESULTS: There were 57 total eligible subjects. In the Asymmetry group, dAnt and dAnt_MF on the non-deviated side were significantly longer than the deviated side (p < 0.001). dInf_MF on the non-deviated side was also significantly longer than the deviated side (p = 0.001). Mandibular body length was significantly longer on the non-deviated side (p < 0.001). There was no significant difference in length in the symphysis area (p = 0.623). In the Symmetry group, there was no difference between the left and right sides for all variables. CONCLUSION: In asymmetric patients, there is a difference tendency in the ALIAN between the deviated and non-deviated sides. In patients with mandibular asymmetry, this should be considered during surgery in the anterior mandible.

Effects of resveratrol on bone-healing capacity in the mouse tooth extraction socket.

BACKGROUND AND OBJECTIVE: After tooth extraction, the extraction socket undergoes several steps of soft and hard tissue healing. The healing process of the extraction socket is modulated by a range of signaling factors and biochemical agents. It has been reported that resveratrol, a polyphenolic compound, exhibits various biological effects, including anti-inflammatory, anti-carcinogenic, antioxidant, and anti-aging effects, and protects cardiovascular and bone tissues. In this study, we examined the cellular effects of resveratrol on human periodontal ligament (hPDL) cells and osteoblast-like (MC3T3-E1) cells and evaluated the bone-healing capacity of tooth extraction sockets in mice. MATERIAL AND METHODS: Resveratrol was applied to hPDL and MC3T3-E1 cells to detect cell proliferation and alkaline phosphatase (ALP) activity, and qPCR was employed to understand the gene expression level in vitro. For in vivo experiment, six-week-old C57BL/6 male mice were randomly divided into control (n = 15) and experimental (n = 15) groups and maxillary first molars were extracted by surgery. Experimental groups received 50-µM resveratrol on extraction sockets and analyzed the degree of new bone formation. RESULTS: Treatment of hPDL and MC3T3-E1 cells with resveratrol increased the cell proliferation and ALP activity and enhanced the expression of ALP, BMP-2, BMP-4, and OC genes. Resveratrol enhanced new bone formation in the lingual extraction socket in mice. CONCLUSION: These results suggest that resveratrol increases the cellular physiology of PDL and osteoblast including their proliferation and differentiation and may play an important role in bone-healing capacity after tooth extraction.

Is There a Difference in Condyle Position Changing Pattern Between Deviated and Non-Deviated Sides After Intraoral Vertical Ramus Osteotomy in Facial Asymmetry?

PURPOSE: The position changing pattern of the condyles after intraoral vertical ramus osteotomy (IVRO) on the deviated and non-deviated sides is not clearly known. This study was conducted to evaluate the changes in condylar position after IVRO in patients with facial asymmetry and to compare the deviated and non-deviated sides using computed tomography imaging. MATERIALS AND METHODS: This retrospective cohort study investigated patients with a diagnosis of mandibular prognathism with facial asymmetry who had undergone bilateral IVRO with Le Fort I osteotomy. Condylar positions were recorded on the non-deviated and deviated sides in the midaxial, midsagittal, and coronal planes at 3 time points using 3-dimensional analysis software: preoperatively (T1), at 6 months postoperatively (T2), and at 12 months postoperatively (T3). Linear and angular changes in condyle position were measured and analyzed between T1, T2, and T3. Reliability and comparative analyses were conducted. RESULTS: Thirty-two patients were involved in this study. At T2, the most superior point of the condyle moved to 1.15 ± 0.24 mm (inferiorly) and 0.88 ± 0.23 mm (anteriorly) on the deviated side (P = .0002 and P = .0005, respectively) and to 0.99 ± 0.25 mm (inferiorly) and 1.08 ± 0.34 mm (anteriorly) on the non-deviated side, showing significant differences (P < .0001 and P = .0007, respectively) compared with T1. The condyle position showed a tendency to recover to its original position by T3. However, there were no statistically significant differences between T2 and T3 (P > .05). Furthermore, there were no statistically significant differences between the deviated and non-deviated sides over the entire follow-up period (P > .05). CONCLUSIONS: The condyles did not completely recover to their preoperative positions until 12 months postoperatively. There was no significant difference between the deviated and non-deviated sides in mandibular prognathism with facial asymmetry.

Experimental Animal Model Systems for Understanding Salivary Secretory Disorders.

Salivary secretory disorders are life-disrupting pathologic conditions with a high prevalence, especially in the geriatric population. Both patients and clinicians frequently feel helpless and get frustrated by the currently available therapeutic strategies, which consist mainly of palliative managements. Accordingly, to unravel the underlying mechanisms and to develop effective and curative strategies, several animal models have been developed and introduced. Experimental findings from these models have contributed to answer biological and biomedical questions. This review aims to provide various methodological considerations used for the examination of pathological fundamentals in salivary disorders using animal models and to summarize the obtained findings. The information provided in this review could provide plausible solutions for overcoming salivary disorders and also suggest purpose-specific experimental animal systems.

Signaling Modulations of miR-206-3p in Tooth Morphogenesis.

MicroRNAs (miRNAs) are a class of naturally occurring small non-coding RNAs that post-transcriptionally regulate gene expression in organisms. Most mammalian miRNAs influence biological processes, including developmental changes, tissue morphogenesis and the maintenance of tissue identity, cell growth, differentiation, apoptosis, and metabolism. The miR-206-3p has been correlated with cancer; however, developmental roles of this miRNA are unclear. In this study, we examined the expression pattern and evaluated the developmental regulation of miR-206-3p during tooth morphogenesis using ex-vivo culture method. The expression pattern of miR-206-3p was examined in the epithelium and mesenchyme of developing tooth germ with stage-specific manners. Perturbation of the expression of miR-206-3p clearly altered expression patterns of dental-development-related signaling molecules, including Axin2, Bmp2, Fgf4, Lef1 and Shh. The gene expression complemented with change in cellular events including, apoptosis and proliferation which caused altered crown and pulp morphogenesis in renal-capsule-calcified teeth. Especially, mislocalization of ß-Catenin and SMAD1/5/8 were observed alongside dramatic alterations in the expression patterns of Fgf4 and Shh. Overall, our data suggest that the miR-206-3p regulate the cellular physiology during tooth morphogenesis through modulation of the Wnt, Bmp, Fgf, and Shh signaling pathways to form proper tooth pulp and crown.

Facilitation of Bone Healing Processes Based on the Developmental Function of Meox2 in Tooth Loss Lesion.

In the present study, we examined the bone healing capacity of Meox2, a homeobox gene that plays essential roles in the differentiation of a range of developing tissues, and identified its putative function in palatogenesis. We applied the knocking down of Meox2 in human periodontal ligament fibroblasts to examine the osteogenic potential of Meox2. Additionally, we applied in vivo periodontitis induced experiment to reveal the possible application of Meox2 knockdown for 1 and 2 weeks in bone healing processes. We examined the detailed histomorphological changes using Masson's trichrome staining and micro-computed tomography evaluation. Moreover, we observed the localization patterns of various signaling molecules, including α-SMA, CK14, IL-1ß, and MPO to examine the altered bone healing processes. Furthermore, we investigated the process of bone formation using immunohistochemistry of Osteocalcin and Runx2. On the basis of the results, we suggest that the knocking down of Meox2 via the activation of osteoblast and modulation of inflammation would be a plausible answer for bone regeneration as a gene therapy. Additionally, we propose that the purpose-dependent selection and application of developmental regulation genes are important for the functional regeneration of specific tissues and organs, where the pathological condition of tooth loss lesion would be.

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.

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.