GSK3beta inhibitor-induced dental mesenchymal stem cells regulate ameloblast differentiation.

OBJECTIVES: Epithelial-mesenchymal interactions are extremely important in tooth development and essential for ameloblast differentiation, especially during tooth formation. We aimed to identify the type of mesenchymal cells important in ameloblast differentiation. METHODS: We used two types of cell culture systems with chambers and found that a subset of debtal mesenchimal cells is important for the differentiatiuon of dental spithelial cells into ameloblasts. Further, we induced dental pulp stem cell-like cells from dental pulp stem cells using the small molecule compound BIO ( a GSK-3 inhibitor IX) to clarify the mechanism involved in ameloblast differentiation induced by dental pulp stem cells. RESULTS: The BIO-induced dental pulp cells promoted the expression of mesenchymal stem cell markers Oct3/4 and Bcrp1. Furthermore, we used artificial dental pulp stem cells induced by BIO to identify the molecules expressed in dental pulp stem cells required for ameloblast differentiation. Panx3 expression was induced in the dental pulp stem cell through interaction with the dental epithelial cells. In addition, ATP release from cells increased in Panx3-expressing cells. We also confirmed that ATP stimulation is accepted in dental epithelial cells. CONCLUSIONS: These results showed that the Panx3 expressed in dental pulp stem cells is important for ameloblast differentiation and that ATP release by Panx3 may play a role in epithelial-mesenchymal interaction.

The Retention Effect of Resin-Based Desensitizing Agents on Hypersensitivity-A Randomized Controlled Trial.

Recently, the development of dental materials has increased the availability of various hyperesthesia desensitizers. However, there are no studies on the duration of retreatment in terms of adherence rates. Thus, the adhesion rates of resin-based desensitizers were investigated. We used a conventional desensitizer and a recently developed desensitizer containing calcium salt of 4-methacryloxyethyl trimellitic acid (C-MET) and 10-methacryloyloxydecyl dihydrogen calcium phosphate (MDCP). These colored agents were applied to the surfaces of premolars and molars, and the area was measured from weekly oral photographs. Areas were statistically analyzed and mean values were calculated using 95% confidence intervals. A p-value of <0.05 was considered statistically significant. These rates were significantly higher on the buccal side of the maxilla and lower on the lingual side of the maxilla. In addition, the desensitizer containing C-MET and MDCP displayed significantly higher adhesion rates. It is suggested that this will require monthly follow-ups and reevaluation because both agents cause less than 10% adherence and there is almost no sealing effect after 4 weeks. In addition, the significantly higher adhesion rate of the desensitizer containing C-MET and MDCP indicated that the novel monomer contributed to the improvement in the adhesion ability.

Evaluation of a Hypersensitivity Inhibitor Containing a Novel Monomer That Induces Remineralization-A Case Series in Pediatric Patients.

BACKGROUND: Recently, tooth deformities have been frequently encountered by pediatric dentists. Severe enamel hypomineralization sometimes induces pain such as hyperesthesia, but composite resin restoration is difficult because it often detaches without any cavity preparation. Resin-based hypersensitivity inhibitors for tooth physically seal the dentinal tubules. It was reported that hypersensitivity inhibitor containing novel adhesive monomers forms apatite and induces remineralization in vitro. Therefore, these case series assessed the clinical effects of remineralization and the suppression of hypersensitivity by Bio Coat Ca (Sun Medical, Shiga, Japan). METHODS: After mechanical tooth cleaning was performed, the hypersensitivity inhibitors were applied and cured by light exposure. Changes in hypersensitivity were determined by visual analog scale (VAS). The improvement of hypomineralization was evaluated by the change in color tone based on the digital images of intraoral photographs. RESULTS: After repeated monthly treatments, these cases showed decreased hypersensitivity after the fourth application, while the opaque white and brownish color improved on the seventh application. CONCLUSION: This novel hypersensitivity inhibitor with calcium salt of 4-methacryloxyethyl trimellitic acid (C-MET) and 10-methacryloyloxydecyl dihydrogen calcium phosphate (MDCP) not only suppressed hypersensitivity but also improved cloudiness and brown spots in recently erupted permanent teeth in presented cases.

Mutant GDF5 enhances ameloblast differentiation via accelerated BMP2-induced Smad1/5/8 phosphorylation.

Bone morphogenetic proteins (BMPs) regulate hard tissue formation, including bone and tooth. Growth differentiation factor 5 (GDF5), a known BMP, is expressed in cartilage and regulates chondrogenesis, and mutations have been shown to cause osteoarthritis. Notably, GDF5 is also expressed in periodontal ligament tissue; however, its role during tooth development is unclear. Here, we used cell culture and in vivo analyses to determine the role of GDF5 during tooth development. GDF5 and its associated BMP receptors are expressed at the protein and mRNA levels during postnatal tooth development, particularly at a stage associated with enamel formation. Furthermore, whereas BMP2 was observed to induce evidently the differentiation of enamel-forming ameloblasts, excess GDF5 induce mildly this differentiation. A mouse model harbouring a mutation in GDF5 (W408R) showed enhanced enamel formation in both the incisors and molars, but not in the tooth roots. Overexpression of the W408R GDF5 mutant protein was shown to induce BMP2-mediated mRNA expression of enamel matrix proteins and downstream phosphorylation of Smad1/5/8. These results suggest that mutant GDF5 enhances ameloblast differentiation via accelerated BMP2-signalling.

Expression of versican and ADAMTS during rat tooth eruption.

A disintegrin and metalloprotease with thrombospondin type 1 motifs (ADAMTS) is a family of extracellular proteases and implicated in cleaving proteoglycans, such as aggrecan, versican and brevican. No information is available about expression or localization of these ADAMTSs in teeth. Versican is a large chondroitin sulfate proteoglycan that is present in a variety of connective tissue including dental pulp, dentin, cementum and periodontal ligaments. The present study was designed to investigate expression of ADAMTSs and versican during rat tooth eruption. Rat maxillary first molars in weeks 1, 2, 3, 4 and 6 were examined. The mRNA expression of ADAMTS1, ADAMTS4, ADAMTS5 and versican was localized using in situ hybridization. ADAMTS1, ADAMTS4, ADAMTS5 and versican were expressed in dental pulp cells, odontoblasts, cementoblasts, cementocytes, periodontal ligament cells, osteoblasts and osteocytes. The temporal and spatial expression pattern in these cellular phenotypes was comparable among ADAMTSs and versican. The present study suggests that dental pulp cells, odontoblasts, cementoblasts, cementocytes, periodontal ligament cells, osteoblasts and osteocytes may be involved in both production and degradation of versican with secreting ADAMTS1, ADAMTS4 and ADAMTS5.

Expression of extracellular matrix molecules, MMPs and TIMPs in alveolar bone, cementum and periodontal ligaments during rat tooth eruption.

Tooth eruption involves extensive degradation and reorganization of extracellular matrix (ECM) components. It is not known how ECM-degrading enzymes are coordinated with each other or how they are regulated in the event. The present study was designed to investigate mRNA expression of inhibitors of metalloproteinases (TIMPs) in comparison with matrix metalloproteinases (MMPs) as well as ECM molecules during rat first molar eruption using in situ hybridization. We also examined how TIMPs are involved in the process of tooth eruption, root formation, cementogenesis and alveolar bone remodelling. Expressions of type-I collagen, osteocalcin, MMPs 2 and 8, and TIMPs 1, 2 and 3 were shown in osteoblasts, osteocytes, cementoblasts, cementocytes and periodontal ligament fibroblasts, and the concomitant high expressions of the ECM molecules, MMPs and TIMPs in alveolar bone, cementum and periodontal ligaments were identified in the middle of first molar eruption. The remodelling of ECM in these periodontal tissues might be regulated through balance among the production of ECM molecules, the degradation of ECM by MMPs and the inhibition of MMPs by TIMPs during tooth eruption.