• Journal of Periodontal and Implant Science
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• v.32 no.1
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• pp.1-12
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• 2002

The periodontal ligament(PDL) is a unique tissue that is crucial for tooth function. However, little is known of the molecular mechanisms controlling PDL function. PDL-specific protein;PDLs22 had been previously identified as a novel protein isolated from cultured human PDL fibroblasts using subtraction hybridization between human gingival fibroblasts and PDL fibroblasts. The aim of this study was to examine the expression pattern and tissue localization of PDLs22 protein in embryonic and various postnatal stages of developing mouse using immunohistochemical staining. Embryos (E18) and postnatal (P1, P4, P5, P15, P18) were decapitated and the heads were fixed overnight in a freshly prepared solution of 4% paraformaldehyde. Some specimens were decalcified for $2{\sim}4$ weeks in a solution containing 10% of the disodium salt of ethylenediamine-tetraacetic acid (EDTA). Next, tissues were dehydrated, embedded in paraffin and sectioned serially at $6{\mu}m$ in thickness. Polyclonal antiserum raised against PDLs22 peptides, ISNKYLVKRQSRD, were made. The localization of PDLs22 in tissues was detected by polyclonal antibody against PDLs22 by means of immunohistochemical staining. The results were as follows; 1. Expression of PDLs22 protein was not detected in the tooth germ of bud and cap stage. 2. At the late bell stage and root formation stage, strong expression of PDLs22 protein was observed in developing tooth follicle, osteoblast-like cells, and subodontoblastic cells in the tooth pulp, but not in gingival fibroblasts, ameloblasts and odontoblasts of tooth germ 3. In erupted tooth, PDLs22 protein was intensely expressed in PDL and osteoblast-like cells of alveolar bone, but not in gingival fibroblasts, mature osteocytes and adjacent salivary glands. 4. In the developing alveolar bone and mid-palatal suture, expression of PDLs22 protein was seen in undifferentiated mesenchymal cells and osteoblast-like cells of developing mid-palatal suture, but not in mature osteocytes and chondrocytes. These results suggest that PDLs22 protein may play an important role in the differentiation of undifferentiated mesenchymal cells in the bone marrow and PDL cells, which can differentiate into multiple cell types including osteoblasts, cementoblasts, and PDL fibroblasts. However, more researches should be performed to gain a better understanding of the exact function of PDLs22 protein which related to the PDL cell differentiation.

• Journal of dental hygiene science
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• v.10 no.5
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• pp.395-401
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• 2010

Over expression of TGF-${\beta}1$ revealed the same phenotype as NFI-C deficient mouse. It has been reported that NFI-C deficient mice demonstrated abnormal odontoblast differentiation and aberrant dentin formation during root development. In the present study, in order to investigate the histological differences between wild type (WT) mouse and NFI-C deficient mouse, we compared morphological characteristics and smad4 expression between those mice. Hematoxyline-eosin (H-E) staining was used to investigate morphological changes and immunohistochemistry was also performed to observe the Smad4 expression pattern. In H-E staining, incisor of NFI-C deficient mouse showed an open area in the lingual root, irregular odontoblasts and osteodentin. Also, NFI-C deficient mouse showed short root and osteodentin in molar. In addition, Smad4 protein was strongly expressed in NFI-C deficient mouse compared with wild type. These findings suggest that NFI-C deficiency affects odontoblast differentiation and result in the formation of abnormal roots. Therefore, balancing between NFI-C and TGF-${\beta}$ signaling including Smad4 is important for the regulation of normal odontoblast differentiation and dentin formation.

• Journal of Periodontal and Implant Science
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• v.36 no.2
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• pp.385-396
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• 2006

For the regeneration of periodontal tissues, the microenvironment for new attachment of connective tissue fibers should be provided, At this point of view, cementum formation in root surface plays a key role for this new attachment. This study was performed to figure out which factor promotes differentiation of cementoblast Considering anatomical structure of tooth, we selected the cells which may affect the differentiation of cementoblast - Ameloblast, OD11&MDPC23 for odontoblasts, NIH3T3 for fibroblsts and MG63 for osteoblasts. And OCCM30 was selected for cementoblast cell line. Then, the cell lines were cultured respectively and transferred the conditioned media to OCCM30. To evaluate the result, Alizarin red S stain was proceeded for evaluation of mineralization. The subjected mRNA genes are bone sialoprotein(BSP), alkaline phosphate(ALP) , osteocalcin(OC), type I collagen(Col I), osteonectin(SPARC ; secreted protein acidic and rich in cysteine). Expression of the gene were analysed by RT-PCR, The results were as follows: 1. For alizarin red S staining, control OCCM30 didn't show any mineralized red nodules until 14 days. But red nodules started to appear from about 4 days in MDPC-OCCM30 & OD11-OCCM30. 2. For results of RT-PCR, ESP mRNAs of control-OCCM30 and others were expressed from 14 days, but in MDPC23-OCCM30 & OD11-OCCM30 from 4 days. Like this, the gene expression of MDPC23-OCCM30 & OD11-OCCM30 were detected much earlier than others. 3. For confirmation of odontoblast effect on cementoblast, conditioned media of osteoblasts(MG63) which is mineralized by producing matrix vesicles didn't affect on the mineralized nodule formation of cementoblasts(OCCM30). This suggest the possibility that cementoblast mineralization is regulated by specific factor in dentin matrix protein rather than matrix vesicles. Therefore, we proved that the dentin/odontoblast promotes differentiation/mineralization of cementoblasts. This new approach might hole promise as diverse possibilities for the regeneration of tissues after periodontal disease.

• Anatomy & Biological Anthropology
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• v.26 no.1
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• pp.41-49
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• 2013

Dental epithelial and mesenchymal cells that form the teeth undergo dynamic changes in cell cycle during tooth development and morphogenesis. Although proliferation has been known as a key event during odontogenesis, the cell cycle phases and their relations with the complicated molecular mechanisms of tooth development are not fully understood yet. This study comparatively examined the expression patterns of Ki-67, cyclin A, and cyclin D1 during tooth development in the mouse incisor and molar in order to identify the cell-cycle characteristics during odontogenesis. We found that Ki-67 and cyclin A were expressed in the proliferating cells in the dental epithelial and mesenchymal tissues at the bud, cap and bell stages. Cycln D1 showed distinct expression in the incisor odontoblast region and the enamel knot, in which Ki-67 nor cyclin A was expressed. Our results provide specific information on the cell cycle phases during tooth development that may provide clues to relate them with the complex odontogenic mechanisms. Furthermore, we suggest that our findings enlightened the previous studies on the incisor odontoblasts and the enamel knot during tooth development.

• Journal of the korean academy of Pediatric Dentistry
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• v.46 no.2
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• pp.219-225
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• 2019

The purpose of this study was to investigate the odontoblast gene expression related to the subculture speed of supernumerary dental pulp stem cells (sDPSCs). The stem cell is undifferentiated cells which has the ability to differentiate into various cells. Specific stimulation or environment induces cell differentiation, and these differentiation leads to bone or muscle formation. 20 sDPSCs were obtained from 20 children under aseptic condition. During the culture through the 10th passage, the third passage cells which showed short subculture period and 10th passage cells which showed long subculture period were earned. Each cell was divided into differentiated group and non-differentiated group. Quantitative real-time polychain reaction (q-RT-PCR) was performed for each group. The genes related to odontoblast differentiation, Alkaline Phosphatase (ALP), Osteocalcin (OCN), Osteonectin (ONT), Dentin sialophosphoprotein (DSPP) and Dentin matrix acidic phosphoprotein 1 (DMP-1), were measured. Differentiated cells showed more gene expression levels. Undifferentiated cells showed higher gene expression level in 10th passages but differentiated cells showed higher gene expression level in 3rd passages. Cells that showed faster subculture period showed relatively lower gene expression level except for OCN and DSPP.

• International Journal of Oral Biology
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• v.46 no.2
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• pp.67-73
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• 2021

This study summarizes the recent cutting-edge approaches for dentin regeneration that still do not offer adequate solutions. Tertiary dentin is formed when odontoblasts are directly affected by various stimuli. Recent preclinical studies have reported that stimulation of the Wnt/β-catenin signaling pathway could facilitate the formation of reparative dentin and thereby aid in the structural and functional development of the tertiary dentin. A range of signaling pathways, including the Wnt/β-catenin pathway, is activated when dental tissues are damaged and the pulp is exposed. The application of small molecules for dentin regeneration has been suggested as a drug repositioning approach. This study reviews the role of Wnt signaling in tooth formation, particularly dentin formation and dentin regeneration. In addition, the application of the drug repositioning strategy to facilitate the development of new drugs for dentin regeneration has been discussed in this study.

• Restorative Dentistry and Endodontics
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• v.32 no.2
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• pp.95-101
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• 2007

This study was performed to verify the possibility of MTA and calcium sulfate as a pulp capping agent through comparing the dental pulp response in dogs after capping with MTA, calcium sulfate, and calcium hydroxide. 24 teeth of 2 dogs, 8 month old, were used in this study. Under general anesthesia, cervical cavities were prepared and pulp was exposed with sterilized #2 round bur in a high speed handpiece. MTA calcium hydroxide, and calcium sulfate were applied on the exposed pulp. Then the coronal openin,fs were sealed with IRM and light-cured composite. Two months after treatment, the animals were sacrificed. The extracted teeth were fixed in 10% neutral-buffered formalin solution and were decalcified in formic acid-sodium citrate. They were prepared for histological examination in the usual manner. The sections were stained with haematoxylin and eosin. In MTA group, a hard tissue bridges formation and newly formed odontoblasts layer was observed. There was no sign of pulp inflammatory reaction in pulp tissue. In calcium hydroxide group, there was no odontoblast layer below the dentin bridge. In pulpal tissue, chronic inflammatory reaction with variable intensity and extension occurred in all samples. In calcium sulfate group, newly formed odontoblast layer was observed below the bridge. Mild chronic inflammation with a few neutrophil infiltrations was observed on pulp tissue. These results suggest that MTA is more biocompatible on pulp tissue than calcium hydroxide or calcium sulfate.

• Restorative Dentistry and Endodontics
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• v.30 no.5
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• pp.423-430
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• 2005

Ameloblasts are responsible for the formation and maintenance of enamel which is an epithelially derived protective covering for teeth. Ameloblast differentiation is controlled by sequential epithelial-mesenchymal interactions. However, little is known about the differentiation and maturation mechanisms. OD314 was firstly identified from odontoblasts by subtraction between odontoblast/pulp cells and osteoblast/dental papilla cells, even though OD314 protein was also expressed in ameloblast during tooth formation. In this study, to better understand the biological function of OD314 during amelogenesis, we examined expression of the OD314 mRNA and protein in various stages of ameloblast differentiation using in-situ hybridization and immunohistochemistry. The results were as follows : 1. The ameloblast showed 4 main morphological and functional stages referred to as the presecretory, secretory, smooth-ended, and ruffle-ended. 2. OD314 mRNA was expressed in secretory ameloblast and increased according to the maturation of the cells. 3. OD314 protein was not expressed in presecretory ameloblast but expressed in secretory ameloblast and maturative ameloblast. OD314 protein was distributed in entire cytoplasm of secretory ameloblast. However, OD314 was localized at the proxiamal and distal portion of the cytoplasm of smooth-ended and ruffle-ended ameloblast. These results suggest that OD314 may play important roles in the ameloblast differentiation and maturation.

• The korean journal of orthodontics
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• v.31 no.5 s.88
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• pp.525-534
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• 2001

Bone remodeling in response to force requires coordinated actions of osteoblasts, osteoclasts, osteocytes, and periodontal ligament cells. Coordination among these cells may be mediated, in part, by cell-to-cell communication via gap junctions. This study was designed to evaluate the expression of gap junction, connection 43 In periodontal tissue during the experimental movement of rat's incisors, by LSAB(labelled streptavidine biotin) immunohistochemical staining for connexin 43. Twenty seven Sprague-Dawley rats were divided into a control group(3 rats), and 6 experimental groups(24 rats) where 75g of force was applied from helical springs across the maxillary incisors. Rats of experimental groups were sacrificed at 12 hours, 1, 4, 7, 14 and 28 days after force application, respectively. And the tissues of a control group and experimental groups were studied immunohistochemically. The results were as follows : 1. In control group, the expression of connexin 43 was rare in gingiva, dentin, cementum, periodontal ligament, and bone cells. 2. In experimental group, the expression of connexin 43 was increased in pulp, periodontal ligament, osteoblasts, and osteoclasts, comparing to that in control. And it was rare in gingiva, dentin, and odontoblasts regardless of the duration of force application, which was not different from that of control group. 3. The expression of connexin 43 in pulp of experimental group began to increase in 4-day after force application and got to the highest degree at 7-day. And it decreased after 14-day to be similar to that of control group at 28-day. 4. The expression of connexin 43 in periodontal ligament was noted in small capillaries adjacent to alveolar bone, showing higher intensity of immunolabelling after 4-day And it was stronger in the pressure side than in tension side of periodontal ligament. After 7-day, decrease in connexin 43 expression was observed. 5. The expression of connexin 43 in alveolar bone began to increase 1-day, reached to the highest degree at 4-day, and decreased at 7-day. And the expression in osteoclasts was more than that in osteoblasts or osteocyte at 7-day.