• International Journal of Oral Biology
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• 제33권4호
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• pp.187-195
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• 2008

Reparative dentine formation requires newly differentiated odontoblast-like cells. Therefore, identification of the molecule that stimulates the odontogenic differentiation of precursor cells in the tooth pulp will be helpful for the development of strategies to repair damaged pulp. In this study, we examined the effect of N-acetylcysteine (NAC) on the odontogenic differentiation of MDPC-23 cells, a mouse odontoblast-like cell line derived from dental papilla, and primary cultured rat dental papilla cells (RDPCs). NAC (1-30 mM) suppressed production of reactive oxygen species in MDPC-23 cells in a dose-dependent manner. Although 5 to 20 mM NAC did not alter MDPC-23 cell proliferation, 1 or 30 mM NAC significantly inhibited it. NAC enhanced mineralized nodule formation and the expression of several odontoblast differentiation-associated genes in both RDPCs and MDPC-23. This NAC stimulatory effect was significant, even at concentrations lower than 1 mM. However, NAC did not stimulate expression of bone morphogenetic protein-2, -4, or -7, which are known to enhance odontogenic differentiation. Since reactive oxygen species are also involved in the pulp toxicity of resin-based restorative materials, these results suggest that NAC may be a promising candidate for supplementation of dental restorative materials in order to enhance reparative dentine formation.

• Applied Microscopy
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• 제43권1호
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• pp.27-33
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• 2013

Inflammatory cells express the inflammatory cytokines and growth factors induced by lipopolysaccharide (LPS). Odontoblasts are located at the pulp-dentin interface and extend their cell processes far into the dentin where they are the first cells to encounter microorganisms or their products. Therefore, this study examined the expression of some growth factors related to the signal pathway, such as growth factor receptor binding protein 2 (Grb2)-Ras in odontoblast-like dental pulp cells, after a treatment with LPS. After 60 minutes, the mRNA and protein expression levels of Grb2 and Ras were higher in the LPS-treated cells than in the control cells. The level of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) mRNA expression was increased significantly to a level similar to that of Grb2 and Ras at 60 minutes. The platelet-derived growth factor-AA (PDGF-AA) mRNA level was expressed strongly in the odontoblast like dental pulp cells without an association with LPS stimulation. Scanning electron microscopy revealed many extensions of the cytoplasmic processes and the number of processes increased gradually at 30, 60 and 90 minutes after LPS stimulation. From these results VEGF and bFGF expression might be induced through the Grb2-Ras signal transduction pathway in LPS treated odontoblasts.

• Restorative Dentistry and Endodontics
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• 제35권4호
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• pp.273-284
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• 2010

본 연구의 목적은 흰 쥐(rat)를 이용하여 미성숙 단계의 치아를 발치 후 즉시 재식 하였을 때 나타나는 치수의 치유 양상을 조직학적으로 관찰하고자 하는 것이다. 생후 4주된 암컷 Sprague-Dawley 계 흰쥐의 상악 우측 제1대구치를 발거 후 원래의 치조골와 내로 위치시켰다. 재식 후 3일째부터 국소적으로 치수 내 염증 소견이 관찰되었으나, 치근 부위에서는 이미 치수의 재혈관화 및 치유가 진행되고 있는 소견이 관찰되었다. 재식 후 5일째부터는 odontoblast-like cell이 관찰되기 시작하였다. 삼차 상아질의 형성은 재식 후 1주째부터 관찰되기 시작하였으며, 2주째부터는 확실히 관찰할 수 있었다. Odontoblast-like cell 및 삼차상아질 형성은 4주째까지도 계속 관찰되었다. 재식 후 4주째에는 bone-like tissue 및 cementum-like tissue이 형성되었음을 관찰하였다. 본 실험을 통하여 흰 쥐 치아 재식 시 석회화 과정은 초기에는 삼차상아질 침착에 의해서 진행되나, 시간이 경과하면서 점차 bone-like tissue 또는 cementum-like tissue가 차지하는 비율이 증가하는 것을 관찰하였다.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제33권4호
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• pp.301-307
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• 2011

Purpose: If the tooth structure is damaged, then it is impossible to regenerate the tooth. The materials used to restore the tooth structure are not related to the composition of the tooth. The materials used to restore the structure can't replace the natural tooth because they just fill the defective structure. Calcium phosphate cement remineralizes the dentin and almost replaces the natural tooth, but there are some disadvantages. We conducted basic tests with Biomimetic CPC (Bio-CPC) to make sure of the possibility of the biomaterial to remineralize the defective tooth structure. Methods: In this study, the bioactivity and biocompatibility of Bio-CPC were evaluated for its potential value as the bio-material for regeneration of damaged tooth structure by conducting a cell toxicity assay (WST-1 assay), a cytokinesis-block micronucleus assay, a chromosomal aberration test, total RNA extraction and RT-PCR on MDPC-23 mouse odontoblast-like cells. Results: The in vitro cytotoxicity test showed that the Bio-CPC was fairly cytocompatible for the MDPC-23 mouse odontoblast-like cells. Conclusion: Bio-CPC has a possibility to be a new biomaterial and further study of Bio-CPC is needed.

• International Journal of Oral Biology
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• 제31권3호
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• pp.73-79
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• 2006

Tissue stem cells are used for the regenerative medicine. In previous study we observed hard tissue formation of human dental pulp-derived cells using alginate scaffold. In this study, we explore the ability to differentiate of the 13th passage cells with glycerol 2-phosphate disodium salt hydrate (${\beta}-GP$) which accelerate calcification. Reverse transcriptase Polymerase Chain Reaction (RT-PCR), transplants using alginate scaffold and histological examination were performed. We observed the expression of DSPP mRNA on day 10 cultured cells with ${\beta}-GP$. In conclusion, the 13th passage cells still have an ability to differentiate into odontoblast-like cells and alginate supports the differentiation of cultured cells in the transplants.

• Applied Microscopy
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• 제16권2호
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• pp.1-13
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• 1986

The purpose of this study was to investigate the morphology and intercellular junctions of the odontoblast of dental pulp in the rat incisor by means of the freeze fracture electron microscopy. Twenty male Sprague-Dawley rats weighing $150{\sim}200g$ were used. After being anesthetized by an intraperitoneal injection of 0.5 ml sodium pentobarbital per kg in body weight(60 mg/ml) the animals were perfused with 2.5% glutaraldehyde-2% paraformaldehyde fixative in 0.1 M cacodylate buffer, pH 7.2 through the ascending aorta for one hour. The incisors were carefully extracted from the jaws and demineralized by suspending them in 0.1 M EDTA in 3% glutaraldehyde (pH 7.2) for two weeks. After demineralization, the specimens were obtained from the portion divided into five equal parts. For freeze-fracture replication, demineralized tissues were infiltrated for several hours with 10%, 25% glycerol in 0.1M cacodylate buffer as a cryoprotectant and then frozen in liquid Freon 22 and stored in liquid nitrogen. Fracturing and replication were done in Balzers BAF 400D high-vacuum freeze-fracture apparatus at $-120^{\circ}C$ under routine $5X10^{-7}$ Torr vacuum. The tissue was immediately replicated with platinum unidirectionally at $45^{\circ}$ angle and reinforced with carbon at $90^{\circ}$ angle unidirectionally or by using a rotary stage. The replication process was monitored by a quartz-crystal device. The replicas were immersed in 100% methanol overnight. The tissue was then digested from the replica by clorox (laundry bleach), placed into 5% EDTA, and washed repeatedly with distilled water. The replicas were picked up on 0.3% formvar-coated 75 mesh grids and examined in the JEOL 100B electron microscope. The results were as follows; 1. Both in thin sections and freeze-fracture replicas, three types of intercellular junctions were recognizable in the plasma membrane of odontoblast: gap junction, tight junction and desmosome-like junction. 2. The nuclear pores were evenly distributed over the nuclear envelope. The pore complex formed a ring about 70 nm in diameter. 3. Gap junctions were found between odontoblasts as well as odontoblasts and neighbouring pulp cells (fibroblast, subodontoblastic cell process, nerve-like fibre). Gap junctions, which were round, ellipsoid and pear-shaped and 600 nm in diameter, were observed in the odontoblast. 4. Numerous round and ellipsoid gap junctions could be frequently seen on the plasma membranes in cell body and apical part of the odontoblasts. On the P face, the junctions were recognized as a cluster of closely packed particles, measuring about 9 nm in diameter, and on the E face, the junctions were recognized as a shallow grooves.

• 대한치과보존학회:학술대회논문집
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• 대한치과보존학회 2003년도 제120회 추계학술대회 제 5차 한ㆍ일 치과보존학회 공동학술대회
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• pp.550-550
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• 2003

IL-1${\alpha}$ and TNF-${\alpha}$ play an important role in initiating and coordinating the cellular events that make up the immune response to infection. The purpose of this study was to examine the effects of proinflammatory cytokines on mineralization and HO-1 protein expression in the human pulp cells. Human pulp cell cultures between the fifth and sixth passage were used in this study. Alkaline phosphatase and osteonectin were selected as markers for mineralization that is, odontoblast-like differentiation.(omitted)

• 한국치위생학회지
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• 제17권3호
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• pp.441-448
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• 2017

Objectives: The purpose of this study was to examine the cell proliferation and expression of alkaline phosphatase (ALP) during the differentiation of murine odontoblast-like cells (MDPC-23) by Cimicifuga rhizoma extract. Cimicifuga rhizoma extract was prepared using 70% ethanol. Then, the cells were treated with 25, 50, 100, 150, and $200{\mu}g$ of Cimicifuga rhizoma extract. Methods: We determined the Cimicifuga rhizoma effects of MDPC-23 using WST-1 (water soluble tetrazolium salt-1) assay, ALP activity assay and histochemical staining. Results: $25-200{\mu}g$ of Cimicifuga rhizoma extract did not inhibit the growth of MDPC-23 cells; $100{\pm}0$, $100{\pm}3.29$, $99{\pm}4.86$, $98{\pm}3.80$, $98{\pm}1.73$, $99{\pm}5.05%$ (p<0.794). $50{\mu}g$ of Cimicifuga rhizoma extract stimulated ALP activity on MDPC-23; $5.1{\pm}0.20units/{\mu}{\ell}$ (p<0.001). Conclusions: It was proven that Cimicifuga rhizoma promoted differentiation of MDPC- 23 cells.

• Journal of Periodontal and Implant Science
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• 제36권2호
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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.

• The Korean Journal of Physiology and Pharmacology
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• 제26권1호
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• pp.37-45
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• 2022

The aim of the present study was to investigate the physiological role of nicotinamide phosphoribosyltransferase (NAMPT) associated with odontogenic differentiation during tooth development in mice. Mouse dental papilla cell-23 (MDPC-23) cells cultured in differentiation media were stimulated with the specific NAMPT inhibitor, FK866, and Visfatin (NAMPT) for up to 10 days. The cells were evaluated after 0, 4, 7, and 10 days. Cell viability was measured using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. The mineralization assay was performed by staining MDPC-23 cells with Alizarin Red S solution. After cultivation, MDPC-23 cells were harvested for quantitative PCR or Western blotting. Analysis of variance was performed using StatView 5.0 software (SAS Institute Inc., Cary, NC, USA). Statistical significance was set at p < 0.05. The expression of NAMPT increased during the differentiation of murine odontoblast-like MDPC-23 cells. Furthermore, the up-regulation of NAMPT promoted odontogenic differentiation and accelerated mineralization through an increase in representative odontoblastic biomarkers, such as dentin sialophosphoprotein, dentin matrix protein-1, and alkaline phosphatase in MDPC-23 cells. However, treatment of the cells with the NAMPT inhibitor, FK866, attenuated odontogenic differentiation, as evidenced by the suppression of odontoblastic biomarkers. These data indicate that NAMPT regulated odontoblastic differentiation through the regulation of odontoblastic biomarkers. The increase in NAMPT expression in odontoblasts was closely related to the formation of the extracellular matrix and dentin via the Runx signaling pathway. Therefore, these data suggest that NAMPT is a critical regulator of odontoblast differentiation during tooth development.