• Restorative Dentistry and Endodontics
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• v.12 no.2
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• pp.73-80
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• 1987

The present study was designed to help elucidate the effect of glass ionomer cements on the exposed dental pulp by means of histologic examination. A total of 40 cavities of class V were prepared on the teeth of 4 dogs with exposure of 1mm in diameter on the bases of them. 20 cavities were filled with glass ionomer cement as the experimental group and the other 20 cavities were filled with zinc oxide eugenol cement as the control group. The dogs were sacrificed at one, two, three, and four weeks after filling, and the specimens were routinely prepared and stained with Hematoxylin-Eosin. The obtained microscopic findings were as follows: Inflammatory cell infiltrations were observed in control in 1 week, which decreased markedly with time. In all control groups, hemorrhage around exposed pulp tissue and coagulation change of pulp were observed. Secondary dentin formation and thickened predentin were observed in 4 week cases, and the recovery of pulp tissue was favorable on the whole. Inflammatory cell infiltration was observed in all GIC groups. Proliferation of blood vessel and congestion were observed with coagulation changes around the exposed pulp tissue. Secondary dentin formation and thickened predentin were observed in 3 weeks. In the experimental 4 week case, secondary dentin formation was evident. On the whole, pulpal irritation of glass ionomer cement was relatively severe. Recovery of pulp tissue in GIC groups was less favorable compared with that of ZOE groups.

• Journal of the korean academy of Pediatric Dentistry
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• v.35 no.1
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• pp.30-38
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• 2008

Dental pulp cells are assumed to possess the capacity to elaborate both bone and dentin matrix under the pathological conditions following tooth injury. The purpose of this study is to examine the effects of mineral trioxide aggregate (MTA) on various gene expression regarding dentinogenesis and cell viability assay in cultured primary human dental pulp cells. The author also examined the effects of this material on cellular alkaline phosphatase activity as a potential indicator of dentinogenesis. For gene expression on MTA, reverse transcriptase polymerase chain reaction was performed using primer sets of glyceraldehyde-3-phosphate dehydrogenase, type I collagen, alkaline phosphatase(ALP), osteonectin, and dentin sialoprotein after 2 and 4 days. Cell viability assay showed that the proportion of MTA-treated pulp cells which had been exposed for 5 days to MTA was higher than that of the control cells. Among the genes investigated in this study, ALP and osteonectin(SPARC) were increased in MTA treated group than in control. These findings suggest that this dental pulp culture system may be useful in the future as a model for studying the mechanisms underlying dentin regeneration after the treatment with MTA. Exposure to MTA material would not induce cytotoxic response in the dental pulp cells. In addition, MTA could influence the behavior of human pulp cells by increasing the ALP activity and SPARC synthesis.

• Restorative Dentistry and Endodontics
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• v.37 no.3
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• pp.142-148
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• 2012

Objectives: We analyzed gene-expression profiles after 14 day odontogenic induction of human dental pulp cells (DPCs) using a DNA microarray and sought candidate genes possibly associated with mineralization. Materials and Methods: Induced human dental pulp cells were obtained by culturing DPCs in odontogenic induction medium (OM) for 14 day. Cells exposed to normal culture medium were used as controls. Total RNA was extracted from cells and analyzed by microarray analysis and the key results were confirmed selectively by reverse-transcriptase polymerase chain reaction (RT-PCR). We also performed a gene set enrichment analysis (GSEA) of the microarray data. Results: Six hundred and five genes among the 47,320 probes on the BeadChip differed by a factor of more than two-fold in the induced cells. Of these, 217 genes were upregulated, and 388 were down-regulated. GSEA revealed that in the induced cells, genes implicated in Apoptosis and Signaling by wingless MMTV integration (Wnt) were significantly upregulated. Conclusions: Genes implicated in Apoptosis and Signaling by Wnt are highly connected to the differentiation of dental pulp cells into odontoblast.

• Journal of Dental Rehabilitation and Applied Science
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• v.25 no.2
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• pp.191-200
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• 2009

Dental Pulp Stem Cells are superior to other types of adult stem cell. Because of teeth are easy to access and are extracted throughout life. A supernumerary tooth is an important clinical problem found in various populations of the world. The incidence of supernumerary teeth varies depending on the literature source. Pediatric dentists are routinely extracted them. However, no studies have been reported regarding Dental Pulp stem cells in a supernumerary tooth, and we failed to note that a valuable source of human stem cell. Herein, we tried to show that a supernumerary tooth contains cells that display the characteristic features of stem cells.

• International Journal of Oral Biology
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• v.37 no.4
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• pp.167-173
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• 2012

This study investigated the genes involved in the differentiation of odontoblasts derived from human dental pulp stem cells (hDPSCs). hDPSCs isolated from human tooth pulp were validated by fluorescence activated cell sorting (FACS). After odontogenic induction, hDPSCs were analyzed investigated by Alizaline red-S staining, ALP assay, ALP staining and RT-PCR. Differential display-polymerase chain reaction (DD-PCR) was performed to screen differentially expressed genes involved in the differentiation of hDPSCs. By FACS analysis, the stem cell markers CD24 and CD44 were found to be highly expressed in hDPSCs. When hDPSCs were treated with agents such as ${\beta}$-glycerophosphate (${\beta}$-GP) and ascorbic acid (AA), nodule formation was exhibited within six weeks. The ALP activity of hDPSCs was found to elevate over time, with a detectable up-regulation at 14 days after odontogenic induction. RT-PCR analysis revealed that dentin sialophosphoprotein (DSPP) and osteocalcin (OC) expression had increased in a time-dependent manner in the induction culture. Through the use of DD-PCR, several genes were differentially detected following the odontogenic induction. These results suggest that these genes may possibly be linked to a variety of cellular process during odontogenesis. Furthermore, the characterization of these regulated genes during odontogenic induction will likely provide valuable new insights into the functions of odontoblasts.

• Applied Microscopy
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• v.43 no.1
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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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• v.41 no.4
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• pp.283-295
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• 2016

Objectives: In this study, we characterized human dental pulp cells (HDPCs) obtained by different culture methods to establish the most suitable methodology for dental tissue engineering and regenerative endodontic applications. Materials and Methods: HDPCs were isolated by the outgrowth method (HDPCs-OG), the enzymatic digestion method (collagenase/dispase/trypsin, HDPCs-ED), or the combination of both methods (HDPCs-Combined). The expression of mesenchymal stem cell markers (CD105, CD90, and CD73) was investigated. In vitro differentiation capacities of HDPCs into adipogenic, osteogenic, and chondrogenic lineages were compared. Differentiation markers were analyzed by quantitative reverse-transcription polymerase chain reaction (RT-PCR) and western blotting. Results: Our data indicated that whole HDPCs-ED, HPDCs-OG, and HDPCs-Combined could be differentiated into adipogenic, chrondrogenic, and osteogenic cell types. However, we found that the methods for isolating and culturing HDPCs influence the differentiation capacities of cells. HDPCs-OG and HDPCs-ED were preferably differentiated into adipogenic and osteogenic cells, respectively. Differentiation markers shown by RT-PCR and western blotting analysis were mostly upregulated in the treated groups compared with the control groups. Conclusions: Our findings confirmed that cell populations formed by two different culture methods and the combined culture method exhibited different properties. The results of this study could provide an insight into regenerative endodontic treatment using HDPCs.

• The Journal of Advanced Prosthodontics
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• v.7 no.1
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• pp.21-26
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• 2015

PURPOSE. To evaluate the cytotoxicity of temporary luting cements on bovine dental pulp-derived cells (bDPCs). MATERIALS AND METHODS. Four different temporary cements were tested: Rely X Temp E (3M ESPE), Ultratemp (Ultradent), GC Fuji Temp (GC), and Rely X Temp NE (3M ESPE). The materials were prepared as discs and incubated in Dulbecco's modified eagle's culture medium (DMEM) for 72 hours according to ISO 10993-5. A real-time cell analyzer was used to determine cell vitality. After seeding $200{\mu}L$ of the cell suspensions into the wells of a 96-well plate, the bDPCs were cured with bioactive components released by the test materials and observed every 15 minutes for 98 hours. One-way ANOVA and Tukey-Kramer tests were used to analyze the results of the proliferation experiments. RESULTS. All tested temporary cements showed significant decreases in the bDPCs index. Rely X Temp E, GC Fuji Temp, and Rely X Temp NE were severely toxic at both time points (24 and 72 hours) (P<.001). When the cells were exposed to media by Ultratemp, the cell viability was similar to that of the control at 24 hours (P>.05); however, the cell viability was significantly reduced at 72 hours (P<.001). Light and scanning electron microscopy examination confirmed these results. CONCLUSION. The cytotoxic effects of temporary cements on pulpal tissue should be evaluated when choosing cement for luting provisional restorations.

• International Journal of Oral Biology
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• v.31 no.3
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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.

• Journal of Oral Medicine and Pain
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• v.9 no.1
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• pp.127-138
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• 1984

The author had tried to identify the sex from single tooth by detecting F-body of Y-chromosome in the nucleus of the dental pulp cells of 70 persons aged from 4 to 61 years under a fluorescent miscroscope. The results were as follows : 1. In the cell nuclei of male and female dental pulp at refrigeratory, the rate of F-body appearancd ranged 42-86%(average 61.06%) in male, while it was 0-6%(average 1.86%) in female, indicationg that male could be distinctly differentiated from female by F-body. 2. With male and female dental pulp puterfide by leaving in at room temperature, the rate of F-body appearance ranged 35-58%(average 48.20%) in male, 1-3%(average1.70%) in female, indicating that it was possible to distinguish male and female by F-body. 3. Even in heat-treated male teeth at $100^{\circ}C$,10 mins, the rate of F-body appearace proved to be 32-56%(averaged 42.50%), also indicating the possibility of identifying male. 4. When detecting of F-body in process of time, the rate of F-body appearance did not show major charges. 5. It was reaffirmed that F-body detection method was a positive determination method of male.