• Journal of the korean academy of Pediatric Dentistry
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• v.46 no.3
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• pp.337-342
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• 2019

The aim of this study was to analyze cells from human dental pulp tissue of impacted supernumerary teeth as stem cells with flow cytometry. Human dental pulp cells from 15 supernumerary teeth were identified their characteristics as stem cells by expression of mesenchymal stem cell markers through flow cytometry analysis at passage 3 and passage 10. Cluster of differentiation (CD) 73, CD 90, CD 34, CD 45 and STRO-1 cell surface markers were used to figure out characteristics of dental pulp stem cells from supernumerary teeth. At passage 3, the cell population showed positive expression of CD 73, CD90 and STRO-1, lacked expression of CD 34 and CD 45. At passage 10, CD 73, CD 90 and STRO-1 showed positive expression while CD 34 and CD 45 showed negative expression. This study indicated that dental pulp stem cells of supernumerary teeth had the properties of mesenchymal stem cells at both early and late passage. Impacted supernumerary teeth could be considered as a noble source of stem cells because of rapid growth and maintaining characteristics of stem cells until late passage.

• Restorative Dentistry and Endodontics
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• v.46 no.2
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• pp.26.1-26.15
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• 2021

Objectives: The aim of the present systematic review was to investigate the cryopreservation process of dental pulp mesenchymal stromal cells and whether cryopreservation is effective in promoting cell viability and recovery. Materials and Methods: This systematic review was developed in accordance with the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) statement and the research question was determined using the population, exposure, comparison, and outcomes strategy. Electronic searches were conducted in the PubMed, Cochrane Library, Science Direct, LILACS, and SciELO databases and in the gray literature (dissertations and thesis databases and Google Scholar) for relevant articles published up to March 2019. Clinical trial studies performed with dental pulp of human permanent or primary teeth, containing concrete information regarding the cryopreservation stages, and with cryopreservation performed for a period of at least 1 week were included in this study. Results: The search strategy resulted in the retrieval of 185 publications. After the application of the eligibility criteria, 21 articles were selected for a qualitative analysis. Conclusions: The cryopreservation process must be carried out in 6 stages: tooth disinfection, pulp extraction, cell isolation, cell proliferation, cryopreservation, and thawing. In addition, it can be inferred that the use of dimethyl sulfoxide, programmable freezing, and storage in liquid nitrogen are associated with a high rate of cell viability after thawing and a high rate of cell proliferation in both primary and permanent teeth.

• Restorative Dentistry and Endodontics
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• v.40 no.4
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• pp.290-298
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• 2015

Objectives: Sodium hypochlorite (NaOCl) is an excellent bactericidal agent, but it is detrimental to stem cell survival, whereas intracanal medicaments such as calcium hydroxide ($Ca[OH]_2$) promote the survival and proliferation of stem cells. This study evaluated the effect of sequential NaOCl and $Ca(OH)_2$ application on the attachment and differentiation of dental pulp stem cells (DPSCs). Materials and Methods: DPSCs were obtained from human third molars. All dentin specimens were treated with 5.25% NaOCl for 30 min. DPSCs were seeded on the dentin specimens and processed with additional 1 mg/mL $Ca(OH)_2$, 17% ethylenediaminetetraacetic acid (EDTA) treatment, file instrumentation, or a combination of these methods. After 7 day of culture, we examined DPSC morphology using scanning electron microscopy and determined the cell survival rate with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. We measured cell adhesion gene expression levels after 4 day of culture and odontogenic differentiation gene expression levels after 4 wk using quantitative real-time polymerase chain reaction. Results: DPSCs did not attach to the dentin in the NaOCl-treated group. The gene expression levels of fibronectin-1 and secreted phosphoprotein-1 gene in both the $Ca(OH)_2$- and the EDTA-treated groups were significantly higher than those in the other groups. All $Ca(OH)_2$-treated groups showed higher expression levels of dentin matrix protein-1 than that of the control. The dentin sialophosphoprotein level was significantly higher in the groups treated with both $Ca(OH)_2$ and EDTA. Conclusions: The application of $Ca(OH)_2$ and additional treatment such as EDTA or instrumentation promoted the attachment and differentiation of DPSCs after NaOCl treatment.

• Restorative Dentistry and Endodontics
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• v.45 no.1
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• pp.3.1-3.10
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• 2020

Objectives: This study investigated the indirect effect of calcium-enriched mixture (CEM) cement and mineral trioxide aggregate (MTA), as 2 calcium silicate-based hydraulic cements, on human dental pulp stem cells (hDPSCs) through different dentin thicknesses. Materials and Methods: Two-chamber setups were designed to simulate indirect pulp capping (IPC). Human molars were sectioned to obtain 0.1-, 0.3-, and 0.5-mm-thick dentin discs, which were placed between the 2 chambers to simulate an IPC procedure. Then, MTA and CEM were applied on one side of the discs, while hDPSCs were cultured on the other side. After 2 weeks of incubation, the cells were removed, and cell proliferation, morphology, and attachment to the discs were evaluated under scanning electron microscopy (SEM). Energy-dispersive X-ray (EDXA) spectroscopy was performed for elemental analysis. Alkaline phosphatase (ALP) activity was assessed quantitatively. The data were analyzed using the Kruskal-Wallis and Mann-Whitney tests. Results: SEM micrographs revealed elongated cells, collagen fibers, and calcified nucleations in all samples. EDXA verified that the calcified nucleations consisted of calcium phosphate. The largest calcifications were seen in the 0.1-mm-thick dentin subgroups. There was no significant difference in ALP activity across the CEM subgroups; however, ALP activity was significantly lower in the 0.1-mm-thick dentin subgroup than in the other MTA subgroups (p < 0.05). Conclusions: The employed capping biomaterials exerted biological activity on hDPSCs, as shown by cell proliferation, morphology, and attachment and calcific precipitations, through 0.1- to 0.5-mm-thick layers of dentin. In IPC, the bioactivity of these endodontic biomaterials is probably beneficial.

• 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.

• The Journal of the Korean dental association
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• v.9 no.4
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• pp.157-160
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• 1971

To compare the effects of various pulp capping agents that are usually applied to human pulp tissue, adult dogs were bred for a certain period and each capping agent was applied experimentally to pulp tissue after vital pulpotomy. Histological observations are as follows. 1) In comparison between methods of vital pulpotomy, one and two appointment method, different courses of healing were observed. In one appointment method, the granulation tissue formation at the amputation sur face of pulp tissue had a tendency to be transformed to scar tissue formation. In two appointment method, more transformation than that of one appointment method from scar tissue to dentin matrix formation were observed. 2) Histologic changes that have appeared in pulp tissue are a) fixation at outer layer b) degeneration at middle layer c) hyperemia and round cell infiltration at inner layer 3) With use of formocresol mixed zinc oxide powder in two appointment method complete formation of dentin matrix were observed. 4) Among the methods and aagents described above formocresol mixed zinc oxide powder in two appointment method appeared to be relatively effective.

• Journal of the korean academy of Pediatric Dentistry
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• v.40 no.3
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• pp.185-193
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• 2013

Mineral trioxide aggregate (MTA) has recently been used as a pulpotomy medicament for primary molars. The aim of this study was to evaluate and compare the proliferation and differentiation potential of dental pulp stromal cells of permanent teeth and deciduous teeth cultured on MTA-coated surface. Human dental pulp stromal cells were obtained from human permanent premolars and deciduous teeth and cultured on MTA-coated culture plates. The cells were subjected to proliferation assay and cell cycle analysis. Their differentiation potential was evaluated by analysing changes in the mRNA expressions of runt-related transcriptional factor 2 (Runx2) and alkaline phosphatase (ALP). Morphological changes of cells in direct contact with MTA were observed using scanning electron microscopy (SEM). The proliferation rates, distribution of cell cycles and mRNA expression patterns of Runx2 and ALP were similar in both types of pulpal cells. SEM observations revealed that both types changed into more dendrite-like cells. On the surface of MTA, human dental pulp stromal cells from deciduous and permanent teeth were able to both proliferate and differentiate into cells that induce mineralization. MTA is suitable as a biocompatible pulpotomy medicament for primary teeth.

• 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.

• Restorative Dentistry and Endodontics
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• v.48 no.4
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• pp.39.1-39.23
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• 2023

Objectives: This study aimed to investigate the effectiveness of different topical/systemic agents in reducing the damage caused by bleaching gel to pulp tissue or cells. Materials and Methods: Electronic searches were performed in July 2023. In vivo and in vitro studies evaluating the effects of different topical or systemic agents on pulp inflammation or cytotoxicity after exposure to bleaching agents were included. The risk of bias was assessed. Results: Out of 1,112 articles, 27 were included. Nine animal studies evaluated remineralizing/anti-inflammatories agents in rat molars subjected to bleaching with 35%-38% hydrogen peroxide (HP). Five of these studies demonstrated a significant reduction in inflammation caused by HP when combined with bioglass or MI Paste Plus (GC America), or following KF-desensitizing or Otosporin treatment (n = 3). However, orally administered drugs did not reduce pulp inflammation (n = 4). Cytotoxicity (n = 17) was primarily assessed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay on human dental pulp cells and mouse dental papilla Cell-23 cells. Certain substances, including sodium ascorbate, butein, manganese chloride, and peroxidase, were found to reduce cytotoxicity, particularly when applied prior to bleaching. The risk of bias was high in animal studies and low in laboratory studies. Conclusions: Few in vivo studies have evaluated agents to reduce the damage caused by bleaching gel to pulp tissue. Within the limitations of these studies, it was found that topical agents were effective in reducing pulp inflammation in animals and cytotoxicity. Further analyses with human pulp are required to substantiate these findings.

• 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.