• Biomaterials and Biomechanics in Bioengineering
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• v.2 no.2
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• pp.61-69
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• 2015

Undifferentiated stem cells are being studied to obtain information on the therapeutic potential of isolates that are produced. Dental Pulp Stem Ccell (DPSC) may provide an abundant supply of highly proliferative, multipotent Mesenchymal Stem Cells (MSC), which are now known to be capable of regenerating a variety of human tissues including bone and other dental structures. Many factors influence DPSC quality and quantity, including the specific methods used to isolate, collect, concentrate, and store these isolates once they are removed. Ancillary factors, such as the choice of media, the selection of early versus late passage cells, and cryopreservation techniques may also influence the differentiation potential and proliferative capacity of DPSC isolates. This literature review concludes that due to the delicate nature of DPSC, more research is needed for dental researchers and clinicians to more fully explore the feasibility and potential for isolating and culturing DPSCs extracted from adult human teeth in order to provide more accurate and informed advice for this newly developing field of regenerative medicine.

• Biomaterials and Biomechanics in Bioengineering
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• v.3 no.2
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• pp.105-114
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• 2016

The purpose of this study is to evaluate the effects of cryopreservation on dental pulp-derived stem cells (DPSC) viability over a period of three years. Dental pulp-derived stem cells were isolated and cultured from thirty-one healthy teeth. DPSC isolates were assessed for doubling-time and baseline viability prior to cryopreservation and were assessed again at three time points; one week (T1), 18 months (T2), and 36 months (T3). DPSC can be grouped based on their observed doubling times; slow (sDT), intermediate (iDT), and rapid (rDT). Viability results demonstrated all three types of DPSC isolates (sDT, iDT and rDT) exhibit time-dependent reductions in viability following cryopreservation, with the greatest reduction observed among sDT-DPSCs and the smallest observed among the rDT-DPSC isolates. Cryopreserved DPSCs demonstrate time-dependent reductions in cellular viability. Although reductions in viability were smallest at the initial time point (T1) and greatest at the final time point (T3), these changes were markedly different among DPSC isolates with similar doubling times (DTs). Furthermore, the analysis of various DPSC biomarkers - including both intracellular and cell surface markers, revealed differential mRNA expression. More specifically, the relative high expression of Sox-2 was only found only among the rDT isolates, which was associated with the smallest reduction in viability over time. The expression of Oct4 and NANOG were also higher among rDT isolates, however, expression was comparatively lower among the sDT isolates that had the highest reduction in cellular viability over the course of this study. These data may suggest that some biomarkers, including Sox-2, Oct4 and NANOG may have some potential for use as biomarkers that may be associated with either higher or lower cellular viability over long-term storage applications although more research will be needed to confirm these findings.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.36 no.1
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• pp.7-15
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• 2010

Complex human tissues harbor stem cells and precursor cells, which are responsible for tissue development or repair. Recently, dental tissues such as dental pulp, periodontal ligament (PDL), dental follicle have been identified as easily accessible sources of undifferentiated cells. These tissues contain mesenchymal stem cells that can be differentiate into bone, cartilage, fat or muscle by exposing them to specific growth conditions. In this study, the authors procured the stem cell from pulp, PDL, and dental follicle and differentiate them into osteoblast and examine the bone induction capacity. Dental pulp stem cell (DPSC), periodontal ligament stem cell (PDLSC), and dental follicle precursor cell (DFPC) were obtained from human 3rd molar and cultured. Each cell was analyzed for presence of stem cell by fluorescence activated cell sorter (FACs) against CD44, CD105 and CD34, CD45. Each stem cell was cultured, expanded and grown in an osteogenic culture medium to allow formation of a layer of extracellular bone matrix. Osteogenic pathway was checked by alizarin red staining, alkaline phosphatase (ALP) activity test and RT-PCR for ALP and osteocalcin (OCN) gene expression. According to results from FACs, mesenchymal stem cell existed in pulp, PDL, and dental follicle. As culturing with bone differentiation medium, stem cells were differentiated to osteoblast like cell. Compare with stem cell from pulp, PDL and dental follicle-originated stem cell has more osteogenic effect and it was assumed that the character of donor cell was able to affect on differential potency of stem cell. From this article, we are able to verify the pulp, PDL, and dental follicle from extracted tooth, and these can be a source of osteoblast and stem cell for tissue engineering.

• International Journal of Stem Cells
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• v.15 no.4
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• pp.384-394
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• 2022

Background and Objectives: Dental pulp stem cells (DPSCs) play an important role in the repair of tooth injuries. Electrogenic sodium bicarbonate cotransporter 1 (NBCe1) is a Na⁺-coupled HCO₃^- transporter encoded by the solute carrier 4A4 (SLC4A4) gene and plays a crucial role in maintaining the pH of DPSCs. Our previous research confirmed that NBCe1 is highly expressed in odontoblasts during the development of the tooth germ. Therefore, in this study, we aimed to investigate the effect of NBCe1 on odontogenic differentiation of DPSCs and further clarify the underlying mechanisms. Methods and Results: DPSCs were isolated and identified, and the selective NBCe1 inhibitor S0859 was used to treat DPSCs. We used a cell counting Kit-8 assay to detect cell proliferative ability, and intracellular pH was assessed using confocal microscopy. Odontogenic differentiation of DPSCs was analyzed using real-time PCR and Alizarin Red S staining, and the NF-κB pathway was assessed using western blotting. Our results indicated that 10 µM S0859 was the optimal concentration for DPSC induction. Intracellular pH was decreased upon treatment with S0859. The mRNA expressions of DSPP, DMP1, RUNX2, OCN, and OPN were upregulated in the NBCe1 inhibited group compared to the controls. Moreover, NBCe1 inhibition significantly activated the NF-κB pathway, and a NF-κB inhibitor reduced the effect of NBCe1 on DPSC differentiation. Conclusions: NBCe1 inhibition significantly promotes odontogenic differentiation of DPSCs, and this process may be regulated by activating the NF-κB signaling pathway.

• Journal of Biosystems Engineering
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• v.38 no.2
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• pp.113-120
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• 2013

Purpose: Stem cells provide new opportunities in the regenerative medicine for human or animal tissue regeneration. In this study, we report an efficient method for the modulating behaviors of electro-active stem cells by micro-electric current stimulation (mES) without using chemical agents, such as serum or induction chemicals. Methods: Dental pulp stem cells (DPSCs) were cultured on the tissue culture dish in the mES system. To find a suitable mES condition to promote the DPSC functions, the response surface analysis was used. Results: We found that a working micro-current of 38 ${\mu}A$ showed higher DPSC proliferation compared with other working conditions. The mES altered the expressions of intracellular and extracellular proteins compared to those in unstimulated cells. The mES with 38 ${\mu}A$ significantly increased osteogenesis of DPSCs compared with ones without mES. Conclusions: Our findings indicate that mES may induce DPSC proliferation and differentiation, resulting in applying to DPSCs-based human or animal tissue regeneration.

• The Korean Journal of Physiology and Pharmacology
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• v.28 no.5
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• pp.423-433
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• 2024

Dental pulp stem cells (DPSCs) are a type of adult stem cell present in the dental pulp tissue. They possess a higher proliferative capacity than bone marrow mesenchymal stem cells. Their ease of collection from patients makes them well-suited for tissue engineering applications, such as tooth and nerve regeneration. Chios gum mastic (CGM), a resin extracted from the stems and leaves of Pistacia lentiscus var. Chia, has garnered attention for its potential in tissue regeneration. This study aims to confirm alterations in cell proliferation rates and induce differentiation in human DPSCs (hDPSCs) through CGM treatment, a substance known for effectively promoting odontogenic differentiation. Administration of CGM to hDPSC cells was followed by an assessment of cell survival, proliferation, and odontogenic differentiation through protein and gene analysis. The study revealed that hDPSCs exhibited low sensitivity to CGM toxicity. CGM treatment induced cell proliferation by activating cell-cycle proteins through the Wnt/β-catenin pathway. Additionally, the study demonstrated that CGM enhances alkaline phosphatase activation by upregulating the expression of collagen type I, a representative matrix protein of dentin. This activation of markers associated with odontogenic and bone differentiation ultimately facilitated the mineralization of hDPSCs. This study concludes that CGM, as a natural substance, fosters the cell cycle and cell proliferation in hDPSCs. Furthermore, it triggers the transcription of odontogenic and osteogenic markers, thereby facilitating odontogenic differentiation.

• 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.

• Journal of the korean academy of Pediatric Dentistry
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• v.42 no.4
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• pp.312-318
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• 2015

The aim of this study was to investigate the possibility of the supernumerary teeth for the stem cell source in dentistry. The Real Time Quantitative Reverse Transcription Polymerase Chain Reaction (Real Time qRT-PCR) method was used to evaluate the differentiation toward the odontoblast of the supernumerary dental pulp stem cells (sDPSCs). Supernumerary dental pulp stem cells were obtained from 3 children (2 males and 1 female, age 7 to 9) diagnosed that the eruption of permanent teeth was disturbed by supernumerary teeth. The common genes for odontoblasts are alkaline phosphatase (ALP), osteocalcin (OC), osteonectin (ON), dentin matrix acidic phosphoprotein 1 (DMP-1), dentin sialophosphoprotein (DSPP). The sDPSCs were treated for 0 days, 8 days and 14 days with additives and then Real Time qRT-PCR was performed in intervals of 0 days, 8 days and 14 days. The alizarin-red solution staining was performed to visualize the stained color for the degree of calcification at 7 days, 14 days, 21 days and 28 days after treating additives to the sDPSCs. From the result of the Real Time qRT-PCR, the manifestation exhibit maximum value at 8 days after additive treatment and shifted to a decrease trend at 14 days. Alizarin-red solution staining exhibit light results at 7 days after staining and generalized dark result at 14 days. Consequently, in studies with sDPSCs, appropriate treatment time of additives for Real Time qRT-PCR is 8 days. Also, a suitable period of Alizarin-red solution staining is 14 days.

• Journal of the korean academy of Pediatric Dentistry
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• v.44 no.3
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• pp.350-357
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• 2017

The purpose of this study was to evaluate the difference of differentiation potential in each passage of dental pulp stem cells from supernumerary tooth (sDPSCs). The sDPSCs were obtained from a healthy 6-year-old male patient under the guidelines and got the informed consent. Cells were cultured until passage number 16 and divided into two groups; 1 - 8 passages as a young group and 9 - 16 passages as an old group. It was taken $2.25{\pm}0.46days$ in a young group and $3.25{\pm}0.46days$ in an old group to propagate cells of each passage until confluence and there were statistically significant differences between two groups (p < 0.05). In every passage, cell morphology was observed with microscope and evaluated the capacity to form high levels of minerals by alizarin red solution staining after treating differentiation medium. Fibroblast-like, spindle shaped, elongated cells and a few nodules were found in uninduced cultures of passage number 1, 8 and 9. But at 16 passage culture, cell size became larger and broader and observed with more nodules. After inducing differentiation, mineralized nodules were detected at the first passage of 7th day culture whereas at the 8 passage culture, nodules were seen clearly at 14th day culture. In addition, the amount of mineralized nodules were remarkably decreased after passage 9. From the data presented in this study, it is recommended to use sDPSCs of passage number within 8 for utilizing as stem cells.