• Restorative Dentistry and Endodontics
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• 제31권3호
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• pp.203-214
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• 2006

치수는 상아질로 둘러싸인 간엽조직으로 다양한 세포와 기저 물질들로 구성되어 있으며 혈관과 신경조직이 분포되어 있다. 치수의 염증은 조직의 분해를 야기하며 이는 Matrix Metalloproteinase에 의해 세포 외 기질의 분해가 촉진되어 병적인 과정을 거치게 된다. 이에 Lipopolysaccharide에 의한 MMP와 inflammatory cytokine의 유도와 peroxisome proliferator-activated receptors (PPAR)에 의한 염증매개 물질의 조절에 대해 알아보고자 하였다. 사람의 치수세포를 다양한 LPS농도에 노출시킨 후 24시간째 MMP-2, MMP-9의 변화를 보고 LPS에 의해 자극된 치수세포에서 ICAM-1, VCAM-1, $IL-1{\beta},\;TNF-{\alpha}$의 분비가 증가됨을 알 수 있었다. 또한 Adenovirus $PPAR{\gamma}\;(Ad/PPAR{\gamma})$와 $PPAR{\gamma}$ agonist인 rosiglitazone를 LPS로 자극된 치수세포에 처리하였을 때 48시간째 MMPs와 Adhesion molecules, cytokines의 감소를 확인하였다. 이로써 사람의 치수세포에서 $PPAR{\gamma}$가 가지는 항 염증효과에 대해 지속적 인 연구가 필요할 것으로 사료된다.

• 대한소아치과학회지
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• 제40권3호
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• pp.185-193
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• 2013

최근 유구치의 치수절단술 약제로 MTA의 임상 적용이 문헌들에서 보고된 바 있으나 MTA 표면에서 일어나는 유치 치수 세포의 반응에 대한 시험관내 연구는 많이 보고되지 않았다. 이번 연구의 목적은 유치 및 영구치에서 유래한 치수기질세포가 경화된 MTA 표면에서 나타내는 증식 및 분화 능력을 비교 평가하는 것이었다. 사람 영구치와 유치 치수 조직에서 분리된 치수기질세포를 경화된 MTA 표면에서 배양 후 세포증식율과 세포주기를 검사하였으며, 정량적 역전사 중합효소 연쇄반응(RT-PCR)을 사용하여 분화양상을 분석하였다. Runt-related transcription factor 2(Runx2)와 alkaline phosphatase(ALP)가 정량적 RT-PCR의 표지자로 사용되었고, MTA 표면에서 증식된 치수기질세포의 형태학적 변화를 주사전자현미경 하에서 관찰하였다. 영구치와 유치의 치수기질세포군은 세포증식률, 세포주기 분포 및 mRNA 발현 양상에 있어서 차이를 보이지 않았으며, 주사전자현미경 상에서 두 군 모두 수지상 형태를 나타내었다. MTA 상에서 관찰된 유치와 영구치의 치수기질세포의 비슷한 증식력 및 광화를 유도하는 세포로의 분화능은 유치의 치수절단술 제재로 MTA가 생체친화적으로 적합함을 보여준다.

• Molecules and Cells
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• 제38권7호
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• pp.604-609
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• 2015

The active metabolite of vitamin D such as $1{\alpha}$,25-dihydroxyvitamin ($D_3(1{\alpha},25(OH)_2D_3)$ is a well-known key regulatory factor in bone metabolism. However, little is known about the potential of vitamin D as an odontogenic inducer in human dental pulp cells (HDPCs) in vitro. The purpose of this study was to evaluate the effect of vitamin $D_3$ metabolite, $1{\alpha},25(OH)_2D_3$, on odontoblastic differentiation in HDPCs. HDPCs extracted from maxillary supernumerary incisors and third molars were directly cultured with $1{\alpha},25(OH)_2D_3$ in the absence of differentiation-inducing factors. Treatment of HDPCs with $1{\alpha},25(OH)_2D_3$ at a concentration of 10 nM or 100 nM significantly upregulated the expression of dentin sialophosphoprotein (DSPP) and dentin matrix protein1 (DMP1), the odontogenesis-related genes. Also, $1{\alpha},25(OH)_2D_3$ enhanced the alkaline phosphatase (ALP) activity and mineralization in HDPCs. In addition, $1{\alpha},25(OH)_2D_3$ induced activation of extracellular signal-regulated kinases (ERKs), whereas the ERK inhibitor U0126 ameliorated the upregulation of DSPP and DMP1 and reduced the mineralization enhanced by $1{\alpha},25(OH)_2D_3$. These results demonstrated that $1{\alpha},25(OH)_2D_3$ promoted odontoblastic differentiation of HDPCs via modulating ERK activation.

• 대한치과보존학회:학술대회논문집
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• 대한치과보존학회 2003년도 제120회 추계학술대회 제 5차 한ㆍ일 치과보존학회 공동학술대회
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• pp.602-602
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• 2003

I. Objectives It is reported that there are complements and immunogloblins in serum from dental pulp in dentinal tubules, and it is thought that dental caries bacteria is opsonized by these serum ingredients, and it is presented by dendritic cells(DCs) in dental pulp. So, we examined whether a maturational difference of DCs occured when S. mutans was opsonized. II. Materials and Methods PBMC was divided from normal human peripheral blood and collected CD14 positive cells by magnetic beads system. Adherent cells were incubated in 5% FCS-RPMI medium included GM-CSF, IL-4 for seven days.(omitted)

• 대한소아치과학회지
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• 제35권1호
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• pp.30-38
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• 2008

치아 치수 세포는 치아 손상에 따르는 병리적인 상황에서 골과 상아질 기질을 형성하는 능력을 가진 것으로 생각된다. 본 연구에서는 MTA가 사람 치수세포의 성장에 미치는 영향과 상아질 형성에 관여하는 유전자의 발현을 유도하는지를 알아보고자 하였다. 또한 상아질 형성의 잠재적 지표인 alkaline phosphatase(ALP) activity에 미치는 영향을 평가하였다. 유전자 발현 검사를 위해 glyceraldehyde-3-phosphate dehydrogenase, type I collagen, alkaline phosphatase, osteonectin(SPARC), and dentin sialoprotein primer set을 이용하여 MTA 처리 2일과 4일 후 reverse transcriptase polymerase chain reaction(RT-PCR)을 시행하였다. cell viability assay(세포 생존력 측정) 에서 5일간 MTA에 노출된 치수 세포의 비율이 대조군보다 높았다. 대조군에 비해 MTA를 처리한 군에서 ALP와 SPARC가 증가되었다. 이상의 결과를 종합하여 보면, 이 연구에 사용한 dental pulp culture system은 MTA를 포함한 치과재료의 처리 후 치수세포의 성장과 분화 그리고 상아질 형성 유도 기전을 연구하는 데 유용한 모델로 사용할 수 있다. MTA 처리는 사람 치수세포에 세포독성을 유도하지 않으며, ALP 활성도와 유전자 발현 그리고 osteonectin (SPARC) 유전자 발현을 증가시켜 수복상아질을 형성할 것으로 사료된다.

• Restorative Dentistry and Endodontics
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• 제40권3호
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• pp.223-228
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• 2015

Objectives: The aim of this study was to investigate the expression of 7 different sirtuin genes (SIRT1-SIRT7) in human dental pulp cells (HDPCs), and to determine the role of SIRTs in the odontoblastic differentiation potential of HDPCs. Materials and Methods: HDPCs were isolated from freshly extracted third molar teeth of healthy patients and cultulred in odontoblastic differentiation inducing media. Osteocalcin (OCN) and dentin sialophosphoprotein (DSPP) expression was analyzed to evaluate the odontoblastic differentiation of HDPCs by reverse transcription-polymerase chain reaction (RT-PCR), while alizarin red staining was used for the mineralization assay. To investigate the expression of SIRTs during odontoblastic differentiation of HDPCs, real time PCR was also performed with RT-PCR. Results: During the culture of HDPCs in the differentiation inducing media, OCN, and DSPP mRNA expressions were increased. Mineralized nodule formation was also increased in the 14 days culture. All seven SIRT genes were expressed during the odontogenic induction period. SIRT4 expression was increased in a time-dependent manner. Conclusions: Our study identified the expression of seven different SIRT genes in HDPCs, and revealed that SIRT4 could exert an influence on the odontoblast differentiation process. Further studies are needed to determine the effects of other SIRTs on the odontogenic potential of HDPCs.

• Restorative Dentistry and Endodontics
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• 제38권4호
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• pp.227-233
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• 2013

Objectives: The purpose of the study was to evaluate human dental pulp response to pulpotomy with calcium hydroxide (CH), mineral trioxide aggregate (MTA), and calcium enriched mixture (CEM) cement. Materials and Methods: A total of nine erupted third molars were randomly assigned to each pulpotomy group. The same clinician performed full pulpotomies and coronal restorations. The patients were followed clinically for six months; the teeth were then extracted and prepared for histological assessments. The samples were blindly assessed by an independent observer for pulp vitality, pulp inflammation, and calcified bridge formation. Results: All patients were free of clinical signs/symptoms of pulpal/periradicular diseases during the follow up period. In CH group, one tooth had necrotic radicular pulp; other two teeth in this group had vital uninflamed pulps with complete dentinal bridge formation. In CEM cement and MTA groups all teeth had vital uninflamed radicular pulps. A complete dentinal bridge was formed beneath CEM cement and MTA in all roots. Odontoblast-like cells were present beneath CEM cement and MTA in all samples. Conclusions: This study revealed that CEM cement and MTA were reliable endodontic biomaterials in full pulpotomy treatment. In contrast, the human dental pulp response to CH might be unpredictable.

• Natural Product Sciences
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• 제19권1호
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• pp.54-60
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• 2013

Rhus verniciflua Stokes (Anacadiaceae) is a plant that is native to East Asian countries, such as Korea, China, and Japan, and it has been found to exert various biological activities including antioxidative, anti-aggregatory, anti-inflammatory, anti-mutagenic, and apoptotic effects. Sulfuretin is one of the major flavonoid component isolated from the heartwood of R. verniciflua. Reactive oxygen species (ROS), produced via dental adhesive bleaching agents and pulpal disease, can cause oxidative stress. In the present study, we isolated sulfuretin from R. verniciflua and demonstrated that sulfuretin possesses cytoprotective effects against hydrogen peroxide ($H_2O_2$)-induced dental cell death. $H_2O_2$ is a representative ROS and causes cell death through necrosis in human dental pulp (HDP) cells. $H_2O_2$-induced cytotoxicity and production of ROS were blocked in the presence of sulfuretin, and these effects were dose dependent. Sulfuretin also increased heme oxygenase-1 (HO-1) protein expression. In addition, to determine whether sulfuretin-induced HO-1 expression mediated this cytoprotective effect, HDP cells were cotreated with sulfuretin in the absence or presence of SnPP, an inhibitor of HO activity. Sulfuretin-dependent HO-1 expression was required for suppression of $H_2O_2$-induced HDP cell death and ROS generation. These results indicate that sulfuretin-dependent HO-1 expression was required for the inhibition of $H_2O_2$-induced cell death and ROS generation. In addition, sulfuretin may be used to prevent functional dental cell death and thus may be useful as a pulpal disease agent.

• Restorative Dentistry and Endodontics
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• 제41권4호
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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.

• Biomaterials and Biomechanics in Bioengineering
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• 제3권2호
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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.