• Molecules and Cells
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• 제39권2호
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• pp.156-162
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• 2016

Estrogen receptor ${\alpha}$ (ER-${\alpha}$), which is involved in bone metabolism and breast cancer, has been shown to have transcriptional targets. Dlx3 is essential for the skeletal development and plays an important role in osteoblast differentiation. Various osteogenic stimulators and transcription factors can induce the protein expression of Dlx3. However, the regulatory function of ER-${\alpha}$ in the Dlx3 mediated osteogenic process remains unknown. Therefore, we investigated the regulation of Dlx3 and found that ER-${\alpha}$ is a positive regulator of Dlx3 transcription in BMP2-induced osteoblast differentiation. We also found that ER-${\alpha}$ interacts with Dlx3 and increases its transcriptional activity and DNA binding affinity. Furthermore, we demonstrated that the regulation of Dlx3 activity by ER-${\alpha}$ is independent of the ligand (estradiol) binding domain. These results indicate that Dlx3 is a novel target of ER-${\alpha}$, and that ER-${\alpha}$ regulates the osteoblast differentiation through modulation of Dlx3 expression and/or interaction with Dlx3.

• International Journal of Oral Biology
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• 제38권4호
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• pp.161-167
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• 2013

Human dental pulp stem cells (DPSCs) are multi-potent mesenchymal stem cells that have several differentiation potentials. An understanding of thetissues that differentiate from these cells can provide insights for future regenerative therapeutics and tissue engineering strategies. The mesiodens is the most frequent form of supernumerary tooth from which DPSCs can differentiate into several lineages similar to cells from normal deciduous teeth. Recently, it has been shown that nanoscale structures can affect stem cell differentiation. In our presentstudy, we investigated the effects of a 250-nm nanoscale ridge/groove pattern array on the osteogenic and adipogenic differentiation of dental pulp cells from mesiodenscontaining human DPSCs. To this end, the expression of lineage specific markers after differentiation induction was analyzed by lineage specific staining and RT-PCR. The nanoscale pattern arrayed surface showed apositive effect on the adipogenic differentiation of DPSCs. There was no difference between nanoscale pattern arrayed surface and conventional surface groups onosteogenic differentiation. In conclusion, the nanoscale ridge/groove pattern arrayed surface can be used to enhance the adipogenic differentiation of DPSCs derived from mesiodens. This finding provides an improved understanding of the effects of topography on cell differentiation as well as the potential use of supernumerary tooth in regenerative dental medicine.

• Journal of Periodontal and Implant Science
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• 제41권1호
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• pp.30-43
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• 2011

Purpose: Under different culture conditions, periodontal ligament (PDL) stem cells are capable of differentiating into cementoblast-like cells, adipocytes, and collagen-forming cells. Several previous studies reported that because of the stem cells in the PDL, the PDL have a regenerative capacity which, when appropriately triggered, participates in restoring connective tissues and mineralized tissues. Therefore, this study analyzed the genes involved in mineralization during differentiation of human PDL (hPDL) cells, and searched for candidate genes possibly associated with the mineralization of hPDL cells. Methods: To analyze the gene expression pattern of hPDL cells during differentiation, the hPDL cells were cultured in two conditions, with or without osteogenic cocktails (${\beta}$-glycerophosphate, ascorbic acid and dexamethasone), and a DNA microarray analysis of the cells cultured on days 7 and 14 was performed. Reverse transcription-polymerase chain reaction was performed to validate the DNA microarray data. Results: The up-regulated genes on day 7 by hPDL cells cultured in osteogenic medium were thought to be associated with calcium/iron/metal ion binding or homeostasis (PDE1A, HFE and PCDH9) and cell viability (PCDH9), and the down-regulated genes were thought to be associated with proliferation (PHGDH and PSAT1). Also, the up-regulated genes on day 14 by hPDL cells cultured in osteogenic medium were thought to be associated with apoptosis, angiogenesis (ANGPTL4 and FOXO1A), and adipogenesis (ANGPTL4 and SEC14L2), and the down-regulated genes were thought to be associated with cell migration (SLC16A4). Conclusions: This study suggests that when appropriately triggered, the stem cells in the hPDL differentiate into osteoblasts/cementoblasts, and the genes related to calcium binding (PDE1A and PCDH9), which were strongly expressed at the stage of matrix maturation, may be associated with differentiation of the hPDL cells into osteoblasts/cementoblasts.

• 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.

• 폴리머
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• 제30권3호
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• pp.196-201
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• 2006

온도감응성 고분자인 폴리 (에틸렌 글리콜)을 기본으로 하는 다이블록 및 트리블록 폴리에스테르 공중합체들은 비독성과 생체적합성 그리고 생분해성 특징 때문에 조직공학 분야 및 주사제형의 약물전달체에서 많은 응용이 이루어지고 있다. 본 연구에서는 다이블록 공중합체를 이용한 솔-젤 전이 현상을 갖는 고분자를 평균분자량 750 g/mole의 메톡시 폴리 (에틸렌 글리콜)과 카프로락톤을 실온에서 HCl $Et_2O$ 존재 하에서 개환중합을 통하여 합성하였다. 이 공중합체를 이용하여 in vivo상에서 골수간엽줄기세포와 골유도물질인 덱사메타손의 존재 여부에 따라 골분화의 가능성을 조사하였다. 조직을 파라핀으로 고정시켜 슬라이드를 제조한 후 hematoxylin & eosin, 본쿠사 및 오스테오칼신 염색을 통하여 골형성 정도를 확인하였다. 결론적으로 덱사메타손이 함유된 젤이 골형성에 도움을 주지만 젤만으로는 강한 골형성을 유도할 수 없으므로 줄기세포나 다른 골형성 유도재료를 사용하면 우수하게 골형성 효과를 가져올 것이라고 판단되었다.

• Molecules and Cells
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• 제41권5호
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• pp.476-485
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• 2018

Although tectorigenin (TG), a major compound in the rhizome of Belamcanda chinensis, is conventionally used for the treatment of inflammatory diseases, its effects on osteogenesis and osteoclastogenesis have not been reported. The objective of this study was to investigate the effects and possible underlying mechanism of TG on in vitro osteoblastic differentiation and in vivo bone formation, as well as in vitro osteoclast differentiation and in vivo bone resorption. TG promoted the osteogenic differentiation of primary osteoblasts and periodontal ligament cells. Moreover, TG upregulated the expression of the BMP2, BMP4, and Smad-4 genes, and enhanced the expression of Runx2 and Osterix. In vivo studies involving mouse calvarial bone defects with ${\mu}CT$ and histologic analysis revealed that TG significantly increased new bone formation. Furthermore, TG treatment inhibited osteoclast differentiation and the mRNA levels of osteoclast markers. In vivo studies of mice demonstrated that TG caused the marked attenuation of bone resorption. These results collectively demonstrated that TG stimulated osteogenic differentiation in vitro, increased in vivo bone regeneration, inhibited osteoclast differentiation in vitro, and suppressed inflammatory bone loss in vivo. These novel findings suggest that TG may be useful for bone regeneration and treatment of bone diseases.

• 대한소아치과학회지
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• 제29권3호
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• pp.345-353
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• 2002

Bone morphogenetic proteins(BMPs)는 형태형성 및 세포 분화동안 다양한 조절 역할을 담당하는 신호전달 인자이다. 시상두개봉합부 발생시 BMPs와 그 수용체의 역할을 분석하기 위해, in situ hybridization방법을 이용하여 태생 15일에서 18일 시상두개봉합부에서 그 발현 양상을 분석하였다. BMP-2와 BMP-3은 태생 15일부터 osteogenic front와 두정골에서 발현을 보였으며 태생 16일부터 모낭에서 발현이 시작되었다. BMP-4는 osteogenic front에서 강하게 발현되었으며, 간엽조직 및 두정골에서 약하게 발현되었다. BMP-5는 모낭에서 발현되었다. 이전 연구에서 BMP-6는 비후된 연골세포에서 발현된다고 보고되었으나 본 연구에서는 발현되지 않았다. BMP-7은 태생기에 두정골에서 발현되었다. BMPR-IB는 osteogenic front의 외측 가장자리에서 발현되었으나, BMPR-IA는 발현되지 않았다. 이런 결과를 종합해 볼 때, 두개봉합부 초기 형태발생시 BMP-4는 미분화 간엽세포로부터 골아세포로 commit되는 초기단계에 중요한 역할을 하며, BMP-2와 BMP-3는 전구 골아세포에서 골아세포로의 분화과정에, BMP-7은 좀 더 분화가 진행된 골아세포 및 골의 분화 유지에 중요하며, type I 수용체 중 BMPR-IB가 BMP들의 신호전달에 중요한 역할을 함을 예측 할 수 있다. 결론적으로 BMP 신호전달은 다양한 BMP 리간드들과 그 수용체들에 의해 골아세포 분화 전반에 걸쳐 관여하고 있음을 시사한다.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제39권4호
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• pp.150-155
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• 2013

Objectives: The objective of this study is to determine the adequate concentration and to evaluate the osteogenic potential of simvastatin in human maxillary sinus membrane-derived stem cells (hSMSC). Materials and Methods: Mesenchymal stem cells derived from the human maxillary sinus membrane were treated with various concentrations of simvastatin. The adequate concentration of simvastatin for osteogenic induction was determined using bone morphogenetic protein (BMP-2). The efficacy of osteogenic differentiation of simavastatin was verified using osteocalcin mRNA, and the mineralization efficacy of hSMSCs and simvastatin treatment was compared with alkaline phosphatase and von Kossa staining. Results: Expression of BMP-2 mRNA and protein was observed after three days and was dependent on the concentration of simvastatin. Expression of osteocalcin mRNA was observed after three days in the $1.0{\mu}M$ simvastatin-treated group. Mineralization was observed after three days in the simvastatin-treated group. Conclusion: These results suggest that simvastatin induces the osteogenic potential of mesenchymal stem cells derived from the human maxillary sinus membrane mucosa.

• BMB Reports
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• 제48권6호
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• pp.319-323
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• 2015

Mesenchymal stem cells (MSCs) are multipotent stem cells capable of differentiating into adipocytes, osteoblasts, or chondrocytes. A mutually inhibitory relationship exists between osteogenic and adipogenic lineage commitment and differentiation. Such cell fate decision is regulated by several signaling pathways, including Wnt and bone morphogenetic protein (BMP). Accumulating evidence indicates that microRNAs (miRNAs) act as switches for MSCs to differentiate into either osteogenic or adipogenic lineage. Different miRNAs have been reported to regulate a master transcription factor for osteogenesis, such as Runx2, as well as molecules in the Wnt or BMP signaling pathway, and control the balance between osteoblast and adipocyte differentiation. Here, we discuss recent advancement of the cell fate decision of MSCs by miRNAs and their targets. [BMB Reports 2015; 48(6): 319-323]

• The Korean Journal of Physiology and Pharmacology
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• 제25권3호
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• pp.197-206
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• 2021

Carnosol is a phenolic diterpene phytochemical found in rosemary and sage with reported anti-microbial, anti-oxidant, anti-inflammatory, and anti-carcinogenic activities. This study aimed to investigate the effect of carnosol on the lineage commitment of mouse bone marrow-derived mesenchymal stem cells (mBMSCs) into osteoblasts and adipocytes. Interestingly, carnosol stimulated the early commitment of mBMSCs into osteoblasts in dose-dependent manner as demonstrated by increased levels of alkaline phosphatase activity and Alizarin red staining for matrix mineralization. On the other hand, carnosol significantly suppressed adipogenesis of mBMSCs and downregulated both early and late markers of adipogenesis. Carnosol showed to induce osteogenesis in a mechanism mediated by activating BMP signaling pathway and subsequently upregulating the expression of BMPs downstream osteogenic target genes. In this context, treatment of mBMSCs with LDN-193189, BMPR1 selective inhibitor showed to abolish the stimulatory effect of carnosol on BMP2-induced osteogenesis. In conclusion, our data identified carnosol as a novel osteoanabolic phytochemical that can promote the differentiation of mBMSCs into osteoblasts versus adipocytes by activating BMP-signaling.