scholarly journals Hexosamine-Induced TGF-βSignaling and Osteogenic Differentiation of Dental Pulp Stem Cells Are Dependent on N-Acetylglucosaminyltransferase V

2015 ◽  
Vol 2015 ◽  
pp. 1-11 ◽  
Author(s):  
Yi-Jane Chen ◽  
Chung-Chen Yao ◽  
Chien-Hsun Huang ◽  
Hao-Hueng Chang ◽  
Tai-Horng Young
Keyword(s):  
Stem Cells ◽  
Osteogenic Differentiation ◽  
Dental Pulp ◽  
Critical Role ◽  
Dental Pulp Stem Cells ◽  
Receptor Type ◽  
Marker Genes ◽  
Type I ◽  
Differentiation Potential ◽  
Matrix Deposition

Glycans of cell surface glycoproteins are involved in the regulation of cell migration, growth, and differentiation. N-acetyl-glucosaminyltransferase V (GnT-V) transfers N-acetyl-d-glucosamine to formβ1,6-branched N-glycans, thus playing a crucial role in the biosynthesis of glycoproteins. This study reveals the distinct expression of GnT-V in STRO-1 and CD-146 double-positive dental pulp stem cells (DPSCs). Furthermore, we investigated three types of hexosamines and their N-acetyl derivatives for possible effects on the osteogenic differentiation potential of DPSCs. Our results showed that exogenous d-glucosamine (GlcN), N-acetyl-d-glucosamine (GlcNAc), d-mannosamine (ManN), and acetyl-d-mannosamine (ManNAc) promoted DPSCs’ early osteogenic differentiation in the absence of osteogenic supplements, but d-galactosamine (GalN) or N-acetyl-galactosamine (GalNAc) did not. Effects include the increased level of TGF-βreceptor type I, activation of TGF-βsignaling, and increased mRNA expression of osteogenic differentiation marker genes. The hexosamine-treated DPSCs showed an increased mineralized matrix deposition in the presence of osteogenic supplements. Moreover, the level of TGF-βreceptor type I and early osteogenic differentiation were abolished in the DPSCs transfected with siRNA for GnT-V knockdown. These results suggest that GnT-V plays a critical role in the hexosamine-induced activation of TGF-βsignaling and subsequent osteogenic differentiation of DPSCs.

scholarly journals Bone Regeneration Potential of Human Dental Pulp Stem Cells Derived from Elderly Patients and Osteo-Induced by a Helioxanthin Derivative

2020 ◽  
Vol 21 (20) ◽  
pp. 7731
Author(s):  
Marika Sato ◽  
Yoko Kawase-Koga ◽  
Daiki Yamakawa ◽  
Yasuyuki Fujii ◽  
Daichi Chikazu
Keyword(s):  
Stem Cells ◽  
Elderly Patients ◽  
Osteogenic Differentiation ◽  
Dental Pulp ◽  
Young Patients ◽  
Dental Pulp Stem Cells ◽  
Proliferative Potential ◽  
Regenerative Therapy ◽  
Differentiation Potential ◽  
Human Dental Pulp

Human dental pulp stem cells (DPSCs) have high clonogenic and proliferative potential. We previously reported that a helioxanthin derivative (4-(4-methoxyphenyl)pyrido[40,30:4,5]thieno[2–b]pyridine-2-carboxamide (TH)) enhances osteogenic differentiation of DPSCs derived from young patients. However, in the clinical field, elderly patients more frequently require bone regenerative therapy than young patients. In this study, we examined and compared the osteogenic differentiation potential of TH-induced DPSCs from elderly patients and young patients to explore the potential clinical use of DPSCs for elderly patients. DPSCs were obtained from young and elderly patients and cultured in osteogenic medium with or without TH. We assessed the characteristics and osteogenic differentiation by means of specific staining and gene expression analyses. Moreover, DPSC sheets were transplanted into mouse calvarial defects to investigate osteogenesis of TH-induced DPSCs by performing micro-computed tomography (micro-CT). We demonstrated that osteogenic conditions with TH enhance the osteogenic differentiation marker of DPSCs from elderly patients as well as young patients in vitro. In vivo examination showed increased osteogenesis of DPSCs treated with TH from both elderly patients and young patients. Our results suggest that the osteogenic differentiation potential of DPSCs from elderly patients is as high as that of DPSCs from young patients. Moreover, TH-induced DPSCs showed increased osteogenic differentiation potential, and are thus a potentially useful cell source for bone regenerative therapy for elderly patients.

scholarly journals Bone regeneration potential of human dental pulp stem cells derived from elderly patients and induced by a helioxanthin derivative

2020 ◽  
Author(s):  
Marika Sato ◽  
YOKO KAWASE-KOGA ◽  
Daiki Yamakawa ◽  
Yasuyuki Fujii ◽  
Daichi Chikazu
Keyword(s):  
Stem Cells ◽  
Elderly Patients ◽  
Osteogenic Differentiation ◽  
Dental Pulp ◽  
Young Patients ◽  
Dental Pulp Stem Cells ◽  
Proliferative Potential ◽  
Regenerative Therapy ◽  
Differentiation Potential ◽  
Human Dental Pulp

Abstract Background: Human dental pulp stem cells (DPSCs) have the ability to differentiate into multiple lineage cell types including adipogenic, neurogenic, and osteogenic cells. Dental pulp stem cells can be easily collected from extracted teeth and are now considered to be a type of mesenchymal stem cell with higher clonogenic and proliferative potential than bone marrow stem cells. Previous studies have described the osteogenic ability of DPSCs that were isolated from only young patients. However, in fact, elderly patients more frequently require bone regenerative therapy—for example, in alveolar bone defects resulting from periodontal disease—than young patients. We previously reported that 4-(4-methoxyphenyl)pyrido[40,30:4,5]thieno[2,3-b]pyridine-2-carboxamide (TH), a helioxanthin derivative, induces osteogenic differentiation of DPSCs derived from young patients. However, the effect of TH on the osteogenic differentiation potential of DPSCs derived from elderly patients remains unknown. Therefore, the present study aimed to examine and compare the osteogenic differentiation potential of TH-induced DPSCs from elderly patients and young patients to explore the potential clinical use of DPSCs for elderly patients.Methods: Dental pulp stem cells were obtained from the dental pulp of teeth of healthy young patients (18-39 years old) and healthy elderly patients (40-67 years old) and cultured in regular medium and osteogenic medium with or without TH. We assessed the morphological characteristics, proliferation, and osteogenic differentiation of DPSCs from both young and elderly patients. Moreover, DPSC sheets were transplanted into mouse calvarial defects to investigate osteogenesis of TH-induced DPSCs in vivo.Results: We demonstrated that osteogenic conditions with TH induce the osteogenic differentiation and osteogenesis of DPSCs from elderly patients as well as young patients more than osteogenic conditions without TH. Conclusions: Our results suggested that the osteogenic differentiation potential of DPSCs from elderly patients is as high as that of DPSCs from young patients. Moreover, TH-induced DPSCs show increased osteogenic differentiation potential, and they can be a useful cell source for bone regenerative therapy for not only young patients but also elderly patients.

scholarly journals The Osteogenic Differentiation of Human Dental Pulp Stem Cells through G0/G1 Arrest and the p-ERK/Runx-2 Pathway by Sonic Vibration

2021 ◽  
Vol 22 (18) ◽  
pp. 10167
Author(s):  
Won Lee ◽  
Su-Rak Eo ◽  
Ju-Hye Choi ◽  
Yu-Mi Kim ◽  
Myeong-Hyun Nam ◽  
...  
Keyword(s):  
Stem Cells ◽  
Osteogenic Differentiation ◽  
Dental Pulp ◽  
Calcium Content ◽  
Dental Pulp Stem Cells ◽  
The Other ◽  
G1 Arrest ◽  
Type I ◽  
Mrna Levels ◽  
Human Dental Pulp

Mechanical/physical stimulations modulate tissue metabolism, and this process involves multiple cellular mechanisms, including the secretion of growth factors and the activation of mechano-physically sensitive kinases. Cells and tissue can be modulated through specific vibration-induced changes in cell activity, which depend on the vibration frequency and occur via differential gene expression. However, there are few reports about the effects of medium-magnitude (1.12 g) sonic vibration on the osteogenic differentiation of human dental pulp stem cells (HDPSCs). In this study, we investigated whether medium-magnitude (1.12 g) sonic vibration with a frequency of 30, 45, or 100 Hz could affect the osteogenic differentiation of HDPSCs. Their cell morphology changed to a cuboidal shape at 45 Hz and 100 Hz, but the cells in the other groups were elongated. FACS analysis showed decreased CD 73, CD 90, and CD 105 expression at 45 Hz and 100 Hz. Additionally, the proportions of cells in the G0/G1 phase in the control, 30 Hz, 45 Hz, and 100 Hz groups after vibration were 60.7%, 65.9%, 68.3%, and 66.7%, respectively. The mRNA levels of osteogenic-specific markers, including osteonectin, osteocalcin, BMP-2, ALP, and Runx-2, increased at 45 and 100 Hz, and the ALP and calcium content was elevated in the vibration groups compared with those in the control. Additionally, the western blotting results showed that p-ERK, BSP, osteoprotegerin, and osteonectin proteins were upregulated at 45 Hz compared with the other groups. The vibration groups showed higher ALP and calcium content than the control. Vibration, especially at 100 Hz, increased the number of calcified nodes relative to the control group, as evidenced by von Kossa staining. Immunohistochemical staining demonstrated that type I and III collagen, osteonectin, and osteopontin were upregulated at 45 Hz and 100 Hz. These results suggest that medium magnitude vibration at 45 Hz induces the G0/G1 arrest of HDPSCs through the p-ERK/Runx-2 pathway and can serve as a potent stimulator of differentiation and extracellular matrix production.

scholarly journals Multilineage Differentiation Potential of Human Dental Pulp Stem Cells—Impact of 3D and Hypoxic Environment on Osteogenesis In Vitro

2020 ◽  
Vol 21 (17) ◽  
pp. 6172
Author(s):  
Anna Labedz-Maslowska ◽  
Natalia Bryniarska ◽  
Andrzej Kubiak ◽  
Tomasz Kaczmarzyk ◽  
Malgorzata Sekula-Stryjewska ◽  
...  
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Osteogenic Differentiation ◽  
Dental Pulp ◽  
Dental Pulp Stem Cells ◽  
Osteogenic Potential ◽  
Stem Cell Population ◽  
Differentiation Potential ◽  
Human Dental Pulp

Human dental pulp harbours unique stem cell population exhibiting mesenchymal stem/stromal cell (MSC) characteristics. This study aimed to analyse the differentiation potential and other essential functional and morphological features of dental pulp stem cells (DPSCs) in comparison with Wharton’s jelly-derived MSCs from the umbilical cord (UC-MSCs), and to evaluate the osteogenic differentiation of DPSCs in 3D culture with a hypoxic microenvironment resembling the stem cell niche. Human DPSCs as well as UC-MSCs were isolated from primary human tissues and were subjected to a series of experiments. We established a multiantigenic profile of DPSCs with CD45−/CD14−/CD34−/CD29+/CD44+/CD73+/CD90+/CD105+/Stro-1+/HLA-DR− (using flow cytometry) and confirmed their tri-lineage osteogenic, chondrogenic, and adipogenic differentiation potential (using qRT-PCR and histochemical staining) in comparison with the UC-MSCs. The results also demonstrated the potency of DPSCs to differentiate into osteoblasts in vitro. Moreover, we showed that the DPSCs exhibit limited cardiomyogenic and endothelial differentiation potential. Decreased proliferation and metabolic activity as well as increased osteogenic differentiation of DPSCs in vitro, attributed to 3D cell encapsulation and low oxygen concentration, were also observed. DPSCs exhibiting elevated osteogenic potential may serve as potential candidates for a cell-based product for advanced therapy, particularly for bone repair. Novel tissue engineering approaches combining DPSCs, 3D biomaterial scaffolds, and other stimulating chemical factors may represent innovative strategies for pro-regenerative therapies.

scholarly journals Characterisation of proliferation, differentiation potential, and gene expression among clonal cultures of human dental pulp cells

2019 ◽  
Author(s):  
Tomoko Kobayashi ◽  
Daisuke Torii ◽  
Takanori Iwata ◽  
Yuichi Izumi ◽  
Masanori Nasu ◽  
...  
Keyword(s):  
Gene Expression ◽  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Dental Pulp ◽  
Dental Pulp Stem Cells ◽  
Marker Genes ◽  
Differentiation Potential ◽  
Candidate Marker ◽  
Cell Clones ◽  
Human Dental Pulp

Abstract Background: Mesenchymal stem cells are a highly promising source of cells for regeneration therapy because of their multilineage differentiation potential. However, distinct markers for mesenchymal stem cells are not well-established. To identify new candidate marker genes for multipotent human dental pulp stem cells, we analysed the characteristics and gene expression profiles of cell clones obtained from a single dental pulp specimen.Results: Fifty colony-forming single cell-derived clones were isolated from a single dental pulp specimen. These clones varied in their proliferation abilities and surface marker (STRO-1 and CD146) expression patterns, as well as their odontogenic, adipogenic, and chondrogenic differentiation potentials. Four clones maintained their original differentiation potentials during long-term culture. Gene expression profile analysis of five representative clones identified 1227 genes that were related to multipotency. Ninety of these 1227 genes overlapped with genes reportedly involved in ‘stemness or differentiation’. Based on the predicted locations of expressed protein products and large changes in expression levels, 14 of the 90 genes were selected as candidate dental pulp stem cell markers, particularly in relation to their multipotency characteristics.Conclusions: This characterisation of cell clones obtained from a single specimen of human dental pulp provided information regarding new candidate marker genes for multipotent dental pulp stem cells, which could facilitate efficient analysis or enrichment of multipotent stem cells.

scholarly journals Human Dental Pulp Stem Cells Exhibit Osteogenic Differentiation Potential

2020 ◽  
Vol 15 (1) ◽  
pp. 229-236
Author(s):  
Sadia Awais ◽  
Samira Shabbir Balouch ◽  
Nabeela Riaz ◽  
Mahmood S Choudhery
Keyword(s):  
Stem Cells ◽  
Osteogenic Differentiation ◽  
Dental Pulp ◽  
Histological Analysis ◽  
Morphological Changes ◽  
Dental Pulp Stem Cells ◽  
Osteogenic Potential ◽  
Proliferative Potential ◽  
Differentiation Potential ◽  
Human Dental Pulp

AbstractBone regeneration after trauma, pathologic and surgical procedures is considered a major medical challenge. Due to limitations in using conventional approaches, cell based regenerative strategies may provide an alternative option to address such issues. In the current study, we sought to determine the osteogenic potential of dental pulp stem cells (DPSCs) isolated from impacted 3rd molars. DPSCs were isolated from human dental pulp tissue (n=6) using explant culture. Growth characteristics of DPSCs were determined using plating efficiency, and the number and time of population doublings. After characterization, DPSCs were induced to differentiate into osteoblasts and were assessed using polymerase chain reactions (PCR) and histological analysis. Results indicated that DPSCs can be isolated from impacted human third molars, and that DPSCs exhibited typical fibroblastic morphology and excellent proliferative potential. In addition, morphological changes, histological analysis and expression of lineage specific genes confirmed osteogenic differentiation of DPSCs. In conclusion, DPSCs isolated from impacted 3rd molars have high proliferative potential and ability to differentiate into osteoblasts.

scholarly journals Comparative evaluation of osteogenic differentiation potential of stem cells derived from dental pulp and exfoliated deciduous teeth cultured over granular hydroxyapatite based scaffold

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Manal Nabil Hagar ◽  
Farinawati Yazid ◽  
Nur Atmaliya Luchman ◽  
Shahrul Hisham Zainal Ariffin ◽  
Rohaya Megat Abdul Wahab
Keyword(s):  
Stem Cells ◽  
Osteogenic Differentiation ◽  
Dental Pulp ◽  
3D Culture ◽  
Permanent Teeth ◽  
Deciduous Teeth ◽  
Osteogenic Potential ◽  
Control Group ◽  
Rt Pcr ◽  
Differentiation Potential

Abstract Background Mesenchymal stem cells isolated from the dental pulp of primary and permanent teeth can be differentiated into different cell types including osteoblasts. This study was conducted to compare the morphology and osteogenic potential of stem cells from exfoliated deciduous teeth (SHED) and dental pulp stem cells (DPSC) in granular hydroxyapatite scaffold (gHA). Preosteoblast cells (MC3T3-E1) were used as a control group. Methodology The expression of stemness markers for DPSC and SHED was evaluated using reverse transcriptase-polymerase chain reaction (RT-PCR). Alkaline phosphatase assay was used to compare the osteoblastic differentiation of these cells (2D culture). Then, cells were seeded on the scaffold and incubated for 21 days. Morphology assessment using field emission scanning electron microscopy (FESEM) was done while osteogenic differentiation was detected using ALP assay (3D culture). Results The morphology of cells was mononucleated, fibroblast-like shaped cells with extended cytoplasmic projection. In RT-PCR study, DPSC and SHED expressed GAPDH, CD73, CD105, and CD146 while negatively expressed CD11b, CD34 and CD45. FESEM results showed that by day 21, dental stem cells have a round like morphology which is the morphology of osteoblast as compared to day 7. The osteogenic potential using ALP assay was significantly increased (p < 0.01) in SHED as compared to DPSC and MC3T3-E1 in 2D and 3D cultures. Conclusion gHA scaffold is an optimal scaffold as it induced osteogenesis in vitro. Besides, SHED had the highest osteogenic potential making them a preferred candidate for tissue engineering in comparison with DPSC.

scholarly journals Development of novel gene carrier using modified nano hydroxyapatite derived from equine bone for osteogenic differentiation of dental pulp stem cells

2021 ◽  
Vol 6 (9) ◽  
pp. 2742-2751
Author(s):  
Myung Chul Lee ◽  
Hoon Seonwoo ◽  
Kyoung Je Jang ◽  
Shambhavi Pandey ◽  
Jaewoon Lim ◽  
...  
Keyword(s):  
Stem Cells ◽  
Osteogenic Differentiation ◽  
Dental Pulp ◽  
Dental Pulp Stem Cells ◽  
Gene Carrier ◽  
Novel Gene ◽  
Equine Bone
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