Proliferative Capacity and Differentiation Potential of Isolated Postnatal Human Dental Pulp Stem Cells in Diabetic Patients

The purpose of this study was to investigate the effects of mesoporous bioactive glass nanoparticle (MBN)/graphene oxide (GO) composites on the mineralization ability and differentiation potential of human dental pulp stem cells (hDPSCs). MBN/GO composites were synthesized using the sol-gel method and colloidal processing to enhance the bioactivity and mechanical properties of MBN. Characterization using FESEM, XRD, FTIR, and Raman spectrometry showed that the composites were successfully synthesized. hDPSCs were then cultured directly on the MBN/GO (40:1 and 20:1) composites in vitro. MBN/GO promoted the proliferation and alkaline phosphatase (ALP) activity of hDPSCs. In addition, qRT-PCR showed that MBN/GO regulated the mRNA levels of odontogenic markers (dentin sialophosphoprotein (DSPP), dentine matrix protein 1 (DMP-1), ALP, matrix extracellular phosphoglycoprotein (MEPE), bone morphogenetic protein 2 (BMP-2), and runt-related transcription factor 2 (RUNX-2)). The mRNA levels of DSPP and DMP-1, two odontogenesis-specific markers, were considerably upregulated in hDPSCs in response to growth on the MBN/GO composites. Western blot analysis revealed similar results. Alizarin red S staining was subsequently performed to further investigate MBN/GO-induced mineralization of hDPSCs. It was revealed that MBN/GO composites promote odontogenic differentiation via the Wnt/β-catenin signaling pathway. Collectively, the results of the present study suggest that MBN/GO composites may promote the differentiation of hDPSCs into odontoblast-like cells, and potentially induce dentin formation.

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Isolation of Human Dental Pulp Stem Cells and Their Characteristics Before and After Cryopreservation

Problems of Cryobiology and Cryomedicine ◽

10.15407/cryo31.01.058 ◽

2021 ◽

Vol 31 (1) ◽

pp. 58-69

Author(s):

Svitlana Mazur ◽

◽

Olena Rogulska ◽

Olena Revenko ◽

Nataliya Volkova ◽

...

Keyword(s):

Stem Cells ◽

Dental Pulp ◽

Third Molar ◽

Tooth Germ ◽

Multipotent Mesenchymal Stromal Cells ◽

Dental Pulp Stem Cells ◽

Slow Cooling ◽

Differentiation Potential ◽

Before And After ◽

Human Dental Pulp

Dental pulp stem cells (DPSCs) from human third molar tooth germ (wisdom tooth) were isolated using a collagenasebased enzymatic method, the obtained cells were analyzed as for morphology in monolayer culture, immunophenotype, proliferation and differentiation potential before and after cryopreservation. In this study, we showed that based on morphological features, surface markers profile and differentiation potential, the isolated DPSCs corresponded to multipotent mesenchymal stromal cells. DPSCs cryopreservation by slow cooling (1 °С / min) down to –80°C with subsequent immersion into liquid nitrogen in cryoprotectant-free culture medium led to cell death. Cryopreservation using the same protocol in the presence of 10% dimethyl sulfoxide (DMSO) and 20% serum ensured (82 ± 6)% cell viability; while metabolic and proliferative activity, as well as the ability to differentiate into the osteo- and adipogenic lineages of cryopreserved DPSCs were similar to their non-cryopreserved counterparts.

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Effect of isolation methodology on stem cell properties and multilineage differentiation potential of human dental pulp stem cells

Cell and Tissue Research ◽

10.1007/s00441-013-1630-x ◽

2013 ◽

Vol 353 (1) ◽

pp. 65-78 ◽

Cited By ~ 109

Author(s):

P. Hilkens ◽

P. Gervois ◽

Y. Fanton ◽

J. Vanormelingen ◽

W. Martens ◽

...

Keyword(s):

Stem Cells ◽

Stem Cell ◽

Dental Pulp ◽

Dental Pulp Stem Cells ◽

Multilineage Differentiation ◽

Differentiation Potential ◽

Human Dental Pulp

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Effects of Helioxanthin Derivative-Treated Human Dental Pulp Stem Cells on Fracture Healing

International Journal of Molecular Sciences ◽

10.3390/ijms21239158 ◽

2020 ◽

Vol 21 (23) ◽

pp. 9158

Author(s):

Daiki Yamakawa ◽

Yoko Kawase-Koga ◽

Yasuyuki Fujii ◽

Yuki Kanno ◽

Marika Sato ◽

...

Keyword(s):

Stem Cells ◽

Fracture Healing ◽

Efficient Method ◽

Dental Pulp ◽

Fracture Site ◽

Dental Pulp Stem Cells ◽

Differentiation Potential ◽

Normal Medium ◽

Human Dental Pulp

Bone defects affect patients functionally and psychologically and can decrease quality of life. To resolve these problems, a simple and efficient method of bone regeneration is required. Human dental pulp stem cells (DPSCs) have high proliferative ability and multilineage differentiation potential. In our previous study, we reported a highly efficient method to induce osteogenic differentiation using DPSC sheets treated with a helioxanthin derivative (4-(4-methoxyphenyl)pyrido[40,30:4,5]thieno[2,3-b]pyridine-2-carboxamide (TH)) in a mouse calvarial defect model. However, the localization of the DPSCs after transplantation remains unknown. Therefore, in this study, we investigated the localization of transplanted DPSCs in a mouse fracture model. DPSCs were collected from six healthy patients aged 18–29 years, cultured in normal medium (NM), osteogenic medium (OM), or OM with TH, and fabricated them into cell sheets. To evaluate the efficacy of fracture healing using DPSCs treated with OM+TH, and to clarify the localization of the transplanted DPSC sheets in vivo, we transplanted OM+TH-treated DPSC sheets labeled with PKH26 into mouse tibiae fractures. We demonstrated that transplanted OM+TH-treated DPSCs sheets were localized to the fracture site and facilitated bone formation. These results indicated that transplanted OM+TH-treated DPSCs were localized at fracture sites and directly promoted fracture healing.

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

International Journal of Molecular Sciences ◽

10.3390/ijms21207731 ◽

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.

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

10.21203/rs.3.rs-16130/v1 ◽

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.

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

International Journal of Molecular Sciences ◽

10.3390/ijms21176172 ◽

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.

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