Safety use of human dental pulp stem cells for cell-based therapies: No tumor and neither teratoma formation in vivo

Tracking transplanted stem cells is necessary to clarify cellular properties and improve transplantation success. In this study, we investigate the effects of fluorescent superparamagnetic iron oxide particles (SPIO) (Molday ION Rhodamine-B™, MIRB) on biological properties of human dental pulp stem cells (hDPSCs) and monitor hDPSCs in vitro and in vivo using magnetic resonance imaging (MRI). Morphological analysis showed that intracellular MIRB particles were distributed in the cytoplasm surrounding the nuclei of hDPSCs. 12.5–100 μg/mL MIRB all resulted in 100% labeling efficiency. MTT showed that 12.5–50 μg/mL MIRB could promote cell proliferation and MIRB over 100 μg/mL exhibited toxic effect on hDPSCs. In vitro MRI showed that 1 × 106cells labeled with various concentrations of MIRB (12.5–100 μg/mL) could be visualized. In vivo MRI showed that transplanted cells could be clearly visualized up to 60 days after transplantation. These results suggest that 12.5–50 μg/mL MIRB is a safe range for labeling hDPSCs. MIRB labeled hDPSCs cell can be visualized by MRI in vitro and in vivo. These data demonstrate that MIRB is a promising candidate for hDPSCs tracking in hDPSCs based dental pulp regeneration therapy.

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Simvastatin Induces the Odontogenic Differentiation of Human Dental Pulp Stem Cells In Vitro and In Vivo

Journal of Endodontics ◽

10.1016/j.joen.2008.11.024 ◽

2009 ◽

Vol 35 (3) ◽

pp. 367-372 ◽

Cited By ~ 53

Author(s):

Yosuke Okamoto ◽

Wataru Sonoyama ◽

Mitsuaki Ono ◽

Kentaro Akiyama ◽

Takuo Fujisawa ◽

...

Keyword(s):

Stem Cells ◽

Dental Pulp ◽

Dental Pulp Stem Cells ◽

Odontogenic Differentiation ◽

Human Dental Pulp

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Establishment of xenogeneic serum-free culture methods for handling human dental pulp stem cells using clinically oriented in-vitro and in-vivo conditions

Stem Cell Research & Therapy ◽

10.1186/s13287-017-0761-5 ◽

2018 ◽

Vol 9 (1) ◽

Cited By ~ 12

Author(s):

Mai Mochizuki ◽

Taka Nakahara

Keyword(s):

Stem Cells ◽

Dental Pulp ◽

Dental Pulp Stem Cells ◽

Culture Methods ◽

Serum Free ◽

Human Dental Pulp

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Magnetic Resonance Imaging of Human Dental Pulp Stem Cells in Vitro and in Vivo

Cell Transplantation ◽

10.3727/096368912x657774 ◽

2013 ◽

Vol 22 (10) ◽

pp. 1813-1829 ◽

Cited By ~ 24

Author(s):

T. Struys ◽

A. Ketkar-Atre ◽

P. Gervois ◽

C. Leten ◽

P. Hilkens ◽

...

Keyword(s):

Magnetic Resonance Imaging ◽

Stem Cells ◽

Magnetic Resonance ◽

Dental Pulp ◽

Dental Pulp Stem Cells ◽

Resonance Imaging ◽

Human Dental Pulp

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LncRNA H19 Promotes Odontoblastic Differentiation of Human Dental Pulp Stem Cells by Regulating miR-140-5p and BMP-2/FGF9

10.21203/rs.3.rs-16133/v1 ◽

2020 ◽

Author(s):

Jialin Zhong ◽

Xinran Tu ◽

Yuanyuan Kong ◽

Liyang Guo ◽

Baishun Li ◽

...

Keyword(s):

Stem Cells ◽

Dental Pulp ◽

Dental Pulp Stem Cells ◽

Biological Role ◽

Luciferase Reporter ◽

Qrt Pcr ◽

Odontoblastic Differentiation ◽

Human Dental Pulp

Abstract Background: Increasing evidence has revealed that long non-coding RNAs (lncRNAs) exert critical roles in biological mineralization. As a critical process for dentin formation, odontoblastic differentiation is regulated by complex signaling networks. The present study aimed to investigate the biological role and regulatory mechanisms of lncRNA-H19 (H19) in regulating the odontoblastic differentiation of human dental pulp stem cells (hDPSCs). Methods: We performed lncRNA microarray assay to reveal the expression patterns of lncRNAs involved in odontoblastic differentiation. H19 was identified and verified by qRT-PCR. The gain- and loss-of-function studies were performed to investigate the biological role of H19 in regulating odontoblastic differentiation of hDPSCs in vitro and in vivo. Odontoblastic differentiation was evaluated through qRT-PCR, Western blot and Alizarin Red S staining. Bioinformatics analysis identified that H19 could directly interact with miR-140-5p, which was further verified by luciferase reporter assay. After overexpression of miR-140-5p in hDPSCs, odontoblastic differentiation was determined. Moreover, the potential target genes of miR-140-5p were investigated and the biological functions of BMP-2 and FGF9 in hDPSCs were verified. Co-transfection experiments were conducted to validate miR-140-5p was involved in H19-mediated odontoblastic differentiation in hDPSCs.Results: The expression of H19 was significantly up-regulated in hDPSCs undergoing odontoblastic differentiation. Overexpression of H19 stimulated odontoblastic differentiation in vitro and in vivo, whereas down-regulation of H19 revealed the opposite effect. H19 binds directly to miR-140-5p and overexpression of miR-140-5p inhibited odontoblastic differentiation of hDPSCs. H19 acted as a miR-140-5p sponge, resulting in regulated the expression of BMP-2 and FGF9. Overexpression of H19 abrogated the inhibitory effect of miR-140-5p on odontoblastic differentiation.Conclusion: Our data revealed that H19 plays a positive regulatory role in odontoblastic differentiation of hDPSCs through miR-140-5p/BMP-2/FGF9 axis, suggesting that H19 may be a stimulatory regulator of odontogenesis.

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The use of human dental pulp stem cells for in vivo bone tissue engineering: A systematic review

Journal of Tissue Engineering ◽

10.1177/2041731417752766 ◽

2018 ◽

Vol 9 ◽

pp. 204173141775276 ◽

Cited By ~ 43

Author(s):

Alessander Leyendecker Junior ◽

Carla Cristina Gomes Pinheiro ◽

Tiago Lazzaretti Fernandes ◽

Daniela Franco Bueno

Keyword(s):

Systematic Review ◽

Tissue Engineering ◽

Stem Cells ◽

Bone Tissue Engineering ◽

Bone Tissue ◽

Dental Pulp ◽

Dental Pulp Stem Cells ◽

Deciduous Teeth ◽

Human Dental Pulp

Dental pulp represents a promising and easily accessible source of mesenchymal stem cells for clinical applications. Many studies have investigated the use of human dental pulp stem cells and stem cells isolated from the dental pulp of human exfoliated deciduous teeth for bone tissue engineering in vivo. However, the type of scaffold used to support the proliferation and differentiation of dental stem cells, the animal model, the type of bone defect created, and the methods for evaluation of results were extremely heterogeneous among these studies conducted. With this issue in mind, the main objective of this study is to present and summarize, through a systematic review of the literature, in vivo studies in which the efficacy of human dental pulp stem cells and stem cells from human exfoliated deciduous teeth (SHED) for bone regeneration was evaluated. The article search was conducted in PubMed/MEDLINE and Web of Science databases. Original research articles assessing potential of human dental pulp stem cells and SHED for in vivo bone tissue engineering, published from 1984 to November 2017, were selected and evaluated in this review according to the following eligibility criteria: published in English, assessing dental stem cells of human origin and evaluating in vivo bone tissue formation in animal models or in humans. From the initial 1576 potentially relevant articles identified, 128 were excluded due to the fact that they were duplicates and 1392 were considered ineligible as they did not meet the inclusion criteria. As a result, 56 articles remained and were fully analyzed in this systematic review. The results obtained in this systematic review open new avenues to perform bone tissue engineering for patients with bone defects and emphasize the importance of using human dental pulp stem cells and SHED to repair actual bone defects in an appropriate animal model.

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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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