Study on Culture of Human Dental Pulp Stem Cells to Apply in Tissue Engineering

2011 ◽  
Vol 11 ◽  
pp. 13-20 ◽  
Author(s):  
Tran Le Bao Ha ◽  
Doan Nguyen Vu ◽  
To Minh Quan ◽  
Ngoc Phan Kim ◽  
Hung Hoang Tu ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Stem Cells ◽  
Dental Pulp ◽  
Dental Pulp Stem Cells ◽  
Dental Tissue ◽  
Dentin Surface ◽  
Human Dental Pulp ◽  
Dental Tissue Engineering ◽  
High Level

Dental pulp cell research might open a promising application in tooth tissue regeneration. The aim of this study is to establish a protocol for in vitro culture the human dental pulp stem cells to apply in tissue engineering. Human premolar and impacted third molars were collected and disinfected. Dental pulp fragments were cultured with Dulbecco's Modified Eagle Medium: Nutrient Mixture F-12 (DMEM/F12) medium supplemented with 10% Fetal Bovine Serum (FBS). Dental pulp stem cells (DPSCs) were identified using proliferation assay, RT-PCR and flow cytometry. Growth of DPSCs on dentin surface was assessed by MTT assay. The study showed that we successfully isolated, cultured and characterized dental pulp cells by outgrowth method. Cultured population of cells expressed in high level of Oct4, CD146, CD90, CD44. DPSCs proliferated on chemically and mechanically treated dentin surface. This research provides important information and a basis for further investigations to establish dental tissue engineering protocols.

Average cell viability levels of human dental pulp stem cells: an accurate combinatorial index for quality control in tissue engineering

Cytotherapy ◽  
2013 ◽  
Vol 15 (4) ◽  
pp. 507-518 ◽  
Author(s):  
Miguel Angel Martin-Piedra ◽  
Ingrid Garzon ◽  
Ana Celeste Oliveira ◽  
Camilo Andres Alfonso-Rodriguez ◽  
Maria Carmen Sanchez-Quevedo ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Stem Cells ◽  
Quality Control ◽  
Cell Viability ◽  
Dental Pulp ◽  
Dental Pulp Stem Cells ◽  
Average Cell ◽  
Human Dental Pulp

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

2018 ◽  
Vol 9 ◽  
pp. 204173141775276 ◽  
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.

scholarly journals The Histone Deacetylase Inhibitor (MS-275) Promotes Differentiation of Human Dental Pulp Stem Cells into Odontoblast-Like Cells Independent of the MAPK Signaling System

2020 ◽  
Vol 21 (16) ◽  
pp. 5771
Author(s):  
Eun-Cheol Lee ◽  
Yu-Mi Kim ◽  
Han-Moi Lim ◽  
Ga-Eun Ki ◽  
Young-Kwon Seo
Keyword(s):  
Stem Cells ◽  
Histone Deacetylase ◽  
Dental Pulp ◽  
Epigenetic Modification ◽  
Cell Types ◽  
Mapk Signaling ◽  
Dental Pulp Stem Cells ◽  
Signaling System ◽  
Dental Tissue ◽  
Human Dental Pulp

The role of dental pulp stem cells (DPSCs) in dental tissue regeneration is gaining attention because DPSCs can differentiate into odontoblasts and other specialized cell types. Epigenetic modification has been found to play an important role in cell differentiation and regulation, among which histone deacetylase (HDAC) is involved in suppressing genes by removing histone acetyl groups. The use of HDAC inhibitor to control this is increasing and has been widely studied by many researchers. This study aimed to induce differentiation by causing epigenetic changes in odontoblast-related genes and the MAPK signaling pathway in human dental pulp stem cells. Western blot and immunofluorescence staining showed increased expression of DMP-1, ALP, DSPP, and RUNX2 compared to the control. However, activation of the MAPK signaling system was similar to but slightly different from the expression of odontoblast-related proteins. After 3 days, as shown by MTT and LDH assays, proliferation decreased overall, but cytotoxicity decreased at only a specific concentration. We confirmed that there was no change in mRNA expression of caspase 3 or 9 using real-time PCR. In addition, flow cytometry analysis confirmed that differentiation occurred due to the decrease in the expression of the CD73 and CD146. Although overall proliferation was reduced due to the G2/M inhibition of the cell cycle, the expression of BCL-2 protected the cells from cell death. Overall, cell proliferation decreased in response to MS-275, but it did not induce cytotoxicity in 5 nM and 10 nM concentration and induces differentiation into odontoblast-like cells.

Differential mineralization of human dental pulp stem cells on diverse polymers

2018 ◽  
Vol 63 (3) ◽  
pp. 261-269 ◽  
Author(s):  
Christian Apel ◽  
Patricia Buttler ◽  
Jochen Salber ◽  
Anandhan Dhanasingh ◽  
Sabine Neuss
Keyword(s):  
Tissue Engineering ◽  
Stem Cells ◽  
Hyaluronic Acid ◽  
Dental Pulp ◽  
Dental Pulp Stem Cells ◽  
Specific Cell ◽  
Alizarin Red ◽  
Alp Activity ◽  
Human Dental Pulp ◽  
Mineralized Matrix

Abstract In tissue engineering, biomaterials are used as scaffolds for spatial distribution of specific cell types. Biomaterials can potentially influence cell proliferation and extracellular matrix formation, both in positive and negative ways. The aim of the present study was to investigate and compare mineralized matrix production of human dental pulp stem cells (DPSC), cultured on 17 different well-characterized polymers. Osteogenic differentiation of DPSC was induced for 21 days on biomaterials using dexamethasone, L-ascorbic-acid-2-phosphate, and sodium β-glycerophosphate. Success of differentiation was analyzed by quantitative RealTime PCR, alkaline phosphatase (ALP) activity, and visualization of calcium accumulations by alizarin red staining with subsequent quantification by colorimetric method. All of the tested biomaterials of an established biomaterial bank enabled a mineralized matrix formation of the DPSC after osteoinductive stimulation. Mineralization on poly(tetrafluoro ethylene) (PTFE), poly(dimethyl siloxane) (PDMS), Texin, LT706, poly(epsilon-caprolactone) (PCL), polyesteramide type-C (PEA-C), hyaluronic acid, and fibrin was significantly enhanced (p<0.05) compared to standard tissue culture polystyrene (TCPS) as control. In particular, PEA-C, hyaluronic acid, and fibrin promoted superior mineralization values. These results were confirmed by ALP activity on the same materials. Different biomaterials differentially influence the differentiation and mineralized matrix formation of human DPSC. Based on the present results, promising biomaterial candidates for bone-related tissue engineering applications in combination with DPSC can be selected.

scholarly journals Vasculogenesis from Human Dental Pulp Stem Cells Grown in Matrigel with Fully Defined Serum-Free Culture Media

Biomedicines ◽  
2020 ◽  
Vol 8 (11) ◽  
pp. 483
Author(s):  
Jon Luzuriaga ◽  
Jon Irurzun ◽  
Igor Irastorza ◽  
Fernando Unda ◽  
Gaskon Ibarretxe ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Stem Cells ◽  
Cell Therapy ◽  
Dental Pulp ◽  
Culture Media ◽  
Dental Pulp Stem Cells ◽  
Adult Brain ◽  
Serum Free ◽  
Human Dental Pulp

The generation of vasculature is one of the most important challenges in tissue engineering and regeneration. Human dental pulp stem cells (hDPSCs) are some of the most promising stem cell types to induce vasculogenesis and angiogenesis as they not only secrete vascular endothelial growth factor (VEGF) but can also differentiate in vitro into both endotheliocytes and pericytes in serum-free culture media. Moreover, hDPSCs can generate complete blood vessels containing both endothelial and mural layers in vivo, upon transplantation into the adult brain. However, many of the serum free media employed for the growth of hDPSCs contain supplements of an undisclosed composition. This generates uncertainty as to which of its precise components are necessary and which are dispensable for the vascular differentiation of hDPSCs, and also hinders the transfer of basic research findings to clinical cell therapy. In this work, we designed and tested new endothelial differentiation media with a fully defined composition using standard basal culture media supplemented with a mixture of B27, heparin and growth factors, including VEGF-A165 at different concentrations. We also optimized an in vitro Matrigel assay to characterize both the ability of hDPSCs to differentiate to vascular cells and their capacity to generate vascular tubules in 3D cultures. The description of a fully defined serum-free culture medium for the induction of vasculogenesis using human adult stem cells highlights its potential as a relevant innovation for tissue engineering applications. In conclusion, we achieved efficient vasculogenesis starting from hDPSCs using serum-free culture media with a fully defined composition, which is applicable for human cell therapy purposes.

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