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Effect of EDTA on Attachment and Differentiation of Dental Pulp Stem Cells

 Nan-Sim Pang  ;  Seung Jong Lee  ;  Euiseong Kim  ;  Dong Min Shin  ;  Sung Won Cho  ;  Wonse Park  ;  Xianglan Zhang  ;  Il-Young Jung 
 JOURNAL OF ENDODONTICS, Vol.40(6) : 811-817, 2014 
Journal Title
Issue Date
Adolescent ; Alkaline Phosphatase/analysis ; Calcification, Physiologic/drug effects ; Cell Adhesion/drug effects ; Cell Count ; Cell Culture Techniques ; Cell Differentiation/drug effects ; Cells, Cultured ; Dental Pulp/cytology* ; Dentin/drug effects ; Edetic Acid/pharmacology* ; Extracellular Matrix Proteins/analysis ; Fibronectins/analysis ; Humans ; Microscopy, Electron, Scanning ; Odontoblasts/drug effects ; Odontoblasts/physiology ; Osteoblasts/drug effects ; Osteocalcin/analysis ; Phosphoproteins/analysis ; Real-Time Polymerase Chain Reaction/methods ; Root Canal Irrigants/pharmacology* ; Sialoglycoproteins/analysis ; Stem Cells/drug effects* ; Stem Cells/physiology ; Time Factors ; Young Adult
Cell attachment ; EDTA ; cell differentiation ; dental pulp stem cells ; regenerative endodontics
INTRODUCTION: In regenerative endodontics, it is believed that EDTA induces odontoblast differentiation by releasing growth factors from the dentin matrix. The aim of this study was to evaluate the effect of EDTA on the attachment and differentiation of dental pulp stem cells (DPSCs). We also investigated whether the behavioral changes of DPSCs could be caused by biochemical components released from EDTA-treated dentin.
METHODS:Cells were obtained from human third molars, and the stem-like nature of the cells was investigated by flow cytometric analysis. DPSCs were seeded on EDTA-treated and untreated dentin slices. After 3 days of culture, cell attachment was evaluated by cell density, fibronectin 1 gene expression level using quantitative real-time polymerase chain reaction, and scanning electron microscopy. After 21 days of culture, the expression of differentiation genes was investigated by quantitative real-time polymerase chain reaction, and calcification was observed using alizarin red S staining. To investigate the EDTA-induced growth factor release, DPSCs were cultured with or without direct contact with the EDTA-treated dentin surface.
RESULTS:After 3 days of culture, both the cell density and fibronectin expression level were significantly higher in the EDTA-treated dentin group. After 3 weeks, the DPSCs on the EDTA-treated dentin surfaces showed higher expression levels of dentin sialophosphoprotein and dentin matrix protein 1, whereas the DPSCs cultured without direct contact with the EDTA-treated dentin surfaces did not exhibit these findings.
CONCLUSIONS:Our results showed that EDTA induced cell attachment and odontoblastic/osteoblastic differentiation, which was observed only in the group in which the DPSCs were placed in direct contact with the EDTA-treated dentin surfaces. These findings suggest that EDTA is beneficial for achieving successful outcomes in regenerative endodontics.
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2. College of Dentistry (치과대학) > Research Institute (부설연구소) > 1. Journal Papers
2. College of Dentistry (치과대학) > Dept. of Advanced General Dentistry (통합치의학과) > 1. Journal Papers
2. College of Dentistry (치과대학) > Dept. of Conservative Dentistry (보존과학교실) > 1. Journal Papers
2. College of Dentistry (치과대학) > Dept. of Oral Biology (구강생물학교실) > 1. Journal Papers
Yonsei Authors
Kim, Eui Seong(김의성) ORCID logo https://orcid.org/0000-0003-2126-4761
Park, Wonse(박원서) ORCID logo https://orcid.org/0000-0002-2081-1156
Pang, Nan Sim(방난심) ORCID logo https://orcid.org/0000-0003-4265-673X
Shin, Dong Min(신동민) ORCID logo https://orcid.org/0000-0001-6042-0435
Lee, Seung Jong(이승종)
Zhang, Xiang Lan(장향란)
Jung, Il Young(정일영) ORCID logo https://orcid.org/0000-0001-8972-2664
Cho, Sung Won(조성원) ORCID logo https://orcid.org/0000-0001-7505-9769
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