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

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• 作者：Nan-Sim Pang ; DDS ; MS ; PhD^&lowast ; ; Seung Jong Lee ; DDS ; MS ; PhD^&dagger ; ; Euiseong Kim ; DDS ; MS ; PhD^&dagger ; ; Dong Min Shin ; DDS ; MS ; PhD^&Dagger ; ; Sung Won Cho ; DDS ; MS ; PhD^&Dagger ; ; Wonse Park ; DDS ; MS ; PhD^&lowast ; ; Xianglan Zhang ; MD ; PhD^§ ; ; ^|| ; Il-Young Jung ; DDS ; MS ; PhD^&dagger ; ; ^{juen@yuhs.ac" class="auth_mail" title="E-mail the corresponding author}
• 关键词：Cell attachment ; cell differentiation ; dental pulp stem cells ; EDTA ; regenerative endodontics
• 刊名：Journal of Endodontics
• 出版年：2014
• 出版时间：June 2014
• 年：2014
• 卷：40
• 期：6
• 页码：811-817
• 全文大小：1309 K

文摘

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.