title: Effect of EDTA on attachment and differentiation of dental pulp stem cells.
abstract: BACKGROUND
  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.
authors.affiliation: Department of Advanced General Dentistry, College of Dentistry, Yonsei University, Seoul, Korea.
authors.forename: Nan-Sim
authors.initials: NS
authors.lastname: Pang
authors.affiliation: Department of Conservative Dentistry, College of Dentistry, Yonsei University, Seoul, Korea.
authors.forename: Seung Jong
authors.initials: SJ
authors.lastname: Lee
authors.affiliation: Department of Conservative Dentistry, College of Dentistry, Yonsei University, Seoul, Korea.
authors.forename: Euiseong
authors.initials: E
authors.lastname: Kim
authors.affiliation: Department of Oral Biology, College of Dentistry, Yonsei University, Seoul, Korea.
authors.forename: Dong Min
authors.initials: DM
authors.lastname: Shin
authors.affiliation: Department of Oral Biology, College of Dentistry, Yonsei University, Seoul, Korea.
authors.forename: Sung Won
authors.initials: SW
authors.lastname: Cho
authors.affiliation: Department of Advanced General Dentistry, College of Dentistry, Yonsei University, Seoul, Korea.
authors.forename: Wonse
authors.initials: W
authors.lastname: Park
authors.affiliation: Department of Oral Cancer Research Institute, College of Dentistry, Yonsei University, Seoul, Korea; Department of Pathology, Yanbian University Hospital, Yanji City, Jilin Province, China.
authors.forename: Xianglan
authors.initials: X
authors.lastname: Zhang
authors.affiliation: Department of Conservative Dentistry, College of Dentistry, Yonsei University, Seoul, Korea. Electronic address: juen@yuhs.ac.
authors.forename: Il-Young
authors.initials: IY
authors.lastname: Jung
mesh_terms: D000293:Adolescent; D000469:Alkaline Phosphatase / Q000032:analysis; D002113:Calcification, Physiologic / Q000187:drug effects; D002448:Cell Adhesion / Q000187:drug effects; D002452:Cell Count; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000187:drug effects; D002478:Cells, Cultured; D003782:Dental Pulp / Q000166:cytology*; D003804:Dentin / Q000187:drug effects; D004492:Edetic Acid / Q000494:pharmacology*; D016326:Extracellular Matrix Proteins / Q000032:analysis; D005353:Fibronectins / Q000032:analysis; D006801:Humans; D008855:Microscopy, Electron, Scanning; D009804:Odontoblasts / Q000187:drug effects / Q000502:physiology; D010006:Osteoblasts / Q000187:drug effects; D015675:Osteocalcin / Q000032:analysis; D010750:Phosphoproteins / Q000032:analysis; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods; D012388:Root Canal Irrigants / Q000494:pharmacology*; D012795:Sialoglycoproteins / Q000032:analysis; D013234:Stem Cells / Q000187:drug effects* / Q000502:physiology; D013997:Time Factors; D055815:Young Adult; D000099311:Dentin Sialophosphoprotein
chemical_list: D000469:Alkaline Phosphatase; D004492:Edetic Acid; D016326:Extracellular Matrix Proteins; D005353:Fibronectins; D015675:Osteocalcin; D010750:Phosphoproteins; D012388:Root Canal Irrigants; D012795:Sialoglycoproteins; D000099311:Dentin Sialophosphoprotein; C513309:DMP1 protein, human; C067625:FN1 protein, human
keywords: Cell attachment; EDTA; cell differentiation; dental pulp stem cells; regenerative endodontics