title: Effects of epicatechin, a crosslinking agent, on human dental pulp cells cultured in collagen scaffolds.
abstract: OBJECTIVE
  The purpose of this study was to investigate the biological effects of epicatechin (ECN), a crosslinking agent, on human dental pulp cells (hDPCs) cultured in collagen scaffolds.
  
  
  METHODS
  To evaluate the effects of ECN on the proliferation of hDPCs, cell counting was performed using optical and fluorescent microscopy. Measurements of alkaline phosphatase (ALP) activity, alizarin red staining, and real-time polymerase chain reactions were performed to assess odontogenic differentiation. The compressive strength and setting time of collagen scaffolds containing ECN were measured. Differential scanning calorimetry was performed to analyze the thermal behavior of collagen in the presence of ECN.
  
  
  RESULTS
  Epicatechin increased ALP activity, mineralized nodule formation, and the mRNA expression of dentin sialophosphoprotein (DSPP), a specific odontogenic-related marker. Furthermore, ECN upregulated the expression of DSPP in hDPCs cultured in collagen scaffolds. Epicatechin activated the extracellular signal-regulated kinase (ERK) and the treatment with an ERK inhibitor (U0126) blocked the expression of DSPP. The compressive strength was increased and the setting time was shortened in a dose-dependent manner. The number of cells cultured in the ECN-treated collagen scaffolds was significantly increased compared to the cells in the untreated control group.
  
  
  CONCLUSIONS
  Our results revealed that ECN promoted the proliferation and differentiation of hDPCs. Furthermore, the differentiation was regulated by the ERK signaling pathway. Changes in mechanical properties are related to cell fate, including proliferation and differentiation. Therefore, our study suggests the ECN treatment might be desirable for dentin-pulp complex regeneration.
authors.affiliation: Institute of Oral Bioscience, School of Dentistry, Chonbuk National University, Jeonju, Korea.
authors.forename: Eun-su
authors.initials: ES
authors.lastname: Lim
authors.affiliation: Institute of Oral Bioscience, School of Dentistry, Chonbuk National University, Jeonju, Korea.
authors.forename: Myung-Jin
authors.initials: MJ
authors.lastname: Lim
authors.affiliation: Institute of Oral Bioscience, School of Dentistry, Chonbuk National University, Jeonju, Korea.
authors.forename: Kyung-San
authors.initials: KS
authors.lastname: Min
authors.affiliation: Institute of Oral Bioscience, School of Dentistry, Chonbuk National University, Jeonju, Korea.
authors.forename: Young-Sun
authors.initials: YS
authors.lastname: Kwon
authors.affiliation: Department of Conservative Dentistry, School of Dentistry, Chonnam National University, Gwangju, Korea.
authors.forename: Yun-Chan
authors.initials: YC
authors.lastname: Hwang
authors.affiliation: Institute of Oral Bioscience, School of Dentistry, Chonbuk National University, Jeonju, Korea.
authors.forename: Mi-Kyung
authors.initials: MK
authors.lastname: Yu
authors.affiliation: Department of Conservative Dentistry, School of Dentistry, Dankook University, Cheonan, Korea.
authors.forename: Chan-Ui
authors.initials: CU
authors.lastname: Hong
authors.affiliation: Institute of Oral Bioscience, School of Dentistry, Chonbuk National University, Jeonju, Korea.
authors.forename: Kwang-Won
authors.initials: KW
authors.lastname: Lee
mesh_terms: D000469:Alkaline Phosphatase / Q000032:analysis; D000704:Analysis of Variance; D015153:Blotting, Western; D002152:Calorimetry, Differential Scanning; D002392:Catechin / Q000494:pharmacology*; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D003094:Collagen / Q000494:pharmacology*; D003432:Cross-Linking Reagents / Q000494:pharmacology*; D003782:Dental Pulp / Q000166:cytology* / Q000187:drug effects*; D048049:Extracellular Signal-Regulated MAP Kinases / Q000032:analysis; D015870:Gene Expression; D006801:Humans; D060888:Real-Time Polymerase Chain Reaction; D015203:Reproducibility of Results; D013997:Time Factors; D054457:Tissue Scaffolds / Q000737:chemistry*
chemical_list: D003432:Cross-Linking Reagents; D002392:Catechin; D003094:Collagen; D048049:Extracellular Signal-Regulated MAP Kinases; D000469:Alkaline Phosphatase