title: Role of the extracellular signal-regulated kinase 1/2 pathway in driving tricalcium silicate-induced proliferation and biomineralization of human dental pulp cells in vitro.
abstract: BACKGROUND
  The aim of this study was to investigate the role of the extracellular signal-regulated kinase 1/2 (ERK1/2) pathway in regulating tricalcium silicate (C3S)-driven proliferation and biomineralization of human dental pulp cells (hDPCs) in vitro.
  
  
  METHODS
  Human DPCs were cultured in C3S-containing medium and compared with untreated controls. Cell viability was measured by the methyl-thiazol-tetrazolium assay. Biomineralization was assessed by staining calcium deposits on the extracellular matrix with von Kossa and alizarin red S stains. Phosphorylated ERK1/2 was evaluated by immunoblotting. The ERK1/2 inhibitor U0126 was used to assess the role of this pathway on stage of the cell cycle and mineralization-dependent gene expressions of hDPCs by using flow cytometry and real-time polymerase chain reaction, respectively. Data were analyzed by analysis of variance followed by the Student-Newman-Keuls post hoc test, with significance set at P < .05.
  
  
  RESULTS
  The viability and biomineralization of hDPCs were promoted by C3S extracts (P < .05). Phosphorylated ERK1/2 strongly appeared after hDPCs were cultured in the C3S extracts for 30 minutes. Moreover, inhibition of the ERK1/2 pathway in C3S-treated hDPCs decreased proliferation and the expression of mineralization-dependent genes, including collagen type I, dentin sialophosphoprotein, osteopontin, and osteocalcin (P < .05).
  
  
  CONCLUSIONS
  C3S stimulated the proliferation and biomineralization of hDPCs in vitro, with the ERK1/2 pathway playing a key role in the regulation of these effects.
authors.affiliation: Department of General Dentistry, Shanghai 9th People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai Key Laboratory of Stomatology, Shanghai, People's Republic of China.
authors.forename: Rong
authors.initials: R
authors.lastname: Du
authors.forename: Tiantian
authors.initials: T
authors.lastname: Wu
authors.forename: Wenjuan
authors.initials: W
authors.lastname: Liu
authors.forename: Lifen
authors.initials: L
authors.lastname: Li
authors.forename: Long
authors.initials: L
authors.lastname: Jiang
authors.forename: Weiwei
authors.initials: W
authors.lastname: Peng
authors.forename: Jiang
authors.initials: J
authors.lastname: Chang
authors.forename: Yaqin
authors.initials: Y
authors.lastname: Zhu
mesh_terms: D000293:Adolescent; D002070:Butadienes / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism; D017610:Calcium Compounds / Q000494:pharmacology*; D018929:Cell Culture Techniques; D049109:Cell Proliferation; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D024042:Collagen Type I / Q000187:drug effects; D003782:Dental Pulp / Q000166:cytology / Q000187:drug effects* / Q000201:enzymology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005109:Extracellular Matrix / Q000378:metabolism; D016326:Extracellular Matrix Proteins / Q000187:drug effects; D048049:Extracellular Signal-Regulated MAP Kinases / Q000037:antagonists & inhibitors; D006801:Humans; D020935:MAP Kinase Signaling System / Q000187:drug effects / Q000502:physiology*; D009570:Nitriles / Q000494:pharmacology; D015675:Osteocalcin / Q000187:drug effects; D053495:Osteopontin / Q000187:drug effects; D010750:Phosphoproteins / Q000187:drug effects; D057872:Pulp Capping and Pulpectomy Agents / Q000494:pharmacology*; D012795:Sialoglycoproteins / Q000187:drug effects; D017640:Silicates / Q000494:pharmacology*; D014074:Tooth Calcification / Q000187:drug effects* / Q000502:physiology; D055815:Young Adult; D000099311:Dentin Sialophosphoprotein
chemical_list: D002070:Butadienes; D002118:Calcium; D017610:Calcium Compounds; D024042:Collagen Type I; D004791:Enzyme Inhibitors; D016326:Extracellular Matrix Proteins; D048049:Extracellular Signal-Regulated MAP Kinases; D009570:Nitriles; D015675:Osteocalcin; D053495:Osteopontin; D010750:Phosphoproteins; D057872:Pulp Capping and Pulpectomy Agents; D012795:Sialoglycoproteins; D017640:Silicates; D000099311:Dentin Sialophosphoprotein; C113580:U 0126; C506393:tricalcium silicate
keywords: Biomineralization; extracellular signal-regulated kinase 1/2 pathway; human dental pulp cells; proliferation; tricalcium silicate