title: Biodentine induces human dental pulp stem cell differentiation through mitogen-activated protein kinase and calcium-/calmodulin-dependent protein kinase II pathways.
abstract: BACKGROUND
  Biodentine (Septodont, Saint-Maur-des-Fossès, France), a new tricalcium silicate cement formulation, has been introduced as a bioactive dentine substitute to be used in direct contact with pulp tissue. The aim of this study was to investigate the response of human dental pulp stem cells (hDPSCs) to the material and whether mitogen-activated protein kinase (MAPK), nuclear factor-kappa B (NF-κB), and calcium-/calmodulin-dependent protein kinase II (CaMKII) signal pathways played a regulatory role in Biodentine-induced odontoblast differentiation.
  
  
  METHODS
  hDPCs obtained from impacted third molars were incubated with Biodentine. Odontoblastic differentiation was evaluated by alkaline phosphatase activity, alizarin red staining, and quantitative real-time reverse-transcriptase polymerase chain reaction for the analysis of messenger RNA expression of the following differentiation gene markers: osteocalcin (OCN), dentin sialophosprotein (DSPP), dentin matrix protein 1 (DMP1), and bone sialoprotein (BSP). Cell cultures in the presence of Biodentine were exposed to specific inhibitors of MAPK (U0126, SB203580, and SP600125), NF-κB (pyrrolidine dithiocarbamate), and CaMKII (KN-93) pathways to evaluate the regulatory effect on the expression of these markers and mineralization assay.
  
  
  RESULTS
  Biodentine significantly increased alkaline phosphatase activity and mineralized nodule formation and the expression of OCN, DSPP, DMP1, and BSP. The MAPK inhibitor for extracellular signal-regulated kinase 1/2 (U0126) and Jun N-terminal kinase (SP600125) significantly decreased the Biodentine-induced mineralized differentiation of hDPSCs and OCN, DSPP, DMP1, and BSP messenger RNA expression, whereas p38 MAPK inhibitors (SB203580) had no effect. The CaMKII inhibitor KN-93 significantly attenuated and the NF-κB inhibitor pyrrolidine dithiocarbamate further enhanced the up-regulation of Biodentine-induced gene expression and mineralization.
  
  
  CONCLUSIONS
  Biodentine is a bioactive and biocompatible material capable of inducing odontoblast differentiation of hDPSCs. Our results indicate that this induction is regulated via MAPK and CaMKII pathways.
authors.affiliation: Department of Operative Dentistry and Endodontics, Fourth Military Medical University, Xian, China.
authors.forename: Zhirong
authors.initials: Z
authors.lastname: Luo
authors.affiliation: Department of Endodontics, University of Pennsylvania, Philadelphia, Pennsylvania.
authors.forename: Meetu R
authors.initials: MR
authors.lastname: Kohli
authors.affiliation: Department of Operative Dentistry and Endodontics, Fourth Military Medical University, Xian, China.
authors.forename: Qing
authors.initials: Q
authors.lastname: Yu
authors.affiliation: Department of Endodontics, University of Pennsylvania, Philadelphia, Pennsylvania.
authors.forename: Syngcuk
authors.initials: S
authors.lastname: Kim
authors.affiliation: Department of Operative Dentistry and Endodontics, Fourth Military Medical University, Xian, China.
authors.forename: Tiejun
authors.initials: T
authors.lastname: Qu
authors.affiliation: Department of Operative Dentistry and Endodontics, Fourth Military Medical University, Xian, China. Electronic address: hewenxi@fmmu.edu.cn.
authors.forename: Wen-xi
authors.initials: WX
authors.lastname: He
mesh_terms: D000293:Adolescent; D000328:Adult; D000469:Alkaline Phosphatase / Q000032:analysis; D000873:Anthracenes / Q000494:pharmacology; D001596:Benzylamines / Q000494:pharmacology; D002070:Butadienes / Q000494:pharmacology; D017610:Calcium Compounds / Q000494:pharmacology*; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2 / Q000037:antagonists & inhibitors / Q000187:drug effects*; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000187:drug effects; D003782:Dental Pulp / Q000166:cytology* / Q000187:drug effects; D016326:Extracellular Matrix Proteins / Q000032:analysis; D006801:Humans; D007093:Imidazoles / Q000494:pharmacology; D058574:Integrin-Binding Sialoprotein / Q000032:analysis; D020935:MAP Kinase Signaling System / Q000187:drug effects*; D016328:NF-kappa B / Q000037:antagonists & inhibitors / Q000187:drug effects; D009570:Nitriles / Q000494:pharmacology; D009804:Odontoblasts / Q000187:drug effects; D015675:Osteocalcin / Q000032:analysis; D010750:Phosphoproteins / Q000032:analysis; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D057872:Pulp Capping and Pulpectomy Agents / Q000494:pharmacology*; D011725:Pyridines / Q000494:pharmacology; D011759:Pyrrolidines / Q000494:pharmacology; D012795:Sialoglycoproteins / Q000032:analysis; D015398:Signal Transduction / Q000187:drug effects*; D017640:Silicates / Q000494:pharmacology*; D013234:Stem Cells / Q000187:drug effects*; D013449:Sulfonamides / Q000494:pharmacology; D013859:Thiocarbamates / Q000494:pharmacology; D055815:Young Adult; D000099311:Dentin Sialophosphoprotein; D000096926:Benzenesulfonamides
chemical_list: D000469:Alkaline Phosphatase; D000873:Anthracenes; D001596:Benzylamines; D002070:Butadienes; D017610:Calcium Compounds; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2; D016326:Extracellular Matrix Proteins; D007093:Imidazoles; D058574:Integrin-Binding Sialoprotein; D016328:NF-kappa B; D009570:Nitriles; D015675:Osteocalcin; D010750:Phosphoproteins; D047428:Protein Kinase Inhibitors; D057872:Pulp Capping and Pulpectomy Agents; D011725:Pyridines; D011759:Pyrrolidines; D012795:Sialoglycoproteins; D017640:Silicates; D013449:Sulfonamides; D013859:Thiocarbamates; D000099311:Dentin Sialophosphoprotein; C513309:DMP1 protein, human; C113580:U 0126; C072105:KN 93; D000096926:Benzenesulfonamides; C432165:pyrazolanthrone; C020972:pyrrolidine dithiocarbamic acid; C506393:tricalcium silicate; C093642:SB 203580
keywords: Biodentine; biosilicate cement; calcium-/calmodulin-dependent protein kinase II; human dental pulp stem cells; mitogen-activated protein kinase pathway; nuclear factor-kappa B pathway; pathway