title: Gene expression and cytokine release during odontogenic differentiation of human dental pulp stem cells induced by 2 endodontic biomaterials.
abstract: BACKGROUND
  Mineral trioxide aggregate (MTA) and calcium-enriched mixture (CEM) have shown osteogenic/cementogenic/dentinogenic activities; however, their mechanism of action is not fully understood. We aimed to evaluate the effect of these biomaterials on odontogenic differentiation of human dental pulp stem cells (DPSCs).
  
  
  METHODS
  Flow cytometry with stem cell markers for the confirmation of stemness and homogeneity was first performed. Then isolated DPSCs were seeded on prepared discs of MTA, CEM, differentiation medium (DM), and growth medium (GM) and incubated up to 14 days. Concentrations of transforming growth factor-β1, bone morphogenetic protein (BMP)2, BMP4, and fibroblast growth factor 4 were measured at each interval using an enzyme-linked immunosorbent assay reader. Gene expression of dentin sialophosphoprotein, dentin matrix protein 1, and the cytokines were evaluated by reverse-transcription polymerase chain reaction. To evaluate the cell morphology, scanning electron micrographs were taken; mineralization potential was evaluated using alizarin red S staining.
  
  
  RESULTS
  Scanning electron micrographs showed that DPSCs spread/adhered/proliferated similarly on MTA and CEM. On day 14, alizarin red S staining confirmed that mineralization occurred in all groups except GM. Expressions of dentin matrix protein 1 and dentin sialophosphoprotein genes were similar in the CEM, MTA, and DM groups; they were significantly higher compared with the GM group (P < .05). A greater amount of transforming growth factor-β1 gene was expressed in MTA compared with the other groups (P < .05). However, the expression of fibroblast growth factor 4 and BMP2 genes was significantly greater in the CEM group (P < .05). In all the tested groups, the expression of BMP4 was less than GM (P < .01); however, CEM and DM were similar but more than MTA (P < .05). Concentrations of protein product detected using an enzyme-linked immunosorbent assay reader confirmed these gene expressions.
  
  
  CONCLUSIONS
  MTA and CEM can induce osteo-/odontogenic-like phenotype differentiation of human DPSCs; however, they stimulate different gene expressions and growth factor release.
authors.affiliation: Iranian Center for Endodontic Research, Research Institute of Dental Sciences, Shahid Beheshti University of Medical Sciences, Tehran, Iran; Dental Research Center, Research Institute of Dental Sciences, Shahid Beheshti University of Medical Sciences, Tehran, Iran. Electronic address: saasgary@yahoo.com.
authors.forename: Saeed
authors.initials: S
authors.lastname: Asgary
authors.affiliation: Department of Biology and Anatomical Sciences, Faculty of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
authors.forename: Hamid
authors.initials: H
authors.lastname: Nazarian
authors.affiliation: Dental Research Center, Research Institute of Dental Sciences, Shahid Beheshti University of Medical Sciences, Tehran, Iran; Department of Oral and Maxillofacial Surgery, Dental School, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
authors.forename: Arash
authors.initials: A
authors.lastname: Khojasteh
authors.affiliation: Iranian Center for Endodontic Research, Research Institute of Dental Sciences, Shahid Beheshti University of Medical Sciences, Tehran, Iran; Endodontic Department, Dental School, Tehran University of Medical Sciences, Tehran, Iran.
authors.forename: Noushin
authors.initials: N
authors.lastname: Shokouhinejad
mesh_terms: D017607:Aluminum Compounds / Q000494:pharmacology; D001672:Biocompatible Materials / Q000494:pharmacology*; D055396:Bone Morphogenetic Protein 2 / Q000187:drug effects; D055415:Bone Morphogenetic Protein 4 / Q000187:drug effects; D002113:Calcification, Physiologic / Q000187:drug effects; D017610:Calcium Compounds / Q000494:pharmacology; D002448:Cell Adhesion / Q000187:drug effects; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D048430:Cell Shape / Q000187:drug effects; D002478:Cells, Cultured; D025021:Cementogenesis / Q000187:drug effects; D016207:Cytokines / Q000187:drug effects*; D003782:Dental Pulp / Q000166:cytology*; D003810:Dentinogenesis / Q000187:drug effects; D004338:Drug Combinations; D016326:Extracellular Matrix Proteins / Q000187:drug effects; D051519:Fibroblast Growth Factor 4 / Q000187:drug effects; D006801:Humans; D008855:Microscopy, Electron, Scanning; D009805:Odontogenesis / Q000187:drug effects*; D010087:Oxides / Q000494:pharmacology; D010750:Phosphoproteins / Q000187:drug effects; D012387:Root Canal Filling Materials / Q000494:pharmacology*; D012795:Sialoglycoproteins / Q000187:drug effects; D012823:Silicate Cement / Q000494:pharmacology; D017640:Silicates / Q000494:pharmacology; D013234:Stem Cells / Q000187:drug effects*; D053773:Transforming Growth Factor beta1 / Q000187:drug effects; D000099311:Dentin Sialophosphoprotein
chemical_list: D017607:Aluminum Compounds; D001672:Biocompatible Materials; D055396:Bone Morphogenetic Protein 2; D055415:Bone Morphogenetic Protein 4; D017610:Calcium Compounds; D016207:Cytokines; D004338:Drug Combinations; D016326:Extracellular Matrix Proteins; D051519:Fibroblast Growth Factor 4; D010087:Oxides; D010750:Phosphoproteins; D012387:Root Canal Filling Materials; D012795:Sialoglycoproteins; D012823:Silicate Cement; D017640:Silicates; D053773:Transforming Growth Factor beta1; D000099311:Dentin Sialophosphoprotein; C525718:BMP2 protein, human; C525809:BMP4 protein, human; C513309:DMP1 protein, human; C496445:FGF4 protein, human; C086631:mineral trioxide aggregate; C031293:calcium silicate
keywords: Biomaterials; CEM cement; calcium-enriched mixture; dental pulp capping; differentiation; human; mineral trioxide aggregate; pulp regeneration; stem cell