title: An assessment of the overexpression of BMP-2 in transfected human osteoblast cells stimulated by mineral trioxide aggregate and Biodentine.
abstract: OBJECTIVE
  To evaluate the effect of MTA and Biodentine on viability, osteogenic differentiation and BMP-2 expression in osteogenic cells.
  
  
  METHODS
  Saos-2 cells were used as a model of osteoblastic cells. Overexpression of BMP-2 was induced by transfection of a CMV-driven plasmid construct including the human BMP-2 coding sequence, and stably transfected cells were selected. Cell viability was assessed by the mitochondrial dehydrogenase enzymatic (MTT) assay. The bioactivity of the materials was evaluated by the alkaline phosphatase (ALP) assay and detection of calcium deposits with alizarin red staining (ARS). The gene expression of BMP-2 and ALP was quantified with real-time PCR. Statistical analysis was performed with analysis of variance and Bonferroni or Tukey post-test (α = 0.05).
  
  
  RESULTS
  Viability tests revealed that MTA and Biodentine were not cytotoxic at the higher dilution (1 : 8) to BMP-2-transfected cells. MTA and Biodentine exhibited the highest ALP activity when compared to the Saos-BMP-2-unexposed control group (P < 0.05). Cell exposure to Biodentine and MTA had a significant stimulatory effect on the formation of mineralized nodules (P < 0.05). The highest increase in BMP-2 gene expression was observed after 3 days of BMP-2-transfected cells exposure to MTA and Biodentine in non-osteogenic medium in relation to Saos-BMP-2-unexposed control cells (P < 0.05). Exposure of cells to MTA in osteogenic medium for 1 day increased ALP gene expression by approximately 1.3-fold in relation to Saos-BMP-2-unexposed control cells (P < 0.05).
  
  
  CONCLUSIONS
  Both MTA and Biodentine showed biocompatibility and bioactivity in Saos-BMP-2 overexpressing cells. Biodentine had a significantly greater effect on mineralization than MTA. Both MTA and Biodentine enhanced BMP-2 mRNA expression in the transfected system. Both MTA and Biodentine are suitable materials to improve osteoblastic cell mineralization.
authors.affiliation: Department of Restorative Dentistry, Dental School of São Paulo State University, Araraquara, São Paulo.
authors.forename: E M
authors.initials: EM
authors.lastname: Rodrigues
authors.affiliation: Department of Restorative Dentistry, Dental School of São Paulo State University, Araraquara, São Paulo.
authors.forename: A L
authors.initials: AL
authors.lastname: Gomes-Cornélio
authors.affiliation: Department of Genetic and Evolution, Laboratory of Plant Biotechnology, Federal University of São Carlos, São Carlos, São Paulo.
authors.forename: A
authors.initials: A
authors.lastname: Soares-Costa
authors.affiliation: Cellular Biology Department, Institute of Biological Sciences, University of Brasília, Brasília, Distrito Federal.
authors.forename: L P
authors.initials: LP
authors.lastname: Salles
authors.affiliation: Department of Diagnosis and Surgery, Dental School of São Paulo State University, Araraquara, São Paulo, Brazil.
authors.forename: M
authors.initials: M
authors.lastname: Velayutham
authors.affiliation: Department of Diagnosis and Surgery, Dental School of São Paulo State University, Araraquara, São Paulo, Brazil.
authors.forename: C
authors.initials: C
authors.lastname: Rossa-Junior
authors.affiliation: Department of Restorative Dentistry, Dental School of São Paulo State University, Araraquara, São Paulo.
authors.forename: J M
authors.initials: JM
authors.lastname: Guerreiro-Tanomaru
authors.affiliation: Department of Restorative Dentistry, Dental School of São Paulo State University, Araraquara, São Paulo.
authors.forename: M
authors.initials: M
authors.lastname: Tanomaru-Filho
mesh_terms: D000469:Alkaline Phosphatase / Q000378:metabolism; D017607:Aluminum Compounds / Q000494:pharmacology*; D015153:Blotting, Western; D055396:Bone Morphogenetic Protein 2 / Q000378:metabolism*; D017610:Calcium Compounds / Q000494:pharmacology*; D002454:Cell Differentiation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D004338:Drug Combinations; D006801:Humans; D008422:Materials Testing; D010006:Osteoblasts / Q000378:metabolism*; D010087:Oxides / Q000494:pharmacology*; D060888:Real-Time Polymerase Chain Reaction; D017640:Silicates / Q000494:pharmacology*; D014162:Transfection
chemical_list: D017607:Aluminum Compounds; C525718:BMP2 protein, human; D055396:Bone Morphogenetic Protein 2; D017610:Calcium Compounds; D004338:Drug Combinations; D010087:Oxides; D017640:Silicates; C086631:mineral trioxide aggregate; C506393:tricalcium silicate; D000469:Alkaline Phosphatase
keywords: 
  MTA
  ; Biodentine; bioactivity; bone morphogenetic protein-2; cytotoxicity