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An assessment of the overexpression of BMP ‐2 in transfected human osteoblast cells stimulated by mineral trioxide aggregate and Biodentine
Author(s) -
Rodrigues E. M.,
GomesCornélio A. L.,
SoaresCosta A.,
Salles L. P.,
Velayutham M.,
RossaJunior C.,
GuerreiroTanomaru J. M.,
TanomaruFilho M.
Publication year - 2017
Publication title -
international endodontic journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.988
H-Index - 119
eISSN - 1365-2591
pISSN - 0143-2885
DOI - 10.1111/iej.12745
Subject(s) - viability assay , transfection , alkaline phosphatase , mineral trioxide aggregate , osteoblast , microbiology and biotechnology , mtt assay , bone morphogenetic protein 2 , chemistry , andrology , cell growth , biology , cell , in vitro , medicine , biochemistry , gene , dentistry , enzyme
Aim To evaluate the effect of MTA and Biodentine on viability, osteogenic differentiation and BMP ‐2 expression in osteogenic cells. Methodology 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.