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An assessment of the overexpression of BMP‐2 in transfected human osteoblast cells stimulated by mineral trioxide aggregate and Biodentine
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...
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| Published in: | International endodontic journal 2017-12, Vol.50 (S2), p.e9-e18 |
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| container_end_page | e18 |
| container_issue | S2 |
| container_start_page | e9 |
| container_title | International endodontic journal |
| container_volume | 50 |
| creator | Rodrigues, E. M. Gomes‐Cornélio, A. L. Soares‐Costa, A. Salles, L. P. Velayutham, M. Rossa‐Junior, C. Guerreiro‐Tanomaru, J. M. Tanomaru‐Filho, M. |
| description | 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 |
| doi_str_mv | 10.1111/iej.12745 |
| format | article |
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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.</description><identifier>ISSN: 0143-2885</identifier><identifier>EISSN: 1365-2591</identifier><identifier>DOI: 10.1111/iej.12745</identifier><identifier>PMID: 28109163</identifier><language>eng</language><publisher>England: Wiley Subscription Services, Inc</publisher><subject>Alkaline phosphatase ; Alkaline Phosphatase - metabolism ; Aluminum Compounds - pharmacology ; bioactivity ; Biocompatibility ; Biodentine ; Biological activity ; Blotting, Western ; Bone Morphogenetic Protein 2 - metabolism ; bone morphogenetic protein‐2 ; Calcium ; Calcium Compounds - pharmacology ; Cell Differentiation - drug effects ; Cell Survival - drug effects ; Cells, Cultured ; Cytotoxicity ; Drug Combinations ; Endodontics ; Gene expression ; Humans ; Materials Testing ; Mineralization ; Mitochondria ; MTA ; Nodules ; Osteoblasts ; Osteoblasts - metabolism ; Oxides - pharmacology ; Real-Time Polymerase Chain Reaction ; Silicates - pharmacology ; Statistical analysis ; Transfection</subject><ispartof>International endodontic journal, 2017-12, Vol.50 (S2), p.e9-e18</ispartof><rights>2017 International Endodontic Journal. Published by John Wiley & Sons Ltd</rights><rights>2017 International Endodontic Journal. Published by John Wiley & Sons Ltd.</rights><rights>Copyright © 2017 International Endodontic Journal. Published by John Wiley & Sons Ltd</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c3555-9c95b654bd352be8641d7adb202531226855b37c7a9d4eac392b75360e492ee13</citedby><cites>FETCH-LOGICAL-c3555-9c95b654bd352be8641d7adb202531226855b37c7a9d4eac392b75360e492ee13</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27924,27925</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/28109163$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Rodrigues, E. M.</creatorcontrib><creatorcontrib>Gomes‐Cornélio, A. L.</creatorcontrib><creatorcontrib>Soares‐Costa, A.</creatorcontrib><creatorcontrib>Salles, L. P.</creatorcontrib><creatorcontrib>Velayutham, M.</creatorcontrib><creatorcontrib>Rossa‐Junior, C.</creatorcontrib><creatorcontrib>Guerreiro‐Tanomaru, J. M.</creatorcontrib><creatorcontrib>Tanomaru‐Filho, M.</creatorcontrib><title>An assessment of the overexpression of BMP‐2 in transfected human osteoblast cells stimulated by mineral trioxide aggregate and Biodentine</title><title>International endodontic journal</title><addtitle>Int Endod J</addtitle><description>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.</description><subject>Alkaline phosphatase</subject><subject>Alkaline Phosphatase - metabolism</subject><subject>Aluminum Compounds - pharmacology</subject><subject>bioactivity</subject><subject>Biocompatibility</subject><subject>Biodentine</subject><subject>Biological activity</subject><subject>Blotting, Western</subject><subject>Bone Morphogenetic Protein 2 - metabolism</subject><subject>bone morphogenetic protein‐2</subject><subject>Calcium</subject><subject>Calcium Compounds - pharmacology</subject><subject>Cell Differentiation - drug effects</subject><subject>Cell Survival - drug effects</subject><subject>Cells, Cultured</subject><subject>Cytotoxicity</subject><subject>Drug Combinations</subject><subject>Endodontics</subject><subject>Gene expression</subject><subject>Humans</subject><subject>Materials Testing</subject><subject>Mineralization</subject><subject>Mitochondria</subject><subject>MTA</subject><subject>Nodules</subject><subject>Osteoblasts</subject><subject>Osteoblasts - metabolism</subject><subject>Oxides - pharmacology</subject><subject>Real-Time Polymerase Chain Reaction</subject><subject>Silicates - pharmacology</subject><subject>Statistical analysis</subject><subject>Transfection</subject><issn>0143-2885</issn><issn>1365-2591</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2017</creationdate><recordtype>article</recordtype><recordid>eNp1kL1OwzAUhS0EglIYeAFkiYkh4J84iUeo-FURDDBHdnxbXCVxsROgGw_AwDPyJLgU2LjLle757jnSQWiPkiMa59jC7IiyPBVraEB5JhImJF1HA0JTnrCiEFtoO4QZIUQQTjfRFisokTTjA_R-0mIVAoTQQNthN8HdI2D3DB5e5z6erWuX19Obu8-3D4Ztizuv2jCBqgODH_tGRT104HStQocrqOuAQ2ebvlZLQi9wY1vwqo6P1r1aA1hNpx6mUcaqNfjUOhOzI7SDNiaqDrD7s4fo4fzsfnSZjG8vrkYn46TiQohEVlLoTKTacME0FFlKTa6MZoQJThnLCiE0z6tcSZOCqrhkOhc8I5BKBkD5EB2sfOfePfUQunLmet_GyJLKLMsLSXMWqcMVVXkXgodJOfe2UX5RUlIuey9j7-V375Hd_3HsdQPmj_wtOgLHK-DF1rD436m8OrteWX4B3emOrg</recordid><startdate>201712</startdate><enddate>201712</enddate><creator>Rodrigues, E. M.</creator><creator>Gomes‐Cornélio, A. L.</creator><creator>Soares‐Costa, A.</creator><creator>Salles, L. P.</creator><creator>Velayutham, M.</creator><creator>Rossa‐Junior, C.</creator><creator>Guerreiro‐Tanomaru, J. M.</creator><creator>Tanomaru‐Filho, M.</creator><general>Wiley Subscription Services, Inc</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QP</scope><scope>K9.</scope></search><sort><creationdate>201712</creationdate><title>An assessment of the overexpression of BMP‐2 in transfected human osteoblast cells stimulated by mineral trioxide aggregate and Biodentine</title><author>Rodrigues, E. M. ; Gomes‐Cornélio, A. L. ; Soares‐Costa, A. ; Salles, L. P. ; Velayutham, M. ; Rossa‐Junior, C. ; Guerreiro‐Tanomaru, J. M. ; Tanomaru‐Filho, M.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c3555-9c95b654bd352be8641d7adb202531226855b37c7a9d4eac392b75360e492ee13</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2017</creationdate><topic>Alkaline phosphatase</topic><topic>Alkaline Phosphatase - metabolism</topic><topic>Aluminum Compounds - pharmacology</topic><topic>bioactivity</topic><topic>Biocompatibility</topic><topic>Biodentine</topic><topic>Biological activity</topic><topic>Blotting, Western</topic><topic>Bone Morphogenetic Protein 2 - metabolism</topic><topic>bone morphogenetic protein‐2</topic><topic>Calcium</topic><topic>Calcium Compounds - pharmacology</topic><topic>Cell Differentiation - drug effects</topic><topic>Cell Survival - drug effects</topic><topic>Cells, Cultured</topic><topic>Cytotoxicity</topic><topic>Drug Combinations</topic><topic>Endodontics</topic><topic>Gene expression</topic><topic>Humans</topic><topic>Materials Testing</topic><topic>Mineralization</topic><topic>Mitochondria</topic><topic>MTA</topic><topic>Nodules</topic><topic>Osteoblasts</topic><topic>Osteoblasts - metabolism</topic><topic>Oxides - pharmacology</topic><topic>Real-Time Polymerase Chain Reaction</topic><topic>Silicates - pharmacology</topic><topic>Statistical analysis</topic><topic>Transfection</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Rodrigues, E. M.</creatorcontrib><creatorcontrib>Gomes‐Cornélio, A. L.</creatorcontrib><creatorcontrib>Soares‐Costa, A.</creatorcontrib><creatorcontrib>Salles, L. P.</creatorcontrib><creatorcontrib>Velayutham, M.</creatorcontrib><creatorcontrib>Rossa‐Junior, C.</creatorcontrib><creatorcontrib>Guerreiro‐Tanomaru, J. M.</creatorcontrib><creatorcontrib>Tanomaru‐Filho, M.</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Calcium & Calcified Tissue Abstracts</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><jtitle>International endodontic journal</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Rodrigues, E. M.</au><au>Gomes‐Cornélio, A. L.</au><au>Soares‐Costa, A.</au><au>Salles, L. P.</au><au>Velayutham, M.</au><au>Rossa‐Junior, C.</au><au>Guerreiro‐Tanomaru, J. M.</au><au>Tanomaru‐Filho, M.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>An assessment of the overexpression of BMP‐2 in transfected human osteoblast cells stimulated by mineral trioxide aggregate and Biodentine</atitle><jtitle>International endodontic journal</jtitle><addtitle>Int Endod J</addtitle><date>2017-12</date><risdate>2017</risdate><volume>50</volume><issue>S2</issue><spage>e9</spage><epage>e18</epage><pages>e9-e18</pages><issn>0143-2885</issn><eissn>1365-2591</eissn><abstract>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.</abstract><cop>England</cop><pub>Wiley Subscription Services, Inc</pub><pmid>28109163</pmid><doi>10.1111/iej.12745</doi><tpages>10</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | Alkaline phosphatase Alkaline Phosphatase - metabolism Aluminum Compounds - pharmacology bioactivity Biocompatibility Biodentine Biological activity Blotting, Western Bone Morphogenetic Protein 2 - metabolism bone morphogenetic protein‐2 Calcium Calcium Compounds - pharmacology Cell Differentiation - drug effects Cell Survival - drug effects Cells, Cultured Cytotoxicity Drug Combinations Endodontics Gene expression Humans Materials Testing Mineralization Mitochondria MTA Nodules Osteoblasts Osteoblasts - metabolism Oxides - pharmacology Real-Time Polymerase Chain Reaction Silicates - pharmacology Statistical analysis Transfection |
| title | An assessment of the overexpression of BMP‐2 in transfected human osteoblast cells stimulated by mineral trioxide aggregate and Biodentine |
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