Cytotoxicity and bioactivity of various pulpotomy materials on stem cells from human exfoliated primary teeth

<jats:title>Abstract</jats:title><jats:sec><jats:title>Aims</jats:title><jats:p>To investigate the cytotoxicity and bioactivity of several pulpotomy materials: Biodentine (Septodont, Saint‐Maur‐des‐Fosses, France) <jats:styled-content style="fixed-case">MTA</jats:styled-content> (Angelus, Londrina, <jats:styled-content style="fixed-case">PR</jats:styled-content>, Brazil), Theracal <jats:styled-content style="fixed-case">LC</jats:styled-content> (Bisco Inc., Schamburg, <jats:styled-content style="fixed-case">IL</jats:styled-content>,<jats:styled-content style="fixed-case"> USA</jats:styled-content>) and <jats:styled-content style="fixed-case">IRM</jats:styled-content> (Dentsply DeTrey GmbH, Konstanz, Germany), after contact with stem cells isolated from human exfoliated primary teeth (<jats:styled-content style="fixed-case">SHED</jats:styled-content>s).</jats:p></jats:sec><jats:sec><jats:title>Methodology</jats:title><jats:p><jats:styled-content style="fixed-case">SHED</jats:styled-content>s were cultured in the presence of the eluates of various pulpotomy materials for 24, 48 and 72 h. Cell viability was determined by mitochondrial dehydrogenase enzymatic (<jats:styled-content style="fixed-case">MTT</jats:styled-content>) assay. Apoptosis and changes in cell phenotype were evaluated by flow cytometry. Also, an <jats:italic>in vitro</jats:italic> scratch wound‐healing assay was used to determine their effects on cell migration. To assess cell morphology and attachment to the different pulpotomy materials, <jats:styled-content style="fixed-case">SHED</jats:styled-content>s were directly seeded onto the material surfaces and analysed by scanning electron microscopy (<jats:styled-content style="fixed-case">SEM</jats:styled-content>). Finally, the deposition of a calcified matrix in presence of these materials was verified by Alizarin Red staining. Statistical analysis was performed with analysis of variance and Bonferroni or Tukey post‐test (α = 0.05).</jats:p></jats:sec><jats:sec><jats:title>Results</jats:title><jats:p>Cell viability in the presence of Biodentine eluates was significantly higher to that obtained using complete medium alone (control; <jats:italic>P</jats:italic> < 0.01) and was also significantly higher than using <jats:styled-content style="fixed-case">MTA</jats:styled-content> Angelus from 48 h of incubation (<jats:italic>P</jats:italic> < 0.01). However, Theracal <jats:styled-content style="fixed-case">LC</jats:styled-content> and <jats:styled-content style="fixed-case">IRM</jats:styled-content> were associated with low rates of cell viability (<jats:italic>P</jats:italic> < 0.001). Similar results were obtained in an apoptosis assay. In addition, <jats:styled-content style="fixed-case">SHED</jats:styled-content>s maintained their mesenchymal phenotype in all conditions although their capacity to migrate was higher in the presence of Biodentine. <jats:styled-content style="fixed-case">SEM</jats:styled-content> studies revealed a suitable proliferation rate, cell spreading and attachment, especially when using Biodentine and <jats:styled-content style="fixed-case">MTA</jats:styled-content> Angelus discs. Finally, Biodentine eluates significantly induced calcified matrix deposition from 7 days of culture (<jats:italic>P</jats:italic> < 0.01).</jats:p></jats:sec><jats:sec><jats:title>Conclusions</jats:title><jats:p>Biodentine exhibited better cytocompatibility and bioactivity than <jats:styled-content style="fixed-case">MTA</jats:styled-content> Angelus, Theracal <jats:styled-content style="fixed-case">LC</jats:styled-content> and <jats:styled-content style="fixed-case">IRM</jats:styled-content>.</jats:p></jats:sec>