Three-dimensional culture of dental pulp stem cells in direct contact to tricalcium silicate cements

Clin Oral Investig. 2016 Mar;20(2):237-46. doi: 10.1007/s00784-015-1515-3. Epub 2015 Jul 1.

Abstract

Objectives: Calcium silicate cements are biocompatible dental materials applicable in contact with vital tissue. The novel tricalcium silicate cement Biodentine™ offers properties superior to commonly used mineral trioxide aggregate (MTA). Objective of this study was to evaluate its cytocompatibility and ability to induce differentiation and mineralization in three-dimensional cultures of dental pulp stem cells after direct contact with the material.

Materials and methods: Test materials included a new tricalcium silicate (Biodentine™, Septodont, Saint-Maur-des-Fossés, France), MTA (ProRoot® MTA, DENSPLY Tulsa Dental Specialities, Johnson City, TN, USA), glass ionomer (Ketac™ Molar Aplicap™, 3M ESPE, Seefeld, Germany), human dentin disks and polystyrene. Magnetic activated cell sorting for to the surface antigen STRO-1 was performed to gain a fraction enriched with mesenchymal stem cells. Samples were allowed to set and dental pulp stem cells in collagen carriers were placed on top. Scanning electron microscopy of tricalcium silicate cement surfaces with and without cells was conducted. Cell viability was measured for 14 days by MTT assay. Alkaline phosphatase activity was evaluated (days 3, 7, and 14) and expression of mineralization-associated genes (COL1A1, ALP, DSPP, and RUNX2) was quantified by real-time quantitative PCR. Nonparametric statistical analysis for cell viability and alkaline phosphatase data was performed to compare different materials as well as time points (Mann-Whitney U test, α = 0.05).

Results: Cell viability was highest on tricalcium silicate cement, followed by MTA. Viability on glass ionomer cement and dentin disks was significantly lower. Alkaline phosphatase activity was lower in cells on new tricalcium silicate cement compared to MTA, whereas expression patterns of marker genes were alike.

Conclusions: Increased cell viability and similar levels of mineralization-associated gene expression in three-dimensional cell cultures on the novel tricalcium silicate cement and mineral trioxide aggregate indicate that the material is cytocompatible and bioactive.

Clinical relevance: The tested new tricalcium silicate cement confirms its suitability as an alternative to MTA in vital pulp therapy.

Keywords: Collagen type I; Dental pulp; Mineral trioxide aggregate; Multipotent stem cells; Three-dimensional cell culture; Tricalcium silicate.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alkaline Phosphatase / metabolism
  • Biocompatible Materials / pharmacology
  • Calcium Compounds / pharmacology*
  • Cell Differentiation
  • Cell Survival
  • Cells, Cultured
  • Dental Cements / pharmacology*
  • Dental Pulp / cytology*
  • Dentin / drug effects*
  • Glass Ionomer Cements / pharmacology
  • Humans
  • Materials Testing
  • Microscopy, Electron, Scanning
  • Polystyrenes
  • Real-Time Polymerase Chain Reaction
  • Silicates / pharmacology*
  • Stem Cells / drug effects*

Substances

  • Biocompatible Materials
  • Calcium Compounds
  • Dental Cements
  • Glass Ionomer Cements
  • Polystyrenes
  • Silicates
  • tricalcium silicate
  • Alkaline Phosphatase