"Biological responses of two calcium-silicate-based cements on a tissue-engineered 3D organotypic deciduous pulp analogue"

Dent Mater. 2024 May;40(5):e14-e25. doi: 10.1016/j.dental.2024.02.024. Epub 2024 Mar 1.

Abstract

Objectives: The biological responses of MTA and Biodentine™ has been assessed on a three-dimensional, tissue-engineered organotypic deciduous pulp analogue.

Methods: Human endothelial (HUVEC) and dental mesenchymal stem cells (SHED) at a ratio of 3:1, were incorporated into a collagen I/fibrin hydrogel; succeeding Biodentine™ and MTA cylindrical specimens were placed in direct contact with the pulp analogue 48 h later. Cell viability/proliferation and morphology were evaluated through live/dead staining, MTT assay and Scanning Electron Microscopy (SEM), and expression of angiogenic, odontogenic markers through real time PCR.

Results: Viable cells dominated at day 3 after treatment presenting typical morphology, firmly attached within the hydrogel structures, as shown by live/dead staining and SEM images. MTT assay at day 1 presented a significant increase of cell proliferation in Biodentine™ group. Real-time PCR showed significant upregulation of odontogenic markers DSPP, BMP-2 (day 3,6), RUNX2, ALP (day 3) in contact with Biodentine™ compared to MTA and the control, whereas MTA promoted significant upregulation of DSPP, BMP-2, RUNX2, Osterix (day 3) and ALP (day 6) compared to the control. MSX1 presented downregulation in both experimental groups. Expression of angiogenic markers VEGFa and ANGPT-1 at day 3 was significantly upregulated in contact with Biodentine™ and MTA respectively, while the receptors VEGFR1, VEGFR2 and Tie-2, as well as PECAM-1 were downregulated.

Significance: Both calcium silicate-based materials are biocompatible and exert positive angiogenic and odontogenic effects, although Biodentine™ during the first days of culture, seems to induce higher cell proliferation and provoke a more profound odontogenic and angiogenic response from SHED.

Keywords: Angiogenic differentiation; Deciduous teeth; Dental pulp analogue; Odontogenic differentiation; Silicate-based cements; Tissue-engineering; Vital pulp therapy.

MeSH terms

  • Aluminum Compounds / chemistry
  • Aluminum Compounds / pharmacology
  • Calcium Compounds* / chemistry
  • Calcium Compounds* / pharmacology
  • Cell Proliferation* / drug effects
  • Cell Survival / drug effects
  • Cells, Cultured
  • Dental Cements / pharmacology
  • Dental Pulp* / cytology
  • Dental Pulp* / drug effects
  • Drug Combinations*
  • Human Umbilical Vein Endothelial Cells / drug effects
  • Humans
  • Mesenchymal Stem Cells / drug effects
  • Microscopy, Electron, Scanning
  • Oxides / chemistry
  • Oxides / pharmacology
  • Real-Time Polymerase Chain Reaction
  • Silicates* / chemistry
  • Silicates* / pharmacology
  • Tissue Engineering* / methods
  • Tooth, Deciduous / cytology

Substances

  • mineral trioxide aggregate
  • tricalcium silicate
  • calcium silicate