Biological interactions of a calcium silicate based cement (Biodentine™) with Stem Cells from Human Exfoliated Deciduous teeth.

Objective: To investigate the biological interactions of a calcium silicate based cement (Biodentine™) with Stem Cells from Human Exfoliated Deciduous teeth (SHED), focusing on viability/proliferation, odontogenic differentiation, biomineralization and elemental release/exchange.

Methods: Biodentine™ specimens were used directly or for eluate preparation at serial dilutions (1:1-1:64). SHED cultures were established from deciduous teeth of healthy children. Viability/proliferation and morphological characteristics were evaluated by live/dead fluorescent staining, MTT assay and Scanning Electron Microscopy. Odontogenic differentiation by qRT-PCR, biomineralization by Alizarin red S staining, while ion elution by Inductively Coupled Plasma-Optical Emission Spectrometry (ICP-OES).

Results: SHED effectively attached within the crystalline surface of Biodentine™ specimens acquiring a spindle-shaped phenotype. Statistically significant stimulation of cell proliferation was induced at day 3 by eluates in dilutions from 1:16 to 1:64. Differential, concentration- and time-dependent expression patterns of odontogenic genes were observed under non-inductive and inductive (osteogenic) conditions, with significant up-regulation of DSPP and Runx2 at higher dilutions and a peak in expression of BMP-2, BGLAP and MSX-2 at 1:8 dilution on day 7. Progressive increase in mineralized tissue formation was observed with increasing dilutions of Biodentine™ eluates. ICP-OES indicated that Biodentine™ absorbed Ca, Mg and P ions from culture medium, while releasing Si and Sr ions from its backbone.

Significance: Biodentine™ interacts through elemental release/uptake with the cellular microenvironment, triggering odontogenic differentiation and biomineralization in a concentration-dependent manner. These results reveal a promising strategy for application of the calcium silicate based cement (Biodentine™) for vital pulp therapies of deciduous teeth in Paediatric Dentistry.