Biocompatibility assessment of resin-based cements on vascularized dentin/pulp tissue-engineered analogues

• Hadjichristou Christina
• Papachristou Eleni
• Vereroudakis Emmanouil
• Chatzinikolaidou Maria
• About Imad
• Koidis Petros
• Bakopoulou Athina

Objectives. A three-dimensional (3D) dentin/pulp tissue analogue, resembling the human natural tissue has been engineered in an in vitro setup, aiming to assess the cytocompatibility of resin-based dental restorative cements. Methods. Stem Cells from Apical Papilla (SCAP) and Human Umbilical Vein Endothelial Cells (HUVEC) were embedded in Collagen-I/Fibrin hydrogels at 1:3 ratio within 24-well plates. Hanging culture inserts were placed over the hydrogels, housing an odontoblast-like cell layer and a human treated-dentin barrier. Shear modulus of the hydrogels at 3.5 and 5 mg/ml was evaluated by dynamic mechanical analysis. Eluates of two resin-based cements, a dual-cure-(Breeze TM , Pentron: Cement-1/C1), and a self-adhesive cement (SpeedCEMplus TM , Ivoclar-Vivadent: Cement-2/C2) were applied into the dentin/pulp tissue analogue after prestimulation with LPS. Cytocompatibility was assessed by MTT assay, live/dead staining and real-time PCR analysis. Results. Both hydrogel concentrations showed similar shear moduli to the natural pulp until day (D) 7, while the 5 mg/ml-hydrogel substantially increased stiffness by D14. Both cements caused no significant toxicity to the dentin/pulp tissue analogue. C1 induced stimulation (p < 0.01) of cell viability (158 ± 3%, 72 h), while pre-stimulation with LPS attenuated this effect. C2 (±LPS) caused minor reduction of viability (15-20%, 24 h) that recovered at 72 h for the LPS+ group. Both cements caused upregulation of VEGF, ANGP-1, and downregulation of the respective receptors VEGFR-2 and Tie-1.