2.6. Cell culture
Donor DPSCs (passage 6-7) were cultured in Alpha modification of Eagle’s medium (α-MEM) supplemented with 10% FBS, 1% penicillin (100 U/mL)/streptomycin (100 µg/mL), 100 μM L-ascorbic acid 2-phosphate, and 2 mM L-glutamine at 37 °C in 5% CO2 humidified atmosphere. Cells reaching 80–90% confluency were harvested using 0.05% (w/v) trypsin/ethylenediaminetetraacetic acid (EDTA) solution and seeded onto scaffolds in 24-well culture plates (100 µL/well) at 1, 2, 4, 8, or 16×104 cells/sample. The scaffolds were incubated for 3 hours at 37 °C with 5% CO2 to allow diffusion of cells through the network of pores before adding culture media (550 µL/well). DPSCs cultured on the surface of the well plate (i.e. No scaffolds) were used as 2D control.