Method: Human DPSC from extracted adult healthy teeth were used in this study. All cells were between 3rd and 5th passages. Hydrogel stiffness was varied by changing the volume ratio of HA:Gn dilutions. The PEGDA concentration remained constant. Human DPSC were embedded in hydrogels mixed in varying concentrations of thiolated HA and Gn, following PEGDA addition. The cell slurry was seeded in 24-well inserts and supplemented with 20% serum containing growth medium. The cells were analyzed for cell viability, adhesion and proliferation. In parallel, hDPSC grown on 2D PEGDA-HA-Gn films were examined.
Results: Our studies demonstrated that hDPSC embedded in 3D PEGDA HA-Gn scaffolds enhanced cell viability. Furthermore, after 14 days of culture cell proliferation significantly increased. Our investigations to address the optimal ratio of PEGDA-HA:Gn revealed that the concentrations of 1:3 ratio of HA-Gelatin significantly increased hDPSC proliferation, either grown on 2D scaffold or embedding in 3D membranes.
Conclusions: We conclude from our findings that PEGDA-HA-Gn membranes facilitate cell proliferation and adhesion, mainly in the presence of ECM proteins. Therefore, employing appropriate and optimal ratios of the matrix components are crucial for DPSC longevity and viability, ensuing dental-pulp regeneration. Future studies are in progress to assess the contribution of fibronectin, in place of gelatin to facilitate hDPSC viability, proliferation and differentiation to an osteo-/odontogenic lineage.