Odontoblastic differentiation of dental pulp stem cells from healthy and carious teeth on an original PCL-based 3D scaffold

Aims: To isolate and characterize dental pulp stem cells (DPSCs) obtained from carious and healthy mature teeth extracted when conservative treatment was not possible or for orthodontic reasons; to evaluate the ability of DPSCs to colonize, proliferate and differentiate into functional odontoblast-like cells when cultured onto a polycaprolactone cone made by jet-spraying and prototyped into a design similar to a gutta-percha cone. Methodology: DPSCs were obtained from nine carious and 12 healthy mature teeth. Then cells were characterized by flow cytometry and submitted to multidifferentiation to confirm their multipotency. These DPSCs were then cultured on a polycaprolactone cone in an odontoblastic differentiation medium. Cell proliferation, colonization of the biomaterial and functional differentiation of cells were histologically assessed. For the characterization, a t-Student test was used to compare the two groups. Results: In all cell cultures, characterization highlighted a mesenchymal stem cell phenotype (CD105+, CD90+, CD73+, CD11b-, CD34-, CD45-, HLA-DR-). No significant differences were found between cultures obtained from carious and healthy mature teeth. DPSCs from both origins were able to differentiate into osteocytes, adipocytes and chondrocytes. Cell colonization was observed both on the surface and in the thickness of polycaprolactone cones as well as a mineralized pericellular matrix deposit composed of type I collagen, alkaline phosphatase, osteocalcin and dentin sialophosphoprotein. Conclusions: DPSCs were isolated from both carious and healthy mature teeth. They were able to colonize and proliferate within a polycaprolactone cone and could be differentiated into functional odontoblast-like cells