Tissue Engineering and Biotechnology applyed to the development of calcium-containing macroporous scaffolds with surface nano-topography to modulate pulp-dentin complex regeneration

Abstract

The development of porous scaffolds with modified surface topography is of great interest on Dentistry, since this substrate features a potential to induce pulp dentin complex regeneration, stimulating the migration of residente Dental Pulp Stem cells (DPSCs) to pulp exposed area. Therefore, the aim of this project is to combine differente Tissue Engineering and Biotechnology techniques in order to obtain calcium-containing scaffolds with an interconnected porous network and surface nanotopography to stimulate the odontogenic phenotype in DPSCs, inducing the dentinogenesis process. For that, scaffolds with different compositions with smooth or nano-globular topography. The phisico-chemistry properties, such as morphology, degradability, elasticity modulus and compressive strength will be characterized. Then, the DPSCs will be obtained by enzymatic desagregation of pulp tissue from sound third molars, followed by stem cells markers characterization by immunofluorescence. The cells will be seeded onto scaffolds surface to evaluate the viability, proliferati, migration, adhesion and spreed, as well as the gene expression and mineralized matrix deposition, which are related with odontoblastic phenotype expression and cell differentiation. Finally, to simulate the in vivo conditions on the laboratorial situation, 3D cell culture models associated with dentin discs adapted to artificial pulp chamber with simulated pulp pressure (pAPC) will be used to evaluate the potential of the scaffold as a cell free Tissue Engineering system. The scaffolds and 3D culture will be evaluated to acess the cell viability, adhesion and spread, and the expression of an odontoblastic phenotype up to 21 days. Numerical data obtained by the laboratorial assays will be subjected to specific statistical analysis. (AU)