Development of chitosan hydrogel scaffold loaded with secretomes of dental pulp stem cells

Abstract

When the vitality of the dental pulp is lost due to trauma or infectious diseases, it is necessary to perform an endodontic treatment, which results in non-vital teeth with loss of mechanical and sensorial characteristics. This fact culminates in more fragile teeth, subjected to fractures. In order to circumvent this problem, the regeneration of dental pulp tissue based on tissue engineering with endogenous stem cells seems promising. Moreover, scaffolds, growth factors and chemokine are essential elements in the dental pulp engineering. In previous study, a regenerative endodontic procedure was investigated, based on the elements of the tissue engineering in which we performed the complete removal of the dental pulp in molars of rodents. Such study was conducted with the use of EDTA into the root canal to release growth factors and the homing of stem cells of the periapical tissue as well as blood clot, resulting in a pulp-like tissue. Nonetheless, this tissue formation occurred in few root canals, probably due to the instability of the blood clot; low level of stem cells survival inside the root canal as well as the influence of the avascular environment. Therefore, an investigation of scaffolds associated with growth factors and chemokine has been carried out to stimulate the cellular viability, migration and angiogenesis as to improve the tissue engineering. The objectives of the study aim at developing an injectable scaffold loaded with stem cells secretomes of the dental pulp culminating in migration; viability, proliferation, cellular differentiation and angiogenesis. Firstly, the stem cells from the dental pulp will be isolated, characterized by flow cytometry and cultivated for secretomes of conditioned media obtention. Secondly, the chitosan hydrogel will load with secretomes of the stem cells and quantification of the vascular endothelial growth factor and the chemokine stromal derived factor will be verified. Lastly, the kinetic release of VEGF and SDF-1 within 21 days through ELISA will be investigated. (AU)