Chitosan-calcium scaffolds for regeneration of pulp-dentin complex: chemotactic potential analysis in realist model - in vitro study

• Grant number: 17/24198-5
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: March 01, 2018
• End date: February 28, 2019
• Field of knowledge: Health Sciences - Dentistry - Dental Materials
• Principal Investigator: Josimeri Hebling Costa
• Grantee: Deise Isabela Moreira dos Anjos
• Host Institution: Faculdade de Odontologia (FOAr). Universidade Estadual Paulista (UNESP). Campus de Araraquara. Araraquara , SP, Brazil
• Associated research grant: 16/15674-5 - Association of tissue engineering techniques for mineralized tissue regeneration under degenerative inflammatory stimulus: analysis on 3D-culture perfusion bioreactor and animal inflammatory models, AP.JP

Abstract

The regeneration of pulp-dentin complex employing macroporous scaffolds associated with a mineral phase capable of inducing osteoblastic/odontoblastic phenotype expression in dental pulp stem cells (DPSCs) has been considered as a promising therapy in Dentistry, mainly because of the structural similarity with dentin matrix. Thereby, the aim of this study is to evaluate the bioactive potential of a macroporous chitosan-scaffold containing calcium, by using an artificial pulp chamber with simulated pulp pressure (pAPCs) associated with 3D-cultured DPSCs. For this purpose, a 2% chitosan solution will be prepared in association with 1% calcium rich suspension, and then applied on teflon molds followed by phase-separation at low temperature, to obtain the macroporous scaffold. After that, DPSCs will be obtained by enzymatic disaggregation of pulp tissue from sound third molars, followed by stem cells markers characterization by immunofluorescence. The cells will be seeded in 3D-culture matrix in direct contact with dentin discs adapted to pAPCs, and the scaffolds will be placed on central perforations on the discs. This set will be connected to a 20 cm column of culture medium for the establishment of simulated pulp pressure. The presence of live cells (live/dead) and cell adhesion and spread (F-Actin) will be evaluated on scaffold and 3D-culture after 1,7 and 14 days of cell culture. Data will be evaluated by qualitative analysis. (AU)