Application of tissue engineering for pulp-dentin complex regeneration: evaluation of a chitosan-calcium scaffold on in vitro simulated pulp exposure model

• Grant number: 18/09378-0
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: September 01, 2018
• End date: October 31, 2020
• Field of knowledge: Health Sciences - Dentistry - Dental Materials
• Principal Investigator: Diana Gabriela Soares dos Passos
• Grantee: Isabela Sanches Pompeo da Silva
• Host Institution: Faculdade de Odontologia de Bauru (FOB). Universidade de São Paulo (USP). Bauru , 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 application of tissue engineering for neo-tissue-genesis by modulating local precursor cells has gained the attention on Medicine and Dentistry, aiming the development of effective biological therapies. Within this concept, the present study aims to evaluate the potential of an experimental chitosan scaffold to modulate chemotaxis and odontogenic differentiation of dental pulp stem cells (DPSCs), by using an alternative study model of pulp exposure in vitro. For that, a 2% chitosan solution will be formulated and 1% calcium rich suspension will be added. This solution will be distributed into teflon molds and subjected to phase separation at low temperatures, obtaining the porous scaffolds. A tri-dimensional culture of DPSCs will be stablished under dentin discs adapted to artificial pulp chambers with 20 cm.H2O pressure, simulating pulp exposure condition in vitro. The scaffolds will be placed on dentin discs' perforations in direct contact with the 3D culture, and this set will be incubated for 14 days. The scaffolds surface will be subjected analysis of cell viability (live/dead assay) and dentin sialoprotein (DSP) expression by immunofluorescence assay, in order to evaluate the chemotaxis of cells with odontoblastic phenotype. The 3D cell culture will be evaluated in regard of cell viability (live/dead assay) and gene expression of DSPP (real time PCR). Data will be subjected to statistic assays and qualitative analysis.