Indirect three-dimensional printing as a strategy to modulate the interconnectivity of chitosan-calcium scaffolds

Abstract

The use of alloplastic scaffolds for the modulation of local precursor cells has been increasingly highlighted in tissue engineering techniques, suggesting a promising application in the medical and dental fields. An important aspect in of a scaffold are the characteristics of its porosity, such as its organization, hierarchy and interconnectivity between the pores. Therefore, the aim of this study is to develop macro-porous chitosan-calcium scaffolds containing a network of oriented macro-channels obtained using 3D printed molds, in order to favor cell infiltration and tissue neo-deposition. For this, molds with different internal geometries will be used and produced in a CAD/CAM system through 3D printing that, associated with bublling-effect technology, will result in different micro and macro-porosity frameworks in the chitosan-calcium scaffolds. The scaffolds will be physiochemically characterized through analysis of morphology and topography (scanning electron microscopy), degree of interconnectivity (X-ray CT), porosity and chemical composition (FTIR). For in vitro biological characterization, a primary culture of human dental pulp cells will be used. Cell viability (Live/Dead), adhesion and spreading (F-actin), alkaline phosphatase activity (end-point assay), and calcium quantification (Alizarin Red) will be performed after seeding the cells onto biomaterials surface. Data will be evaluated quantitatively and qualitatively.(AU)