Bioactive multilayer scaffolds for tissue engineering: in vitro osteogenesis analysis

Abstract

Scaffolds (three-dimensional porous architectures for cell growth) enables the regeneration of different types of tissues by allowing cell migration, proliferation, adhesion, and differentiation, and can be applied in dentistry, bone regeneration and wound healing. The combination of electrospinning and additive manufacturing techniques in order to generate a scaffold composed of nanofiber and hydrogel may allow obtaining a porous structure with optimized bioactive properties through the incorporation of 45B5 borate glass (46.1 B2O3 - 26.9 CaO - 24.4 NaO - 2.6 P2O5, mol%). In this context, the aim of this proposal will be to evaluate the influence of the multilayer cellular scaffold with hierarchical pore structure on osteogenesis in vitro. The scaffolds will be initially produced with Poly(lactic acid) (PLA) nanofibers containing 45B5 glass (5% m/v) by electrospinning technique. They will then be 3D printed using sodium alginate hydrogel on the surface of the produced nanofibers. The bioactive glass 45B5 will be evaluated by X-ray diffraction (XRD), and the nanofibers and hydrogels will be individually characterized by scanning electron microscopy (SEM) and Fourier-transform infrared spectroscopy (FTIR). After production, the multilayer cellular scaffolds will be evaluated by SEM and subjected to in vitro osteogenesis analysis. In vitro biological tests will be performed using MG63 and HFF1 cell lines. The cells will be cultured with the scaffolds to assess the influence of the biomaterials on the activity and differentiation of osteoblasts and fibroblasts, using cell interaction assays, cytotoxicity tests, total protein content evaluation, cellular morphology analysis, and mineralization nodule formation for the MG63 osteoblastic lineage. Initially, the data will be statistically analyzed using the normality test to apply the appropriate statistical test. The adopted significance level will be the conventional value of 5%. With the execution of this research project, it is expected to obtain a cellular scaffold with a bioactive and porous structure with optimized osteogenic properties.