Abstract
Replicating the intricate architecture of the extracellular matrix (ECM) is an actual challenge in the field of bone tissue engineering. In the present research study, calcium alginate/cellulose nanofibrils-based 3D printed scaffolds, double-reinforced with chitosan/polyethylene oxide electrospun nanofibers (NFs) and graphene oxide (GO) were prepared using the 3D printing technique. The porous matrix was provided by the calcium alginate, while the anisotropy degree and mechanical properties were ensured by the addition of fillers with different sizes and shapes (CNFs, NFs, GO), similar to the components naturally found in bone ECM. Surface morphology and 3D internal microstructure were analyzed using scanning electron microscopy (SEM) and micro-computed tomography (μ-CT), which evidenced a synergistic effect of the reinforcing and functional fibers addition, as well as of the GO sheets that seem to govern materials structuration. Also, the nanoindentation measurements showed significant differences in the elasticity and viscosity modulus, depending on the measurement point, this supported the anisotropic character of the scaffolds. In vitro assays performed on MG-63 osteoblast cells confirmed the biocompatibility of the calcium alginate-based scaffolds and highlighted the osteostimulatory and mineralization enhancement effect of GO. In virtue of their biocompatibility, structural complexity similar with the one of native bone ECM, and biomimetic mechanical characteristics (e.g. high mechanical strength, durotaxis), these novel materials were considered appropriate for specific functional needs, like guided support for bone tissue formation.