Enzymatic degradation of chitosan scaffold supporting 3D neuronal networks

•Chitosan microbeads were degraded by chitosanase and β-glucosidase.•The 2D neuronal network was not affected by the two enzymes.•The scaffold supporting 3D neuronal network was partially enzymatically degraded.•The functional activity of 3D network was not modified by scaffold degradation. The degradation of cell seeded 3D scaffolds is an important step both for the in vivo and in vitro use of biomimetic matrices mimicking the extracellular matrix. In this respect, we investigated the possibility of enzymatically degrading a chitosan microbeads scaffold supporting a mature 3D neuronal network. To this purpose, the enzymes chitosanase and β-glycosidase were used. As a first step, their impact on the development of a mature 2D neuronal and their degradative action on chitosan microbeads were evaluated. Then, the degradation protocol was used to treat chitosan microbeads supporting a 3D neuronal network. It was observed that the degradation was slowed down by the presence of the cells. Finally, the electrophysiological activity of 3D neuronal networks before and after the scaffold degradation was found to be not modified.