To recapitulate the heterogeneous complexity of tissues in the human body with synthetic mimics of the extracellular matrix (ECM), it is important to develop methods that can easily allow the selective functionalization of defined spatial domains. Here, a facile method is introduced to functionalize microfibrillar meshes with different reactive groups able to bind biological moieties in a one-step reaction. The resulting scaffolds prove to selectively support a differential neurite growth after being seeded with dorsal root ganglia. Considering the general principles behind the method developed, this is a promising strategy to realize enhanced biomimicry of native ECM for different regenerative medicine applications.