Sulfated glycosaminoglycans (sGAGs) are vital molecules of the extracellular matrix (ECM) of the nervous system known to regulate proliferation, migration, and differentiation of neurons mainly through binding relevant growth factors. Alginate sulfate (AlgSulf) mimics sGAGs and binds growth factors such as basic fibroblast growth factor (FGF‐2). Here, thin films of biotinylated AlgSulf (b‐AlgSulfn) are engineered with sulfation degrees (DS = 0.0 and 2.7) and the effect of polysaccharide concentration on FGF‐2 and nerve growth factor (β‐NGF) binding and subsequent primary neural viability and neurite outgrowth is assessed. An increase in b‐AlgSulfn concentration results in higher FGF‐2 and β‐NGF binding as demonstrated by greater frequency and dissipation shifts measured with quartz crystal microbalance with dissipation monitoring (QCM‐D). Primary neurons seeded on the 2D b‐AlgSulfn films maintain high viability comparable to positive controls grown on poly‐d‐lysine. Neurons grown in 3D AlgSulf hydrogels (DS = 0.8) exhibit a significantly higher viability, neurite numbers and mean branch length compared to neurons grown in nonsulfated controls. Finally, a first step is made toward constructing 3D neuronal networks by controllably patterning neurons encapsulated in AlgSulf into an alginate carrier. The substrates and neural networks developed in the current study can be used in basic and applied neural applications.

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