Silk fibroin (SF) is a biocompatible and slowly biodegradable material with excellent mechanical properties and huge potential for use as biofunctional interface in electronic devices that aim to stimulate and control neural network activity and peripheral nerve repair. It is shown that SF films act as material interfaces that support the adherence and neurite outgrowth of dorsal root ganglion (DRG) neurons and preserve neuronal functions. Silk films preserve the capability of neuronal cells to fire and DRG neurons on silk films retain the intracellular free Ca2+ concentration ([Ca2+]i) response to capsaicin, a typical noxious stimulus for this neuronal culture model. It is also demonstrated that nerve growth factor (NGF)-functionalized silk films promote neurite outgrowth and modulate functional properties of DRG neurons. The results show that silk preserves DRG neuronal physiology and is a promising biomaterial platform for the future development of devices with goals including functional recovery of injured neurons, neurite functional outgrowth in vitro, or functional electrostimulation in vivo.