Crosslinked and bundled actin filaments form networks that are essential for the mechanical properties of living cells. Reconstituted actin networks have been extensively studied not only as a model system for the cytoskeleton, but also to understand the interplay between microscopic structure and macroscopic viscoelastic properties of network-forming soft materials. These constitute a broad class of materials with countless applications in science and industry. So far, it has been widely assumed that reconstituted actin networks represent equilibrium structures. Here, we show that fully polymerized actin/fascin bundle networks exhibit surprising age-dependent changes in their viscoelastic properties and spontaneous dynamics, a feature strongly reminiscent of out-of-equilibrium, or glassy, soft materials. Using a combination of rheology, confocal microscopy and space-resolved dynamic light scattering, we demonstrate that actin networks build up stress during their formation and then slowly relax towards equilibrium owing to the unbinding dynamics of the crosslinking molecules.
