The presently disclosed subject matter provides a scalable and electrostretching approach for generating hydrogel microfibers exhibiting uniaxial alignment from aqueous polymer solutions. Such hydrogel microfibers can be generated from a variety of water-soluble natural polymers or synthetic polymers. The hydrogel microfibers can be used for controlled release of bioactive agents. The internal uniaxial alignment exhibited by the presently disclosed hydrogel fibers provides improved mechanical properties to hydrogel microfibers, and contact guidance cues and induces alignment for cells seeded on or within the hydrogel microfibers.