Fabricating tissue architecture-mimicking scaffolds is one of the major challenges in the field of tissue engineering. Electrospun nanofibers have been considered as potent techniques for fabricating fibrous scaffolds biomimicking extracellular frameworks. Therapeutic agent-incorporated nanofibrous meshes have widely served as excellent substrates for adhesion, proliferation and differentiation. Many drugs, proteins and nucleic acids were incorporated into the scaffolds for regeneration of skin, musculoskeletal, neural and vascular tissue engineering in aims to control the release of the therapeutic agents. In the current article, we focus on introducing various fabrication techniques for electrospun nanofiber-based scaffolds and subsequent functionalization of nanofibers for therapeutic purposes. We also detail how the therapeutic nanofibrous meshes can be employed in the field of tissue engineering.