One-dimensional (1-D) nanostructures have attracted enormous research interest due to their unique physicochemical properties and wide application potential. These 1-D nanofibers are being increasingly applied to biomedical fields owing to their high surface area-to-volume ratio, high porosity, and the ease of tuning their structures, functionalities, and properties. Many biomedical nanofiber reviews have focused on tissue engineering and drug delivery applications but have very rarely discussed their use as wound dressings. However, nanofibers have enormous potential as wound dressings and other clinical applications that could have wide impacts on the treatment of wounds. Herein, the authors review the main fabrication methods of nanofibers as well as requirements, strategies, and recent applications of nanofibers, and provide perspectives of the challenges and opportunities that face multifunctional nanofibers for active therapeutic applications.