Electrospinning has been widely used as a nanofiber fabrication technique. Its simple process, cost effectiveness and versatility have appealed to materials scientists globally. Pristine polymeric nanofibers or composite nanofibers with dissimilar morphologies and multidimensional assemblies ranging from one dimension (1D) to three dimensions (3D) can be obtained from electrospinning. Critically, these as-prepared nanofibers possessing high surface area to volume ratio, tunable porosity and facile surface functionalization present numerous possibilities for applications, particularly in biomedical field. This review gives us an overview of some recent advances of electrospinning-based nanomaterials in biomedical applications such as antibacterial mats, patches for rapid hemostasis, wound dressings, drug delivery systems, as well as tissue engineering. We further highlight the current challenges and future perspectives of electrospinning-based nanomaterials in the field of biomedicine.