Electrospun polymeric nanofibers are essential in various fields for various applications because of their unique properties. Their features are similar to extracellular matrices, which suggests them for applications in healthcare fields, such as antimicrobials, tissue engineering, drug delivery, wound healing, bone regeneration, and biosensors. This review focuses on the synthesis of electrospun polymeric nanofibers, their surface modification, and their biomedical applications. Nanofibers can be fabricated from both natural and synthetic polymers and their composites. Even though they mimic extracellular matrices, their surface features (physicochemical characteristics) are not always capable of fulfilling the purpose of the target application. Therefore, they need to be improved via surface modification techniques. Both needle-based and needleless electrospinning are thoroughly discussed. Various techniques and setups employed in each method are also reviewed. Furthermore, pre-and postspinning modification approaches for electrospun nanofibers, including instrument design and the modification features for targeted biomedical applications, are also extensively discussed. In this way, the remarkable potential of electrospun polymeric nanofibers can be highlighted to reveal future research directions in this dynamic field.