A wide array of biomedical applications, extending from the fabrication of implant materials to targeted drug delivery, can be attributed to polymers. The utilization of chemical monomers to form polymers, such as polypropylene, polystyrene, and polyethylene, can provide high mechanical stability to them and they can be utilized for diverse electronic or thermal applications. However, certain chemical-based synthetic polymers are toxic to humans, animals, plants, and microbial cells. Thus, biopolymers have been introduced as an alternative to make them utilizable for biomedical applications. Even though biopolymers possess beneficial biomedical applications, they are not stable in biological fluids and exhibit toxicity in certain cases. Recent advances in nanotechnology have expanded its applicational significance in various domains, especially in the evolution of biopolymers to transform them into nanoparticles for numerous biomedical applications. In particular, biopolymers are fabricated as nanofibers to enhance their biological properties and to be utilized for exclusive biomedical applications. The aim of this review is to present an overview of various biopolymer nanofibers and their distinct synthesis approaches. In addition, the medical applications of biopolymer nanofibers, including antimicrobial agents, drug delivery systems, biosensor production, tissue engineering, and implant fabrication, are also discussed.