Over the last few decades the electrospinning process has been extensively acknowledged in the biomedical fields for its ability to manufacture a nanofibrous architecture composed of natural polymers (biopolymers) and/or synthetic polymers for various medical uses. As compared to synthetic polymers, the biopolymers are more biocompatible, causing less immunogenic effects and toxicities to the body. The biopolymer-based electrospun nanofibrous materials have widely been used for dressing wounds, tissue engineering, regenerative medicine, and drug delivery. In some cases, biopolymers are electrospun together with synthetic polymers to obtain composite nanofibers with optimal characteristics that are critical for their functionality. This chapter reviews the current electrospinning settings to produce nanofibers. This is followed by various medical applications of biopolymer-based electrospun nanofibers in tissue engineering, drug delivery, biosensors, and stem cell research.