Nanofibers are an important material for regenerative medicine as they have a commensurate morphology to that of the macromolecular matrix that supports and houses the growth of cells and tissues within the body. Electrospinning is widely used to fabricate non-woven structures on the nanoscale and the versatility of the technique has widened the application of nanofibers. This is due to ease of extending nanofiber functionality through the incorporation of active materials both during and after electrospinning. Recent developments in electrospinning devices, such as needle-free systems, have reinvigorated research as these advances now allow fabrication of nanofibers at commercial scales. The process of electrospinning has a number of operating parameters that are adjusted in optimisation to achieve ideal fibres and a multitude of instrument configurations can be adopted to achieve the required manufacture. The innate properties of nanofibers, such as high surface area to volume ratio, have many proven benefits for regenerative medicine and the chapter examines these before discussing how functionality can be further improved. Numerous materials can be incorporated in the manufacture of electrospun mats, however when choosing materials for regenerative medicine, biocompatibility and biodegradability are the dominant functionalities that are required.