Research in polymer nanofibers has undergone significant progress in the last one decade. One of the main driving forces for this progress is the increasing use of these polymer nanofibers for biomedical and biotechnological applications. This article presents a review on the latest research advancement made in the use of polymer nanofibers for applications such as tissue engineering, controlled drug release, wound dressings, medical implants, nanocomposites for dental restoration, molecular separation, biosensors, and preservation of bioactive agents.