Engineered scaffolds function to supplement or replace injured, missing, or compromised tissue or organs. The current direction in this research area is to create scaffolds that mimic the structure and function of the native extracellular matrix (ECM). It is believed that the fabrication of a scaffold that has both structural integrity and allows for normal cellular function and interaction will bring scaffolds closer to clinical relevance. Nanotechnology innovations have aided in the development of techniques for the production of nanofiber scaffolds. The three major processing techniques, self-assembly, phase separation, and electrospinning, produce fibers that rival the size of those found in the native ECM. However, the simplicity, versatility, and scalability of electrospinning make it an attractive processing method that can be used to reproduce aspects of the complexity that characterizes the native ECM. Novel electrospinning strategies include alterations of scaffold composition and architecture, along with the addition and encapsulation of cells, pharmaceuticals and growth factors within the scaffold. This article reviews the major nanofiber fabrication technologies as well as delves into recent significant contributions to the conception of a meaningful and practical electrospun scaffold. (C) 2009 John Wiley & Sons, Inc. WIREs Nanomed Nanobiotechnol 2010 2 20-34