A scaffold is a three-dimensional (3D) structure having extracellular matrix (ECM) mimicking properties both chemically and biologically. Ideally, scaffold matrices should: support cell growth and maintenance; provide appropriate mechanical support; have a degradation rate in synchrony with defect healing rate; and facilitate effective nutrient transfer, gas exchange (i.e., O2 and CO2), metabolic waste removal, and signal transduction. In addition to physical and spatial cues, the scaffold itself can be the carrier of signaling biomolecules, thus emphasizing the need for scaffold-based tissue engineering. In this chapter, we review the fundamental requirements and considerations to enhance the design and manufacture of scaffolds, analyze the use of tissue engineering scaffolds in terms of biomaterials structure and cell material interactions; and also review traditional and advanced scaffold fabrication methods. The challenges of these scaffold-based regenerative methods are also discussed. © 2015 Elsevier Inc. All rights reserved.