Endothelial cell adhesion and migration is fundamental to a number of physiologic processes, including vascular development and angiogenesis. It has been investigated in a variety of contexts, including tumorigenesis, wound healing, tissue engineering, and biomaterial design. The chemical and mechanical extracellular environments are critical regulators of these processes, affecting integrin-matrix binding, cell adhesion strength, and cell migration. Understanding the synergy between matrix chemistry and mechanics will ultimately lead to precise control over adhesion and migration. Moreover, a better understanding of endothelial cell adhesion is critical for development of therapeutics and biomaterials for the treatment of endothelial cell dysfunction and the progression of vascular disease. This chapter will focus on the specific interactions between endothelial cells and the extracellular matrix that mediate adhesion and migration. Several engineering methods used to probe and quantify endothelial cell adhesion and migration will be discussed.