The formation of a stable vascular network in a scaffold is one of the most challenging tasks in tissue engineering and regenerative medicine. Despite the common use of porous scaffolds in these applications, little is known about the effect of pore size on the neovascularization in these scaffolds. Herein is fabricated poly(D, L-lactide-co-glycolide) inverse opal scaffolds with uniform pore sizes of 79, 147, 224, and 312 μm in diameter and which are then used to systematically study neovascularization in vivo. Histology analyses reveal that scaffolds with small pores (<200 μm) favor the formation of vascular networks with small vessels at high densities and poor penetration depth. By contrast, scaffolds with large pores (>200 μm) favor the formation of vascular networks with large blood vessels at low densities and deep penetration depth. Based on the different patterns of vessel ingrowth as regulated by the pore size, a model is proposed to describe vascularization in a 3D porous scaffold, which can potentially serve as a guideline for future design of porous scaffolds.