In general, the design of the scaffold should imitate certain advantageous properties of native extracellular matrix (ECM) in order to operate as a temporary ECM for cells. From this aspect, a biomimetic scaffold was prepared by using poly (vinyl alcohol) (PVA) and carrageenan (CAR), in which axially oriented pore structure can be formed through a facile unidirectional freeze-thaw method. We examined the feasibility of this oriented scaffold, which has better physico-chemical properties compared to non-oriented scaffold fabricated by conventional method. The microenvironment of this oriented scaffold could imitate biochemical and physical cues of natural cartilage ECM for guiding spatial organization and proliferation of cells in vitro, indicating its potential in cartilage repair strategy. Furthermore, the biocompatibility of the scaffold in vivo was demonstrated in a subcutaneous rat model, which revealed uniform infiltration and survival of newly formed tissue into oriented scaffold after 4 weeks, with only a minimal inflammatory response being observed over the course of the experiments. These results together indicated that the present biomimetic scaffold with oriented microarchitecture could be a promising candidate for cartilage tissue engineering.