A highly aligned pattern of carbon nanofibers (CNF) on polycarbonate urethane (PCU) for tissue engineering applications was created by placing a CNF-ethanol solution in 30 microm width copper grid grooves on top of PCU. In vitro results provided the first evidence that fibroblasts and vascular smooth muscle cells selectively adhered to the PCU regions. However, endothelial cells did not display a preference for adhesion to the CNF compared with PCU regions. Previous studies have shown selective adhesion of osteoblasts (bone-forming cells) on CNF compared with PCU regions. Thus, the present results suggest that CNF aligned on PCU may be useful substrates for the control of spatial cell attachment, criteria useful for the design of a wide range of tissue engineering materials, from orthopedic to vascular.