The extracellular matrix (ECM), with its multilayer fiber structure, regulates diverse functions including cell proliferation, migration, differentiation and tissue regeneration effects. To mimic and replace the native ECM, the structures and properties of three single-layer fabric substrates including warp/weft-knitted and woven fabrics were analyzed, then two-layer warp/weft-knitted composite fabrics prepared by polyurethane (PU) bonding, and woven composite fabrics prepared by polycaprolactone (PCL)/collagen solution bonding or PU bonding, were studied. After PCL/collagen solution bonding or PU bonding, properties such as pore diameter, air permeability, stress and the contact angle of composite fabrics decreased by some degree, while fiber diameter, thickness and the thermal conductivity of composite fabrics increased. In combination with fiber diameter, pore diameter and physical properties, we know that warp- or weft-knitted composite fabrics are ideal scaffolda for potential applications in nerve, myocardium and tendon tissue engineering.