Novel materials have been recently developed for coping with various environmental factors. Generally, to improve the thermal comfort to humans in cold environments, securing an air layer is important. Therefore, this study analyzed the thermal properties of 3D spacer technical materials, 3D printed using thermoplastic polyurethane, according to the structural changes. Four 3D spacer technical material structures were designed with varying pore size and thickness. These samples were moved into a cold climate chamber (temperature 5 ± 1 °C, relative humidity (60 ± 5)%, wind velocity ≤0.2 m/s) and placed on a heating plate set to 30 °C. The surface and internal temperatures were measured after 0, 10, 20, and 30 min and then 10 min after turning off the heating plate. When heat was continuously supplied, the 3D spacer technical material with large pores and a thick air layer showed superior insulation among the materials. However, when no heat was supplied, the air gap thickness dominantly affected thermal insulation, regardless of the pore size. Hence, increasing the air gap is more beneficial than increasing the pore size. Notably, we found that the air gap can increase insulation efficiency, which is of importance to the new concept of 3D printing an interlining.