Thermoplastic poly(ether/ester) elastomer (TPEE) has great potential as a mattress material due to its high resilience, breathability, and light weight. This study aimed to evaluate the feasibility of TPEE-3D fibrous material (T3DF), a three-dimensional block material made of TPEE fibers randomly aligned and loop-connected, for mattress application. After testing the compression properties of T3DF, the effects of T3DF structural layers on mattress firmness were investigated. The results showed that T3DF had good energy absorption capacity, broad indentation hardness range (126.94–333.82 N), and high compression deflection coefficient (2.79–4.39). The thickness and density of T3DF were the main factors influencing mattress firmness, and the impact of thickness was more significant (p < 0.05). Owing to the hard and soft segments contained in TPEE, T3DF could be used for both the padding and core layers of the mattress. The hardness value and Dsurface of the mattress with a T3DF padding layer increased with T3DF density but decreased with T3DF thickness. Moreover, the hardness value and Dsurface of the mattress with a T3DF core layer increased with T3DF density, while with T3DF thickness, its Dsurface increased and Dbottom decreased. Therefore, the thick and low-density T3DF padding layer could improve the comfort of the mattress surface, a thin T3DF core layer could satisfy both the softer surface and the firmer bottom of the mattress.