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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 (T_3DF), a three-dimensional block material made of TPEE fibers randomly aligned and loop-connected, for mattress application. After testing the compression properties of T_3DF, the effects of T_3DF structural layers on mattress firmness were investigated. The results showed that T_3DF 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 T_3DF were the main factors influencing mattress firmness, and the impact of thickness was more significant (<i>p</i> < 0.05). Owing to the hard and soft segments contained in TPEE, T_3DF could be used for both the padding and core layers of the mattress. The hardness value and <inline-formula><math xmlns="http://www.w3.org/1998/Math/MathML" display="inline"><semantics><mrow><msub><mrow><mi>D</mi></mrow><mrow><mi>s</mi><mi>u</mi><mi>r</mi><mi>f</mi><mi>a</mi><mi>c</mi><mi>e</mi></mrow></msub></mrow></semantics></math></inline-formula> of the mattress with a T_3DF padding layer increased with T_3DF density but decreased with T_3DF thickness. Moreover, the hardness value and <inline-formula><math xmlns="http://www.w3.org/1998/Math/MathML" display="inline"><semantics><mrow><msub><mrow><mi>D</mi></mrow><mrow><mi>s</mi><mi>u</mi><mi>r</mi><mi>f</mi><mi>a</mi><mi>c</mi><mi>e</mi></mrow></msub></mrow></semantics></math></inline-formula> of the mattress with a T_3DF core layer increased with T_3DF density, while with T_3DF thickness, its <inline-formula><math xmlns="http://www.w3.org/1998/Math/MathML" display="inline"><semantics><mrow><msub><mrow><mi>D</mi></mrow><mrow><mi>s</mi><mi>u</mi><mi>r</mi><mi>f</mi><mi>a</mi><mi>c</mi><mi>e</mi></mrow></msub></mrow></semantics></math></inline-formula> increased and <inline-formula><math xmlns="http://www.w3.org/1998/Math/MathML" display="inline"><semantics><mrow><msub><mrow><mi>D</mi></mrow><mrow><mi>b</mi><mi>o</mi><mi>t</mi><mi>t</mi><mi>o</mi><mi>m</mi></mrow></msub></mrow></semantics></math></inline-formula> decreased. Therefore, the thick and low-density T_3DF padding layer could improve the comfort of the mattress surface, a thin T_3DF core layer could satisfy both the softer surface and the firmer bottom of the mattress.