Conventional experimental methods for studying cardiac muscle in vitro often do not expose the tissue preparations to a mechanical impedance that resembles the in vivo hemodynamic impedance dictated by the arterial system. That is, the afterload in work‑loop contraction is conventionally simplified to be constant throughout muscle shortening, and at a magnitude arbitrarily defined. This conventional afterload does not capture the time‑varying interaction between the left ventricle and the arterial system. We have developed a contraction protocol for isolated tissue experiments that allows the afterload to be described within a Windkessel framework that captures the mechanics of the large arteries. We aim to compare the energy expenditure of cardiac muscle undergoing the two contraction protocols: conventional versus Windkessel loading. Isolated rat left‑ventricular trabeculae were subjected to the two force-length work‑loop contractions. Mechanical work and heat liberation were assessed, and mechanical efficiency quantified, over wide ranges of afterloads or peripheral resistances. Both extent of shortening and heat output were unchanged between protocols, but peak shortening velocity was 39.0 % lower and peak work output was 21.8 % greater when muscles contracted against the Windkessel afterload than against the conventional isotonic afterload. The greater work led to a 25.2 % greater mechanical efficiency. Our findings demonstrate that the mechanoenergetic performance of cardiac muscles in vitro may have been previously constrained by the conventional, arbitrary, loading method. A Windkessel loading protocol, by contrast, unleashes more cardiac muscle mechanoenergetic potential, where the slower shortening increases efficiency in performing mechanical work.