A 6-Minute Limb Function Assessment for Therapeutic Testing in Experimental Peripheral Artery Disease Models, 2025, Victoria R. Palzkill et al

Highlights
Therapeutic development for PAD has been largely stagnant for the past 2 decades, and promising preclinical targets have failed to translate into clinical practice.

Preclinical models of HLI are widely used; however, most studies rely on limb perfusion recovery as a primary outcome measure—a stark contrast to the 6-minute walk test used clinically.

Herein, a 6-minute limb function test has been developed for preclinical PAD models. The test involves repeated isotonic muscle contractions coupled with continuous measurement of muscle blood flow, allowing quantification of total muscular work and perfusion flux.

The 6-minute limb function test demonstrates superior effect sizes compared to assessments of limb perfusion recovery or muscle strength and identifies therapeutic benefit in scenarios where limb perfusion recovery could not.

Summary
In this study, we present a novel 6-minute limb function test that allows for the congruent assessment of muscular performance and hemodynamics in preclinical models of peripheral artery disease. Using several experimental conditions, the results demonstrate the superior efficacy of the 6-minute limb function test to detect differences in the response to hindlimb ischemia across several interventions, including where traditional perfusion recovery, capillary density, and muscle strength measures were unable to detect interventional differences, thus allowing for more rigorous assessment of preclinical therapies before clinical translation.
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