Effects of life-long physical activity on tendon function have been investigated in cross-sectional studies, but these are at risk of "survivorship" bias. Here, we investigate if life-long side-specific loading is associated with greater cross-sectional area (CSA), mechanical properties, cell density (DNA content) and collagen cross-link composition of the male human patellar tendon (PT), in vivo. Nine seniors and six young male life-long elite badminton players and fencers were included. CSA of the PT obtained by 3-tesla MRI, and ultrasonography-based bilateral PT mechanics were assessed. Collagen fibril characteristics, enzymatic cross-links, non-enzymatic glycation (autofluorescence), collagen and DNA content were measured biochemically in PT biopsies. The elite athletes had a ≥15% side-to-side difference in maximal knee extensor strength, reflecting chronic unilateral sport-specific loading patterns. The PT CSA was greater on the lead extremity compared with the non-lead extremity (17 %, p=0.0001). Furthermore, greater tendon stiffness (18 %, p=0.0404) together with lower tendon stress (22 %, p=0.0005) and tendon strain (18 %, p=0.0433) were observed on the lead extremity. No effects were demonstrated from side-to-side for glycation, enzymatic cross-link, collagen, and DNA content (50%, p=0.1160). Moreover, tendon fibril density was 87±28 fibrils/μm2 on the lead extremity and 68±26 fibrils/μm2 on the non-lead extremity (28%, p=0.0544). Tendon fibril diameter was 86±14 nm on the lead extremity and 94±14 nm on the non-lead extremity (-9%, p=0.1076). These novel data suggest that life-long side-specific loading in males yields greater patellar tendon size and stiffness possibly with concomitant greater fibril density but without changes of collagen cross-link composition.