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It was hypothesized that interference screw insertion significantly increases graft tension. Methods Ten fresh frozen human cadaveric femurs were utilized to compare graft tension at three different preloading conditions (2 N, 5 N, 10 N) using a tensile testing machine (Admet Inc., Norwood, MA). Each preloading condition was analyzed with varying graft sizes (5-8 mm), tunnel diameters (7-9 mm), and interference screw sizes (7-9 mm). Non-parametric statistical analysis was utilized to compare testing conditions among each other. Results Graft tension significantly increased after interference screw insertion by 100% to 552%, with 2 N preload showing the greatest increase (p < 0.001). Grafts with a larger diameter (7-8 mm) had a significantly greater increase in tension than smaller grafts (5-6 mm), regardless of preloading conditions (p < 0.001). Interference screw size had no influence on graft tension (n.s.). A graft-tunnel interference (tunnel diameter-graft diameter) fit of 0 mm and 1 mm significantly increased graft tension for each preloading condition when compared to a slightly looser fit of >= 2 mm (p < 0.05). Conclusion Femoral interference screw insertion significantly increases graft tension in MPFL reconstruction even in low preloading conditions, with graft size and graft-tunnel interference fit having a considerably effect on graft tension. 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