PURPOSE: This study aimed to quantify the effect of interference screw insertion on MPFL graft tension when securing the femoral attachment after patellar fixation. 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. Surgeons should be aware of the inadvertent increases in graft tension even in low preloading conditions to mitigate the risk of graft overtensioning.