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Background: Reliable fixation of the soft hamstring grafts in ACL reconstruction has been reported as problematic., Hypothesis: The biomechanical properties of patellar tendon (PT) grafts fixed with biodegradable screws (PTBS) are superior compared to quadrupled hamstring grafts fixed with BioScrew (HBS) or Suture-Disc fixation (HSD)., Study Design: Controlled laboratory study with roentgen stereometric analysis (RSA)., Methods: Ten porcine specimens were prepared for each group. In the PT group, the bone plugs were fixed with a 7 x 25 mm BioScrew. In the hamstring group, four-stranded tendon grafts were anchored within a tibial tunnel of 8 mm diameter either with a 7 x 25 mm BioScrew or eight polyester sutures knotted over a Suture-Disc. The grafts were loaded stepwise, and micromotion of the graft inside the tibial tunnel was measured with RSA., Results: Hamstring grafts failed at lower loads (HBS: 536 N, HSD 445 N) than the PTBS grafts (658 N). Stiffness in the PTBS group was much greater compared to the hamstring groups (3500 N/mm versus HBS = 517 N/mm and HSD = 111 N/mm). Irreversible graft motion after graft loading with 200 N was measured at 0.03 mm (PTBS), 0.38mm (HBS), and 1.85mm (HSD). Elasticity for the HSD fixation was measured at 0.67 mm at 100 N and 1.32 mm at 200 N load., Conclusion: Hamstring graft fixation with BioScrew and Suture-Disc displayed less stiffness and early graft motion compared to PTBS fixation. Screw fixation of tendon grafts is superior to Suture-Disc fixation with linkage material since it offers greater stiffness and less graft motion inside the tibial tunnel., Clinical Relevance: Our results revealed graft motion for hamstring fixation with screw or linkage material at loads that occur during rehabilitation. This, in turn, may lead to graft laxity.