Background: There is little data in the literature regarding the preparation methods of the intra-articular portion of quadruple hamstring-tendon grafts for anterior cruciate ligament (ACL) reconstruction. The aim of this study was to compare the biomechanical properties of a sutured transplant to that of a non-sutured transplant. The hypothesis was that adding stitches to the intra-articular portion of the graft increased its resistance. Method: A comparative cadaveric study was carried out on five pairs of knees. The average age of the cadavers was 68 years. The exclusion criterion was past knee surgery. In the Sutured Group (SG) two stitches were made on the grafts. No stitches were made on the grafts of the Non-sutured Group (NSG). A tensile failure test was carried out using an Instron® loading machine. The maximal load to failure and stiffness were recorded and we observed the mode of failure for each graft. Statistical analysis was performed using the Wilcoxon rank sum test. Level of significance was set at p < 0.05. Results: The hypothesis proposed was not confirmed; adding stitches to the intra-articular portion of the four-strand hamstring-tendon graft does not increase its biomechanical properties. The maximal load to failure was 233.5 N ± 40.6 (186.7–274.5 N) for the NSG, 19.6% higher than for the SG which was 195.2 N ± 42.9 (139.0–238.2 N). Nevertheless, the difference observed was not statistically significant (p = 0.188). The stiffness of the grafts for the NSG was 23.5 N/mm ± 5.3 (17.8–29 N/mm) and 19.7 N/mm ± 5.5 (13.2–24.7 N/mm) for the SG grafts. Overall stiffness values for the NSG were 19% higher than those of the SG; however, the results were not statistically significant (p = 0.438). The failure mode was a rupture at the fixation point except for one sample from the SG which failed at an intra-articular stitch. Conclusion: Whilst the initial hypothesis was not verified, nevertheless, the maximal loads to failure and stiffness were approximately 20% higher when there were no intra-articular stitches compared to the situation where stitches were added to the intra-articular portion of the graft. This was a cadaveric pilot study and, therefore, whilst we are not able to extend our results to clinical practice, the outcomes would indicate the need for further development of this and related protocols deriving from the question of whether there is weakening the graft when adding stitches to its mid-substance. These results remain to be confirmed by further research.