Background: Increased tibial slope has been shown to lead to higher rates of anterior cruciate ligament graft failure. A slope-decreasing osteotomy can reduce in situ anterior cruciate ligament force and may mitigate this risk. However, how this procedure may affect the length change behavior of the medial ligamentous structures is unknown. Purpose/Hypothesis: The purpose of this study was to examine the effect of anterior slope–modifying osteotomies on the medial ligamentous structures. It was hypothesized that (1) decreasing the tibial slope would lead to shortening of the superficial medial collateral ligament (sMCL), (2) while the fibers of the posterior oblique ligament (POL) would be unaffected. Study Design: Descriptive laboratory study. Methods: Eight fresh-frozen cadaveric knee specimens underwent anatomic dissection to precisely identify the medial ligamentous structures. The knees were mounted in a custom-made kinematics rig with the quadriceps muscle and iliotibial tract loaded. An anterior slope–modifying osteotomy was performed and fixed using an external fixator, which allowed modification of the wedge height between −15 and +10 mm in 5-mm increments. Threads were mounted between pins positioned at the anterior, middle, and posterior parts of the tibial and femoral attachments of the sMCL and POL. For different tibial slope modifications, length changes between the tibiofemoral pin combinations were recorded using a rotary encoder as the knee was flexed between 0° and 120°. Results: All sMCL fiber regions shortened with slope reduction ( P < .001) and lengthened with slope increase ( P < .001), with the anterior sMCL fibers more affected than the posterior sMCL fibers. A 15-mm anterior closing-wedge high tibial osteotomy (ACWHTO) resulted in a 6.9% ± 3.0% decrease in the length of the anterior sMCL fibers compared with a 3.6% ± 2.3% decrease for the posterior sMCL fibers. A 10-mm anterior opening-wedge high tibial osteotomy (AOWHTO) increased anterior sMCL fiber length by 5.9% ± 2.3% and posterior sMCL fiber length by 1.6% ± 1.0%. The POL fibers were not significantly affected by a slope-modifying osteotomy. Conclusion: Tibial slope–modifying osteotomies changed the length change pattern of the sMCL such that an AOWHTO increased whereas an ACWHTO decreased the sMCL strain. This effect was most pronounced for the anterior fibers of the sMCL. The length change pattern of the POL remained unaffected by slope-modifying osteotomy. Clinical Relevance: Surgeons should be aware that anterior tibial slope–modifying osteotomies affect the biomechanics of the sMCL. After an extensive ACWHTO, patients may develop a medial or anteromedial instability, while an AOWHTO may overconstrain the medial compartment.