OBJECTIVES: This in vitro study compares the femorotibial shear in canine stifles with intact and transected cranial cruciate ligaments (CrCL), before and after tibial tuberosity advancement (TTA) by measuring the distance between origin and insertion of the cruciate ligaments. METHODS: Radiodense markers were inserted into bones at the attachment sites of the cruciate ligaments in sixteen cadaveric stifles of adult dogs. Each stifle was then mounted in a testing apparatus. The distances between the markers were measured on mediolateral radiographs, performed on each stifle under three different situations: intact, after CrCL transection, and after performing a TTA. Stifles were loaded to create a constant tibiofemoral reaction force by maintaining the load parallel to the patellar ligament. Radiographs were taken in a preloaded and loaded state in the intact stifle and only in a loaded state after CrCL transection, and after performing a TTA. RESULTS: Loading the stifle joints after transection of the CrCL resulted in a mean lengthening of the CrCL marker distance of 22.4%. Loading the transected CrCL stifles after performing a TTA resulted in a mean shortening of the CrCL marker distance by 3.0% compared to the loaded intact condition. CLINICAL SIGNIFICANCE: This study demonstrates that, in loaded stifles with transected CrCLs, TTA causes a caudal shift in the cranial shear force, counteracting cranial subluxation of the tibia.