Cartilage morphology change is an important biomarker for the progression of osteoarthritis. The purpose of this study was to assess the accuracy of in vivo cartilage thickness measurements from MR image-based 3D cartilage models using a laser scanning method and to test if the accuracy changes with cartilage thickness. Three-dimensional tibial cartilage models were created from MR images (in-plane resolution of 0.55 mm and thickness of 1.5 mm) of osteoarthritic knees of ten patients prior to total knee replacement surgery using a semi-automated B-spline segmentation algorithm. Following surgery, the resected tibial plateaus were laser scanned and made into 3D models. The MR image and laser-scan based models were registered to each other using a shape matching technique. The thicknesses were compared point wise for the overall surface. The linear mixed-effects model was used for statistical test. On average, taking account of individual variations, the thickness measurements in MRI were overestimated in thinner regions. The cartilage thicker than 2.5 mm was accurately predicted in MRI, though the thick cartilage in the central regions was underestimated. The accuracy of thickness measurements in the MRI-derived cartilage models systemically varied according to native cartilage thickness.