PurposeWe aimed to investigate the relationship between quantitative evaluation by compositional MRI, including T1ρ, and histological and macroscopic assessments, to verify the validity of compositional MRI, and examine the relationship between compositional MRI evaluation reconstructed in three dimensions (3D) and histological and visual assessments. MethodTwenty-seven patients with knee osteoarthritis underwent T1ρ and T2 magnetic resonance imaging (MRI). Histological and gross tissue evaluations were performed on the excised bone sections of total knee arthroplasty. Semi-quantitative histological evaluation of tissue changes were assessed by measuring the optical density of digitally captured safranin O-stained and Collagen type II antibody-stained paraffin sections. Macroscopic cartilage severity was determined on a 5-grade scale (G0–G5). T1ρ and T2 values (3D and 2D), and their correlation with each of these parameters were investigated. Results3D T1 ρ is negatively correlated with histological evaluations and positively correlated with visual assessments. Only 3D T1ρ values correlated with histological quantitative evaluation (Safranin-O staining; r = −0.53, P = 0.014, Collagen type II antibody staining; r = −0.60, P = 0.019). 2D T1ρ and 3D, 2D T2 values did not correlate with histological evaluation results. Macroscopic cartilage severity grade correlated with all T1ρ and T2 values (3D T1ρ; r = 0.61, P < 0.001, 2D T1ρ; r = 0.52, P < 0.001, 3D T2; r = 0.33, P = 0.045, 2D T2; r = 0.41, P = 0.01). Conclusions3D T1ρ mapping reflects the changes in the molecular structure of the cartilage matrix that occur in arthropathic changes and may be an effective tool for detecting cartilage degeneration.