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We have developed a new integrated approach for quantitative computed tomography of the knee in order to quantify bone mineral density (BMD) and subchondral bone structure. The present framework consists of image acquisition and reconstruction, 3-D segmentation, determination of anatomic coordinate systems, and reproducible positioning of analysis volumes of interest (VOI). Novel segmentation algorithms were developed to identify growth plates of the tibia and femur and the joint space with high reproducibility. Five different VOIs with varying distance to the articular surface are defined in the epiphysis. Each VOI is further subdivided into a medial and a lateral part. In each VOI, BMD is determined. In addition, a texture analysis is performed on a high-resolution computed tomography (CT) reconstruction of the same CT scan in order to quantify subchondral bone structure. Local and global homogeneity, as well as local and global anisotropy were measured in all VOIs. Overall short-term precision of the technique was evaluated using double measurements of 20 osteoarthritic cadaveric human knees. Precision errors for volume were about 2–3% in the femur and 3–5% in the tibia. Precision errors for BMD were about 1–2% lower. Homogeneity parameters showed precision errors up to about 2% and anisotropy parameters up to about 4%.