Abstract: Magnetic resonance imaging (MRI) is a common clinical practice to visualize defects and to distinguish different tissue types and pathologies in the human body. So far, MRI data have not been used to model and generate a patient-specific design of multilayered tissue substitutes in the case of interfacial defects. For orthopedic cases that require highly individual surgical treatment, implant fabrication by additive manufacturing holds great potential. Extrusion-based techniques like 3D plotting allow the spatially defined application of several materials, as well as implementation of bioprinting strategies. With the example of a typical multi-zonal osteochondral defect in an osteochondritis dissecans (OCD) patient, this study aimed to close the technological gap between MRI analysis and the additive manufacturing process of an implant based on different biomaterial inks. A workflow was developed which covers the processing steps of MRI-based defect identification, segmentation, modeling, implant design adjustment, and implant generation. A model implant was fabricated based on two biomaterial inks with clinically relevant properties that would allow for bioprinting, the direct embedding of a patient’s own cells in the printing process. As demonstrated by the geometric compatibility of the designed and fabricated model implant in a stereolithography (SLA) model of lesioned femoral condyles, a novel versatile CAD/CAM workflow was successfully established that opens up new perspectives for the treatment of multi-zonal (osteochondral) defects. Graphic abstract: [Figure not available: see fulltext.]