Spontaneous tissue healing of focal osteochondral defects does not provide tissue with the same properties as the native one. Tissue engineering aims to repair these injured tissues. One of the pitfalls is the difficulty of reproducing a full implant considering this tissue’s composition and architecture (non-calcified cartilage, calcified cartilage, and subchondral bone (SB)). Newly developed implants must integrate the complexity of this tissue and the pivot zone with the SB to promote successful grafting. The multi-layered biomaterial will be obtained by bio-extrusion’s 3D printing. This process enables the implant to be printed in a single piece and allows the bio-ink composition to be topographically adapted. To better prepare the composition of the bio-inks, we decided to characterize the organization and composition of this osteochondral junction (OJ) from patients undergoing knee surgery. The modified Mankin’s score (MS) provides some evidence that the organization of the cartilage and SB does not correlate with the pathological evolution of OJ. However, a correlation is evident between the Kellgren-Lawrence score and the MS. In the context of reproducing the pivot zone, the results indicate that a detailed analysis of OJ is required to design bio-inks to produce a suitable zone for treating focal osteochondral defects.