Articular cartilage is generally known to be a complex tissue with multiple layers. Each layer has different composition, structure, and mechanical properties, making regeneration after knee joint defects a troubling clinical problem. A novel integrated stratified decalcified bone matrix (SDBM) scaffold with different stiffness to mimic the mechanical properties of articular cartilage is presented herein. This SDBM scaffold was modified using fibrinogen (Fg) (Fg + SDBM) to enhance its vascularization ability and improve its repair efficiency for osteochondral defects of knee joints. A Fg + SDBM scaffold with different elastic modulus in each layer (high-stiffness DBM (HDBM) layer, 174.208 ± 44.330 MPa (Fg + HDBM); medium-stiffness DBM (MDBM) layer, 21.214 ± 6.922 MPa (Fg + MDBM); and low-stiffness DBM (LDBM) layer, 0.678 ± 0.269 MPa (Fg + LDBM)) was constructed by controlling the stratified decalcification time with layered embedding paraffin (0, 3, and 5 days). The low-and medium-stiffness layers of the Fg + SDBM scaffold remarkably promoted the cartilage differentiation of bone marrow mesenchymal stem cells in vitro. Subcutaneous transplantation and rabbit knee joint osteochondral defect repair experiments revealed that the low-and medium-stiffness layers of the Fg + SDBM scaffold exhibited wonderful cartilage capacity, whereas the high-stiffness layer of Fg + SDBM scaffold exhibited good osteogenesis ability. Furthermore, this scaffold could promote blood vessel formation in subchondral bone area. This study presents a feasible strategy for osteochondral regeneration of defective knee joints, which is of great clinical value for tissue repair.
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