Osteochondral (OC) defects pose an enormous challenge with no entirely satisfactory repair strategy to date. Herein, a 3D printed gradient hydrogel scaffold with a similar structure to that of OC tissue is designed, involving a pure hydrogel‐based top cartilage layer, an intermediate layer for calcified cartilage with 40% (w w−1) nanohydroxyapatite (nHA) and 60% (w w−1) hydrogel, and a 70/30% (w w−1) nHA/hydrogel‐based bottom subchondral bone layer. This study is conducted to evaluate the efficacy of the scaffold with nHA gradients in terms of its ability to promote OC defect repair. The fabricated composites are evaluated for physicochemical, mechanical, and biological properties, and then implanted into the OC defects in 56 rats. Overall, bone marrow stromal cells (BMSCs)‐loaded gradient scaffolds exhibit superior repair results as compared to other scaffolds based on gross examination, micro‐computed tomography (micro‐CT), as well as histologic and immunohistochemical analyses, confirming the ability of this novel OC graft to facilitate simultaneous regeneration of cartilage‐subchondral bone.
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