Objective To explore the feasibility of achieving stable in vivo cartilage regeneration using demineralized bone matrix-gelatin （DBM-GT） composite scaffolds combined with BMSC-chondrocyte co-culture technology. Methods  DBM-GT composite scaffolds were prepared via gelatin freeze-drying cross-linking technology， and bone marrow mesenchymal stem cells （BMSCs） were seeded to determine cell loading efficiency. Live/dead staining of scaffold-loaded cells was performed to evaluate the role of chondrocytes in regulating BMSC proliferation. Subsequently， a mixture of BMSCs and chondrocytes at a 1∶4 ratio was seeded onto the composite scaffold as the experimental group， while chondrocytes alone were seeded as the control group. After 5 days of in vitro culture， both groups were implanted subcutaneously into nude mice. Post-harvest histological examination was conducted to assess differences in cartilage regeneration between the experimental and control groups. Results The DBM-GT scaffold exhibited superior porous structure compared to the DBM scaffold. Live/ dead staining and DNA quantitative analysis demonstrated that chondrocytes effectively promoted BMSC proliferation. No significant difference in cartilage regeneration was observed between the experimental group （BMSC-chondrocyte co-culture） and the control group （chondrocyte-only） in vivo. Conclusion The DBM-GT gelatin scaffold can be successfully 

Decalcified bone matri, Gelatin, Scaffold material, Co-culture, Cartilage regeneration