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Because cartilage has limited potential for self-repair, tissue engineering is expected to replace the present therapies for damaged cartilage, such as total knee arthroplasty. However, scaffolds suitable for cartilage tissue engineering have not been established. We synthesized a novel porous scaffold, a collagen sponge incorporating a hydroxyapatite/chondroitinsulfate composite (pCol-HAp/ChS), containing materials which resemble extracellular matrices in bone and cartilage tissues. In this report, the physical, mechanical and biological properties of the scaffold are compared with those of a collagen sponge (pCol) and pCol incorporating a hydroxyapatite composite (pCol-HAp). HAp/ChS had smaller crystals and a larger total surface area than HAp. SEM images of the three materials showed pCol-HAp/ChS to have the roughest surface. The mechanical properties suggest that pCol-HAp/ChS and pCol/HAp are similar, and superior to pCol. Seeding experiments showed a uniform distribution of mesenchymal stem cells (MSCs) in pCol-HAp/ChS and pCol/HAp. Histochemical staining after 2 weeks of culture revealed pCol-HAp/ChS to be the most chondrogenic. From these results, pCol-HAp/ChS is expected to be a candidate for a scaffold for cartilage tissue engineering in place of collagen sponge.