Porous scaffolds of alkaline-soluble collagen including nanocomposite particles of chondroitin sulfate and low crystalline hydroxyapatite for cartilage regeneration were fabricated by freeze-drying and thermal dehydration treatments; porous collagen scaffolds were also synthesized as a reference. The scaffolds were cross-linked using glutaraldehyde (GA) vapor treatment in order to enhance biodegradable resistance. Microstructural observation with scanning electron microscope indicated that the scaffolds with and without GA cross-linkage had open pores between 130 to 200 μm in diameter and well-interconnected pores of 10 to 30 μm even after cross-linkage. In vitro biodegradable resistance to collagenase was significantly enhanced by GA cross-linking of the scaffolds. All these results suggest that the GA cross-linked scaffolds consisting of collagen, chondroitin sulfate, and low crystalline hydroxyapatite have suitable microporous structures and long-term biochemical stability for cartilage tissue engineering.