2017 Volume 50 Issue 2 Pages 142-148
Chitosan has been widely utilized as a porous scaffold material for cartilage regeneration. During the preparation process of chitosan scaffolds, toxic chemical cross-linkers, such as glutaraldehyde, have been used for cross-linking of chitosan molecules. We previously developed chemical cross-linker-free chitosan sponges formed only by freeze-thawing an aqueous solution of chitosan-gluconic acid conjugate (CG). In the present study, we applied the CG cryosponges as scaffolds for cartilage tissue engineering. CG cryosponges with low gluconic acid content showed a defined porous structure and high stability in cell culture medium supplemented with fetal bovine serum that originally contained a low level of lysozyme, a hydrolysis enzyme for chitosan. A favorable mean pore size of the cryosponges for cartilage regeneration (100–300 µm) was accomplished by freezing the CG solution at −30°C. Proliferation and glycosaminoglycan production of bovine chondrocytes were facilitated in plain CG cryosponges compared with those cross-linked with glutaraldehyde. These results demonstrated that the CG cryosponges were promising scaffolds for cartilage tissue engineering.