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Toxic Chemical Cross-linker-free Cryosponges Made from Chitosan-Gluconic Acid Conjugate for Chondrocyte Culture

  • Takei Takayuki
    Department of Chemical Engineering, Graduate School of Science and Engineering, Kagoshima University
  • Yoshitomi Hiroki
    Department of Chemical Engineering, Graduate School of Science and Engineering, Kagoshima University
  • Fukumoto Kohei
    Department of Chemical Engineering, Graduate School of Science and Engineering, Kagoshima University
  • Danjo So
    Department of Chemical Engineering, Graduate School of Science and Engineering, Kagoshima University
  • Yoshinaga Takuma
    Division of Clinical Application, Nanpuh Hospital
  • Nishimata Hiroto
    Department of Gastroenterology, Nanpuh Hospital
  • Yoshida Masahiro
    Department of Chemical Engineering, Graduate School of Science and Engineering, Kagoshima University

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Abstract

<p>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.</p>

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