Evaluation of Chitosan-alginate-hyaluronate Complexes Modified by an RGD-containing Protein as Tissue-engineering Scaffolds for Cartilage Regeneration

Hsu, Shan-hui
Whu, Shu Wen
Hsieh, Shu-Chih
Tsai, Ching-Lin
Chen, David Chanhen
Tan, Tai-Sheng

^*Department of Chemical Engineering, National Chung Hsing University, Taichung, Taiwan
†Department of Chemistry, National Chung Hsing University, Taichung, Taiwan
‡Department of Orthopaedics, National Taiwan University Hospital, Taipei, Taiwan
§Graduate Institute of Veterinary Microbiology, National Chung Hsing University, Taichung, Taiwan
#Department of Orthopaedics, Taichung Veterans General Hospital, Taichung, Taiwan

Address Correspondence and reprint requests to Dr. Shan-hui Hsu, Department of Chemical Engineering, National Chung Hsing University, 250 Kuo Kung Road, Taichung, Taiwan 402. E-mail: [email protected]

Acknowledgments: IRC were kindly supplied by Drs. W. Horton and T. Liang at the Gerontology Research Center, National Institute of Aging, National Institutes of Health, USA. Sodium hyaluronate were kindly supplied by Hong Ru Yang Corporation of Taiwan, R.O.C. This study was funded by National Science Council of Taiwan, R.O.C., via a grant (NSC89-2320-B-002-160-M08), and was conducted in the Center of Tissue Engineering and Stem Cells Research (TESC), National Chung Hsing University.

Received July 2003; revised February 2004.

Artificial Organs 28(8):p 693-703, August 2004.

In this study, a series of natural biodegradable materials in the form of chitosan (C)-alginate (A)-hyaluronate (H) complexes are evaluated as tissue-engineering scaffolds. The weight ratio of C/A is 1: 1 or 1: 2. Sodium hyaluronate is mixed in 2%. The complexes can be cast into films or fabricated as scaffolds. Their surface can be further modified by an Arg-Gly-Asp (RGD)-containing protein, a cellulose-binding domain-RGD (R). Cytocompatibility tests of the films are conducted using immortalized rat chondrocyte (IRC) as well as primary articular chondrocytes harvested from rabbits. The neocartilage formation in cell-seeded scaffolds is examined in vitro as well as in rabbits, where the scaffolds are implanted into the defect-containing joints. The results from cytocompatibility tests demonstrate that R enhances cell attachment and proliferation on C-A and C-A-H complex films. Complex C1A1HR (C: A = 1: 1 with H and R) has better performance than the other formulation. Cells retain their spherical morphology on all C-A and C-A-H complexes. The in vitro evaluation of the seeded scaffolds indicates that the C1A1HR complex is the most appropriate for 3-D culture, manifested by the better cell growth as well as higher glycosaminoglycan and collagen contents. When the chondrocyte scaffolds are implanted into rabbit knee cartilage defects, partial repair is observed after 1 month in C1A1HR as well as in C1A1 (C: A = 1: 1 without H and R) scaffolds. The defects are completely repaired in 6 months when C1A1HR constructs are implanted. It is concluded that C1A1HR is a potential tissue-engineering scaffold for cartilage regeneration.