Use of Neonatal Chondrocytes for Cartilage Tissue Engineering

  • KANG SUN WOONG (Department of Chemical Engineering, Hanyang University) ;
  • PARK JUNG HO (Department of Orthopeadic Surgery, Ansan Hospital, College of Medicine, Korea University) ;
  • KIM BYUNG SOO (Department of Chemical Engineering, Hanyang University)
  • 발행 : 2005.04.01

초록

Transplantation of cultured chondrocytes can regenerate cartilage tissues in cartilage defects in humans. However, this method requires a long culture period to expand chondrocytes to a large number of cells for transplantation. In addition, chondrocytes may dedifferentiate during long-term culture. These problems can potentially be overcome by the use of undifferentiated or partially developed cartilage precursor cells derived from neonatal cartilage, which, unlike chondrocytes from adult cartilage, have the capacity for rapid in vitro cell expansion and may retain their differentiated phenotype during long-term culture. The purpose of this study was to compare the cell growth rate and phenotypic modulation during in vitro culture between adult chondrocytes and neonatal chondrocytes, and to demonstrate the feasibility of regenerating cartilage tissues in vivo by transplantation of neonatal chondrocytes expanded in vitro and seeded onto polymer scaffolds. When cultured in vitro, chondrocytes isolated from neonatal (immediately postpartum, 2 h of age) rats exhibited much higher growth rate than chondrocytes isolated from adult rats. After 5 days of culture, more neonatal chondrocytes were in the differentiated state than adult chondrocytes. Cultured neonatal chondrocytes were seeded onto biodegradable polymer scaffolds and transplanted into athymic mice's subcutaneous sites. Four weeks after implantation, neonatal chondrocyte-seeded scaffolds formed white cartilaginous tissues. Histological analysis of the implants with hematoxylin and eosin showed mature and well-formed cartilage. Alcian blue/ safranin-O staining and Masson's trichrome staining indicated the presence of highly sulfated glycosarninoglycans and collagen, respectively, both of which are the major extracellular matrices of cartilage. Immunohistochemical analysis showed that the collagen was mainly type II, the major collagen type in cartilage. These results showed that neonatal chondrocytes have potential to be a cell source for cartilage tissue engineering.

키워드

참고문헌

  1. Benya, P. D., S. R Padilla, and M. E. Nimni. 1978. Independent regulation of collagen types by chondrocytes during the loss of differentiated function in culture. Cell 15: 1313- 1321 https://doi.org/10.1016/0092-8674(78)90056-9
  2. Benya, P. D. and J. D. Shaffer. 1982. Dedifferentiated chondrocytes reexpress the differentiated collagen phenotype when cultured in agarose gels. Cell 30: 215- 224 https://doi.org/10.1016/0092-8674(82)90027-7
  3. Brittberg, M., A. Lindahl, A. Nilsson, C. Ohlsson, O. Isaksson, and L. Peterson. 1994. Treatment of deep cartilage defects in knee with autologous chondrocyte transplantation. N. Engl. J. Med. 331: 889- 895 https://doi.org/10.1056/NEJM199410063311401
  4. Buckwalter, J. A. and H. J. Mankin. 1998. Articular cartilage: Degeneration and osteoarthritis, repair, regeneration, and transplantation. lnstr. Course Lect. 47: 487- 504
  5. Chung, M. I., M. H. Lim, Y. J. Lee, I. H. Kim, I. Y. Kim, J. H. Kim, K. Chang, and H. Kim. 2003. Reduction of ammonia accumulation and improvement of cell viability by expression of urea cycle enzymes in Chinese hamster ovary cells. J. Microbiol. Biotechnol. 13: 217 - 224
  6. Convery, F. R., W. H. Akeson, D. Amiel, M. H. Meyers, and A. Monosov, 1996. Long-term survival of chondrocytes in an osteochondral articular cartilage allograft. A case report. J. Bone Joint Surg. Am. 78: 1082- 1088 https://doi.org/10.2106/00004623-199607000-00013
  7. Dekel, B., T. Burakova, F. D. Arditti, S. Reich-Zeliger, O. Milstein, S. Aviel-Ronen, G. Rechavi, N. Friedman, N. Kaminski, J. H. Passwell, and Y. Reisner. 2003. Human and porcine early kidney precursors as a new source for transplantation. Nat. Med. 9: 53- 60 https://doi.org/10.1038/nsb738
  8. Dominice, J., C. Levasseur, S. Larno, X. Ronot, and M. Adolphe. 1986. Age-related changes in rabbit articular chondrocytes, Mech. Ageing Dev. 37: 231- 240 https://doi.org/10.1016/0047-6374(86)90040-0
  9. Dozin, B., M. Malpeli, L. Camardella, R. Cancedda, and A. Pietrangelo. 2002. Response of young, aged and osteoarthritic human articular cnondrocytes to inflammatory cytokines: Molecular and cellular aspects. Matrix Biology 21: 449-459 https://doi.org/10.1016/S0945-053X(02)00028-8
  10. Fuchs, J. R., S. Terada, D. Hannouche, E. R. Ochoa, J. P. Vacanti, and D. O. Fauza, 2003. Fetal tissue engineering: Chest wall reconstruction. J. Pediatr. Surg. 38: 1188- 1193 https://doi.org/10.1016/S0022-3468(03)00265-3
  11. Ghazavi, M. T., K. P. Pritzker, A. M. Davis, and A. E. Gross. 1997. Fresh osteochondral allografts for post-traumatic osteochondral defects of the knee. J. Bone Joint Surg, Br. 79: 1008- 1013 https://doi.org/10.1302/0301-620X.79B6.7534
  12. Kim, B., S. I. Jeong, S. Cho, J. Nikolovski, D. J. Mooney, S. H. Lee, O. Jeon, T. W. Kim, S. H. Lim, Y. S. Hong, C. Y. Choi, Y. M. Lee, S. H. Kim, and Y. H. Kim. 2003. Tissue engineering of smooth muscle under a mechanically dynamic condition. J. Microbiol. Biotechnol. 13: 841- 845
  13. Mayne, R., M. S. Vail, P. M. Mayne, and E. J. Miller. 1976. Changes in type of collagen synthesized as clones of chick chondrocytes grow and eventually lose division capacity. Proc, Natl. Acad. Sci. USA 73: 1674- 1678
  14. Meyers, M. H., W Akeson, and F. R. Convery. 1989. Resurfacing of the knee with fresh osteochondral allograft. J. Bone Joint Surg. Am. 71: 704- 713 https://doi.org/10.2106/00004623-198971050-00010
  15. Minas, T. and S. Nehrer. 1997. Current concepts in the treatment of articular cartilage defects. Orthopedics 20: 525- 538
  16. Morihara, T., F. Harwood, R. Goerner, Y. Hirasawa, and D. Amiel. 2002. Tissue-engineered repair of osteochondral defects: Effects of the age of donor cells and host tissue. Tissue Eng. 8: 921- 929 https://doi.org/10.1089/107632702320934029
  17. Na, K. and K. Park. 2004. Immobilization of Arg-Gly-Asp (RGD) sequence in sugar-containing copolymer for culturing fibroblast cells. J. Microbiol. Biotechnol. 14: 193- 196
  18. Puelacher, W. C., S. W. Kim, J. P. Vacanti, B. Schloo, D. Mooney, and C. A. Vacanti. 1994. Tissue-engineered growth of cartilage: The effect of varying the concentration of chondrocytes seeded onto synthetic polymer matrices. J. Oral Maxillofac. Surg. 23: 49- 53 https://doi.org/10.1016/S0901-5027(05)80328-5
  19. Rotter, N., L. J. Bonassar, G. Tobias, M. Lebl, A. K. Roy, and C. A. Vacanti. 2002. Age dependence of biochemical and biomechanical properties of tissue-engineered human septal cartilage. Biomaterials 23: 3087- 3094 https://doi.org/10.1016/S0142-9612(02)00031-5
  20. Veilleux, N. H., I. V. Yannas, and M. Spector. 2004. Effect of passage number and collagen type on the proliferative, biosynthetic, and contractile activity of adult canine articular chondrocytes in type I and II collagen-glycosaminoglycan matrices in vitro. Tissue Eng. 10: 119- 127 https://doi.org/10.1089/107632704322791763
  21. von der Mark, K., V. Gauss, H. von der Mark, and P. Muller. 1977. Relationship between cell shape and type of collagen synthesised as chondrocytes lose their cartilage phenotype in culture. Nature 9: 531- 532
  22. Yang, B., S. Jeong, and C. Song. 2002. Hypolipidemic effect of exo- and endo-biopolymers produced from submerged mycelial culture of Ganoderma lucidum in rats. J. Microbiol. Biotechnol. 12: 872- 877