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Cell and Tissue Research

, Volume 311, Issue 3, pp 393–399 | Cite as

Stimulation of type II collagen biosynthesis and secretion in bovine chondrocytes cultured with degraded collagen

Regular Article

Abstract

The functional integrity of articular cartilage is dependent on the maintenance of the extracellular matrix (ECM), a process which is controlled by chondrocytes. The regulation of ECM biosynthesis is complex and a variety of substances have been found to influence chondrocyte metabolism. In the present study we have investigated the effect of degraded collagen on the formation of type II collagen by mature bovine chondrocytes in a cell culture model. The culture medium was supplemented with collagen hydrolysate (CH) and biosynthesis of type II collagen by chondrocytes was compared to control cells treated with native type I and type II collagen and a collagen-free protein hydrolysate. The quantification of type II collagen by means of an ELISA technique was confirmed by immunocytochemical detection as well as by the incorporation of 14C-proline in the ECM after a 48 h incubation. Chondrocytes in the control group were maintained in the basal medium for 11 days. The presence of extracellular CH led to a dose-dependent increase in type II collagen secretion. However, native collagens as well as a collagen-free hydrolysate of wheat proteins failed to stimulate the production of type II collagen in chondrocytes. These results clearly indicate a stimulatory effect of degraded collagen on the type II collagen biosynthesis of chondrocytes and suggest a possible feedback mechanism for the regulation of collagen turnover in cartilage tissue.

Keywords

Collagen hydrolysate Collagen secretion Chondrocyte metabolism Type II collagen Cell culture (Bovine) 

Notes

Acknowledgements

We wish to thank Anja Raabe for her excellent technical assistance.

References

  1. Adyelotte MB, Schmid TR, Greenhill RR, Luchene L, Schumacher BL, Kuettner KE (1991) Synthesis of collagen by cultured bovine chondrocytes derived from different depth of articular cartilage. Trans Orthop Res Soc 16:26Google Scholar
  2. Aigner T, Glückert K, von der Mark K (1997) Activation of fibrillar collagen synthesis and phenotypic modulation of chondrocytes in early human osteoarthritic lesions. Osteoarthritis Cartilage 5:183–189Google Scholar
  3. Aycock RS, Raghow R, Stricklin GP, Seyer JM, Kang AH (1986) Post-transcriptional inhibition of collagen and fibronectin synthesis by a synthetic homolog of a portion of the carboxyl-terminal propeotide of human type I collagen. J Biol Chem 261:14355–14360Google Scholar
  4. Bateman JF, Lamandé SR, Ramshaw JAM (1996) Collagen superfamily. In: Comper WD (ed) Extracellular matrix, vol 2. Harwood Academic, Amsterdam, pp 22–67Google Scholar
  5. Benya PD, Padilla SR, Nimni ME (1987) Independent regulation of collagen types by chondrocytes during the loss of differentiated function in culture. Cell 15:1313–1321Google Scholar
  6. Benz K, Breit S, Lukoschek M, Mau H, Richter W (2002) Molecular analysis of expansion, differentiation, and growth factor treatment of human chondrocytes identifies differentiation markers and growth-related genes. Biochem Biophys Res Commun 293:284–292Google Scholar
  7. Buckwalter JA, Mankin HJ (1998) Articular cartilage: degeneration and osteoarthritis, repair, regeneration, and transplantation. Instr Course Lect 47:487–504Google Scholar
  8. Burstein D, Bashir A, Gray ML (2000) MRI techniques in early stages of cartilage disease. Invest Radiol 35:602–621Google Scholar
  9. Elima K, Vuorio E (1989) Expression of mRNAs for collagen and other matrix components in dedifferentiating and redifferentiating human chondrocytes in culture. FEBS Lett 258:195–198Google Scholar
  10. Eyre DR, Wu JJ, Woods PE (1991) The cartilage collagens: structural and metabolic studies. J Rheumatol 18:49–51Google Scholar
  11. Hardingham TE, Bayliss M (1990) Proteoglycans of articular cartilage: changes in aging and in joint disease. Semin Arthritis Rheum 3:12–33Google Scholar
  12. Huber M, Tratting S, Lintner F (2000) Anatomy, biochemistry, and physiology of articular cartilage. Invest Radiol 35:573–580Google Scholar
  13. Katayama K, Seyer JM, Raghow R, Kang AH (1991) Regulation of extracellular matrix production by chemically synthesized subfragments of type I collagen carboxy propeptide. Biochemistry 30:7097–7104Google Scholar
  14. Lotz M, Blanco FJ, von Kempis J, et al. (1995) Cytokine regulation of chondrocyte functions. J Rheumatol 43:104–108Google Scholar
  15. Mayne R (1989) Collagens: what is their function, are they involved in articular disease. Arthritis Rheum 32:241–246Google Scholar
  16. Mlynarik V, Tratting S (2000) Physicochemical properties of normal articular cartilage and its MR appearance. Invest Radiol 35:589–594Google Scholar
  17. Moskowitz RW (2000) Role of collagen hydrolysate in bone and joint disease. Semin Arthritis Rheum 30:87–99Google Scholar
  18. Muir H (1995) The chondrocyte, architect of cartilage: biomechanics, structure, function and molecular biology of cartilage matrix macromolecules. Bioessays 17:1039–1048Google Scholar
  19. Oesser S, Adam M, Babel W, Seifert J (1999) Oral administration of 14C labeled gelatin hydrolysate leads to an accumulation of radioactivity in cartilage of mice (C57/BL). J Nutr 129:1891–1895Google Scholar
  20. Osborne KD, Trppel SB, Mankin HJ (1989) Growth factor stimulation of adult articular cartilage. J Orthop Res 7:35–42Google Scholar
  21. Pasternak RD, Hubbs SJ, Caccese RG, Marks RL, Conaty JM, DiPasquale G (1986) Interleukin-1 stimulates the secretion of proteoglycan- and collagen-degrading proteases by rabbit articular chondrocytes. Clin Immunol 41:351–367Google Scholar
  22. Phadke K, Nanda S (1983) Secretion of chondrocyte stimulating factor by macrophages as a result of activation with collagen and proteoglycans. Clin Exp Immunol 51:494–500Google Scholar
  23. Qi WN, Scully SP (1997) Extracellular collagen modulates the regulation of chondrocytes by transforming growth factor-β1. J Orthop Res 15:483–490Google Scholar
  24. Rosier RN, O'Keefe RJ (1998) Autocrine regulation of articular cartilage. Instr Course Lect 47:469–475Google Scholar
  25. Roughley PJ, Lee ER (1994) Cartilage proteoglycans: structure and potential function. Microsc Res Technol 28:385–397Google Scholar
  26. Roughley PJ, Nguyen Q, Mort SM (1991) Mechanisms of proteoglycan degradation in human articular cartilage. J Rheum 18:52–54Google Scholar
  27. Sandy PJ, Adams ME, Billingham MEJ, Plaas A, Muir H (1984) In vivo and in vitro stimulation of chondrocyte biosynthetic activity in early experimental osteoarthritis. Arthritis Rheum 27:388–397Google Scholar
  28. Sklatvala J (1986) Tumor necrosis factor-a stimulates resorption and inhibits synthesis of proteoglycan in cartilage. Nature 322:547–549Google Scholar
  29. Trippel SB (1995) Growth factor actions on articular cartilage. J Rheumatol 43:129–132Google Scholar
  30. von der Mark K, Gauss V, von der Mark H, Müller P (1977) Relationship between cell shape and type of collagen synthesised as chondrocytes lose their cartilage phenotype in culture. Nature 267:531–532Google Scholar
  31. Zaucke F, Dinser R, Maurer P, Paulsen M (2001) Cartilage oligomeric matrix protein (COMP) and collagen IX are sensitive markers for the differentiation state of articular primary chondrocytes. Biochem J 258:17–24Google Scholar

Copyright information

© Springer-Verlag 2003

Authors and Affiliations

  1. 1.Surgical Research of the Department of General Surgery and Thoracic Surgery of the University of KielKielGermany

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