Summary
Primary, high density bovine articular chondrocyte (BAC) cultures, stimulated with transforming growth factor-β-1, elaborated a high molecular weight anionic glycoconjugate, kDa 540, which does not contain glycosaminoglycan chains (Chan and Anastassiades, 1996). The effect of exogenously added transforming growth factor-β-1 on the elaboration of the high molecular weight glycoconjugate and of proteoglycans was studied during dedifferentiation of the chondrocytes, utilizing a serial subculture technique under anchorage-dependent conditions, up to four subcultures. The high molecular weight glycoconjugate was detected in the media of all growth-factor-stimulated chondrocyte subcultures, as well as stimulated primary cultures, but not in unstimulated primary cultures or subcultures. By contrast, a large proteoglycan, was only secreted by primary cultures and first subcultures, whether treated with transforming growth factor-β-1 or untreated. This proteoglycan contained mostly chondroitin sulfate chains, whose hydrodynamic size was increased by the addition of transforming growth factor-β-1. Further, the pattern of the proteoglycans appearing in the media of subcultures 2–4 was influenced by the addition of transforming growth factor-β-1, so that while these control subcultures elaborated both the large and small chondroitin sulfate proteoglycans, the equivalent stimulated subcultures elaborated only intermediate sized chondroitin sulfate proteoglycan(s). These results suggest that while dedifferentiation of articular chondrocytes, achieved by subculturing, strongly modulates the effect of exogenously added transforming growth factor-β-1 on the type of proteoglycan elaborated, the process of dedifferentiation does not influence the transforming-growth-factor-β-dependent synthesis of the high molecular weight anionic glycoconjugate.
Similar content being viewed by others
References
Athanassiadis, A.; Anastassiades, T. P. A “coupled” subchondral bone-articular cartilage culture system for the study of proteoglycan metabolism. In Vitro Cell. Dev. Biol. 30A:504–511; 1994.
Bjornsson, S.; Heinegard, D. Isolation and culture techniques of foetal calf chondrocytes. Biochem. J. 198:141–148; 1981.
Chan, K. T. C.; Anastassiades, T. P. Isolation and partial characterization of a high molecular weight anionic glycoconjugate from transforming growth factor-β treated bovine articular chondrocyte cultures. Biochem. Cell Biol. 74:233–240; 1996.
Clark, J. M. The structure of vascular channels in the subchondral plate. J. Anat. 171:105–115; 1990.
Fife, R.; Brandt, K. Identification of a high molecular weight (>400,000) protein in hyaline cartilage. Biochim. Biophys. Acta 802:506–514; 1984.
Galera, P.; Redini, F.; Vivien, D., et al. Effect of transforming growth factor beta (TGF-beta 1) on matrix synthesis by monolayer cultures of rabbit articular chondrocytes during the dedifferentiation process. Exp. Cell Res. 200:379–392; 1992.
Glowacki, J.; Trepman, E.; Folkman, J. Cell shape and phenotypic expression in chondrocytes. Proc. Soc. Exp. Biol. Med. 172:93–98; 1983.
Hedbom, E.; Antonsson, P.; Hjerpe, A., et al. Cartilage matrix proteins: an acidic oligomeric protein (COMP) detected only in cartilage. J. Biol. Chem. 267:6132–6236; 1992.
Howard, S.; Anastassiades, T. Differential effects of bone-associated factors on newly synthesized anionic glycoconjugates by articular chondrocyte cultures from adult and immature bovines. J. Rheumatol. 20:2083–2094; 1993.
Kjellen, L.; Lindahl, U. Proteoglycans structures and interactions. Annu. Rev. Biochem. 60:443–475; 1991.
Kuettner, K.; Pauli, B.; Gall, G., et al. Synthesis of cartilage matrix by mammalian chondrocytes in vitro. I. Isolation, culture characteristics and morphology. J. Cell Biol. 93:743–750; 1982.
Lafeber, F. P.; Vander Kraan, P. M.; Huber-Bruning, O., et al. Osteoarthritic cartilage is more sensitive to transforming growth factor beta than is normal cartilage. Br. J. Rheumatol. 32:281–286; 1993.
Malemud, C. J.; Killeen, W.; Hering, T. M., et al. Enhanced sulfated-proteoglycan core protein synthesis by incubation of chondrocytes with recombinant growth factor-beta 1. J. Cell. Physiol. 149:152–159; 1991.
Mitchell, N.; Shepard, N. The resurfacing of articular cartilage by multiple perforations through the subchondral bone. J. Bone Jt. Surg. 58A:230–233; 1976.
Morales, T. Transforming growth factor-β1 stimulates synthesis of proteoglycan aggregates in calf articular cartilage organ cultures. Arch. Biochem. Biophys. 286:99–106; 1991.
Morales, T.; Roberts, A. Transforming growth factor β regulates the metabolism of proteoglycans in bovine cartilage organ cultures. J. Biol. Chem. 263:12828–12831; 1988.
Muir, H. Structure, biology and pathology of proteoglycans. Biochem. Soc. Trans. 18:787–789; 1990.
Ŕedini, F.; Galera, P.; Mauviel, A., et al. Transforming growth factor β stimulates collagen glycosaminoglycan biosynthesis in cultured rabbit articular chondroctyes. FEBS Lett. 234:172–176; 1988.
Schiltz, J.; Mayne, R.; Holtzer, H. Differences among sulfated proteoglycans synthesized in nonchondrogenic cell, presumptive chondroblasts, and chondroblasts. Proc. Natl. Acad. Sci. USA 76:1251–1264; 1973.
Seyedin, S.; Segarini, P.; Rosen, D., et al. Cartilage-inducing factor-B is a unique protein structurally and functionally related to transforming growth factor-β. J. Biol. Chem. 262:1946–1949; 1987.
Seyedin, S.; Thompson, A.; Bentz, H., et al. Cartilage-inducing factor-A. Apparent identity to transforming growth factor-β. J. Biol. Chem. 261:5693–5695; 1986.
Sokoloff, L. Ageing and degenerative diseases affecting cartilage. In: Hall, B. K., ed. Cartilage. Vol. 3. New York: Academic Press; 1983:109–142.
Sporn, M. B.; Roberts, A. B. What is TGF-β? In: Bock, G. R.; Marsh, J., ed. Clinical applications of TGF-β. Ciba Foundation Symposium, Vol. 157. New York: John Wiley & Sons; 1991:1–6.
Visser, N.; Brand, H.; Jos Vankampen, G., et al. A high molecular weight (>8 × 105) non-collagenous glycoprotein is synthesized by bovine cartilage in vitro. Biochim. Biophys. Acta 1120:308–314; 1992.
Vogel, K. G.; Hernandez, D. J. The effects of transforming growth factor-beta and serum on proteoglycan synthesis by tendon fibrocartilage. Eur. J. Cell Biol. 59:304–313; 1992.
von der Mark, K. Differentiation, modulation and dedifferentiation of chondrocytes. Rheumatology 10:272–315; 1986.
Wateson, A. A method for the determination of the molecular weight and molecular weight-distribution of chondroitin sulfate. J. Chromatogr. 59:87–97; 1971.
Watt, F. Effect of seeding density on stability of the differentiated phenotype of pig articular cartilage chondrocytes in culture. J. Cell Sci. 89:373–378; 1988.
Wozney, R.; Rosen, V.; Celeste, A., et al. Novel regulators of bone formation: molecular clones and activities. Science 325:81–84; 1988.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Chan, C.K.T., Anastassiades, T.P. Anionic glycoconjugates from differentiated and dedifferentiated cultures of bovine articular chondrocytes: Modulation by TGF-β. In Vitro Cell.Dev.Biol.-Animal 34, 492–498 (1998). https://doi.org/10.1007/s11626-998-0084-z
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/s11626-998-0084-z