Directing Chondrogenesis of Primary Chondrocytes by Exposure to Glucose Concentrations

Article Preview

Abstract:

Since articular cartilage is avascular, both nutrient supply and metabolic waste excretion depend on diffusion. However, the major cause of the progression of articular cartilage defect is the poor inherent regenerative capacity of chondrocytes which limits the process of cartilage tissue repair. Creation of nutrient gradients in in vitro cell culture, however, can provide a clue on zonal distributions of cells and glycosaminoglycan synthesis throughout the tissue engineered cartilage. We hypothesized that glucose gradient, in combination with growth factors, could induce differences in matrix distributions for articular cartilage regeneration. Chondrocytes were harvested from bovine cartilage and expanded in monolayers. First, either p0 or p2 chondrocytes were differentiated in serum-free chondrogenic medium containing different glucose concentrations supplemented with TGFβ3/dex or IGF-1under hypoxic or normoxic conditions for 7 days in monolayer. The results indicate that cellular metabolism, cell numbers and glycosaminoglycan (GAG) content increased with increase in glucose concentration in all conditions. Aggrecan (AGC) expression consistently increased with decreasing glucose concentration in both normoxic and hypoxic conditions. COL II and COL I expressions increased with increasing glucose concentration up to 5mmol/L. The expression of COMP increased with increasing glucose concentration under hypoxic conditions and interestingly showed an opposite trend under normoxic conditions. However, comparing the chondrogenic capacity of p0 and p2 cells in the different glucose concentrations did not show differences, but the potential of p2 cells was in general lower compared to p0. Hypoxia had stimulatory effects on matrix production compared to normoxia in both passages. Therefore, supplemented glucose concentration in monolayer could induce differences in matrix production, but the chondrogenic potential remained equal. Therefore, this information could be use to a create gradients through a tissue-engineered cartilage.

You might also be interested in these eBooks

Info:

Pages:

30-42

Citation:

Online since:

July 2015

Export:

Price:

Permissions CCC:

Permissions PLS:

Сopyright:

© 2015 Trans Tech Publications Ltd. All Rights Reserved

Share:

Citation:

* - Corresponding Author

[1] Woodfield, T.B., et al., Polymer scaffolds fabricated with pore-size gradients as a model for studying the zonal organization within tissue-engineered cartilage constructs. Tissue Eng, 2005. 11(9-10): pp.1297-311.

DOI: 10.1089/ten.2005.11.1297

Google Scholar

[2] Mithoefer, K., et al., Chondral resurfacing of articular cartilage defects in the knee with the microfracture technique. Surgical technique. J Bone Joint Surg Am, 2006. 88 Suppl 1 Pt 2: pp.294-304.

DOI: 10.2106/jbjs.f.00292

Google Scholar

[3] Brittberg, M., et al., Treatment of deep cartilage defects in the knee with autologous chondrocyte transplantation. N Engl J Med, 1994. 331(14): pp.889-95.

DOI: 10.1056/nejm199410063311401

Google Scholar

[4] Wong, B.J., et al., Identification of chondrocyte proliferation following laser irradiation, thermal injury, and mechanical trauma. Lasers Surg Med, 2005. 37(1): pp.89-96.

DOI: 10.1002/lsm.20180

Google Scholar

[5] Bobic, V. and J. Noble, Articular cartilage--to repair or not to repair. J Bone Joint Surg Br, 2000. 82(2): pp.165-6.

DOI: 10.1302/0301-620x.82b2.0820165

Google Scholar

[6] Lynn, A.K., et al., Repair of defects in articular joints. Prospects for material-based solutions in tissue engineering. J Bone Joint Surg Br, 2004. 86(8): pp.1093-9.

DOI: 10.1302/0301-620x.86b8.15609

Google Scholar

[7] Redman, S.N., S.F. Oldfield, and C.W. Archer, Current strategies for articular cartilage repair. Eur Cell Mater, 2005. 9: pp.23-32; discussion 23-32.

Google Scholar

[8] Mow, V.C., A. Ratcliffe, and A.R. Poole, Cartilage and diarthrodial joints as paradigms for hierarchical materials and structures. Biomaterials, 1992. 13(2): pp.67-97.

DOI: 10.1016/0142-9612(92)90001-5

Google Scholar

[9] Darling, E.M., J.C. Hu, and K.A. Athanasiou, Zonal and topographical differences in articular cartilage gene expression. J Orthop Res, 2004. 22(6): pp.1182-7.

DOI: 10.1016/j.orthres.2004.03.001

Google Scholar

[10] Aydelotte, M.B., R.R. Greenhill, and K.E. Kuettner, Differences between sub-populations of cultured bovine articular chondrocytes. II. Proteoglycan metabolism. Connect Tissue Res, 1988. 18(3): pp.223-34.

DOI: 10.3109/03008208809016809

Google Scholar

[11] Hunziker, E.B., T.M. Quinn, and H.J. Hauselmann, Quantitative structural organization of normal adult human articular cartilage. Osteoarthritis Cartilage, 2002. 10(7): pp.564-72.

DOI: 10.1053/joca.2002.0814

Google Scholar

[12] Otte, P., Basic cell metabolism of articular cartilage. Manometric studies. Z Rheumatol, 1991. 50(5): pp.304-12.

Google Scholar

[13] Cernanec, J., et al., Influence of hypoxia and reoxygenation on cytokine-induced production of proinflammatory mediators in articular cartilage. Arthritis and rheumatism, 2002. 46(4): pp.968-75.

DOI: 10.1002/art.10213

Google Scholar

[14] Grimshaw, M.J. and R.M. Mason, Bovine articular chondrocyte function in vitro depends upon oxygen tension. Osteoarthritis and cartilage / OARS, Osteoarthritis Research Society, 2000. 8(5): pp.386-92.

DOI: 10.1053/joca.1999.0314

Google Scholar

[15] Gibson, J.S., et al., Oxygen and reactive oxygen species in articular cartilage: modulators of ionic homeostasis. Pflugers Archiv : European journal of physiology, 2008. 455(4): pp.563-73.

DOI: 10.1007/s00424-007-0310-7

Google Scholar

[16] Di Cesare, P.E., et al., Increased degradation and altered tissue distribution of cartilage oligomeric matrix protein in human rheumatoid and osteoarthritic cartilage. J Orthop Res, 1996. 14(6): pp.946-55.

DOI: 10.1002/jor.1100140615

Google Scholar

[17] Murray, R.C., et al., The distribution of cartilage oligomeric matrix protein (COMP) in equine carpal articular cartilage and its variation with exercise and cartilage deterioration. Vet J, 2001. 162(2): pp.121-8.

DOI: 10.1053/tvjl.2001.0590

Google Scholar

[18] Leonard, C.M., et al., Abnormal ambient glucose levels inhibit proteoglycan core protein gene expression and reduce proteoglycan accumulation during chondrogenesis: possible mechanism for teratogenic effects of maternal diabetes. Proc Natl Acad Sci U S A, 1989. 86(24): pp.10113-7.

DOI: 10.1073/pnas.86.24.10113

Google Scholar

[19] Windhaber, R.A., R.J. Wilkins, and D. Meredith, Functional characterisation of glucose transport in bovine articular chondrocytes. Pflugers Arch, 2003. 446(5): pp.572-7.

DOI: 10.1007/s00424-003-1080-5

Google Scholar

[20] Zhao, K., et al., Effect of glucose availability on glucose transport in bovine mammary epithelial cells. Animal, 2012. 6(3): pp.488-93.

DOI: 10.1017/s1751731111001893

Google Scholar

[21] Lane, J.M., C.T. Brighton, and B.J. Menkowitz, Anaerobic and aerobic metabolism in articular cartilage. J Rheumatol, 1977. 4(4): pp.334-42.

Google Scholar

[22] Rajpurohit, R., et al., Adaptation of chondrocytes to low oxygen tension: relationship between hypoxia and cellular metabolism. J Cell Physiol, 1996. 168(2): pp.424-32.

DOI: 10.1002/(sici)1097-4652(199608)168:2<424::aid-jcp21>3.0.co;2-1

Google Scholar

[23] Ysart, G.E. and R.M. Mason, Responses of articular cartilage explant cultures to different oxygen tensions. Biochim Biophys Acta, 1994. 1221(1): pp.15-20.

DOI: 10.1016/0167-4889(94)90210-0

Google Scholar

[24] Johnson, W.E., S. Stephan, and S. Roberts, The influence of serum, glucose and oxygen on intervertebral disc cell growth in vitro: implications for degenerative disc disease. Arthritis Res Ther, 2008. 10(2): p. R46.

DOI: 10.1186/ar2405

Google Scholar