Abstract
The objective of the study was to evaluate the growth-promoting activity of human platelet supernanant on primary chondrocytes in comparison with fetal calf serum (FCS) supplemented cell culture medium. Furthermore, the differentiation potential of platelet supernatant was determined in three-dimensional artificial cartilage tissues of bovine articular chondrocytes. Proliferation of articular and nasal septal chondrocytes was assayed by incorporation of BrdU upon stimulation with ten different batches of human platelet supernatant. On bovine articular chondrocytes, all these batches were at least as growth-promoting as FCS. On nasal septal chondrocytes, nine out of ten batches revealed increased or equivalent mitogenic stimulation compared with medium supplemented with FCS. Three-dimensional culture and subsequent histological analysis of matrix formation were used to determine the differentiation properties of platelet supernatant on articular chondrocytes. Human platelet supernatant failed to induce the deposition of typical cartilage matrix components, whereas differentiation and matrix formation were apparent upon cultivation of articular chondrocytes with FCS. Proliferation assays demonstrated that human platelet supernatant stimulates growth of articular and nasal septal chondrocytes; however, platelet supernatant failed to stimulate articular chondrocytes to redifferentiate in three-dimensional chondrocyte cultures. Therefore platelet lysate may be suitable for chondrocyte expansion, but not for maturation of tissue-engineered cartilage.
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Arevalo-Silva, C. A., Cao, Y., Weng, Y., Vacanti, M., Rodriguez, A., Vacanti, C. A., andEavey, R. D. (2001): ‘The effect of fibroblast growth factor and transforming growth factor-beta on porcine chondrocytes and tissue-engineered autologous elastic cartilage’,Tissue Eng.,7, pp. 81–88
Benya, P. D., andShaffer, J. D. (1982): ‘Dedifferentiated chondrocytes reexpress the differentiated collagen phenotype when cultured in agarose gels’,Cell,30, pp. 215–224
Blanco, F. J., Geng, Y., andLotz, M. (1995): ‘Differentiation-dependent effects of IL-1 and TGF-beta on human articular chondrocyte proliferation are related to inducible nitric oxide synthase expression’,J. Immunol.,154, pp. 4018–4026
Boumediene, K., Vivien, D., Macro, M., Bogdanowicz, P., Lebrun, E., andPujol, J. P. (1995): ‘Modulation of rabbit articular chondrocyte (RAC) proliferation by TGF-beta isoforms’,Cell. Prolif.,28, pp. 221–234
Bradford, M. M. (1976): ‘A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding’,Anal. Biochem.,72, pp. 248–254
Bubel, S., Wilhelm, D., Entelmann, M., Kirchner, H., andKluter, H. (1996): ‘Chemokines in stored platelet concentrates’.Transfusion,36, pp. 445–449
Choi, Y. C., Morris, G. M., Lee, F. S., andSokoloff, L. (1980a): ‘The effect of serum on monolayer cell culture of mammalian articular chondrocytes’,Connect. Tissue Res.,7, pp. 105–112
Choi, Y. C., Morris, G. M., andSokoloff, L. (1980b): ‘Effect of platelet lysate on growth and sulfated glycosaminoglycan synthesis in articular chondrocyte cultures’,Arthritis Rheum.,23, pp. 220–224
Fujimoto, E., Ochi, M., Kato, Y., Mochizuki, Y., Sumen, Y., andIkuta, Y. (1999): ‘Beneficial effect of basic fibroblast growth factor on the repair of full-thickness defects in rabbit articular cartilage’,Arch. Orthop. Trauma Surg.,119, pp. 139–145
Gillogly, S. D., Voight, M., andBlackburn, T. (1998): ‘Treatment of articular cartilage defects of the knee with autologous chondrocyte implantation’,J. Orthop. Sports Phys. Ther.,28, pp. 241–251
Grainger, D. J., Mosedale, D. E., Metcalfe, J. C., Weissberg, P. L., andKemp, P. R. (1995): ‘Active and acid-activatable TGF-beta in human sera, platelets and plasma’,Clin. Chim. Acta,235, pp. 11–31
Gruber, R., Sittinger, M., andBujia, J. (1996): ‘In vitro cultivation of human chondrocytes using autologous human serum supplemented culture medium: minimizing possible risk of infection with pathogens of prion diseases’,Laryngorhinootologie,75, pp. 105–108
Guerne, P. A., Blanco, F., Kaelin, A., Desgeorges, A., andLotz, M. (1995): ‘Growth factor responsiveness of human articular chondrocytes in aging and development’,Arthritis Rheum.,38, pp. 960–968
Guerne, P. A., Sublet, A., andLotz, M. (1994): ‘Growth factor responsiveness of human articular chondrocytes: distinct profiles in primary chondrocytes, subcultured chondrocytes, and fibroblasts’,J. Cell Physiol.,158, pp. 476–484.
Haisch, A., Groger, A., Radke, C., Ebmeyer, J., Sudhoff, H., Grasnick, G., Jahnke, V., Burmester, G. R., andSittinger, M. (2000): ‘Macroencapsulation of human cartilage implants: pilot study with polyelectrolyte complex membrane encapsulation’,Biomaterials,21, pp. 1561–1566
Haisch, A., Schultz, O., Perka, C., Jahnke, V., Burmester, G. R., andSittinger, M. (1996): ‘Tissue engineering of human cartilage tissue for reconstructive surgery using biocompatible resorbable fibrin gel and polymer carriers’,Hno,44, pp. 624–629.
Hart, C. E., Bailey, M., Curtis, D. A., Osborn, S., Raines, E., Ross, R., andForstrom, J. W. (1990): ‘Purification of PDGF-AB and PDGF-BB from human platelet extracts and identification of all three PDGF dimers in human platelets’,Biochemistry,29, pp. 166–172
Jakob, M., Demarteau, O., Schafer, D., Hintermann, B., Dick, W., Heberer, M., andMartin, I. (2001): ‘Specific growth factors during the expansion and redifferentiation of adult human articular chondrocytes enhance chondrogenesis and cartilaginous tissue formation in vitro’,J. Cell. Biochem.,81, 368–377
Klinger, M. H. (1997): ‘Platelets and inflammation’,Anat. Embryol. (Berl.),196, pp. 1–11
Mason, J. M., Breitbart, A. S., Barcia, M., Porti, D., Pergolizzi, R. G., andGrande, D. A. (2000): ‘Cartilage and bone regeneration using gene-enhanced tissue engineering’,Clin. Orthop., pp. S171–178
Mayne, R., Vail, M. S., Mayne, P. M., andMiller, E. J. (1976): ‘Changes in type of collagen synthesized as clones of chick chondrocytes grow and eventually lose division capacity’,Proc. Nat. Acad. Sci. USA,73, pp. 1674–1678
Mohle, R., Green, D., Moore, M. A., Nachman, R. L., andRafh, S. (1997): ‘Constitutive production and thrombin-induced release of vascular endothelial growth factor by human megakaryocytes and platelets’,Proc. Nat. Acad. Sci. USA,94, pp. 663–668
Narczewska, B., Czyrski, J. A., andInglot, A. D. (1985): ‘Properties of purified bovine platelet-derived growth factor stimulating proliferation of human and mouse fibroblasts’,Can. J. Biochem. Cell. Biol.,63, pp. 187–194
Puelacher, W. C., Kim, S. W., Vacanti, J. P., Schloo, B.,Mooney, D., andVacanti, C. A. (1994): ‘Tissue-engineered growth of cartilage: the effect of varying the concentration of chondrocytes seeded onto synthetic polymer matrices’,Int. J. Oral Maxillofac. Surg.,23, pp. 49–53
Quatela, V. C., Sherris, R. N. andRosier, R. N. (1993): ‘The human auricular chondrocyte. Responses to growth factors’,Arch. Otolaryngol. Head Neck Surg.,119, pp. 32–37
Rodriguez, A., Cao, Y. L., Ibarra, C., Pap, S., Vacanti, M., Eavey, R. D., andVacanti, C. A. (1999): ‘Characteristics of cartilage engineered from human pediatric auricular cartilage’,Plast. Reconstr. Surg.,103, pp. 1111–1119
Ross, R., Glomset, J., Kariya, B., andRaines, E. (1978): ‘Role of platelet factors in the growth of cells in culture’,Natl. Cancer Inst. Monogr., pp. 103–108
Rutherford, R. B., andRoss, R. (1976): ‘Platelet factors stimulate fibroblasts and smooth muscle cells quiescent in plasma serum to proliferate’,J. Cell. Biol.,69, pp. 196–203
Sams A. E., andNixon, A. J. (1995): ‘Chondrocyte-laden collagen scaffolds for resurfacing extensive articular cartilage defects’,Osteoarthritis Cartilage,3, pp. 47–59
Schultz, O., Keyszer, G., Zacher, J., Sittinger, M., andBurmester, G. R. (1997): ‘Development ofin vitro model systems for destructive joint diseases: novel strategies for establishing inflammatory pannus’,Arthritis Rheum.,40, pp. 1420–1428
Selvaggi, T. A., Walker, R. E., andFleisher, T. A. (1997): ‘Development of antibodies to fetal calf serum with arthus-like reactions in human immunodeficiency virus-infected patients given syngeneic lymphocyte infusions’,Blood,89, pp. 776–779
Sittinger, M., Bujia, J., Minuth, W. W., Hammer, C., andBurmester, G. R. (1994): ‘Engineering of cartilage tissue using bioresorbable polymer carriers in perfusion culture’,Biomaterials,15, pp. 451–456
van Osch, G. J., van der Veen, S. W., Buma, P., andVerwoerd-Verhoef, H. L. (1998): ‘Effect of transforming growth factor-beta on proteoglycan synthesis by chondrocytes in relation to differentiation stage and the presence of pericellular matrix’,Matrix Biol.,17, pp. 413–424
Vivien, D., Galera, P., Lebrun, E., Loyau, G., andPujol, J. P. (1990): ‘Differential effects of transforming growth factor-beta and epidermal growth factor on the cell cycle of cultured rabbit articular chondrocytes’,J. Cell. Physiol.,143, pp. 534–545
Wakitani, S., Kimura, T., Hirooka, A., Ochi, T., Yoneda, M., Yasui, N., Owaki, H., andOno, K. (1989): ‘Repair of rabbit articular surfaces with allograft chondrocytes embedded in collagen gel’,Bone Joint Surg. Br.,71, pp. 74–80
Weiser, L., Bhargava, M., Attia, E., Torzilli, P. A. (1999): ‘Effect of serum and platelet-derived growth factor on chondrocytes grown in collagen gels’,Tissue Eng.,5, pp. 533–544
Yang, S. Y., Ahn, S. T., Rhie, J. W., Lee, K. Y., Choi, J. H., Lee, B. J., andOh, G. T. (2000): ‘Platelet supernatant promotes proliferation of auricular chondrocytes and formation of chondrocyte mass’,Ann. Plast. Surg.,44, pp. 405–411
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Kaps, C., Loch, A., Haisch, A. et al. Human platelet supernatant promotes proliferation but not differentiation of articular chondrocytes. Med. Biol. Eng. Comput. 40, 485–490 (2002). https://doi.org/10.1007/BF02345083
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DOI: https://doi.org/10.1007/BF02345083