Early Metabolite Levels Predict Long-Term Matrix Accumulation for Chondrocytes in Elastin-like Polypeptide Biopolymer Scaffolds
Publication: Tissue Engineering Part A
Volume 15, Issue Number 8
Abstract
The development of cartilage tissue engineering scaffolds could greatly benefit from methods to evaluate the interactions of cells with scaffolds that are rapid, are nondestructive, and can be carried out at early culture times. Motivated by this rationale, the objective of the current study was to evaluate whether the concentration of metabolites in scaffold–cell cultures at early culture times could predict matrix synthesis in the same samples at longer culture times. Metabolite and matrix synthesis were measured for 16 different formulations of cell-laden elastin–like polypeptide hydrogels. Metabolites were measured at days 4 and 7 of culture, while matrix accumulation was evaluated at day 28. Four of the 16 formulations resulted in molar ratios of lactate:glucose near 2, indicating anaerobic metabolism of glucose, which resulted in collagen:glycosaminoglycan accumulation ratios near those of native tissue. Lactate and pyruvate concentrations were found to significantly correlate with both sulfated glycosaminoglycan and hydroxyproline accumulation, with better fits for the latter. Lactate was found to be the strongest predictor of both matrix components, suggesting that measuring this metabolite at very early culture times may be useful for evaluating the status of tissue engineering constructs in a rapid and nondestructive manner.
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References
1.
Lee R.B.Wilkins R.J.Razaq S.Urban J.P.G. The effect of mechanical stress on cartilage energy metabolismBiorheology391332002. 1. Lee, R.B., Wilkins, R.J., Razaq, S., and Urban, J.P.G. The effect of mechanical stress on cartilage energy metabolism. Biorheology 39, 133, 2002.
2.
Shapiro I.M.Tokuoka T.Silverton S.F.Hall B.K.Newman S.A. Energy metabolism in cartilageCartilage: Molecular AspectsBoca Raton, FLCRC Press199197-130. 2. Shapiro, I.M., Tokuoka, T., and Silverton, S.F. Energy metabolism in cartilage. In: Hall, B.K., and Newman, S.A., eds. Cartilage: Molecular Aspects. Boca Raton, FL: CRC Press, 1991, pp. 97–130.
3.
Stockwell R.A.Hall B.K. Metabolism of cartilageCartilage: Structure, Function, and BiochemistryNew York, NYAcademic Press1983253-280. 3. Stockwell, R.A. Metabolism of cartilage. In: Hall, B.K., ed. Cartilage: Structure, Function, and Biochemistry. New York, NY: Academic Press, 1983, pp. 253–280.
4.
Lee R.B.Urban J.P.G. Evidence for a negative Pasteur effect in articular cartilageBiochem J321951997. 4. Lee, R.B., and Urban, J.P.G. Evidence for a negative Pasteur effect in articular cartilage. Biochem J 321, 95, 1997.
5.
Lehninger A.L. Biochemistrysecond editionNew York, NYWorth Publisher, Inc.1975. 5. Lehninger, A.L. Biochemistry, second edition. New York, NY: Worth Publisher, Inc., 1975.
6.
Lane J.M.Brighton C.T.Menkowitz B.J. Anaerobic and aerobic metabolism in articular cartilageJ Rheumatol43341977. 6. Lane, J.M., Brighton, C.T., and Menkowitz, B.J. Anaerobic and aerobic metabolism in articular cartilage. J Rheumatol 4, 334, 1977.
7.
Otte P. Basic cell-metabolism of articular-cartilage—manometric studiesZ Rheumatol503041991. 7. Otte, P. Basic cell-metabolism of articular-cartilage—manometric studies. Z Rheumatol 50, 304, 1991.
8.
Rajpurohit R.Koch C.J.Tao Z.L.Teixeira C.M.Shapiro I.M. Adaptation of chondrocytes to low oxygen tension: relationship between hypoxia and cellular metabolismJ Cell Physiol1684241996. 8. Rajpurohit, R., Koch, C.J., Tao, Z.L., Teixeira, C.M., and Shapiro, I.M. Adaptation of chondrocytes to low oxygen tension: relationship between hypoxia and cellular metabolism. J Cell Physiol 168, 424, 1996.
9.
Wu M.H.Urban J.P.G.Cui Z.F.Cui Z.Xu X. Effect of extracellular pH on matrix synthesis by chondrocytes in 3D agarose gelBiotechnol Prog234302007. 9. Wu, M.H., Urban, J.P.G., Cui, Z.F., Cui, Z., and Xu, X. Effect of extracellular pH on matrix synthesis by chondrocytes in 3D agarose gel. Biotechnol Prog 23, 430, 2007.
10.
Heywood H.K.Bader D.L.Lee D.A. Glucose concentration and medium volume influence cell viability and glycosaminoglycan synthesis in chondrocyte-seeded alginate constructsTissue Eng1234872006. 10. Heywood, H.K., Bader, D.L., and Lee, D.A. Glucose concentration and medium volume influence cell viability and glycosaminoglycan synthesis in chondrocyte-seeded alginate constructs. Tissue Eng 12, 3487, 2006.
11.
Heywood H.K.Bader D.L.Lee D.A. Rate of oxygen consumption by isolated articular chondrocytes is sensitive to medium glucose concentrationJ Cell Physiol2064022006. 11. Heywood, H.K., Bader, D.L., and Lee, D.A. Rate of oxygen consumption by isolated articular chondrocytes is sensitive to medium glucose concentration. J Cell Physiol 206, 402, 2006.
12.
Obradovic B.Carrier R.L.Vunjak-Novakovic G.Freed L.E. Gas exchange is essential for bioreactor cultivation of tissue engineered cartilageBiotechnol Bioeng631971999. 12. Obradovic, B., Carrier, R.L., Vunjak-Novakovic, G., and Freed, L.E. Gas exchange is essential for bioreactor cultivation of tissue engineered cartilage. Biotechnol Bioeng 63, 197, 1999.
13.
Sandberg L.B.Soskel N.T.Leslie J.G. Elastin structure, biosynthesis, and relation to disease statesN Engl J Med3045661981. 13. Sandberg, L.B., Soskel, N.T., and Leslie, J.G. Elastin structure, biosynthesis, and relation to disease states. N Engl J Med 304, 566, 1981.
14.
Urry D.W. Free-energy transduction in polypeptides and proteins based on inverse temperature transitionsProg Biophys Mol Biol57231992. 14. Urry, D.W. Free-energy transduction in polypeptides and proteins based on inverse temperature transitions. Prog Biophys Mol Biol 57, 23, 1992.
15.
Betre H.Setton L.A.Meyer D.E.Chilkoti A. Characterization of a genetically engineered elastin-like polypeptide for cartilaginous tissue repairBiomacromolecules39102002. 15. Betre, H., Setton, L.A., Meyer, D.E., and Chilkoti, A. Characterization of a genetically engineered elastin-like polypeptide for cartilaginous tissue repair. Biomacromolecules 3, 910, 2002.
16.
Lee J.Macosko C.W.Urry D.W. Elastomeric polypentapeptides cross-linked into matrixes and fibersBiomacromolecules21702001. 16. Lee, J., Macosko, C.W., and Urry, D.W. Elastomeric polypentapeptides cross-linked into matrixes and fibers. Biomacromolecules 2, 170, 2001.
17.
Trabbic-Carlson K.Setton L.A.Chilkoti A. Swelling and mechanical behaviors of chemically cross-linked hydrogels of elastin-like polypeptidesBiomacromolecules45722003. 17. Trabbic-Carlson, K., Setton, L.A., and Chilkoti, A. Swelling and mechanical behaviors of chemically cross-linked hydrogels of elastin-like polypeptides. Biomacromolecules 4, 572, 2003.
18.
Urry D.W.Pattanaik A.Xu J.Woods T.C.McPherson D.T.Parker T.M. Elastic protein-based polymers in soft tissue augmentation and generationJ Biomater Sci Polym Ed910151998. 18. Urry, D.W., Pattanaik, A., Xu, J., Woods, T.C., McPherson, D.T., and Parker, T.M. Elastic protein-based polymers in soft tissue augmentation and generation. J Biomater Sci Polym Ed 9, 1015, 1998.
19.
Lim D.W.Nettles D.L.Setton L.A.Chilkoti A. Rapid crosslinking of elastin-like polypeptides with hydroxymethylphosphines in aqueous solutionBiomacromolecules814632007. 19. Lim, D.W., Nettles, D.L., Setton, L.A., and Chilkoti, A. Rapid crosslinking of elastin-like polypeptides with hydroxymethylphosphines in aqueous solution. Biomacromolecules 8, 1463, 2007.
20.
Nowatzki P.J.Tirrell D.A. Physical properties of artificial extracellular matrix protein films prepared by isocyanate crosslinkingBiomaterials2512612004. 20. Nowatzki, P.J., and Tirrell, D.A. Physical properties of artificial extracellular matrix protein films prepared by isocyanate crosslinking. Biomaterials 25, 1261, 2004.
21.
Lee J.Macosko C.W.Urry D.W. Mechanical properties of cross-linked synthetic elastomeric polypentapeptidesMacromolecules3459682001. 21. Lee, J., Macosko, C.W., and Urry, D.W. Mechanical properties of cross-linked synthetic elastomeric polypentapeptides. Macromolecules 34, 5968, 2001.
22.
Nagapudi K.Brinkman W.T.Leisen J.E.Huang L.McMillan R.A.Apkarian R.P.Conticello V.P.Chaikof E.L. Photomediated solid-state cross-linking of an elastin-mimetic recombinant protein polymerMacromolecules3517302002. 22. Nagapudi, K., Brinkman, W.T., Leisen, J.E., Huang, L., McMillan, R.A., Apkarian, R.P., Conticello, V.P., and Chaikof, E.L. Photomediated solid-state cross-linking of an elastin-mimetic recombinant protein polymer. Macromolecules 35, 1730, 2002.
23.
McHale M.K.Setton L.A.Chilkoti A. Synthesis and in vitro evaluation of enzymatically cross-linked elastin-like polypeptide gels for cartilaginous tissue repairTissue Eng1117682005. 23. McHale, M.K., Setton, L.A., and Chilkoti, A. Synthesis and in vitro evaluation of enzymatically cross-linked elastin-like polypeptide gels for cartilaginous tissue repair. Tissue Eng 11, 1768, 2005.
24.
Bellingham C.M.Lillie M.A.Gosline J.M.Wright G.M.Starcher B.C.Bailey A.J.Woodhouse K.A.Keeley F.W. Recombinant human elastin polypeptides self-assemble into biomaterials with elastin-like propertiesBiopolymers704452003. 24. Bellingham, C.M., Lillie, M.A., Gosline, J.M., Wright, G.M., Starcher, B.C., Bailey, A.J., Woodhouse, K.A., and Keeley, F.W. Recombinant human elastin polypeptides self-assemble into biomaterials with elastin-like properties. Biopolymers 70, 445, 2003.
25.
Dandu R.Megeed Z.Haider M.Cappello J.Ghandehari H.Svenson S. Silk-elastinlike hydrogels: thermal characterization and gene deliveryPolymeric Drug Delivery II: Polymeric Matrices and Drug Particle EngineeringWashington, DCAmerican Chemical Society2006150-168. 25. Dandu, R., Megeed, Z., Haider, M., Cappello, J., and Ghandehari, H. Silk-elastinlike hydrogels: thermal characterization and gene delivery. In: Svenson, S, ed. Polymeric Drug Delivery II: Polymeric Matrices and Drug Particle Engineering, Washington, DC: American Chemical Society, 2006, pp. 150–168.
26.
Nagapudi K.Brinkman W.T.Thomas B.S.Park J.O.Srinivasarao M.Wright E.Conticello V.P.Chaikof E.L. Viscoelastic and mechanical behavior of recombinant protein elastomersBiomaterials2646952005. 26. Nagapudi, K., Brinkman, W.T., Thomas, B.S., Park, J.O., Srinivasarao, M., Wright, E., Conticello, V.P., and Chaikof, E.L. Viscoelastic and mechanical behavior of recombinant protein elastomers. Biomaterials 26, 4695, 2005.
27.
Wu X.Y.Sallach R.Haller C.A.Caves J.A.Nagapudi K.Conticello V.P.Levenston M.E.Chaikof E.L. Alterations in physical cross-linking modulate mechanical properties of two-phase protein polymer networksBiomacromolecules630372005. 27. Wu, X.Y., Sallach, R., Haller, C.A., Caves, J.A., Nagapudi, K., Conticello, V.P., Levenston, M.E., and Chaikof, E.L. Alterations in physical cross-linking modulate mechanical properties of two-phase protein polymer networks. Biomacromolecules 6, 3037, 2005.
28.
Lim D.W.Nettles D.L.Setton L.A.Chilkoti A. In-situ crosslinking of elastin-like polypeptide block copolymers for tissue repairBiomacromolecules92222008. 28. Lim, D.W., Nettles, D.L., Setton, L.A., and Chilkoti, A. In-situ crosslinking of elastin-like polypeptide block copolymers for tissue repair. Biomacromolecules 9, 222, 2008.
29.
Betre H.Ong S.R.Guilak F.Chilkoti A.Fermor B.Setton L.A. Chondrocytic differentiation of human adipose-derived adult stem cells in elastin-like polypeptideBiomaterials27912006. 29. Betre, H., Ong, S.R., Guilak, F., Chilkoti, A., Fermor, B., and Setton, L.A. Chondrocytic differentiation of human adipose-derived adult stem cells in elastin-like polypeptide. Biomaterials 27, 91, 2006.
30.
Lim D.W.Trabbic-Carlson K.MacKay J.A.Chilkoti A. Improved non-chromatographic purification of a recombinant protein by cationic elastin-like polypeptidesBiomacromolecules814172007. 30. Lim, D.W., Trabbic-Carlson, K., MacKay, J.A., and Chilkoti, A. Improved non-chromatographic purification of a recombinant protein by cationic elastin-like polypeptides. Biomacromolecules 8, 1417, 2007.
31.
Meyer D.E.Chilkoti A. Purification of recombinant proteins by fusion with thermally responsive polypeptidesNat Biotechnol1711121999. 31. Meyer, D.E., and Chilkoti, A. Purification of recombinant proteins by fusion with thermally responsive polypeptides. Nat Biotechnol 17, 1112, 1999.
32.
Nettles D.L.Elder S.H.Gilbert J.A. Potential use of chitosan as a cell scaffold material for cartilage tissue engineeringTissue Eng810092002. 32. Nettles, D.L., Elder, S.H., and Gilbert, J.A. Potential use of chitosan as a cell scaffold material for cartilage tissue engineering. Tissue Eng 8, 1009, 2002.
33.
Estes B.T.Diekman B.O.Guilak F. Monolayer cell expansion conditions affect the chondrogenic potential of adipose-derived stem cellsBiotechnol Bioeng999862008. 33. Estes, B.T., Diekman, B.O., and Guilak, F. Monolayer cell expansion conditions affect the chondrogenic potential of adipose-derived stem cells. Biotechnol Bioeng 99, 986, 2008.
34.
Kim Y.J.Sah R.L.Y.Doong J.Y.H.Grodzinsky A.J. Fluorometric assay of DNA in cartilage explants using Hoechst-33258Anal Biochem1741681988. 34. Kim, Y.J., Sah, R.L.Y., Doong, J.Y.H., and Grodzinsky, A.J. Fluorometric assay of DNA in cartilage explants using Hoechst-33258. Anal Biochem 174, 168, 1988.
35.
Farndale R.W.Buttle D.J.Barrett A.J. Improved quantitation and discrimination of sulfated glycosaminoglycans by use of dimethylmethylene blueBiochim Biophys Acta8831731986. 35. Farndale, R.W., Buttle, D.J., and Barrett, A.J. Improved quantitation and discrimination of sulfated glycosaminoglycans by use of dimethylmethylene blue. Biochim Biophys Acta 883, 173, 1986.
36.
Neidert M.R.Lee E.S.Oegema T.R.Tranquillo R.T. Enhanced fibrin remodeling in vitro with TGF-beta 1, insulin and plasmin for improved tissue-equivalentsBiomaterials2337172002. 36. Neidert, M.R., Lee, E.S., Oegema, T.R., and Tranquillo, R.T. Enhanced fibrin remodeling in vitro with TGF-beta 1, insulin and plasmin for improved tissue-equivalents. Biomaterials 23, 3717, 2002.
37.
Dombi G.W.Haut R.C.Sullivan W.G. Correlation of high-speed tensile-strength with collagen content in control and lathyritic rat skinJ Surg Res54211993. 37. Dombi, G.W., Haut, R.C., and Sullivan, W.G. Correlation of high-speed tensile-strength with collagen content in control and lathyritic rat skin. J Surg Res 54, 21, 1993.
38.
VunjakNovakovic G.Freed L.E.Biron R.J.Langer R. Effects of mixing on the composition and morphology of tissue-engineered cartilageAIChE J428501996. 38. VunjakNovakovic, G., Freed, L.E., Biron, R.J., and Langer, R. Effects of mixing on the composition and morphology of tissue-engineered cartilage. AIChE J 42, 850, 1996.
39.
Lee R.B.Urban J.P.G. Functional replacement of oxygen by other oxidants in articular cartilageArthritis Rheum4631902002. 39. Lee, R.B., and Urban, J.P.G. Functional replacement of oxygen by other oxidants in articular cartilage. Arthritis Rheum 46, 3190, 2002.
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Copyright 2009, Mary Ann Liebert, Inc.
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Published in print: August 2009
Published online: 4 February 2009
Accepted: 11 December 2008
Received: 2 August 2008
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