Hydrogel properties influence ECM production by chondrocytes photoencapsulated in poly(ethylene glycol) hydrogels
Stephanie J. Bryant
Department of Chemical Engineering, University of Colorado, Boulder, Colorado 80309-0424
Search for more papers by this authorCorresponding Author
Kristi S. Anseth
Department of Chemical Engineering, University of Colorado, Boulder, Colorado 80309-0424
Howard Hughes Medical Institute, Chevy Chase, Maryland 20815-6789
Department of Chemical Engineering, University of Colorado, Boulder, Colorado 80309-0424Search for more papers by this authorStephanie J. Bryant
Department of Chemical Engineering, University of Colorado, Boulder, Colorado 80309-0424
Search for more papers by this authorCorresponding Author
Kristi S. Anseth
Department of Chemical Engineering, University of Colorado, Boulder, Colorado 80309-0424
Howard Hughes Medical Institute, Chevy Chase, Maryland 20815-6789
Department of Chemical Engineering, University of Colorado, Boulder, Colorado 80309-0424Search for more papers by this authorAbstract
When using hydrogel scaffolds for cartilage tissue engineering, two gel properties are particularly important: the equilibrium water content (q, equilibrium swelling ratio) and the compressive modulus, K. In this work, chondrocytes were photoencapsulated in degrading and nondegrading poly(ethylene glycol)-based hydrogels to assess extracellular matrix (ECM) formation as a function of these gel properties. In nondegrading gels, the glycosaminoglycan (GAG) content was not significantly different in gels when q was varied from 4.2 to 9.3 after 2 and 4 weeks in vitro. However, gels with a q of 9.3 allowed GAGs to diffuse throughout the gels homogenously, but a q ≤ 5.2 resulted in localization of GAGs pericellularly. Interestingly, in the moderately crosslinked gels with a K of 360 kPa, an increase in type II collagen synthesis was observed compared with gels with a higher (960 kPa) and lower (30 kPa) K after 4 weeks. With the incorporation of degradable linkages into the network, gel properties with an initially high K (350 kPa) and final high q (7.9) were obtained, which allowed for increased type II collagen synthesis coupled with a homogenous distribution of GAGs. Thus, a critical balance exists between gel swelling, mechanics, and degradation in forming a functional ECM. © 2001 Wiley Periodicals, Inc. J Biomed Mater Res 59: 63–72, 2002
References
- 1Cao Y, Rodriguez A, Vacanti M, Ibarra C, Arevalo C, Vacanti CA. Comparative study of the use of poly(glycolic acid), calcium alginate and pluronics in the engineering of autologous porcine cartilage. J Biomater Sci Polym Ed 1998; 9: 474–487.
- 2Ting V, Sims CD, Brecht LE, McCarthy JG, Kasabian AK, Connelly PR, Elisseeff J, Gittes GK, Longaker MT. In vitro prefabrication of human cartilage shapes using fibrin glue and human chondrocytes. Ann Plast Surg 1998; 40: 413–421.
- 3Mann BK, Schmedlen RH, West JL. Tethered-TGF-beta increases extracellular matrix production of vascular smooth muscle cells. Biomaterials 2001; 22: 439–444.
- 4Elisseeff J, McIntosh W, Anseth K, Riley S, Ragan P, Langer R. Photoencapsulation of chondrocytes in poly(ethylene oxide)-based semi-interpenetrating networks. J Biomed Mater Res 2000; 51: 164–171.
10.1002/(SICI)1097-4636(200008)51:2<164::AID-JBM4>3.0.CO;2-W CAS PubMed Web of Science® Google Scholar
- 5Elisseeff J, Anseth K, Sims D, McIntosh W, Randolph M, Yaremchuk M, Langer R. Transdermal photopolymerization of poly(ethylene oxide)-based injectable hydrogels for tissue-engineered cartilage. Plast Reconstr Surg 1999; 104: 1014–1022.
- 6Alsberg E, Franceschi RT, Mooney DJ. Bone tissue engineering using injectable hydrogels. J Dent Res 2000; 79: 94.
- 7Akhouayri O, Lafage-Proust MH, Rattner A, Laroche N, Caillot-Augusseau A, Alexandre C, Vico L. Effects of static or dynamic mechanical stresses on osteoblast phenotype expression in three-dimensional contractile collagen gels. J Cell Biochem 2000; 76: 217–230.
- 8Winn SR, Schmitt JM, Buck D, Hu YH, Grainger D, Hollinger JO. Tissue-engineered bone biomimetic to regenerate calvarial critical-sized defects in athymic rats. J Biomed Mater Res 1999; 45: 414–421.
10.1002/(SICI)1097-4636(19990615)45:4<414::AID-JBM17>3.0.CO;2-Z CAS PubMed Web of Science® Google Scholar
- 9Awad HA, Butler DL, Boivin GP, Smith FNL, Malaviya P, Huibregtse B, Caplan AI. Autologous mesenchymal stem cell-mediated repair of tendon. Tissue Eng 1999; 5: 267–277.
- 10Woerly S. Restorative surgery of the central nervous system by means of tissue engineering using NeuroGel implants. Neurosurg Rev 2000; 23: 59–77.
- 11Woerly S, Plant GW, Harvey AR. Neural tissue engineering: From polymer to biohybrid organs. Biomaterials 1996; 17: 301–310.
- 12Bellamkonda R, Ranieri JP, Bouche N, Aebischer P. Hydrogel-based 3-dimensional matrix for neural cells. J Biomed Mater Res 1995; 29: 663–671.
- 13Bryant SJ, Nuttelman CR, Anseth KS. The effects of crosslinking density on cartilage formation in photocrosslinkable hydrogels. In: P Patterson, editor. Biomedical Science Instrumentation. Research Triangle Park, NC: Training and Publications Services; 1999. p 309–314.
- 14Armstrong CG, Mow VC. Variations in the intrinsic mechanical proterties of human articular cartilage with age, degeneration, and water content. J Bone Joint Surg Am 1982; 64: 88–94.
- 15Lee KY, Rowley JA, Eiselt P, Moy EM, Bouhadir KH, Mooney DJ. Controlling mechanical and swelling properties of alginate hydrogels independently by cross-linker type and cross-linking density. Macromolecules 2000; 33: 4291–4294.
- 16Arokoski JPA, Hyttinen MM, Helminen HJ, Jurvelin JS. Biomechanical and structural characteristics of canine femoral and tibial cartilage. J Biomed Mater Res 1999; 48: 99–107.
- 17Metters AT, Anseth KS, Bowman CN. Fundamental studies of a novel, biodegradable PEG–b-PLA hydrogel. Polymer 2000; 41: 3993–4004.
- 18Kim BS, Mooney DJ. Development of biocompatible synthetic extracellular matrices for tissue engineering. Trends Biotechnol 1998; 16: 224–230.
- 19Sawhney AS, Pathak CP, Hubbell JA. Bioerodible hydrogels based on photopolymerized poly(ethylene glycol)-co-poly(alpha-hydroxy acid) diacrylate macromers. Macromolecules 1993; 26: 581–587.
- 20Freed LE, Marquis JC, Nohria A, Emmanual J, Mikos AG, Langer R. Neocartilage formation in vitro and in vivo using cells cultured on synthetic biodegradable polymers. J Biomed Mater Res 1993; 27: 11–23.
- 21Rotter N, Aigner J, Naumann A, Planck H, Hammer C, Burmester G, Sittinger M. Cartilage reconstruction in head and neck surgery: Comparison of resorbable polymer scaffolds for tissue engineering of human septal cartilage. J Biomed Mater Res 1998; 42: 347–356.
10.1002/(SICI)1097-4636(19981205)42:3<347::AID-JBM2>3.0.CO;2-J CAS PubMed Web of Science® Google Scholar
- 22Gugala Z, Gogolewski S. In vitro growth and activity of primary chondrocytes on a resorbable polylactide three-dimensional scaffold. J Biomed Mater Res 2000; 49: 183–191.
10.1002/(SICI)1097-4636(200002)49:2<183::AID-JBM5>3.0.CO;2-D CAS PubMed Web of Science® Google Scholar
- 23Bryant SJ, Nuttelmen CR, Anseth KS. Cytocompatibility of ultraviolet and visible light photoinitiating systems on cultured NIH/3T3 fibroblasts in vitro. J Biomater Sci Polym Ed 2000; 11: 439–457.
- 24Taylor KB, Jeffree GM. A new basic metachromatic dye, I:9-dimethyl methylene blue. Histochem J 1969; 1: 199–204.
- 25Woessner JF. The determination of hydroxyproline in tissue and protein samples containing small proportions of this imino acid. Arch Biochem Biophys 1961; 93: 440–447.
- 26Hollander AP, Heathfield TF, Webber C, Iwata Y, Bourne R, Rorabeck C, Poole AP. Increased damage to type-II collagen in osteoarthritic articular-cartilage detected by a new immunoassay. J Clin Invest 1994; 93: 1722–1732.
- 27Kim YJ, Sah RLY, Doong JYH, Grodzinsky AJ. Fluorometric assay of DNA in cartilage explants using Hoechst-33258. Anal Biochem 1988; 174: 168–176.
- 28Canal T, Peppas NA. Correlation between mesh size and equilibrium degree of swelling of polymeric networks. J Biomed Mater Res 1989; 23: 1183–1193.
- 29Lu SX, Anseth KS. Release behavior of high molecular weight solutes from poly(ethylene glycol)-based degradable networks. Macromolecules 2000; 33: 2509–2515.
- 30Peppas NA. Hydrogels in medicine and pharmacy, 1. Boca Raton, FL: CRC Press; 1986.
- 31Merrill EW, Dennison KA, Sung C. Partitioning and diffusion of solutes in hydrogels of poly(ethylene oxide). Biomaterials 1993; 14: 1117–1126.
- 32Larsen IB, Munksgaard EC. Effect of human saliva on surface degradation of composite resins. Scand J Dent Res 1991; 99: 254–261.
- 33Elliott JE, Anseth JW, Bowman CN. Kinetic modeling of the effect of solvent concentration on primary cyclization during polymerization of multifunctional monomers. Chem Eng Sci 2001; 56: 3173–3184.
- 34Lee DA, Knight MM, Bolton JF, Idowu BD, Kayser MV, Bader DL. Chondrocyte deformation within compressed agarose constructs at the cellular and sub-cellular levels. J Biomech 2000; 33: 81–95.
- 35Buckwalter JA, Rosenberg L. Structural changes during development in bovine fetal epiphyseal cartilage. Coll Relat Res 1983; 3: 498–504.
10.1016/S0174-173X(83)80028-4 Google Scholar
- 36Kaab MJ, Gwynn IA, Notzli HP. Collagen fibre arrangement in the tibial plateau articular cartilage of man and other mammalian species. J Anat 1998; 193: 23–34.
- 37Bryant SJ, Anseth KS. Controlling the spatial distribution of ECM components in degradable PEG hydrogels for tissue engineering cartilage. Submitted for publication.
- 38Smith RL, Rusk SF, Ellison BE, Wessells P, Tsuchiya K, Carter DR, Caler WE, Sandell LJ, Schurman DJ. In vitro stimulation of articular chondrocyte mRNA and extracellular matrix synthesis by hydrostatic pressure. J Orthopaed Res 1996; 14: 53–60.
- 39Holmvall K, Camper L, Johansson S, Kimura JH, Lundgrenakerlund E. Chondrocyte and chondrosarcoma cell integrins with affinity for collagen type-Ii and their response to mechanical stress. Exp Cell Res 1995; 221: 496–503.
- 40Buckwalter JA. Cartilage. In: R Dulbecco, editor. Encyclopedia of human biology. San Diego: Academic Press; 1997. p 431–445.
- 41Metters AT, Bowman CN, Anseth KS. Verification of scaling laws for degrading PLA-b-PEG-b-PLA hydrogels. AICHE J 2001; 47: 1432–1437.