Biomimetics of the extracellular matrix: an integrated three-dimensional fiber-hydrogel composite for cartilage tissue engineering

  • Coburn, Jeannine (Department of Biomedical Engineering, Johns Hopkins University) ;
  • Gibson, Matt (Department of Biomedical Engineering, Johns Hopkins University) ;
  • Bandalini, Pierre Alain (Ecole Polytechnique) ;
  • Laird, Christopher (Department of Biomedical Engineering, Johns Hopkins University) ;
  • Mao, Hai-Quan (Department of Materials Science and Engineering, Johns Hopkins University) ;
  • Moroni, Lorenzo (Department of Biomedical Engineering, Johns Hopkins University) ;
  • Seliktar, Dror (Faculty of Biomedical Engineering, Technion - Israel Institute of Technology) ;
  • Elisseeff, Jennifer (Department of Biomedical Engineering, Johns Hopkins University)
  • Received : 2010.10.22
  • Accepted : 2010.10.29
  • Published : 2011.03.25


The native extracellular matrix (ECM) consists of an integrated fibrous protein network and proteoglycan-based ground (hydrogel) substance. We designed a novel electrospinning technique to engineer a three dimensional fiber-hydrogel composite that mimics the native ECM structure, is injectable, and has practical macroscale dimensions for clinically relevant tissue defects. In a model system of articular cartilage tissue engineering, the fiber-hydrogel composites enhanced the biological response of adult stem cells, with dynamic mechanical stimulation resulting in near native levels of extracellular matrix. This technology platform was expanded through structural and biochemical modification of the fibers including hydrophilic fibers containing chondroitin sulfate, a significant component of endogenous tissues, and hydrophobic fibers containing ECM microparticles.


  1. Awad, H.A., Wickham, M.Q., Leddy, H.A., Gimble, J.M. and Guilak, F. (2004), "Chondrogenic differentiation of adipose-derived adult stem cells in agarose, alginate, and gelatin scaffolds", Biomaterials 25(16), 3211-3222.
  2. Chiara, G. and Ranieri, C. (2009), "Cartilage and bone extracellular matrix", Curr. Pharm. Design, 15(12), 1334- 1348.
  3. Engler, A.J., Sen, S., Sweeney, H.L. and Discher, D.E. (2006), "Matrix elasticity directs stem cell lineage specification", Cell, 126(4), 677-689.
  4. Farndale, R.W., Buttle, D.J. and Barrett, A.J. (1986), "Improved quantitation and discrimination of sulphated glycosaminoglycans by use of dimethylmethylene blue", Biochim. Biophys. Acta. Gen. Sub., 883(2),173-177.
  5. Gong, J.P., Katsuyama, Y., Kurokawa, T. and Osada, Y. (2003), "Double-network hydrogels with extremely high mechanical strength", Adv. Mater., 15(14), 1155-1158.
  6. Gregory, S.S. and Annette, W. (2009), "Interactions between extracellular matrix and growth factors in wound healing", Wound Repair Regen., 17(2), 153-162.
  7. Hannouche, D., Terai, H., Fuchs, J.R., Terada, S., Zand, S., Nasseri, B.A., Petite, H., Sedel, L. and Vacanti, J.P. (2007), "Engineering of implantable cartilaginous structures from bone marrow-derived mesenchymal stem cells", Tissue Eng., 13(1), 87-99.
  8. Janna, K.M., John, T.C., Christopher, G.W., Kristin, E.M. and Marc, E.L. (2007), "Dynamic compression regulates the expression and synthesis of chondrocyte-specific matrix molecules in bone marrow stromal cells", Stem Cells, 25(3), 655-663.
  9. Kim, Y.-J., Sah, R.L.Y., Doong, J.Y.H. and Grodzinsky, A.J. (1988), "Fluorometric assay of DNA in cartilage explants using Hoechst 33258", Anal. Biochem., 174(1), 168-176.
  10. Lee, H.J., Yu, C., Chansakul, T., Hwang, N.S., Varghese, S., Yu, S.M. and Elisseeff, J.H. (2008), "Enhanced chondrogenesis of mesenchymal stem cells in collagen mimetic peptide-mediated microenvironment", Tissue Eng., 14(11), 1843-1851.
  11. Li, Q., Wang, D.A. and Elisseeff, J.H. (2003), "Heterogeneous-phase reaction of glycidyl methacrylate and chondroitin sulfate: Mechanism of ring-opening-transesterification Competition", Macromolecules, 36(7), 2556-2562.
  12. Li, W., Cooper, J.A., Mauck, R.L. and Tuan, R.S. (2006), "Fabrication and characterization of six electrospun poly(a-hydroxy ester)-based fibrous scaffolds for tissue engineering applications", Acta Biomater., 2(4), 377-385.
  13. Marijnissen, W.J., van Osch, G.J., Aigner, J., van der Veen, S.W., Hollander, A.P., Verwoerd-Verhoef, H.L. and Verhaar, J.A. (2002), "Alginate as a chondrocyte-delivery substance in combination with a non-woven scaffold for cartilage tissue engineering", Biomaterials, 23(6), 1511-1517.
  14. Martens, P. and Anseth, K.S. (2000), "Characterization of hydrogels formed from acrylate modified poly(vinyl alcohol) macromers", Polymer, 41(21), 7715-7722.
  15. Moutos, F.T., Freed, L.E. and Guilak, F. (2007), "A biomimetic three-dimensional woven composite scaffold for functional tissue engineering of cartilage", Nat. Mater., 6(2), 162-167.
  16. Moutos, F.T. and Guilak, F. (2010), "Functional properties of cell-seeded three-dimensionally woven poly( $\varepsilon$ - caprolactone) scaffolds for cartilage tissue engineering", Tissue Eng., 16(4), 1291-1301.
  17. Mow, V.C., Ratcliffe, A. and Poole, A.R. (1992), "Carilage and diarthrodial joints as paradigms for hierarchical materials and structures", Biomaterials, 13(2), 67-97.
  18. Nakayama, A., Kakugo, A., Gong, J.P., Osada, Y., Takai, M., Erata T. and Kawano, S. (2004), "High mechanical strength double-network hydrogel with bacterial cellulose", Adv. Funct. Mater., 14(11), 1124-1128.
  19. Place, E.S., Evans, N.D. and Stevens, M.M. (2009), "Complexity in biomaterials for tissue engineering", Nat. Mater., 8(6), 457-470.
  20. Ramadoss, P. and Nagamani, K. (2009), "Behavior of high-strength fiber reinforced concrete plates under inplane and transverse loads", Struct. Eng. Mech., 31(4), 371-382.
  21. Schmidt, O., Mizrahi, J., Elisseeff, J. and Seliktar, D. (2006), "Immobilized fibrinogen in PEG hydrogels does not improve chondrocyte-mediated matrix deposition in response to mechanical stimulation", Biotechnol. Bioeng., 95(6), 1061-1069.
  22. Segawa, K. and Takiguchi, R. (1992), "Ultrastructural alteration of cartilaginous fibril arrangement in the rat mandibular condyle as revealed by high-resolution scanning electron microscopy", Anat. Rec., 234(4), 493-499.
  23. Slivka, M.A., Leatherbury, N.C., Kieswetter, K. and Niederauer, G.G. (2001), "Porous, resorbable, fiberreinforced scaffolds tailored for articular cartilage repair", Tissue Eng., 7(6), 767-780.
  24. Strehin, I., Winnette McIntosh, A., Oliver, S., Afrah, S. and Elisseeff, J. H. (2009), "Synthesis and characterization of a chondroitin sulfate-polyethylene glycol corneal adhesive", J. Cataract Refr. Surg., 35(3), 567-576.
  25. Tsang, K., Cheung, M., Chan, D. and Cheah, K. (2010), "The developmental roles of the extracellular matrix: beyond structure to regulation", Cell Tissue Res., 339(1), 93-110.
  26. Tzezana, R., Zussman, E. and Levenberg, S. (2008), "A layered ultra-porous scaffold for tissue engineering, created via a hydrospinning method", Tissue Eng., 14(4), 281-288.
  27. Vanessa, T., Nathaniel, H., Lorenzo, M., Hyung, B.P., Zijun, Z., Joseph, M., Dror, S. and Jennifer, E. (2007), "Differential response of adult and embryonic mesenchymal progenitor cells to mechanical compression in hydrogels", Stem Cells, 25(11), 2730-2738.
  28. Varghese, S., Hwang, N.S., Canver, A.C., Theprungsirikul, P., Lin, D.W. and Elisseeff, J. (2008), "Chondroitin sulfate based niches for chondrogenic differentiation of mesenchymal stem cells", Matrix Biol., 27(1), 12-21.
  29. Wagenseil, J.E. and Mecham, R.P. (2009), "Vascular extracellular matrix and arterial mechanics", Physiol. Rev., 89(3), 957-989.
  30. Williams, C.G., Kim, T.K., Taboas, A., Malik, A., Manson, P. and Elisseeff, J. (2003), "In Vitro chondrogenesis of bone marrow-derived mesenchymal stem cells in a photopolymerizing hydrogel", Tissue Eng., 9(4), 679-688.
  31. Williams, E.M., Graham, S.S., Akers, S.A., Reed, P.A. and Rushing, T.S. (2010), "Constitutive property behavior of an untra-high-performance concrete with and without steel fibers", Comput.Concrete, 7(2), 191-202.
  32. Winer, J.P., Janmey, P.A., McCormick, M.E. and Funaki, M. (2009), "Bone marrow-derived human mesenchymal stem cells become quiescent on soft substrates but remain responsive to chemical or mechanical stimuli", Tissue Eng., 15(1), 147-154.
  33. Woodfield, T.B.F., Malda, J., de Wijn, J., Péters, F., Riesle, J. and van Blitterswijk, C.A. (2004), "Design of porous scaffolds for cartilage tissue engineering using a three-dimensional fiber-deposition technique", Biomaterials, 25(18), 4149-4161.
  34. Yang, F., Williams, C.G., Wang, D.A., Lee, H., Manson, P.N. and Elisseeff, J. (2005), "The effect of incorporating RGD adhesive peptide in polyethylene glycol diacrylate hydrogel on osteogenesis of bone marrow stromal cells", Biomaterials, 26(30), 5991-5998.

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