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Enzymatic incorporation of bioactive peptides into fibrin matrices enhances neurite extension

Nature Biotechnology volume 18, pages 415–419 (2000)Cite this article

Abstract

Fibrin plays an important role in wound healing and regeneration, and enjoys widespread use in surgery and tissue engineering. The enzymatic activity of Factor XIIIa was employed to covalently incorporate exogenous bioactive peptides within fibrin during coagulation. Fibrin gels were formed with incorporated peptides from laminin and N-cadherin alone and in combination at concentrations up to 8.2 mol peptide per mole of fibrinogen. Neurite extension in vitro was enhanced when gels were augmented with exogenous peptide, with the maximal improvement reaching 75%. When this particular fibrin derivative was evaluated in rats in the repair of the severed dorsal root within polymeric tubes, the number of regenerated axons was enhanced by 85% relative to animals treated with tubes filled with unmodified fibrin. These results demonstrate that it is possible to enhance the biological activity of fibrin by enzymatically incorporating exogenous oligopeptide domains of morphoregulatory proteins.

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Figure 1: The effect of incorporated peptide at 8 mol peptide/mol fibrinogen on neurite length within three-dimensional fibrin matrices without (open bars) and with (hatched bars) soluble peptide added to the culture medium as competitive inhibitor.
Figure 2: The effect of peptide incorporated in a concentration series on neurite length within three-dimensional fibrin matrices.
Figure 3: Cross-sectional images of nerves regenerated in polymer tubes.

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References

  1. Langer, R. & Vacanti, J. Tissue engineering. Science 260, 920–926 (1993).

    Article  CAS  Google Scholar 

  2. Peppas, N. & Langer, R. New challenges in biomaterials. Science 263, 1715–1720 (1994).

    Article  CAS  Google Scholar 

  3. Hubbell, J. Biomaterials in tissue engineering. Biotechnology 13, 565–576 (1995).

    CAS  PubMed  Google Scholar 

  4. Pittman, R.N. & Buettner, H.M. Degradation of extracellular matrix by neuronal proteases. Dev. Neurosci. 11, 361–375 (1989).

    Article  CAS  Google Scholar 

  5. Herbert, C.B., Bittner, G.D. & Hubbell, J.A. Effects of Fibrinolysis on neurite growth from dorsal root ganglia cultured in two- and three-dimensional fibrin gels. J. Comp. Neurol. 365, 380–391 (1996).

    Article  CAS  Google Scholar 

  6. Lander, A. Understanding the molecules of cell contacts. Trends Neurosci. 12, 189–195 (1989).

    Article  CAS  Google Scholar 

  7. Malinda, K.M. & Kleinman, J.K. The laminins. Int. J. Biochem. Cell Biol. 28, 957–959 (1996).

    Article  CAS  Google Scholar 

  8. Luckenbill-Edds, L. Laminin and the mechanism of neuronal outgrowth. Brain Res. Brain Res. Rev. 23, 1–27 (1997).

    Article  CAS  Google Scholar 

  9. Martin, G.R. Laminin and other basement membrane proteins. Ann. Rev. Cell Biol. 3, 57–85 (1987).

    Article  CAS  Google Scholar 

  10. Tashiro, K. et al. The RGD containing site of the mouse laminin A chain is active for cell attachment, migration and neurite outgrowth. J. Cell Physiol. 146, 451–459 (1991).

    Article  CAS  Google Scholar 

  11. Tashiro, K., Sephel, G.E. & Weeks, B. A synthetic peptide containing the IKVAV sequence from the α chain of laminin mediates cell attachment, migration and neurite outgrowth. J. Biol. Chem. 264, 16174–16182 (1989).

    CAS  PubMed  Google Scholar 

  12. Graf, J. et al. Identification of an amino acid sequence in laminin mediating cell attachment, chemotaxis, and receptor binding. Cell 48, 989–996 (1987).

    Article  CAS  Google Scholar 

  13. Kleinman, H.K. et al. Identification of a second site in laminin for promotion of cell adhesion and migration and inhibition of in vivo melanoma lung colonization. Arch. Biochem. Biophys. 272, 39–45 (1989).

    Article  CAS  Google Scholar 

  14. Massia, S.P., Rao, S.S. & Hubbell, J.A. Covalently immobilized laminin peptide Tyr-Ile-Gly-Ser-Arg (YIGSR) supports cell spreading and colocalization of the 67 kilodalton receptor with α-actinin and viniculin. J. Biol. Chem. 268, 8053–8059 (1993).

    CAS  PubMed  Google Scholar 

  15. Liesi, P., Narvanen, A., Soos, J., Sariola, H. & Snounou, G. Identification of a neurite-outgrowth promoting domain using synthetic peptides. FEBS Lett. 244, 141–148 (1989).

    Article  CAS  Google Scholar 

  16. Hynes, R.O. Integrins: versatility, modulation, and signaling in cell adhesion. Cell 69, 11–25 (1992).

    Article  CAS  Google Scholar 

  17. Yamada, K.M. Adhesive recognition sequences. J. Biol. Chem. 266, 12809–12812 (1991).

    CAS  PubMed  Google Scholar 

  18. Zutter, M.M. & Santaro, S.A. Widespread histologic distribution of the α2β1 integrin cell-surface receptor. Am. J. Pathol. 137, 113–120 (1990).

    CAS  PubMed  PubMed Central  Google Scholar 

  19. Letourneau, P.C., Shattuck, T.A., Roche, F.K., Takeichi, M. & Lemmon, V. Nerve growth cone migration onto Schwann cells involves the calcium-dependent adhesion molecule, N-cadherin. Dev. Biol. 138, 430–442 (1990).

    Article  CAS  Google Scholar 

  20. Blaschuk, O.W. Identification of cadherin cell adhesion recognition sequence. Dev. Biol. 139, 227–229 (1990).

    Article  CAS  Google Scholar 

  21. Schense, J.C. & Hubbell, J.A. Cross-linking exogenous bifunctional peptides into fibrin gels with Factor XIIIa. Bioconjug. Chem. 10, 75–81 (1999).

    Article  CAS  Google Scholar 

  22. Sakata, Y. & Aoki, N. Cross-linking of α2-plasmin inhibitor to fibrin by fibrin-stabilizing factor. J. Clin. Invest. 65, 290–297 (1980).

    Article  CAS  Google Scholar 

  23. Ichinose, A., Tamaki, T. & Aoki, N. Factor XIII-mediated cross-linking of NH2−terminal peptide of α2-plasmin inhibitor to fibrin. FEBS Lett. 153, 369–371 (1983).

    Article  CAS  Google Scholar 

  24. Schense, J.C. & Hubbell, J.A. Three-dimensional migration of neurites is mediated by adhesion site density and affinity. J. Biol. Chem. in press (2000).

  25. Sakiyama, S.E., Schense, J.C. & Hubbell, J.A. Incorporation of heparin-binding peptides into fibrin gels enhances neurite extension: an example of designer matrices in tissue engineering. FASEB J. 13, 2214–2224 (1999).

    Article  CAS  Google Scholar 

  26. Sakiyama-Elbert, S.E. & Hubbell, J.A. Development of fibrin derivatives for controlled release of heparin-binding growth factors. J. Control Release 65, 389–402 (2000).

    Article  CAS  Google Scholar 

  27. Palecek, S.P., Loftus, J.C., Ginsberg, M.H., Lauffenburger, D.A. & Horwitz, A.F. Integrin-ligand binding properties govern cell migration speed through cell–substratum adhesiveness. Nature 385, 537–540 (1997).

    Article  CAS  Google Scholar 

  28. McCormack, M.L., Goddard, M., Guenard, V. & Aebischer, P. Comparison of dorsal and ventral spinal root regeneration through semipermeable guidance channels. J. Comp. Neurol. 313, 449–456 (1991).

    Article  CAS  Google Scholar 

  29. Williams, L.R. & Varon, S. Modification of fibrin matrix formation in situ enhances nerve regeneration in silicone chambers. J. Comp. Neurobiol. 231, 209–220 (1985).

    Article  CAS  Google Scholar 

  30. Fields, G. & Noble, R. Solid phase peptide synthesis utilizing 9-fluorenylmethoxycarbonyl amino acids. Intl. J. Pept. Protein Res. 35, 161–214 (1990).

    Article  CAS  Google Scholar 

  31. Varon, S.J., Nomura, J., Perez-Polo, J.R. & Shooter, E.M. The isolation and assay of the nerve growth factor proteins. In Methods of Neurochemistry (Marcel Dekker Inc., ed. Fried, R.) 203–229, New York, 1972).

    Google Scholar 

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Acknowledgements

This work was funded by Grant 31-52261 NFP 38 of the Swiss National Science Foundation.

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Authors and Affiliations

  1. Department of Materials and Institute for Biomedical Engineering, ETH-Zurich and the University of Zurich, Zurich, Switzerland

    Jason C. Schense & Jeffrey A. Hubbell

  2. Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena, 91125, CA

    Jason C. Schense

  3. Division of Surgical Research and Gene Therapy Center, Centre Hospitalier Universitaire Vaudois (CHUV), Lausanne University Medical School, Switzerland

    Jocelyne Bloch & Patrick Aebischer

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  1. Jason C. Schense
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  2. Jocelyne Bloch
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  4. Jeffrey A. Hubbell
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Correspondence to Jeffrey A. Hubbell.

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Schense, J., Bloch, J., Aebischer, P. et al. Enzymatic incorporation of bioactive peptides into fibrin matrices enhances neurite extension. Nat Biotechnol 18, 415–419 (2000). https://doi.org/10.1038/74473

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