Glutaraldehyde as a crosslinking agent for collagen-based biomaterials

The formation of Schiff bases during crosslinking of dermal sheep collagen (DSC) with glutaraldehyde (GA), their stability and their reactivity towards GA was studied. All available free amine groups had reacted with GA to form a Schiff base within 5 min after the start of the reaction under the conditions studied (0.5% (w/w) GA). Before crosslinks are formed the hydrolysable Schiff bases initially present were stabilized by further reaction with GA molecules. An increase in shrinkage temperature (T s) from 56°C for non-crosslinked DSC (N-DSC) to 78°C for GA crosslinked DSC (G-DSC) was achieved after crosslinking for 1 h. From the relationship between the free amine group content and the T s during crosslinking it was concluded that higher GA concentrations and longer reaction times will result in the introduction of pendant-GA-related molecules rather than crosslinks. After 24 h crosslinking an average uptake of 3 GA molecules per reacted amine group was found. No increase in the tensile strength of the materials was observed after crosslinking, which may be a result of formation of crosslinks within the fibres rather than in between fibres. Aligning of the fibres by applying a pre-strain to the samples and subsequent crosslinking yielded materials with an increased tensile strength.

This is a preview of subscription content, access via your institution.


  1. 1.

    M. E. NIMNI, D. T. CHEUNG, B. STRATES, M. KODAMA and K. SKEIKH, J. Biomed. Mater. Res. 21 (1987) 741.

    Google Scholar 

  2. 2.

    M. CHVAPIL, D. GIBEAULT and T. F. WANG, J. Biomed. Mater. Res. 21 (1987) 1383.

    Google Scholar 

  3. 3.

    J. M. MCPHERSON, P. W. LEDGER, S. SAWAMURA, A. CONTI, S. WADE, H. REIHANIAN and D. G. WALLACE, J. Biomed. Mater. Res. 20 (1986) 79.

    Google Scholar 

  4. 4.

    D. T. CHEUNG and M. E. NIMNI, Conn. Tissue Res. 10 (1982) 187.

    Google Scholar 

  5. 5.

    J. M. RUIJGROK, M. E. BOON and J. R.DE WIJN, Histochem. J. 22 (1990) 389.

    Google Scholar 

  6. 6.

    R. LUBIG, P. KUSCH, K. RöPER and H. ZAHN, Monatshefte für Chemie 112 (1981) 1313.

    Google Scholar 

  7. 7.

    U. TOMIMATSU, E. F. JANSEN, W. GAFFIELD and A. C. OLSEN, J. Colloid Interface Sci. 36 (1971) 51.

    Google Scholar 

  8. 8.

    D. T. CHEUNG and M. E. NIMNI, Conn. Tissue Res. 10 (1982) 201.

    Google Scholar 

  9. 9.

    P. M. HARDY, A. C. NICHOLLS and H. N. RYDON, J. Chem. Soc. Perkin I. (1976) 958.

    Google Scholar 

  10. 10.

    K. OKUDA, I. URABE, Y. YAMADA and H. OKADA, J. Ferm. Bioeng. 71 (1991) 100.

    Google Scholar 

  11. 11.

    E. A. WOODROOF, J. Bioengng 2 (1978) 1.

    Google Scholar 

  12. 12.

    T. J. A. JOHNSON, Eur. J. Cell Biol. 45 (1987) 160.

    Google Scholar 

  13. 13.

    P. M. HARDY, G. J. HUGHES and H. N. RYDON, J. Chem. Soc. Perkin I. (1978) 2282.

    Google Scholar 

  14. 14.

    T. M.Van GULIK and P. J. KLOPPER, Neth. J. Surg. 39 (1987) 90.

    Google Scholar 

  15. 15.

    L. L. H. HUANG-LEE, D. T. CHEUNG and M. E.NIMNI, Biomed. Mater. Res. 24 (1990) 1185.

    Google Scholar 

  16. 16.

    R. GILLET and K. GULL, Histochem. 30 (1972) 162.

    Google Scholar 

  17. 17.

    P. J. FLORY and R. R. GARRETT, J. Amer. Chem. Soc. 80 (1958) 4836.

    Google Scholar 

  18. 18.

    Society Leather Trades' Chemists, J. Soc. Leather Trades' Chemists 47 (1963).

  19. 19.

    D. L. GILBERT and S. W. KIM, J. Biomed. Mater. Res. 24 (1990) 1221.

    Google Scholar 

  20. 20.

    C. L. WANG, T. MIYATA, B. WEKSLER, A. L. RUBIN and K. H. STENZEL, Biochim. Biophys. Acta 544 (1978) 555.

    Google Scholar 

  21. 21.

    A. VOS and P. J.VAN VLIMMEREN, J. Soc. Leather Technol. Chem. 57 (1973) 93.

    Google Scholar 

  22. 22.

    Society Leather Trades' Chemists, J. Soc. Leather Trades' Chemists, 44 (1960).

  23. 23.

    E. H. BLAAUW, M. F. JONKMAN and P. O. GERRITS, Acta Morphol. Neerl.-Scand. 25 (1987) 167.

    Google Scholar 

  24. 24.

    R. F. BORCH, M. D. BERNSTEIN and H. D. DURST, J. Amer. Chem. Soc. 93 (1971) 2897.

    Google Scholar 

  25. 25.

    D. T. CHEUNG, N. PERELMAN, E. C. KO and M. E. NIMNI, Conn. Tissue Res. 13 (1985) 109.

    Google Scholar 

  26. 26.

    J. H. BOWES and C. W. CATER, Biochim. Biophys. Acta 168 (1968) 341.

    Google Scholar 

  27. 27.

    S. C. ROE, B. K. MILTHORPE and K. SCHINDHELM, Artif. Org. 14 (1990) 443.

    Google Scholar 

  28. 28.

    C. E. VISSER, A. B. E. VOUTE, J. OOSTING, M. E. BOON and L. P. KOK, Biomaterials 13 (1992) 34.

    Google Scholar 

  29. 29.

    C. A. PEREIRA, J. M. LEE and S. A. HABERER, J. Biomed. Mater. Res. 24 (1990) 345.

    Google Scholar 

  30. 30.

    G. A. ROBERTS and K. E. TAYLOR, Makromol. Chem. 190 (1989) 951.

    Google Scholar 

  31. 31.

    E. HEIDEMANN, J. Soc. Leather Techn. Chem. 66 (1982) 21.

    Google Scholar 

  32. 32.

    B. M. HAINES, J. Amer. Leather Chem. Assoc. 77 (1982) 180.

    Google Scholar 

  33. 33.

    J. VACULIK, Leder Schuhe Lederwaren 3 (1983) 132.

    Google Scholar 

  34. 34.

    P. L. KRONICK and P. R. BUECHLER, J. Amer. Leather Chem. Assoc. 81 (1986) 221.

    Google Scholar 

  35. 35.

    V. MOHANARADHAKRISHNAN, P. L. MUTHIAH and A. HADHANYI, Arzneim.-Forsch. (Drug Res.) 25 (1975) 726.

    Google Scholar 

  36. 36.

    A. HILTNER, J. J. CASSIDY and E. BAER, Ann. Rev. Mater. Sci. 15 (1985) 455.

    Google Scholar 

  37. 37.

    J. M. LEE, D. R. BOUGHNER and D. W. COURTMAN, J. Biomed. Mater. Res. 18 (1984) 79.

    Google Scholar 

  38. 38.

    Y. P. KATO, D. L. CHRISTIANSEN, R. A. HAHN, S. J. SHIEH, J. D. GOLDSTEIN and F. H. SILVER, Biomaterials 10 (1989) 38.

    Google Scholar 

  39. 39.

    J. K. LAW, J. R. PARSONS, F. H. SILVER and A. B. WEISS, J. Biomed. Mater. Res. 23 (1989) 961.

    Google Scholar 

Download references

Author information



Rights and permissions

Reprints and Permissions

About this article

Cite this article

Olde Damink, L.H.H., Dijkstra, P.J., Van Luyn, M.J.A. et al. Glutaraldehyde as a crosslinking agent for collagen-based biomaterials. J Mater Sci: Mater Med 6, 460–472 (1995).

Download citation


  • Tensile Strength
  • Shrinkage
  • Glutaraldehyde
  • Schiff Base
  • Crosslinking Agent