Enzyme-Pretreatment Removes In Vitro Cytoxic Effects of Dermal Sheep Collagen

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Abstract

We investigated the in vitro cytotoxicity of commercially available hexamethylenediisocyanate-crosslinked dermal sheep collagen (HDSC). HDSC was found to induce medium cytotoxic effects, as measured with methylcellulose cell culture. Apart from primary cytotoxicity, due to direct release of (extractable) cytotoxic products, HDSC was found to contain secondary cytotoxicity, possibly released by enzymatic interactions. In this study we found proof for this hypothesis, by exposing extracted HDSC to enzyme-containing medium. Furthermore we observed, that enzymatic pre-treatment can remove all secondary cytotoxic products, possibly due to detachment of pendants, which are still coupled to fragments of collagen molecules. The possibility of enzymatic pretreatment of HDSC, to obtain a non-cytotoxic/biocompatible material, may be important for in vivo applications.

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References

  1. 1.

    Chvaphil, M, Speer, D., Mora, W., Eskelson, C., Effect of tanning agent on tissue reaction to tissue implanted collagen sponge, J. Surg. Res. 35, 402–409, 1983.

    Article  Google Scholar 

  2. 2.

    Van Wachem, P.B., Van Luyn, M.J.A., Koerten, H.K., Olde Damink, L., Ten Hoopen, H., Feijen, J., Nieuwenhuis, P., In vivo degradation of processed dermal sheep collagen evaluated with transmission electron microscopy, Biomaterials, 12, 215–223, 1991.

    Article  Google Scholar 

  3. 3.

    Van Wachem, P.B., Van Luyn, M.J.A., Olde Damink, L., Feijen, J., Nieuwenhuis, P., Tissue interactions with dermal sheep collagen implants: a transmission electron microscopical evaluation, Cells and Materials, 1, 3, 251–263, 1991.

    Google Scholar 

  4. 4.

    Ulreich, J.B., Chvapil, M., A quantitative microassay for in vitro toxicity testing of biomaterials, J. Biomed. Mater. Res., 15, 913–922, 1981.

    CAS  Article  Google Scholar 

  5. 5.

    Van Luyn, M.J.A., Van Wachem, P.B., Olde Damink, L., Ten Hoopen, H., Feijen, J., Nieuwenhuis, P., Methylcellulose culture as a new cytotoxicity test system for biomaterials, J. of Mat. Sci.: Mater. in Med., 2, 142–148, 1991.

    Google Scholar 

  6. 6.

    Van Luyn, M.J.A., Van Wachem, P.B., Olde Damink, L.H.H., Dijkstra, P. J., Feijen, J., Nieuwenhuis, P., Relations between in vitro cytotoxicity and crosslinked dermal sheep collagens, Submitted to J. Biomed. Mater. Res.

  7. 7.

    Van Luyn, M.J.A., Van Wachem, M.H. Jonkman, J., Nieuwenhuis, P., Cytotoxicity testing of wound dressings using methylcellulose cell culture, accepted for publication in Biomaterials, 1991

    Google Scholar 

  8. 8.

    Iscove, N.N. and Schreier, H., In: Clonal Growth of Cells in Semisolid or Viscous Medium, Immunological Methods, Academic Press, 1979.

    Google Scholar 

  9. 9.

    Berode, M., Leuenberger, Ph. and Savolainen, H., Phenotype-dependent Inhibition of Human α1-antitrypsin by 1,6-Hexanediamine in Vitro, Biochem. Int.. 17, 1, 1988.

    Google Scholar 

  10. 10.

    Yano, E., Yoshioka, M. and Koizumi, A., Relationship between chemical structure and cytotoxicity of aliphatic amines examined by a microtiter system with cultured fibroblasts, Jpn. J. Ind. Health, 23, 537–543, 1981.

    CAS  Google Scholar 

  11. 11.

    Johannsen, F.R. and Levinskas, G.J., Toxicological Profile of orally administered 1,6-Hexanediamine in the rat, J.of Applied Toxicol. 7, 4, 259–263, 1987.

    CAS  Article  Google Scholar 

  12. 12.

    Murphy, G., Reynolds, J.J., Current views of collagen degradation, Progress towards understanding the resorption of connective tissues, Bio Essays 2, 55–60, 1985.

    Google Scholar 

  13. 13.

    Yajima, T., Acid phosphatase activity and intracellular collagen degradation by fibroblasts in vitro, Cell and Tissue Research, 245, 253–260, 1986.

    CAS  Article  Google Scholar 

  14. 14.

    Woolley, D.E., Mammalian collagenases, In: Extracellular Matrix Biochemistry, Eds. Piez K.A. and Reddi, A.H., Elsevier, New York, 119–157, 1984.

  15. 15.

    Lazarus, S.L., and Fullmer, H.M., Collagenase production by human dermis in vitro, J. of Invest. Derm., 52, 6, 1969.

    Article  Google Scholar 

  16. 16.

    Harris, E D., Krane, S. M., Collagenases: Specific involement of collagenase in normal physiology and pathology, The New. Eng. J. of Med. 652–661, 1974

    Google Scholar 

  17. 17.

    Kleiman, H. K., Klebe, R. J., and Martin, G.R. Role of Collagenous Matrices in the Adhesion and Growth of Cells, J. Cell Biol., 88, 473–485, 1981.

    Article  Google Scholar 

  18. 18.

    Nimni, M.E., and Harkness, R.,D., Molecular Structure of Collagen, In: Collagen, Vol. 1, Chapter 1, Ed. Nimni, M.E., CRC Press, Inc., Boca Raton Florida, U.S.A., 1988.

  19. 19.

    Fields, G.B., Van Wart, H. E., and Birkedal-Hansen, H., Sequence specificity of human skin fibroblast collagenase, The J. of Biol. Chem., Vol 262, 6221–6226,1987.

    CAS  Article  Google Scholar 

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van Luyn, M.J.A., van Wachem, P.B., Olde Damink, L.H.H. et al. Enzyme-Pretreatment Removes In Vitro Cytoxic Effects of Dermal Sheep Collagen. MRS Online Proceedings Library 252, 167–174 (1991). https://doi.org/10.1557/PROC-252-167

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