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Assessment of cell viability in a three-dimensional enzymatically cross-linked collagen scaffold

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Abstract

Microbial transglutaminase (mTGase) is an enzyme that introduces a covalent bond between peptide bound glutamine and lysine residues. Proteins cross-linked in this manner are often more resistant to proteolytic degradation and show increased tensile strength. This study evaluates the effects of mTGase mediated cross-linking of collagen on the cellular morphology, behaviour and viability of murine 3T3 fibroblasts following their seeding into collagen scaffolds. Additionally, cell mediated scaffold contraction, porosity and level of cross-linking of the scaffold has been analysed using image analysis software, scanning electron microscopy (SEM), colorimetric assays, and Fourier transform infrared spectroscopy (FTIR). We demonstrate that the biocompatibility and cellular morphology, when comparing cultures of fibroblasts integrated in mTGase cross-linked collagen scaffolds with the native collagen counterparts, remained unaffected. It has been also elicited that the structural characteristics of collagen have been preserved while introducing enzymatically resistant covalent bonds.

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References

  1. C. H. LEE, A. SINGLA and Y. LEE, Int. J. Pharm. 221 (2001) 1

    Article  CAS  Google Scholar 

  2. M. D. FERNANDEZ-DIAZ, P. MONTERO and M. C. GOMEZ-GUILLEN, Food Chem. 74 (2001) 161

    Article  CAS  Google Scholar 

  3. W. FRIESS, Eur. J. Pharm. Biopharm. 45 (1998) 113

    Article  CAS  Google Scholar 

  4. D. Y. S. CHAU, R. J. COLLIGHAN, E. A. M. VERDERIO, V. L. ADDY and M. GRIFFIN, Biomaterials 26 (2005) 6518

    Article  CAS  Google Scholar 

  5. B. ECKES, P. ZIGRINO, D. KESSLER, O. HOLTKOTTER, P. SHEPHARD, C. MAUCH and T. KRIEG, Matrix. Biol. 19 (2000) 325

    Article  CAS  Google Scholar 

  6. F. GRINNELL, Trends Cell Biol. 10 (2000) 362

    Article  CAS  Google Scholar 

  7. M. GRIFFIN, R. CASADIO and C. M. BERGAMINI, Biochem. J. 368 (2002) 377

    Article  CAS  Google Scholar 

  8. R. SHARMA, P. C. LORENZEN and K. B. QVIST, Int. Dairy J. 11 (2001) 785

    Article  CAS  Google Scholar 

  9. M. MOTOKI and K. SEGURO, Trends Food Sci. Technol. 9 (1998) 204

    Article  CAS  Google Scholar 

  10. P. GRENARD, S. BRESSON-HADNI, S. EL ALAOUI, M. CHEVALLIER, D. A. VUITTON and S. RICARD-BLUM, J. Hepatol. 35 (2001) 367

    Article  CAS  Google Scholar 

  11. S. ALLEN, J. SOTOS, M. J. SYLTE and C. J. CZUPRYNSKI, Clin. Diagn. Lab. Immunol. 8 (2001) 460

    Article  CAS  Google Scholar 

  12. W. A. BUBNIS and C. M. OFNER 3rd, Anal. Biochem. 207 (1992) 129

    Article  CAS  Google Scholar 

  13. C. M. OFNER 3rd and W. A. BUBNIS, Pharm. Res. 13 (1996) 1821

    Article  CAS  Google Scholar 

  14. A. KAMINSKA and A. SIONKOWSKA, Polym. Degrad. Stab. 51 (1996) 15

    Article  CAS  Google Scholar 

  15. X. H. WANG, D. P. LI, W. J. WANG, Q. L. FENG, F. Z. CUI, Y. X. XU, X. H. SONG and M. VAN DER WERF, Biomaterials 24 (2003) 3213

    Article  CAS  Google Scholar 

  16. D. GOPINATH, M. R. AHMED, K. GOMATHI, K. CHITRA, P. K. SEHGAL and R. JAYAKUMAR, Biomaterials 25 (2004) 1911

    Article  CAS  Google Scholar 

  17. K. W. NG, D. T. LEONG and D. W. HUTMACHER, Tissue Eng. 11 (2005) 182

    Article  CAS  Google Scholar 

  18. P. FRIEDL and E.-B. BRÖCKER, Cell Mol. Life Sci. 57 (2000) 41

    Article  CAS  Google Scholar 

  19. C. A. LAMBERT, A. C. COLIGE, C. MUNAUT, C. M. LAPIERE and B. V. NUSGENS, Matrix Biol. 20 (2001) 397

    Article  CAS  Google Scholar 

  20. F. GRINNELL, Trends Cell Biol. 13 (2003) 264

    Article  CAS  Google Scholar 

  21. M. CHIQUET, A. S. RENEDO, F. HUBER and M. FLUCK, Matrix Biol. 22 (2003) 73

    Article  CAS  Google Scholar 

  22. B. CHEVALLAY, N. ABDUL-MALAK and D. HERBAGE, J. Biomed. Mater. Res. 49 (2000) 448

    Article  CAS  Google Scholar 

  23. T. L. LEE, Y. C. LIN, K. MOCHITATE and F. GRINNELL, J. Cell Sci. 105(Pt 1) (1993) 167

    CAS  Google Scholar 

  24. R. T. PRAJAPATI, B. CHAVALLY-MIS, D. HERBAGE, M. EASTWOOD and R. A. BROWN, Wound Repair Regen. 8 (2000) 226

    Article  CAS  Google Scholar 

  25. M. CHIQUET, Matrix Biol. 18 (1999) 417

    Article  CAS  Google Scholar 

  26. F. GRINNELL, C. H. HO, E. TAMARIZ, D. J. LEE and G. SKUTA, Mol. Biol. Cell 14 (2003) 384

    Article  CAS  Google Scholar 

  27. E. TAMARIZ and F. GRINNELL, Mol. Biol. Cell 13 (2002) 3915

    Article  CAS  Google Scholar 

  28. M. EASTWOOD, V. C. MUDERA, D. A. MCGROUTHER, R. A. BROWN, Cell Motil. Cytoskeleton 40 (1998) 13

    Article  CAS  Google Scholar 

  29. M. M. NOCIARI, A. SHALEV, P. BENIAS and C. RUSSO, J. Immunol. Methods 213 (1998) 157

    Article  CAS  Google Scholar 

  30. L. LORAND and R. M. GRAHAM, Nat. Rev. Mol. Cell Biol. 4 (2003) 140

    Article  CAS  Google Scholar 

  31. K. YOKOYAMA, N. NIO and Y. KIKUCHI, Appl. Microbiol. Biotechnol. 64 (2004) 447

    Article  CAS  Google Scholar 

  32. B. BRODSKY and J. A. M. RAMSHAW, Matrix Biol. 15 (1997) 545

    Article  CAS  Google Scholar 

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Acknowledgements

This experimental work has been funded by The Irish Higher Education Authority’s Programme for Research in third Level Institutions, Enterprise Ireland Research Innovation Fund and EPSRC (grant reference GR/S21755/02). Special thanks are due to Damien O’Halloran and Michael Ball for all the help provided in the laboratory work and Afshin Samali and Eva Szegezdi for their valuable comments on apoptosis evaluation.

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Authors and Affiliations

  1. National Centre for Biomedical Engineering Science, National University of Ireland, Galway, Ireland

    Y. Garcia & A. Pandit

  2. School of Life and Health Sciences, Aston University, Aston Triangle, Birmingham, B4 7ET, UK

    R. Collighan & M. Griffin

  3. Department of Mechanical and Biomedical Engineering, National University of Ireland, Galway, Ireland

    A. Pandit

Authors
  1. Y. Garcia
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  2. R. Collighan
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  3. M. Griffin
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  4. A. Pandit
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Corresponding author

Correspondence to A. Pandit.

Additional information

This work was performed in the National Centre for Biomedical Engineering Science, Orbsen Building, National University of Ireland, Galway, Ireland.

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Garcia, Y., Collighan, R., Griffin, M. et al. Assessment of cell viability in a three-dimensional enzymatically cross-linked collagen scaffold. J Mater Sci: Mater Med 18, 1991–2001 (2007). https://doi.org/10.1007/s10856-007-3091-9

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  • DOI: https://doi.org/10.1007/s10856-007-3091-9

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