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Annals of Biomedical Engineering

, Volume 41, Issue 5, pp 883–893 | Cite as

Non-Destructive Analysis of Extracellular Matrix Development in Cardiovascular Tissue-Engineered Constructs

  • M. Tuemen
  • D. V. A. Nguyen
  • J. Raffius
  • T. C. Flanagan
  • M. Dietrich
  • J. Frese
  • T. Schmitz-Rode
  • S. JockenhoevelEmail author
Article

Abstract

In the field of tissue engineering, there is an increasing demand for non-destructive methods to quantify the synthesis of extracellular matrix (ECM) components such as collagens, elastin or sulphated glycosaminoglycans (sGAGs) in vitro as a quality control before clinical use. In this study, procollagen I carboxyterminal peptide (PICP), procollagen III aminoterminal peptide (PIIINP), tropoelastin and sGAGs are investigated for their potential use as non-destructive markers in culture medium of statically cultivated cell-seeded fibrin gels. Measurement of PICP as marker for type I collagen synthesis, and PIIINP as marker of type III collagen turnover, correlated well with the hydroxyproline content of the fibrin gels, with a Pearson correlation coefficient of 0.98 and 0.97, respectively. The measurement of tropoelastin as marker of elastin synthesis correlated with the amount of elastin retained in fibrin gels with a Pearson correlation coefficient of 0.99. sGAGs were retained in fibrin gels, but were not detectable in culture medium at any time of measurement. In conclusion, this study demonstrates the potential of PICP and tropoelastin as non-destructive culture medium markers for collagen and elastin synthesis. To our knowledge, this is the first study in cardiovascular tissue engineering investigating the whole of here proposed biomarkers of ECM synthesis to monitor the maturation process of developing tissue non-invasively, but for comprehensive assessment of ECM development, these biomarkers need to be investigated in further studies, employing dynamic cultivation conditions and more complex tissue constructs.

Keywords

Extracellular matrix Collagen Elastin Sulphated glycosaminoglycans Biomarkers Non-destructive monitoring 

Notes

Acknowledgments

The present work has been done within a project of the Foerdergemeinschaft Deutsche Kinderherzzentren e.V. and within the Patim project, which is part of the in.nrw-innovation medical technology project and is funded by the European Union (EFRE-programme) and the NRW-Ziel2 programme. Furthermore we would like to thank the Department of Pathology, University Hospital Aachen, Germany. Author Disclosure Statement: No competing financial interests exist.

References

  1. 1.
    Ahmann, K. A., J. S. Weinbaum, S. L. Johnson, and R. T. Tranquillo. Fibrin degradation enhances vascular smooth muscle cell proliferation and matrix deposition in fibrin-based tissue constructs fabricated in vitro. Tissue Eng. A 16:3261–3270, 2010.CrossRefGoogle Scholar
  2. 2.
    Burgeson, R. E., and M. E. Nimni. Collagen types. Molecular structure and tissue distribution. Clin. Orthop. Relat. Res. 282:250–272, 1992.PubMedGoogle Scholar
  3. 3.
    Cholewinski, E., M. Dietrich, T. C. Flanagan, T. Schmitz-Rode, and S. Jockenhoevel. Tranexamic acid—an alternative to aprotinin in fibrin-based cardiovascular tissue engineering. Tissue Eng. A 15:3645–3653, 2009.CrossRefGoogle Scholar
  4. 4.
    Divya, P., and L. K. Krishnan. Glycosaminoglycans restrained in a fibrin matrix improve ECM remodelling by endothelial cells grown for vascular tissue engineering. J. Tissue Eng. Regen. Med. 3:377–388, 2009.PubMedCrossRefGoogle Scholar
  5. 5.
    Engelmayr, Jr., G. C., E. Rabkin, F. W. Sutherland, F. J. Schoen, J. E. Mayer, Jr., and M. S. Sacks. The independent role of cyclic flexure in the early in vitro development of an engineered heart valve tissue. Biomaterials. 26:175–187, 2005.PubMedCrossRefGoogle Scholar
  6. 6.
    Fite, B. Z., M. Decaris, Y. Sun, A. Lam, C. K. Ho, J. K. Leach, and L. Marcu. Noninvasive multimodal evaluation of bioengineered cartilage constructs combining time-resolved fluorescence and ultrasound imaging. Tissue Eng. C Methods. 17:495–504, 2011.CrossRefGoogle Scholar
  7. 7.
    Flanagan, T. C., C. Cornelissen, S. Koch, B. Tschoeke, J. S. Sachweh, T. Schmitz-Rode, and S. Jockenhoevel. The in vitro development of autologous fibrin-based tissue-engineered heart valves through optimised dynamic conditioning. Biomaterials 28:3388–3397, 2007.PubMedCrossRefGoogle Scholar
  8. 8.
    Flanagan, T. C., B. Wilkins, A. Black, S. Jockenhoevel, T. J. Smith, and A. S. Pandit. A collagen-glycosaminoglycan co-culture model for heart valve tissue engineering applications. Biomaterials 27:2233–2246, 2006.PubMedCrossRefGoogle Scholar
  9. 9.
    Gupta, V., J. A. Werdenberg, J. S. Mendez, and K. Jane Grande-Allen. Influence of strain on proteoglycan synthesis by valvular interstitial cells in three-dimensional culture. Acta Biomater. 4:88–96, 2009.CrossRefGoogle Scholar
  10. 10.
    Halme, T., T. Vihersaari, and R. Penttinen. Lysyl oxidase activity and synthesis of desmosines in cultured human aortic cells and skin fibroblasts: comparison of cell lines from control subjects and patients with the Marfan syndrome or other annulo-aortic ectasia. Scand. J. Clin. Lab. Invest. 46:31–37, 1986.PubMedCrossRefGoogle Scholar
  11. 11.
    Hoffman-Kim, D., M. S. Maish, P. M. Krueger, H. Lukoff, A. Bert, T. Hong, and R. A. Hopkins. Comparison of three myofibroblast cell sources for the tissue engineering of cardiac valves. Tissue Eng. 11:288–301, 2005.PubMedCrossRefGoogle Scholar
  12. 12.
    Jensen, L. T., and N. B. Host. Collagen: scaffold for repair or execution. Cardiovasc. Res. 33:535–539, 1997.PubMedCrossRefGoogle Scholar
  13. 13.
    Konig, K., K. Schenke-Layland, I. Riemann, and U. A. Stock. Multiphoton autofluorescence imaging of intratissue elastic fibers. Biomaterials 26:495–500, 2005.PubMedCrossRefGoogle Scholar
  14. 14.
    Kreitz, S., G. Dohmen, S. Hasken, T. Schmitz-Rode, P. Mela, and S. Jockenhoevel. Nondestructive method to evaluate the collagen content of fibrin-based tissue engineered structures via ultrasound. Tissue Eng. C Methods 17:1021–1026, 2011.CrossRefGoogle Scholar
  15. 15.
    Lieber, C. S., D. G. Weiss, and F. Paronetto. Value of fibrosis markers for staging liver fibrosis in patients with precirrhotic alcoholic liver disease. Alcohol. Clin. Exp. Res. 32:1031–1039, 2008.PubMedCrossRefGoogle Scholar
  16. 16.
    Long, J. L., and R. T. Tranquillo. Elastic fiber production in cardiovascular tissue-equivalents. Matrix Biol. 22:339–350, 2003.PubMedCrossRefGoogle Scholar
  17. 17.
    Mori, K., A. Shioi, S. Jono, Y. Nishizawa, and H. Morii. Dexamethasone enhances In vitro vascular calcification by promoting osteoblastic differentiation of vascular smooth muscle cells. Arterioscler. Thromb. Vasc. Biol. 19:2112–2118, 1999.PubMedCrossRefGoogle Scholar
  18. 18.
    Neame, P. J., and J. D. Sandy. Cartilage aggrecan. Biosynthesis, degradation and osteoarthritis. J. Fla Med. Assoc. 81:191–193, 1994.PubMedGoogle Scholar
  19. 19.
    Reddy, G. K., and C. S. Enwemeka. A simplified method for the analysis of hydroxyproline in biological tissues. Clin. Biochem. 29:225–229, 1996.PubMedCrossRefGoogle Scholar
  20. 20.
    Sato, Y., T. Miyamoto, R. Taniguchi, Y. Nishio, T. Kita, H. Fujiwara, and Y. Takatsu. Current understanding of biochemical markers in heart failure. Med. Sci. Monit. 12:RA252–RA264, 2006.PubMedGoogle Scholar
  21. 21.
    Schenke-Layland, K., I. Riemann, F. Opitz, K. Konig, K. J. Halbhuber, and U. A. Stock. Comparative study of cellular and extracellular matrix composition of native and tissue engineered heart valves. Matrix Biol. 23:113–125, 2004.PubMedCrossRefGoogle Scholar
  22. 22.
    Seibel, M. J. Molecular markers of bone turnover: biochemical, technical and analytical aspects. Osteoporos. Int. 11(Suppl 6):S18–S29, 2000.PubMedCrossRefGoogle Scholar
  23. 23.
    Stock, U. A., M. Nagashima, P. N. Khalil, G. D. Nollert, T. Herden, J. S. Sperling, A. Moran, J. Lien, D. P. Martin, F. J. Schoen, J. P. Vacanti, and J. E. Mayer, Jr. Tissue-engineered valved conduits in the pulmonary circulation. J. Thorac. Cardiovasc. Surg. 119:732–740, 2000.PubMedCrossRefGoogle Scholar
  24. 24.
    Tschoeke, B., T. C. Flanagan, S. Koch, M. S. Harwoko, T. Deichmann, V. Ella, J. S. Sachweh, M. Kellomaki, T. Gries, T. Schmitz-Rode, and S. Jockenhoevel. Tissue-engineered small-caliber vascular graft based on a novel biodegradable composite fibrin-polylactide scaffold. Tissue Eng. A 15:1909–1918, 2009.CrossRefGoogle Scholar
  25. 25.
    Weinbaum, J. S., J. Qi, and R. T. Tranquillo. Monitoring collagen transcription by vascular smooth muscle cells in fibrin-based tissue constructs. Tissue Eng. C Methods. 16:459–467, 2010.CrossRefGoogle Scholar
  26. 26.
    Woessner, Jr., J. F. The determination of hydroxyproline in tissue and protein samples containing small proportions of this imino acid. Arch. Biochem. Biophys. 93:440–447, 1961.PubMedCrossRefGoogle Scholar
  27. 27.
    Wu, W. J., B. Vrhovski, and A. S. Weiss. Glycosaminoglycans mediate the coacervation of human tropoelastin through dominant charge interactions involving lysine side chains. J. Biol. Chem. 274:21719–21724, 1999.PubMedCrossRefGoogle Scholar

Copyright information

© Biomedical Engineering Society 2013

Authors and Affiliations

  • M. Tuemen
    • 1
  • D. V. A. Nguyen
    • 1
  • J. Raffius
    • 1
  • T. C. Flanagan
    • 2
  • M. Dietrich
    • 1
  • J. Frese
    • 1
  • T. Schmitz-Rode
    • 1
  • S. Jockenhoevel
    • 1
    Email author
  1. 1.Department of Tissue Engineering and Textile Implants, Applied Medical Engineering, Helmholtz InstituteRWTH Aachen UniversityAachenGermany
  2. 2.School of Medicine and Medical Science, Health Sciences CentreUniversity College DublinDublinIreland

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