Skip to main content
SpringerLink
Log in

Behavior of Human Mesenchymal Stem Cells in Fibrin-Based Vascular Tissue Engineering Constructs

  • Published:
Annals of Biomedical Engineering Aims and scope Submit manuscript

Abstract

A limitation of current tissue engineering vascular graft technology is the provision of an expandable, autologous cell source. By harnessing the multipotency of mesenchymal stem cells (MSC), it is hoped that functional vascular cells can be produced. To date, a range of 2D and 3D environments have been investigated for the manipulation of MSC differentiation pathways. To this end, this study aims to test the hypothesis that MSC seeded in various fibrin gel environments will exhibit evidence of a smooth muscle cell (SMC) phenotype. Initially, a range of cell-seeding densities were screened for 2D and 3D fibrin constructs, where it was observed that a seeding densities of 500,000 cells/mL facilitated gel compaction without degradation or loss in cell viability. Additionally, positive expression of CD49, CD73, CD105 markers and negative expression of hemopoietic stem cell-associated CD34 and CD45 indicated that MSC phenotype was retained within the fibrin gel. Nonetheless, a decrease in the gene expression of α-smooth cell actin and calponin was observed for MSC cultured in static 3D fibrin gels. Although a slight recovery was observed after 24 h mechanical stimulation, the fold-change remained significantly lower than that observed for cells cultured on 2D tissue culture plastic. While MSC differentiation toward a SMC appears possible in both 2D and 3D environments, scaffold architecture and mechanical stimulation undoubtedly play an important role in the creation of a functional SMC phenotype.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7

Abbreviations

MSC:

Mesenchymal stem cell

α-SMA:

α-Smooth cell actin

SMC:

Smooth muscle cell

DMEM:

Dulbecco’s modified Eagle’s medium

TBS:

Tris-buffered saline

TEVG:

Tissue-engineered vascular graft

References

  1. Anderson, J. S., T. M. Price, S. R. Hanson, and L. A. Harker. In vitro endothelialization of small-caliber vascular grafts. Surgery 101(5):577–586, 1987.

    PubMed  CAS  Google Scholar 

  2. Aper, T., O. E. Teebken, G. Steinhoff, and A. Haverich. Use of a fibrin preparation in the engineering of a vascular graft model. Eur. J. Vasc. Endovasc. Surg. 28(3):296–302, 2004.

    Article  PubMed  CAS  Google Scholar 

  3. Barocas, V. H., T. S. Girton, and R. T. Tranquillo. Engineered alignment in media equivalents: Magnetic prealignment and mandrel compaction. J. Biomech. Eng. 120(5):660–666, 1998.

    Article  PubMed  CAS  Google Scholar 

  4. Calderwood, D. A., S. J. Shattil, and M. H. Ginsberg. Integrins and actin filaments: Reciprocal regulation of cell adhesion and signaling. J. Biol. Chem. 275(30):22607–22610, 2000.

    Article  PubMed  CAS  Google Scholar 

  5. Cho, S. W., S. H. Lim, I. K. Kim, Y. S. Hong, S. S. Kim, K. J. Yoo, H. Y. Park, Y. Jang, B. C. Chang, and C. Y. Choi. Small-diameter blood vessels engineered with bone marrow-derived cells. Ann. Surg. 241(3):506–515, 2005.

    Article  PubMed  Google Scholar 

  6. Christman, K. L., A. J. Vardanian, Q. Fang, R. E. Sievers, H. H. Fok, and R. J. Lee. Injectable fibrin scaffold improves cell transplant survival, reduces infarct expansion, and induces neovasculature formation in ischemic myocardium. J. Am. Coll. Cardiol. 44(3):654–660, 2004.

    Article  PubMed  CAS  Google Scholar 

  7. Cox, S., M. Cole, and B. Tawil. Behavior of human dermal fibroblasts in three-dimensional fibrin clots: dependence on fibrinogen and thrombin concentration. Tissue Eng. 10(5–6):942–954, 2004.

    Article  PubMed  CAS  Google Scholar 

  8. Cukierman, E., R. Pankov, D. R. Stevens, and K. M. Yamada. Taking cell-matrix adhesions to the third dimension. Science 294(5547):1708, 2001.

    Article  PubMed  CAS  Google Scholar 

  9. Cummings, C. L., D. Gawlitta, R. M. Nerem, and J. P. Stegemann. Properties of engineered vascular constructs made from collagen, fibrin, and collagen-fibrin mixtures. Biomaterials 25(17):3699–3706, 2004.

    Article  PubMed  CAS  Google Scholar 

  10. DeMali, K. A., K. Wennerberg, and K. Burridge. Integrin signaling to the actin cytoskeleton. Curr. Opin. Cell Biol. 15(5):572–582, 2003.

    Article  PubMed  CAS  Google Scholar 

  11. Emura, M., A. Ochiai, M. Horino, W. Arndt, K. Kamino, and S. Hirohashi. Development of myofibroblasts from human bone marrow mesenchymal stem cells cocultured with human colon carcinoma cells and TGF beta 1. In Vitro Cell. Dev. Biol. Anim. 36(2):77–80, 2000.

    Article  PubMed  CAS  Google Scholar 

  12. Engelmayr, Jr., G. C., V. L. Sales, J. E. Mayer, Jr., and M. S. Sacks. Cyclic flexure and laminar flow synergistically accelerate mesenchymal stem cell-mediated engineered tissue formation: Implications for engineered heart valve tissues. Biomaterials 27(36):6083–6095, 2006.

    Article  PubMed  CAS  Google Scholar 

  13. Gundy, S., G. Manning, E. O’Connell, V. Ellä, M. Sri Harwoko, Y. Rochev, T. J. Smith, and V. Barron. Human coronary artery smooth muscle cell response to a novel PLA textile/fibrin gel composite scaffold. Acta Biomater. 4(6):1734–1744, 2008.

    Article  PubMed  CAS  Google Scholar 

  14. Ho, W., B. Tawil, J. C. Dunn, and B. M. Wu. The behavior of human mesenchymal stem cells in 3D fibrin clots: dependence on fibrinogen concentration and clot structure. Tissue Eng. 12(6):1587–1595, 2006.

    Article  PubMed  CAS  Google Scholar 

  15. Huang, C. Y., M. A. Deitzer, and H. S. Cheung. Effects of fibrinolytic inhibitors on chondrogenesis of bone-marrow derived mesenchymal stem cells in fibrin gels. Biomech. Model. Mechanobiol. 6(1–2):5–11, 2007.

    Article  PubMed  Google Scholar 

  16. Kadletz, M., H. Magometschnigg, E. Minar, G. Konig, M. Grabenwoger, M. Grimm, and E. Wolner. Implantation of in vitro endothelialized polytetrafluoroethylene grafts in human beings: A preliminary report. J. Thorac. Cardiovasc. Surg. 104(3):736–742, 1992.

    PubMed  CAS  Google Scholar 

  17. Kashiwakura, Y., Y. Katoh, K. Tamayose, H. Konishi, N. Takaya, S. Yuhara, M. Yamada, K. Sugimoto, and H. Daida. Isolation of bone marrow stromal cell-derived smooth muscle cells by a human SM22alpha promoter: in vitro differentiation of putative smooth muscle progenitor cells of bone marrow. Circulation 107(16):2078–2081, 2003.

    Article  PubMed  Google Scholar 

  18. Kinner, B., J. M. Zaleskas, and M. Spector. Regulation of smooth muscle actin expression and contraction in adult human mesenchymal stem cells. Exp. Cell Res. 278(1):72–83, 2002.

    Article  PubMed  CAS  Google Scholar 

  19. Kjaergard, H. K., and H. R. Trumbull. Vivostat system autologous fibrin sealant: preliminary study in elective coronary bypass grafting. Ann. Thorac. Surg. 66(2):482–486, 1998.

    Article  PubMed  CAS  Google Scholar 

  20. Kurpinski, K., J. Chu, C. Hashi, and S. Li. Anisotropic mechanosensing by mesenchymal stem cells. Proc. Natl Acad. Sci. USA 103(44):16095–16100, 2006.

    Article  PubMed  CAS  Google Scholar 

  21. Kurpinski, K., J. Park, R. G. Thakar, and S. Li. Regulation of vascular smooth muscle cells and mesenchymal stem cells by mechanical strain. Mol. Cell. Biomech. 3(1):21–34, 2006.

    PubMed  Google Scholar 

  22. Li, S., J. J. Moon, H. Miao, G. Jin, B. P. C. Chen, S. Yuan, Y. Hu, S. Usami, and S. Chien. Signal transduction in matrix contraction and the migration of vascular smooth muscle cells in three-dimensional matrix. J. Vasc. Res. 40(4):378–388, 2003.

    Article  PubMed  CAS  Google Scholar 

  23. Liu, J. Y., D. D. Swartz, H. F. Peng, S. F. Gugino, J. A. Russell, and S. T. Andreadis. Functional tissue-engineered blood vessels from bone marrow progenitor cells. Cardiovasc. Res. 75(3):618–628, 2007.

    Article  PubMed  CAS  Google Scholar 

  24. Livak, K. J., and T. D. Schmittgen. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods 25(4):402–408, 2001.

    Article  PubMed  CAS  Google Scholar 

  25. Murphy, J. M., D. J. Fink, E. B. Hunziker, and F. P. Barry. Stem cell therapy in a caprine model of osteoarthritis. Arthritis Rheum. 48(12):3464–3474, 2003.

    Article  PubMed  Google Scholar 

  26. Nieponice, A., T. M. Maul, J. M. Cumer, L. Soletti, and D. A. Vorp. Mechanical stimulation induces morphological and phenotypic changes in bone marrow-derived progenitor cells within a three-dimensional fibrin matrix. J. Biomed. Mater. Res. A 81(3):523–530, 2007.

    PubMed  Google Scholar 

  27. O’Cearbhaill, E. D., M. Punchard, M. Murphy, F. Barry, P. E. McHugh, and V. Barron. Response of mesenchymal stem cells to the biomechanical environment of the endothelium on a flexible tubular silicone substrate. Biomaterials 29(11):1610, 2007.

    Article  CAS  Google Scholar 

  28. Park, J. S., J. S. Chu, C. Cheng, F. Chen, D. Chen, and S. Li. Differential effects of equiaxial and uniaxial strain on mesenchymal stem cells. Biotechnol. Bioeng. 88(3):359–368, 2004.

    Article  PubMed  CAS  Google Scholar 

  29. Park, J. S., N. F. Huang, K. T. Kurpinski, S. Patel, S. Hsu, and S. Li. Mechanobiology of mesenchymal stem cells and their use in cardiovascular repair. Front. Biosci. 12:5098–5116, 2007.

    Article  PubMed  CAS  Google Scholar 

  30. Pelham, R. J., and Y. Wang. Cell locomotion and focal adhesions are regulated by substrate flexibility. Proc. Natl Acad. Sci. USA 94(25):13661–13665, 1997.

    Article  PubMed  CAS  Google Scholar 

  31. Punchard, M. A., C. Stenson-Cox, E. D. O’Cearbhaill, E. Lyons, S. Gundy, L. Murphy, A. Pandit, P. E. McHugh, and V. Barron. Endothelial cell response to biomechanical forces under simulated vascular loading conditions. J. Biomech. 40(14):3146–3154, 2007.

    Article  PubMed  CAS  Google Scholar 

  32. Reyna, S., D. Ensenat, F. Johnson, H. Wang, A. Schafer, and W. Durante. Cyclic strain stimulates proline transport in vascular smooth muscle cells. Am. J. Hypertens. 17(8):712–717, 2004.

    Article  PubMed  CAS  Google Scholar 

  33. Riha, G. M., P. H. Lin, A. B. Lumsden, Q. Yao, and C. Chen. Review: application of stem cells for vascular tissue engineering. Tissue Eng. 11(9–10):1535–1552, 2005.

    Article  PubMed  CAS  Google Scholar 

  34. Ross, J. J., and R. T. Tranquillo. ECM gene expression correlates with in vitro tissue growth and development in fibrin gel remodeled by neonatal smooth muscle cells. Matrix Biol. 22(6):477–490, 2003.

    Article  PubMed  CAS  Google Scholar 

  35. Simpson, D., H. Liu, T. H. M. Fan, R. Nerem, and S. C. Dudley, Jr. A tissue engineering approach to progenitor cell delivery results in significant cell engraftment and improved myocardial remodeling. Stem Cells 25(9):2350–2357, 2007.

    Article  PubMed  Google Scholar 

  36. Stops, A. J., L. A. McMahon, D. O’Mahoney, P. J. Prendergast, and P. E. McHugh. A finite element prediction of strain on cells in a highly porous collagen-glycosaminoglycan scaffold. J. Biomech. Eng. 130(6):061001, 2008.

    Article  PubMed  CAS  Google Scholar 

  37. Swartz, D. D., J. A. Russell, and S. T. Andreadis. Engineering of fibrin-based functional and implantable small-diameter blood vessels. Am. J. Physiol. Heart Circ. Physiol. 288(3):H1451–H1460, 2005.

    Article  PubMed  CAS  Google Scholar 

  38. Tuan, T. L., A. Song, S. Chang, S. Younai, and M. E. Nimni. In vitro fibroplasia: Matrix contraction, cell growth, and collagen production of fibroblasts cultured in fibrin gels. Exp. Cell Res. 223(1):127–134, 1996.

    Article  PubMed  CAS  Google Scholar 

  39. Wang, D., J. S. Park, J. S. Chu, A. Krakowski, K. Luo, D. J. Chen, and S. Li. Proteomic profiling of bone marrow mesenchymal stem cells upon transforming growth factor beta1 stimulation. J. Biol. Chem. 279(42):43725–43734, 2004.

    Article  PubMed  CAS  Google Scholar 

  40. Ye, Q., G. Zund, P. Benedikt, S. Jockenhoevel, S. P. Hoerstrup, S. Sakyama, J. A. Hubbell, and M. Turina. Fibrin gel as a three dimensional matrix in cardiovascular tissue engineering. Eur. J. Cardiothorac. Surg. 17(5):587–591, 2000.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

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

    Eoin D. O’Cearbhaill, Peter E. McHugh & Valerie Barron

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

    Eoin D. O’Cearbhaill & Peter E. McHugh

  3. Regenerative Medicine Institute, National Centre for Biomedical Engineering Science, National University of Ireland, Galway, Galway, Ireland

    Mary Murphy, Frank Barry & Valerie Barron

Authors
  1. Eoin D. O’Cearbhaill
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Mary Murphy
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Frank Barry
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Peter E. McHugh
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Valerie Barron
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Valerie Barron.

Additional information

Associate Editor Kyriacos A. Athanasiou oversaw the review of this article.

Rights and permissions

Reprints and permissions

About this article

Cite this article

O’Cearbhaill, E.D., Murphy, M., Barry, F. et al. Behavior of Human Mesenchymal Stem Cells in Fibrin-Based Vascular Tissue Engineering Constructs. Ann Biomed Eng 38, 649–657 (2010). https://doi.org/10.1007/s10439-010-9912-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10439-010-9912-x

Keywords

Search

Navigation