Skip to main content
SpringerLink
Log in

The effects of scaffold architecture and fibrin gel addition on tendon cell phenotype

  • Tissue Engineering Constructs and Cell Substrates
  • Published:
Journal of Materials Science: Materials in Medicine Aims and scope Submit manuscript

Abstract

Development of tissue engineering scaffolds relies on careful selection of pore architecture and chemistry of the cellular environment. Repair of skeletal soft tissue, such as tendon, is particularly challenging, since these tissues have a relatively poor healing response. When removed from their native environment, tendon cells (tenocytes) lose their characteristic morphology and the expression of phenotypic markers. To stimulate tendon cells to recreate a healthy extracellular matrix, both architectural cues and fibrin gels have been used in the past, however, their relative effects have not been studied systematically. Within this study, a combination of collagen scaffold architecture, axial and isotropic, and fibrin gel addition was assessed, using ovine tendon-derived cells to determine the optimal strategy for controlling the proliferation and protein expression. Scaffold architecture and fibrin gel addition influenced tendon cell behavior independently in vitro. Addition of fibrin gel within a scaffold doubled cell number and increased matrix production for all architectures studied. However, scaffold architecture dictated the type of matrix produced by cells, regardless of fibrin addition. Axial scaffolds, mimicking native tendon, promoted a mature matrix, with increased tenomodulin, a marker for mature tendon cells, and decreased scleraxis, an early transcription factor for connective tissue. This study demonstrated that both architectural cues and fibrin gel addition alter cell behavior and that the combination of these signals could improve clinical performance of current tissue engineering constructs.

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

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  1. Kastelic J, Galeski A, Baer E. Multicomposite structure of tendon. Connect Tissue Res. 1978;6(1):11–23.

    Article  Google Scholar 

  2. Biewener A. Tendons and ligaments: structure, mechanical behavior and biological function. In: Fratzl P, editor. Collagen. New York: Springer; 2008. p. 269–84.

    Chapter  Google Scholar 

  3. Amiel D, Frank C, Harwood F, Fronek J, Akeson W. Tendons and ligaments a morphological and biochemical comparison. J Orthop Res. 1984;1(3):257–65.

    Article  Google Scholar 

  4. Klatte-Schulz F, Pauly S, Scheibel M, Greiner S, Gerhardt C, Schmidmaier G, Wildemann B. Influence of age on the cell biological characteristics and the stimulation potential of male human tenocyte-like cells. Eur Cells Mater. 2012;24:74–89.

    Google Scholar 

  5. Bayer M, Yeung C-YC, Kadler KE, Qvortrup K, Baar K, Svensson RB, Magnusson SP, Krogsgaard M, Koch M, Kjaer M. The initiation of embryonic-like collagen fibrillogenesis by adult human tendon fibroblasts when cultured under tension. Biomaterials. 2010;31(18):4889–97.

    Article  Google Scholar 

  6. Caliari SR, Harley BAC. The effect of anisotropic collagen-gag scaffolds and growth factor supplementation on tendon cell recruitment, alignment, and metabolic activity. Biomaterials. 2011;32(23):5330–40.

    Article  Google Scholar 

  7. Kapacce Z, Richardson SH, Lu Y, Starborg T, Holmes DF, Baar K, Kadler KE. Tension is required for fibripositor formation. Matrix Biol. 2008;27(4):371–5.

    Article  Google Scholar 

  8. Kalson NS, Holmes DF, Kapacee Z, Otermin I, Lu Y, Ennos RA, Canty-Laird EG, Kadler KE. An experimental model for studying the biomechanics of embryonic tendon: evidence that the development of mechanical properties depends on the actinomyosin machinery. Matrix Biol. 2010;29(8):678–89.

    Article  Google Scholar 

  9. Smithmyer ME, Sawickia LA, Kloxin AM. Hydrogel scaffolds as in vitro models to study fibroblast activation in wound healing and disease. Biomater Sci. 2014;2:634–50.

    Article  Google Scholar 

  10. Ross JJ, Tranquillo R. ECM gene expression correlates with in vitro tissue growth and development in fibrin gel remodeled by neonatal smooth muscle cells. Matrix Biol. 2003;22(6):477–90.

    Article  Google Scholar 

  11. Lesman A, Koffler J, Atlas R, Blinder YJ, Kam Z, Levenberg S. Engineering vessel-like networks within multicellular fibrin-based constructs. Biomaterials. 2011;32(31):7856–69.

    Article  Google Scholar 

  12. Lohse N, Schulz J, Schliephake H. Effect of fibrin on osteogenic differentiation and VEGf expression of bone marrow stromal cells in mineralized scaffolds: a three-dimensional analysis. Eur Cells Mater. 2012;23:413–24.

    Google Scholar 

  13. Pawelec KM, Husmann A, Best SM, Cameron RE. Understanding anisotropy and architecture in ice-templated biopolymer scaffolds. Mater Sci Eng. 2014;37:141–7.

    Article  Google Scholar 

  14. Pawelec KM, Husmann A, Best SM, Cameron RE. A design protocol for tailoring ice-templated scaffold structure. J R Soc Interface. 2014;11:20130958.

    Article  Google Scholar 

  15. Davidenko N, Gibb T, Schuster C, Best SM, Campbell JJ, Watson CJ, Cameron RE. Biomimetic collagen scaffolds with anisotropic pore architecture. Acta Biomater. 2012;8(2):667–76.

    Article  Google Scholar 

  16. Damink L, Dijkstra PJ, van Luyn MJA, van Wachem PB, Nieuwenhuis P, Feijen J. Cross-linking of dermal sheep collagen using a water-soluble carbodiimide. Biomaterials. 1996;17(8):765–73.

    Article  Google Scholar 

  17. Kim Y-J, Sah RL, Doong J-YH, Grodzinsky AJ. Fluorometric assay of DNA in cartilage explants using Hoechst 33258. Anal Biochem. 1988;174(1):168–76.

    Article  Google Scholar 

  18. Farndale R, Buttle DJ, Barrett AJ. Improved quantitation and discrimination of sulphated glycosaminoglycans by use of dimethylmethylene blue. Biochem Biophys Acta. 1986;883(2):173–7.

    Article  Google Scholar 

  19. Venugopal J, Ma LL, Yong T, Ramakrishna S. In vitro study of smooth muscle cells on polycaprolactone and collagen nanofibrous matrices. Cell Biol Int. 2005;29:861–7.

    Article  Google Scholar 

  20. Shukunami C, Takimoto A, Oro M, Hiraki Y. Scleraxis positively regulates the expression of tenomodulin, a differentiation marker of tenocytes. Dev Biol. 2006;298(1):234–47.

    Article  Google Scholar 

  21. Pearlstein E, Gold LI, Garciapardo A. Fibronectin—review of its structure and biological-activity. Mol Cell Biochem. 1980;29(2):103–28.

    Article  Google Scholar 

  22. Friess W. Collagen—biomaterial for drug delivery. Eur J Pharm Biopharm. 1998;45(2):113–36.

    Article  Google Scholar 

  23. Ayad S, Boot-Handford RP, Humphries MJ, Kadler KE, Shuttleworth CA. The extracellular matrix facts book. 2nd ed. New York: Academic Press; 1998.

    Google Scholar 

  24. Schneider PRA, Buhrmann C, Mobasheri A, Matis U, Shakibaei M. Three-dimensional high-density co-culture with primary tenocytes induces tenogenic differentiation in mesenchymal stem cells. J Orthop Res. 2011;29(9):1351–60.

    Article  Google Scholar 

  25. Cserjesi P, Brown D, Ligon KL, Lyons GE, Copeland NG, Gilbert DJ, Jenkins NA, Olson EN. Scleraxis—a basic helix-loop-helix protein that prefigures skeletal formation during mouse embryogenesis. Development. 1995;121(4):1099–110.

    Google Scholar 

  26. Carlberg AL, Tuan RS, Hall DJ. Regulation of scleraxis function by interaction with the BHLH protein E47. Mol Cell Biol Res Commun. 2000;3(2):82–6.

    Article  Google Scholar 

  27. Taylor SE, Vaughan-Thomas A, Clements DN, Pinchbeck G, Macrory LC, Smith RKW, Clegg PD. Gene expression markers of tendon fibroblasts in normal and diseased tissue compared to monolayer and three dimensional culture systems. BMC Musculoskelet Disord. 2009;10(1):27.

    Article  Google Scholar 

  28. Mendias CL, Gumucio JP, Bakhurin KI, Lynch EB, Brooks SV. Physiological loading of tendons induces scleraxis expression in epitenon fibroblasts. J Orthop Res. 2012;30(4):606–12.

    Article  Google Scholar 

  29. Yamana K, Wada H, Takahashi Y, Sato H, Kasahara Y, Kiyoki M. Molecular cloning and characterization of Chm1 l, a novel membrane molecule similar to chondromodulin-I. Biochem Biophys Res Commun. 2001;280(4):1101–6.

    Article  Google Scholar 

  30. Qi J, Dmochowski JM, Banes AN, Tsuzaki M, Bynum D, Patterson M, Creighton A, Gomez S, Tech K, Cederlund A, Banes AJ. Differential expression and cellular localization of novel isoforms of the tendon biomarker tenomodulin. J Appl Physiol. 2012;113(6):861–71.

    Article  Google Scholar 

  31. Kishore V, Uquillas JA, Dubikovsky A, Alshehabat MA, Snyder PW, Breur GJ, Akkus O. In vivo response to electrochemically aligned collagen bioscaffolds. J Biomed Mater Res, Part B. 2012;100B(2):400–8.

    Article  Google Scholar 

  32. Yin Z, Chen X, Chen JL, Shen WL, Nguyen TMH, Gao L, Ouyang HW. The regulation of tendon stem cell differentiation by the alignment of nanofibers. Biomaterials. 2010;31(8):2163–75.

    Article  Google Scholar 

  33. Caliari SR, Weisgerber DW, Ramirez MA, Kelkhoff DO, Harley BAC. The influence of collagen-glycosaminoglycan scaffold relative density and microstructural anisotropy on tenocyte bioactivity and transcriptomic stability. J Mech Behav Biomed Mater. 2012;11:27–40.

    Article  Google Scholar 

  34. Nakayama GR, Caton MC, Nova MP, Parandoosh Z. Assessment of the alamar blue assay for cellular growth and viability in vitro. J Immunol Methods. 1997;204(2):205–8.

    Article  Google Scholar 

  35. Mazzocca AD, Chowaniec D, McCarthy MB, Beitzel K, Cote MP, McKinnon W, Arciero R. In vitro changes in human tenocyte cultures obtained from proximal biceps tendon: multiple passages result in changes in routine cell markers. Knee Surg Sports Traumatol Arthrosc. 2012;20(9):1666–72.

    Article  Google Scholar 

  36. Schweitzer R, Chyung JH, Murtaugh LC, Brent AE, Rosen V, Olson EN, Lassar A, Tabin CJ. Analysis of the tendon cell fate using scleraxis, a specific marker for tendons and ligaments. Development. 2001;128(19):3855–66.

    Google Scholar 

  37. Wang YZ, Kim UJ, Blasioli DJ, Kim HJ, Kaplan DL. In vitro cartilage tissue engineering with 3D porous aqueous-derived silk scaffolds and mesenchymal stem cells. Biomaterials. 2005;26(34):7082–94.

    Article  Google Scholar 

  38. Kishore V, Bullock W, Sun X, Van Dyke WS, Akkus O. Tenogenic differentiation of human MSCs induced by the topography of electrochemically aligned collagen threads. Biomaterials. 2012;33(7):2137–44.

    Article  Google Scholar 

Download references

Acknowledgments

The authors gratefully acknowledge the financial support of the Gates Cambridge Trust, the ERC Advanced Grant 320598 3D-E, and the Technology Strategy Board UK. Special thanks to Nigel Loveridge for help with the statistical analysis and Natalia Davidenko for supplying the freeze drying molds.

Author information

Authors and Affiliations

  1. Cambridge Centre for Medical Materials, Materials Science and Metallurgy Department, University of Cambridge, Cambridge, CB3 0FS, UK

    K. M. Pawelec, S. M. Best & R. E. Cameron

  2. Division of Trauma and Orthopaedic Surgery, University of Cambridge, Cambridge, CB2 2QQ, UK

    R. J. Wardale

Authors
  1. K. M. Pawelec
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. R. J. Wardale
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. S. M. Best
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. R. E. Cameron
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to R. E. Cameron.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Pawelec, K.M., Wardale, R.J., Best, S.M. et al. The effects of scaffold architecture and fibrin gel addition on tendon cell phenotype. J Mater Sci: Mater Med 26, 13 (2015). https://doi.org/10.1007/s10856-014-5349-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1007/s10856-014-5349-3

Keywords

Search

Navigation