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Extruded collagen fibres for tissue engineering applications: effect of crosslinking method on mechanical and biological properties

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Abstract

Reconstituted collagen fibres are promising candidates for tendon and ligament tissue regeneration. The crosslinking procedure determines the fibres’ mechanical properties, degradation rate, and cell–fibre interactions. We aimed to compare mechanical and biological properties of collagen fibres resulting from two different types of crosslinking chemistry based on 1-ethyl-3-(3-dimethyllaminopropyl)carbodiimide (EDC). Fibres were crosslinked with either EDC or with EDC and ethylene-glycol-diglycidyl-ether (EDC/EGDE). Single fibres were mechanically tested to failure and bundles of fibres were seeded with tendon fibroblasts (TFs) and cell attachment and proliferation were determined over 14 days in culture. Collagen type I and tenascin-C production were assessed by immunohistochemistry and dot-blotting. EDC chemistry resulted in fibres with average mechanical properties but the highest cell proliferation rate and matrix protein production. EDC/EGDE chemistry resulted in fibres with improved mechanical properties but with a lower biocompatibility profile. Both chemistries may provide useful structures for scaffolding regeneration of tendon and ligament tissue and will be evaluated for in vivo tendon regeneration in future experiments.

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References

  1. Brooks P. Extensor mechanism ruptures. Orthopedics. 2009;32:9.

    Google Scholar 

  2. Maffulli N, Ajis A, Longo UG, Denaro V. Chronic rupture of tendo Achillis. Foot Ankle Clin. 2007;12:583–96.

    Article  Google Scholar 

  3. Sherman OH, Banffy MB. Anterior cruciate ligament reconstruction: which graft is best? Arthroscopy. 2004;20:974–80.

    Google Scholar 

  4. Spindler KR, Kuhn JE, Freedman KB, Matthews CE, Dittus RS, Harrell FE. Anterior cruciate ligament reconstruction autograft choice: bone-tendon-bone versus hamstring—does it really matter? A systematic review. Am J Sports Med. 2004;32:1986–95.

    Article  Google Scholar 

  5. Shelton WR, Treacy SH, Dukes AD, Bomboy AL. Use of allografts in knee reconstruction: II. Surgical considerations. J Am Acad Orthop Surg. 1998;6:169–75.

    CAS  Google Scholar 

  6. Ventura A, Terzaghi C, Legnani C, Borgo E, Albisetti W. Synthetic grafts for anterior cruciate ligament rupture: 19-year outcome study. Knee. 2010;17:108–13.

    Article  Google Scholar 

  7. Vunjak-Novakovic G, Altman G, Horan R, Kaplan DL. Tissue engineering of ligaments. Annu Rev Biomed Eng. 2004;6:131–56.

    Article  CAS  Google Scholar 

  8. Drexler JW, Powell HM. DHT crosslinking of electrospun collagen. Tissue Eng Part C Methods. 2011;17:9–17.

    Article  CAS  Google Scholar 

  9. Mekhail M, Wong KK, Padavan DT, Wu Y, O’Gorman DB, Wan W. Genipin-cross-linked electrospun collagen fibers. J Biomater Sci Polym Ed. 2010 (Epub ahead of print).

  10. Zeugolis DI, Paul GR, Attenburrow G. Cross-linking of extruded collagen fibers-A biomimetic three-dimensional scaffold for tissue engineering applications. J Biomed Mater Res A. 2009;89A:895–908.

    Article  CAS  Google Scholar 

  11. Carlisle CR, Coulais C, Guthold M. The mechanical stress–strain properties of single electrospun collagen type I nanofibers. Acta Biomater. 2010;6:2997–3003.

    Article  CAS  Google Scholar 

  12. Chen X, Qi YY, Wang LL, Yin Z, Yin GL, Zou XH, Ouyang HW. Ligament regeneration using a knitted silk scaffold combined with collagen matrix. Biomaterials. 2008;29:3683–92.

    Article  CAS  Google Scholar 

  13. McNeil SE, Griffiths HR, Perrie Y. Polycaprolactone fibres as a potential delivery system for collagen to support bone regeneration. Curr Drug Deliv. 2011 (Epub ahead of print).

  14. Zhu Y, Liu T, Song K, Jiang B, Ma X, Cui Z. Collagen-chitosan polymer as a scaffold for the proliferation of human adipose tissue-derived stem cells. J Mater Sci Mater Med. 2009;20:799–808.

    Article  CAS  Google Scholar 

  15. Gigante A, Cesari E, Busilacchi A, Manzotti S, Kyriakidou K, Greco F, Di Primio R, Mattioli-Belmonte M. Collagen I membranes for tendon repair: effect of collagen fiber orientation on cell behavior. J Orthop Res. 2009;27:826–32.

    Article  CAS  Google Scholar 

  16. Lynn AK, Yannas IV, Bonfield W. Antigenicity and immunogenicity of collagen. J Biomed Mater Res B Appl Biomater. 2004;71:343–54.

    Article  CAS  Google Scholar 

  17. Kato YP, Christiansen DL, Hahn RA, Shieh SJ, Goldstein JD, Silver FH. Mechanical-properties of collagen-fibers—a comparison of reconstituted and rat tail tendon fibers. Biomaterials. 1989;10:38–41.

    Article  CAS  Google Scholar 

  18. Caruso AB, Dunn MG. Functional evaluation of collagen fiber scaffolds for ACL reconstruction: cyclic loading in proteolytic enzyme solutions. J Biomed Mater Res A. 2004;69A:164–71.

    Article  CAS  Google Scholar 

  19. Bellincampi LD, Closkey RF, Prasad R, Zawadsky JP, Dunn MG. Viability of fibroblast-seeded ligament analogs after autogenous implantation. J Orthop Res. 1998;16:414–20.

    Article  CAS  Google Scholar 

  20. Gentleman E, Lay AN, Dickerson DA, Nauman EA, Livesay GA, Dee KC. Mechanical characterization of collagen fibers and scaffolds for tissue engineering. Biomaterials. 2003;24:3805–13.

    Article  CAS  Google Scholar 

  21. Koob TJ, Willis TA, Qiu YS, Hernandez DJ. Biocompatibility of NDGA-polymerized collagen fibers. II. Attachment, proliferation, and migration of tendon fibroblasts in vitro. J Biomed Mater Res. 2001;56:40–8.

    Article  CAS  Google Scholar 

  22. Chvapil M, Speer DP, Holubec H, Chvapil TA, King DH. Collagen-fibers as a temporary scaffold for replacement of ACL in goats. J Biomed Mater Res. 1993;27:313–25.

    Article  CAS  Google Scholar 

  23. Dunn MG, Tria AJ, Kato YP, Bechler JR, Ochner RS, Zawadsky JP, Silver FH. Anterior cruciate ligament reconstruction using a composite collagenous prosthesis—a biomechanical and histologic study in rabbits. Am J Sports Med. 1992;20:507–75.

    Article  CAS  Google Scholar 

  24. Cornwell KG, Lei P, Andreadis ST, Pins GD. Crosslinking of discrete self-assembled collagen threads: effects on mechanical strength and cell–matrix interactions. J Biomed Mater Res A. 2007;80A:362–71.

    Article  CAS  Google Scholar 

  25. Wang MC, Pins GD, Silver FH. Collagen fibres with improved strength for the repair of soft tissue injuries. Biomaterials. 1994;15:507–12.

    Article  CAS  Google Scholar 

  26. Sung HW, Huang RN, Huang LLH, Tsai CC. In vitro evaluation of cytotoxicity of a naturally occurring cross-linking reagent for biological tissue fixation. J Biomater Sci Polym Ed. 1999;10:63–78.

    Article  CAS  Google Scholar 

  27. van Wachem PB, Plantinga JA, Wissink MJ, Beernink R, Poot AA, Engbers GH, Beugeling T, van Aken WG, Feijen J, van Luyn MJ. In vivo biocompatibility of carbodiimide-crosslinked collagen matrices: effects of crosslink density, heparin immobilization, and bFGF loading. J Biomed Mater Res. 2001;55:368–78.

    Article  Google Scholar 

  28. Zeeman R, Dijkstra PJ, van Wachem PB, van Luyn MJ, Hendriks M, Cahalan PT, Feijen J. Successive epoxy and carbodiimide cross-linking of dermal sheep collagen. Biomaterials. 1999;20:921–31.

    Article  CAS  Google Scholar 

  29. Kato YP, Dunn MG, Zawadsky JP, Tria AJ, Silver FH. Regeneration of achilles tendon with a collagen tendon prosthesis—results of a one-year implantation study. J Bone Joint Surg Am. 1991;73A:561–74.

    Google Scholar 

  30. Weadock KS, Miller EJ, Keuffel EL, Dunn MG. Effect of physical crosslinking methods on collagen-fiber durability in proteolytic solutions. J Biomed Mater Res. 1996;32:221–6.

    Article  CAS  Google Scholar 

  31. Caruso AB, Dunn MG. Changes in mechanical properties and cellularity during long-term culture of collagen fiber ACL reconstruction scaffolds. J Biomed Mater Res A. 2005;73A:388–97.

    Article  CAS  Google Scholar 

  32. Gentleman E, Livesay GA, Dee KC, Nauman EA. Development of ligament-like structural organization and properties in cell-seeded collagen scaffolds in vitro. Ann Biomed Eng. 2006;34:726–36.

    Article  Google Scholar 

  33. Sung HW, Huang RN, Huang LLH, Tsai CC, Chiu CT. Feasibility study of a natural crosslinking reagent for biological tissue fixation. J Biomed Mater Res. 1998;42:560–7.

    Article  CAS  Google Scholar 

  34. Yu X-x, Wan C-x, Chen H-q. Preparation and endothelialization of decellularised vascular scaffold for tissue-engineered blood vessel. J Mater Sci Mater Med. 2008;19:319–26.

    Article  CAS  Google Scholar 

  35. 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;26:10–27.

    Google Scholar 

  36. Chiquet-Ehrismann R, Tucker RP. Connective tissues: signalling by tenascins. Int J Biochem Cell Biol. 2004;36:1085–9.

    Article  CAS  Google Scholar 

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Acknowledgments

The authors gratefully acknowledge the founding support of the Technology Strategy Board (grant number DT/F006977/1) and of the National Institute for Health Research. Dr D. Enea gratefully acknowledges Prof F. Greco and Dr A. Gigante (Clinica Ortopedica, Polytechnic University of Marche) for their helpfulness and support.

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Authors and Affiliations

  1. Orthopaedic Research Unit, Department of Surgery, Cambridge University, Addenbrooke’s Hospital, Hills Rd, CB2 2QQ, Cambridge, UK

    Davide Enea, John Wardale, Alan Getgood, Roger Brooks & Neil Rushton

  2. Clinica Ortopedica, Polytechnic University of Marche, Ospedali Riuniti Ancona, Via Tronto 10, 60126, Ancona, Italy

    Davide Enea

  3. Department of Veterinary Medicine, University of Cambridge, Madingley Road, CB3 OES, Cambridge, UK

    Frances Henson

  4. Orthomimetics Laboratories, Byron House, Milton Rd, CB4 0WZ, Cambridge, UK

    Simon Kew

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  1. Davide Enea
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  2. Frances Henson
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  3. Simon Kew
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  4. John Wardale
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  5. Alan Getgood
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  6. Roger Brooks
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Correspondence to Davide Enea.

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Enea, D., Henson, F., Kew, S. et al. Extruded collagen fibres for tissue engineering applications: effect of crosslinking method on mechanical and biological properties. J Mater Sci: Mater Med 22, 1569–1578 (2011). https://doi.org/10.1007/s10856-011-4336-1

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  • DOI: https://doi.org/10.1007/s10856-011-4336-1

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