AIMS Bioengineering, 2017, 4(4): 431-445. doi: 10.3934/bioeng.2017.4.431.

Research article

Export file:


  • RIS(for EndNote,Reference Manager,ProCite)
  • BibTex
  • Text


  • Citation Only
  • Citation and Abstract

Tissue engineering approaches to develop decellularized tendon matrices functionalized with progenitor cells cultured under undifferentiated and tenogenic conditions

1 Cell and Tissue Engineering Laboratory, IRCCS Galeazzi Orthopaedic Institute, Milan, Italy
2 Department of Veterinary Medicine (DiMeVet), University of Milan, Milan, Italy
3 Regenerative Medicine Technologies Lab, Ente Ospedaliero Cantonale (EOC), Lugano, Switzerland
4 Swiss Institute of Regenerative Medicine (SIRM), Lugano, Switzerland
§ Daniele D’Arrigo and Marta Bottagisio contributed equally to this work.

Tendon ruptures and retractions with an extensive tissue loss represent a major clinical problem and a great challenge in surgical reconstruction. Traditional approaches consist in autologous or allogeneic grafts, which still have some drawbacks. Hence, tissue engineering strategies aimed at developing functionalized tendon grafts. In this context, the use of xenogeneic tissues represents a promising perspective to obtain decellularized tendon grafts. This study is focused on the identification of suitable culture conditions for the generation of reseeded and functional decellularized constructs to be used as tendon grafts. Equine superficial digital flexor tendons were decellularized, reseeded with mesenchymal stem cells (MSCs) from bone marrow and statically cultured in two different culture media to maintain undifferentiated cells (U-MSCs) or to induce a terminal tenogenic differentiation (T-MSCs) for 24 hours, 7 and 14 days. Cell viability, proliferation, morphology as well as matrix deposition and type I and III collagen production were assessed by means of histological, immunohistochemical and semi-quantitative analyses. Results showed that cell viability was not affected by any culture conditions and active proliferation was maintained 14 days after reseeding. However, seeded MSCs were not able to penetrate within the dense matrix of the decellularized tendons. Nevertheless, U-MSCs synthesized a greater amount of extracellular matrix rich in type I collagen compared to T-MSCs. In spite of the inability to deeply colonize the decellularized matrix in vitro, reseeding tendon matrices with U-MSCs could represent a suitable method for the functionalization of biological constructs, considering also any potential chemoattractant capability of the newly deposed extracellular matrix to recruit resident cells. This bioengineering approach can be exploited to produce functionalized tendon constructs for the substitution of large tendon defects.
  Article Metrics

Keywords decellularized tendon; undifferentiated mesenchymal stem cells (U-MSCs); extracellular matrix; type I collagen; tenogenic differentiation (T-MSCs); functionalized tendon constructs; cell repopulation and static culture

Citation: Daniele D’Arrigo, Marta Bottagisio, Silvia Lopa, Matteo Moretti, Arianna B. Lovati. Tissue engineering approaches to develop decellularized tendon matrices functionalized with progenitor cells cultured under undifferentiated and tenogenic conditions. AIMS Bioengineering, 2017, 4(4): 431-445. doi: 10.3934/bioeng.2017.4.431


  • 1. Andia I, Maffulli N (2015) Muscle and tendon injuries: the role of biological interventions to and assist healing and recovery. Arthroscopy 31: 999–1015.    
  • 2. Bottagisio M, Lovati AB (2017) A review on animal models and treatments for the reconstruction of Achilles and flexor tendons. J Mater Sci Mater Med 28: 1–15.
  • 3. Hopkins C, Fu SC, Chua E, et al. (2016) Critical review on the socio-economic impact of tendinopathy. Asia-Pacific J Sport Med Arthrosc Rehabil Technol 4: 9–20.    
  • 4. Docheva D, Müller SA, Majewski M, et al. (2015) Biologics for tendon repair. Adv Drug Deliv Rev 84: 222–239.    
  • 5. Petrou IG, Grognuz A, Hirt-Burri N, et al. (2014) Cell therapies for tendons: old cell choice for modern innovation. Swiss Med Wkly 144: 1–11.
  • 6. Legrand A, Kaufman Y, Long C, et al. (2017) Molecular biology of flexor tendon healing in relation to reduction of tendon adhesions. J Hand Surg Am 42: 722–726.    
  • 7. Akpancar S, Tatar O, Turgut H, et al. (2016) The current perspectives of stem cell therapy in orthopedic surgery. Arch trauma Res 5: e37976.
  • 8. Wobma H, Vunjak-Novakovic G (2016) Tissue engineering and regenerative medicine 2015: a year in review. Tissue Eng Part B Rev 22: 101–113.    
  • 9. Lovati AB, Bottagisio M, Moretti M (2016) Decellularized and engineered tendons as biological substitutes: a critical review. Stem Cells Int 2016: 7276150.
  • 10. Bottagisio M, Pellegata AF, Boschetti F, et al. (2016) A new strategy for the decellularisation of large equine tendons as biocompatible tendon substitutes. Eur Cells Mater 32: 58–73.
  • 11. Pellegata AF, Bottagisio M, Boschetti F, et al. (2017) Terminal sterilization of equine-derived decellularized tendons for clinical use. Mater Sci Eng C 75: 43–49.    
  • 12. Wang S, Wang Y, Song L, et al. (2017) Decellularized tendon as a prospective scaffold for tendon repair. Mater Sci Eng C 77: 1290–1301.    
  • 13. Yao L, Bestwick CS, Bestwick L, et al. (2006) Phenotypic drift in human tenocyte culture. Tissue Eng 12: 1843–1849.    
  • 14. Schwarz R, Colarusso L, Doty P (1976) Maintenance of differentiation in primary cultures of avian tendon cells. Exp Cell Res 102: 63–71.    
  • 15. Van Eijk F, Saris DBF, Riesle J, et al. (2004) Tissue engineering of ligaments: a comparison of bone marrow stromal cells, anterior cruciate ligament, and skin fibroblasts as cell source. Tissue Eng 10: 893–903.    
  • 16. Hankemeier S, Hurschler C, Zeichen J, et al. (2009) Bone marrow stromal cells in a liquid fibrin matrix improve the healing process of patellar tendon window defects. Tissue Eng Part A 15: 1019–1030.    
  • 17. Shenaq DS, Rastegar F, Petkovic D, et al. (2010) Mesenchymal progenitor cells and their orthopedic applications: forging a path towards clinical trials. Stem Cells Int 2010: 519028.
  • 18. Contreras-Kallens P, Terraza C, Oyarce K, et al. (2017) Mesenchymal stem cells and their immunosuppressive role in transplantation tolerance. Ann Ny Acad Sci: 1–22.
  • 19. Bottagisio M, Lopa S, Granata V, et al. (2017) Different combinations of growth factors for the tenogenic differentiation of bone marrow mesenchymal stem cells in monolayer culture and in fibrin-based three-dimensional constructs. Differentiation 95: 44–53.    
  • 20. Goncalves AI, Rodrigues MT, Lee SJ, et al. (2013) Understanding the role of growth factors in modulating stem cell tenogenesis. PLoS One 8: e83734.    
  • 21. Gaspar D, Spanoudes K, Holladay C, et al. (2014) Progress in cell-based therapies for tendon repair. Adv Drug Deliv Rev 84: 240–256.
  • 22. Spanoudes K, Gaspar D, Pandit A, et al. (2014) The biophysical, biochemical, and biological toolbox for tenogenic phenotype maintenance in vitro. Trends Biotechnol 32: 474–482.    
  • 23. Li VC, Kirschner MW (2014) Molecular ties between the cell cycle and differentiation in embryonic stem cells. Proc Natl Acad Sci USA 111: 9503–9508.    
  • 24. Ozasa Y, Amadio PC, Thoreson AR, et al. (2013) Repopulation of intrasynovial flexor tendon allograft with bone marrow stromal cells: an ex vivo model. Tissue Eng Part A 20: 566–574.
  • 25. Kryger GS, Chong AKS, Costa M, et al. (2007) A comparison of tenocytes and mesenchymal stem cells for use in flexor tendon tissue engineering. J Hand Surgery Am 32: 597–605.
  • 26. Thorfinn J, Saber S, Angelidis IK, et al. (2009) Flexor tendon tissue engineering: temporal distribution of donor tenocytes versus recipient cells. Plast Reconstr Surg 124: 2019–2026.    
  • 27. Taylor SH, Al-Youha S, van Agtmael T, et al. (2011) Tendon is covered by a basement membrane epithelium that is required for cell retention and the prevention of adhesion formation. PLoS One 6: e16337.    
  • 28. Angelidis IK, Thorfinn J, Connolly ID, et al. (2010) Tissue engineering of flexor tendons: The effect of a tissue bioreactor on adipoderived stem cellseeded and fibroblast-seeded tendon constructs. J Hand Surg Am 35: 1466–1472.    
  • 29. Qin TW, Sun YL, Thoreson AR, et al. (2015) Effect of mechanical stimulation on bone marrow stromal cell-seeded tendon slice constructs: a potential engineered tendon patch for rotator cuff repair. Biomaterials 51: 43–50.    
  • 30. Youngstrom DW, Rajpar I, Kaplan DL, et al. (2015) A bioreactor system for in vitro tendon differentiation and tendon tissue engineering. J Orthop Res 33: 911–918.    
  • 31. Schulze-Tanzil G, Al-Sadi O, Ertel W, et al. (2012) Decellularized tendon extracellular matrix-a valuable approach for tendon reconstruction? Cells 1: 1010–1028.    
  • 32. Fitzpatrick LE, McDevitt TC (2015) Cell-derived matrices for tissue engineering and regenerative medicine applications. Biomater Sci 3: 12–24.    
  • 33. Sharma P, Maffulli N (2006) Biology of tendon injury: healing, modeling and remodeling. J Musculoskelet Neuronal Interact 6: 181–190.
  • 34. Herberts CA, Kwa MS, Hermsen HP (2011) Risk factors in the development of stem cell therapy. J Transl Med 9: 29.    


This article has been cited by

  • 1. Giuseppe Talò, Daniele D’Arrigo, Sergio Lorenzi, Matteo Moretti, Arianna B. Lovati, Independent, Controllable Stretch-Perfusion Bioreactor Chambers to Functionalize Cell-Seeded Decellularized Tendons, Annals of Biomedical Engineering, 2019, 10.1007/s10439-019-02257-6
  • 2. Marta Bottagisio, Daniele D’Arrigo, Giuseppe Talò, Matilde Bongio, Marco Ferroni, Federica Boschetti, Matteo Moretti, Arianna B. Lovati, Achilles Tendon Repair by Decellularized and Engineered Xenografts in a Rabbit Model, Stem Cells International, 2019, 2019, 1, 10.1155/2019/5267479

Reader Comments

your name: *   your email: *  

Copyright Info: 2017, Arianna B. Lovati, et al., licensee AIMS Press. This is an open access article distributed under the terms of the Creative Commons Attribution Licese (

Download full text in PDF

Export Citation

Copyright © AIMS Press All Rights Reserved