Advertisement

Formation of Corneal Stromal-Like Assemblies Using Human Corneal Fibroblasts and Macromolecular Crowding

  • Mehmet Gürdal
  • Gülinnaz Ercan
  • Dimitrios I. ZeugolisEmail author
Protocol
  • 52 Downloads
Part of the Methods in Molecular Biology book series (MIMB, volume 2145)

Abstract

Tissue engineering by self-assembly allows for the formation of living tissue substitutes, using the cells’ innate capability to produce and deposit tissue-specific extracellular matrix. However, in order to develop extracellular matrix-rich implantable devices, prolonged culture time is required in traditionally utilized dilute ex vivo microenvironments. Macromolecular crowding, by imitating the in vivo tissue density, dramatically accelerates biological processes, resulting in enhanced and accelerated extracellular matrix deposition. Herein, we describe the ex vivo formation of corneal stromal-like assemblies using human corneal fibroblasts and macromolecular crowding.

Key words

Corneal stroma Tissue engineering by self-assembly Macromolecular crowding Extracellular matrix 

Notes

Acknowledgements

This work has been supported from: Science Foundation Ireland, Career Development Award Programme (grant agreement number: 15/CDA/3629) and Science Foundation Ireland and the European Regional Development Fund (grant agreement number: 13/RC/2073). Mehmet Gürdal was supported by The Scientific and Technological Research Council of Turkey (TÜBİTAK), Science Fellowships and Grant Programmes Department (BİDEB), Programme of 2214-A Ph.D. Research Scholarship for Abroad. The authors have no competing interests.

References

  1. 1.
    Fini ME, Stramer BM (2005) How the cornea heals: cornea-specific repair mechanisms affecting surgical outcomes. Cornea 24(8 Suppl):S2–S11CrossRefGoogle Scholar
  2. 2.
    Radner W, Zehetmayer M, Aufreiter R, Mallinger R (1998) Interlacing and cross-angle distribution of collagen lamellae in the human cornea. Cornea 17(5):537–543CrossRefGoogle Scholar
  3. 3.
    Fini ME (1999) Keratocyte and fibroblast phenotypes in the repairing cornea. Prog Retin Eye Res 18(4):529–551CrossRefGoogle Scholar
  4. 4.
    Hay ED (1980) Development of the vertebrate cornea. Int Rev Cytol 63:263–322CrossRefGoogle Scholar
  5. 5.
    Kumar P, Pandit A, Zeugolis D (2016) Progress in corneal stromal repair: from tissue grafts and biomaterials to modular supramolecular tissue-like assemblies. Adv Mater 28(27):5381–5399.  https://doi.org/10.1002/adma.201503986CrossRefPubMedGoogle Scholar
  6. 6.
    Guillame-Gentil O, Semenov O, Roca AS, Groth T, Zahn R, Voros J, Zenobi-Wong M (2010) Engineering the extracellular environment: strategies for building 2d and 3d cellular structures. Adv Mater 22(48):5443–5462.  https://doi.org/10.1002/adma.201001747CrossRefPubMedGoogle Scholar
  7. 7.
    Peck M, Dusserre N, McAllister TN, L’Heureux N (2011) Tissue engineering by self-assembly. Mater Today 14(5):218–224.  https://doi.org/10.1016/S1369-7021(11)70117-1CrossRefGoogle Scholar
  8. 8.
    Canty EG, Kadler KE (2005) Procollagen trafficking, processing and fibrillogenesis. J Cell Sci 118(Pt 7):1341–1353.  https://doi.org/10.1242/jcs.01731CrossRefPubMedGoogle Scholar
  9. 9.
    Sorushanova A, Delgado L, Wu Z, Shologu N, Kshirsagar A, Raghunath R, Mullen A, Bayon Y, Pandit A, Raghunath M, Zeugolis D (2019) The collagen suprafamily: from biosynthesis to advanced biomaterial development. Adv Mater 31(1):e1801651.  https://doi.org/10.1002/adma.201801651CrossRefPubMedGoogle Scholar
  10. 10.
    Kumar P, Satyam A, Fan X, Collin E, Rochev Y, Rodriguez BJ, Gorelov A, Dillon S, Joshi L, Raghunath M, Pandit A, Zeugolis DI (2015) Macromolecularly crowded in vitro microenvironments accelerate the production of extracellular matrix-rich supramolecular assemblies. Sci Rep 5:8729.  https://doi.org/10.1038/srep08729CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Kumar P, Satyam A, Fan X, Rochev Y, Rodriguez BJ, Gorelov A, Joshi L, Raghunath M, Pandit A, Zeugolis DI (2015) Accelerated development of supramolecular corneal stromal-like assemblies from corneal fibroblasts in the presence of macromolecular crowders. Tissue Eng Part C Methods 21(7):660–670.  https://doi.org/10.1089/ten.TEC.2014.0387CrossRefPubMedGoogle Scholar
  12. 12.
    Graceffa V, Zeugolis D (2019) Carrageenan enhances chondrogenesis and osteogenesis in human bone marrow stem cell culture. Eur Cell Mater 37:310–332.  https://doi.org/10.22203/eCM.v037a19CrossRefPubMedGoogle Scholar
  13. 13.
    Graceffa V, Zeugolis D (2019) Macromolecular crowding as a means to assess the effectiveness of chondrogenic media. J Tissue Eng Regen Med 13(2):217–231.  https://doi.org/10.1002/term.2783CrossRefPubMedGoogle Scholar
  14. 14.
    Cigognini D, Gaspar D, Kumar P, Satyam A, Alagesan S, Sanz-Nogues C, Griffin M, O'Brien T, Pandit A, Zeugolis DI (2016) Macromolecular crowding meets oxygen tension in human mesenchymal stem cell culture - a step closer to physiologically relevant in vitro organogenesis. Sci Rep 6:30746.  https://doi.org/10.1038/srep30746CrossRefPubMedPubMedCentralGoogle Scholar
  15. 15.
    Satyam A, Kumar P, Fan X, Gorelov A, Rochev Y, Joshi L, Peinado H, Lyden D, Thomas B, Rodriguez B, Raghunath M, Pandit A, Zeugolis D (2014) Macromolecular crowding meets tissue engineering by self-assembly: a paradigm shift in regenerative medicine. Adv Mater 26(19):3024–3034.  https://doi.org/10.1002/adma.201304428CrossRefPubMedGoogle Scholar
  16. 16.
    Gaspar D, Fuller K, Zeugolis D (2019) Polydispersity and negative charge are key modulators of extracellular matrix deposition under macromolecular crowding conditions. Acta Biomater 88:197–210.  https://doi.org/10.1016/j.actbio.2019.02.050CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  • Mehmet Gürdal
    • 1
    • 2
    • 3
  • Gülinnaz Ercan
    • 3
    • 4
  • Dimitrios I. Zeugolis
    • 1
    • 2
    Email author
  1. 1.Regenerative, Modular & Developmental Engineering Laboratory (REMODEL)National University of Ireland Galway (NUI Galway)GalwayIreland
  2. 2.Science Foundation Ireland (SFI) Centre for Research in Medical Devices (CÚRAM)National University of Ireland Galway (NUI Galway)GalwayIreland
  3. 3.Faculty of Medicine, Department of Medical BiochemistryEge UniversityIzmirTurkey
  4. 4.Department of Stem Cell, Institute of Health SciencesEge UniversityIzmirTurkey

Personalised recommendations