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Part of the Methods in Molecular Biology book series

3D Stacked Construct: A Novel Substitute for Corneal Tissue Engineering

  • Shrestha Priyadarsini
  • Sarah E. Nicholas
  • Dimitrios Karamichos
Protocol

Abstract

Corneal trauma/injury often results in serious complications including permanent vision loss or loss of visual acuity which demands corneal transplantations or treatment with allogenic graft tissues. There is currently a huge shortage of donor tissue worldwide and the need for human corneal equivalents increases annually. In order to meet such demand the current clinical approach of treating corneal injuries is limited and involves synthetic and allogenic materials which have various shortcomings when it comes to actual transplantations. In this study we introduce the newly developed, next generation of our previously established 3D self-assembled constructs, where multiple constructs are grown and stacked on top of each other without any other artificial product. This new technology brings our 3D in vitro model closer to what is seen in vivo and provides a solid foundation for future studies on corneal biology.

Lipids are known for playing a vital role during metabolism and diseased state of various tissues and Sphingolipids are one such class of lipids which are involved in various cellular mechanisms and signaling processes. The impacts of Sphingolipids that have been documented in several human diseases often involve inflammation, neovascularization, tumorigenesis, and diabetes, but these conditions are not yet thoroughly studied. There is very little information about the exact role of Sphingolipids in the human cornea and future studies aiming at dissecting the mechanisms and pathways involved in order to develop novel therapies. We believe that our novel 3D stacked model can be used to delineate the role of Sphingolipids in the human cornea and provide new insights for understanding and treating various human corneal diseases.

Keywords:

3D constructs Cornea Extra cellular matrix Sphingolipids Stacking 

References

  1. 1.
    Akpek EK, Alkharashi M, Hwang FS, Ng SM, Lindsley K (2014) Artificial corneas versus donor corneas for repeat corneal transplants. Cochrane Database Syst Rev CD009561Google Scholar
  2. 2.
    Chen FM, Liu X (2016) Advancing biomaterials of human origin for tissue engineering. Prog Polym Sci 53:86–168CrossRefPubMedGoogle Scholar
  3. 3.
    Karamichos D, Brown RA, Mudera V (2007) Collagen stiffness regulates cellular contraction and matrix remodeling gene expression. J Biomed Mater Res A 83:887–894CrossRefPubMedGoogle Scholar
  4. 4.
    Griffith LG, Naughton G (2002) Tissue engineering—current challenges and expanding opportunities. Science 295:1009–1014ADSCrossRefPubMedGoogle Scholar
  5. 5.
    Ruberti JW, Zieske JD (2008) Prelude to corneal tissue engineering—gaining control of collagen organization. Prog Retin Eye Res 27:549–577CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Guo X, Hutcheon AE, Melotti SA, Zieske JD, Trinkaus-Randall V et al (2007) Morphologic characterization of organized extracellular matrix deposition by ascorbic acid-stimulated human corneal fibroblasts. Invest Ophthalmol Vis Sci 48:4050–4060CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Karamichos D (2015) Ocular tissue engineering: current and future directions. J Funct Biomater 6:77–80CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Zieske JD (2001) Extracellular matrix and wound healing. Curr Opin Ophthalmol 12:237–241CrossRefPubMedGoogle Scholar
  9. 9.
    Karamichos D, Guo XQ, Hutcheon AE, Zieske JD (2010) Human corneal fibrosis: an in vitro model. Invest Ophthalmol Vis Sci 51:1382–1388CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Priyadarsini S, Sarker-Nag A, Rowsey TG, Ma JX, Karamichos D (2016) Establishment of a 3D in vitro model to accelerate the development of human therapies against corneal diabetes. PLoS One 11:e0168845CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Karamichos D, Hjortdal J (2014) Keratoconus: tissue engineering and biomaterials. J Funct Biomater 5:111–134CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Wilson SL, Yang Y, el Haj AJ (2014) Corneal stromal cell plasticity: in vitro regulation of cell phenotype through cell-cell interactions in a three-dimensional model. Tissue Eng A 20:225–238CrossRefGoogle Scholar
  13. 13.
    Proulx S, Uwamaliya JD, Carrier P, Deschambeault A, Audet C et al (2010) Reconstruction of a human cornea by the self-assembly approach of tissue engineering using the three native cell types. Mol Vis 16:2192–2201PubMedPubMedCentralGoogle Scholar
  14. 14.
    Gonzalez-Andrades M, Alonso-Pastor L, Mauris J, Cruzat A, Dohlman CH et al (2016) Establishment of a novel in vitro model of stratified epithelial wound healing with barrier function. Sci Rep 6:19395ADSCrossRefPubMedPubMedCentralGoogle Scholar
  15. 15.
    Hopkins AM, DeSimone E, Chwalek K, Kaplan DL (2015) 3D in vitro modeling of the central nervous system. Prog Neurobiol 125:1–25CrossRefPubMedGoogle Scholar
  16. 16.
    Schulz S, Beck D, Laird D, Steinberg T, Tomakidi P et al (2014) Natural corneal cell-based microenvironment as prerequisite for balanced 3D corneal epithelial morphogenesis: a promising animal experiment-abandoning tool in ophthalmology. Tissue Eng Part C Meth 20:297–307CrossRefGoogle Scholar
  17. 17.
    Ghezzi CE, Rnjak-Kovacina J, Kaplan DL (2015) Corneal tissue engineering: recent advances and future perspectives. Tissue Eng Part B Rev 21:278–287CrossRefPubMedPubMedCentralGoogle Scholar
  18. 18.
    Karamichos D, Hutcheon AE, Zieske JD (2011) Transforming growth factor-beta3 regulates assembly of a non-fibrotic matrix in a 3D corneal model. J Tissue Eng Regen Med 5:e228–e238CrossRefPubMedPubMedCentralGoogle Scholar
  19. 19.
    Karamichos D, Lakshman N, Petroll WM (2009) An experimental model for assessing fibroblast migration in 3-D collagen matrices. Cell Motil Cytoskeleton 66:1–9CrossRefPubMedPubMedCentralGoogle Scholar
  20. 20.
    Karamichos D, Zareian R, Guo X, Hutcheon AE, Ruberti JW et al (2012) Novel in vitro model for Keratoconus disease. J Funct Biomater 3:760–775CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    Karamichos D, Rich CB, Zareian R, Hutcheon AE, Ruberti JW et al (2013) TGF-β3 stimulates stromal matrix assembly by human corneal keratocyte-like cells. Invest Ophthalmol Vis Sci 54:6612–6619CrossRefPubMedPubMedCentralGoogle Scholar
  22. 22.
    Karamichos D, Hutcheon AE, Zieske JD (2014) Reversal of fibrosis by TGF-beta3 in a 3D in vitro model. Exp Eye Res 124:31–36CrossRefPubMedGoogle Scholar
  23. 23.
    Saika S (2006) TGFbeta pathobiology in the eye. Lab Invest 86:106–115CrossRefPubMedGoogle Scholar
  24. 24.
    Coant N, Sakamoto W, Mao C, Hannun YA (2016) Ceramidases, roles in sphingolipid metabolism and in health and disease. Adv Biol Regul 63:122–131 Google Scholar
  25. 25.
    Brush RS, Tran JT, Henry KR, McClellan ME, Elliott MH et al (2010) Retinal sphingolipids and their very-long-chain fatty acid-containing species. Invest Ophthalmol Vis Sci 51:4422–4431CrossRefPubMedPubMedCentralGoogle Scholar
  26. 26.
    Shea BS, Brooks SF, Fontaine BA, Chun J, Luster AD et al (2010) Prolonged exposure to sphingosine 1-phosphate receptor-1 agonists exacerbates vascular leak, fibrosis, and mortality after lung injury. Am J Respir Cell Mol Biol 43:662–673CrossRefPubMedPubMedCentralGoogle Scholar
  27. 27.
    Shea BS, Tager AM (2012) Sphingolipid regulation of tissue fibrosis. Open Rheumatol J 6:123–129CrossRefPubMedPubMedCentralGoogle Scholar
  28. 28.
    Swaney JS, Moreno KM, Gentile AM, Sabbadini RA, Stoller GL (2008) Sphingosine-1-phosphate (S1P) is a novel fibrotic mediator in the eye. Exp Eye Res 87:367–375CrossRefPubMedGoogle Scholar
  29. 29.
    Watsky MA, Weber KT, Sun Y, Postlethwaite A (2010) New insights into the mechanism of fibroblast to myofibroblast transformation and associated pathologies. Int Rev Cell Mol Biol 282:165–192CrossRefPubMedGoogle Scholar
  30. 30.
    Priyadarsini S, McKay TB, Sarker-Nag A, Allegood J, Chalfant C et al (2016) Complete metabolome and lipidome analysis reveals novel biomarkers in the human diabetic corneal stroma. Exp Eye Res 153:90–100CrossRefPubMedGoogle Scholar
  31. 31.
    French KJ, Schrecengost RS, Lee BD, Zhuang Y, Smith SN et al (2003) Discovery and evaluation of inhibitors of human sphingosine kinase. Cancer Res 63:5962–5969PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  • Shrestha Priyadarsini
    • 1
  • Sarah E. Nicholas
    • 1
  • Dimitrios Karamichos
    • 1
  1. 1.Department of Ophthalmology/Dean McGee Eye InstituteUniversity of Oklahoma Health Sciences CenterOklahoma CityUSA

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