July 2018
Volume 59, Issue 9
Open Access
ARVO Annual Meeting Abstract  |   July 2018
Ultrathin Poly-DL-Lactic Membranes for Corneal Endothelial Transplantation
Author Affiliations & Notes
  • Bert Van den Bogerd
    Laboratory of Ophthalmology, Translational Neurosciences, University of Antwerp, Wilrijk, Belgium
  • Jasper Van Hoorick
    Polymer Chemistry & Biomaterials Group, Department of Organic and Macromolecular Chemistry, Ghent University, Ghent, Belgium
    Brussels Photonics, Department of Applied Physics and Photonics, Free University of Brussels, Brussels, Belgium
  • Jürgen Van Erps
    Brussels Photonics, Department of Applied Physics and Photonics, Free University of Brussels, Brussels, Belgium
  • Hugo Thienpont
    Polymer Chemistry & Biomaterials Group, Department of Organic and Macromolecular Chemistry, Ghent University, Ghent, Belgium
    Brussels Photonics, Department of Applied Physics and Photonics, Free University of Brussels, Brussels, Belgium
  • Sorcha Ní Dhubhghaill
    Department of Ophthalmology, Antwerp University Hospital, Edegem, Belgium
  • carina koppen
    Department of Ophthalmology, Antwerp University Hospital, Edegem, Belgium
  • Peter Dubruel
    Polymer Chemistry & Biomaterials Group, Department of Organic and Macromolecular Chemistry, Ghent University, Ghent, Belgium
  • Sandra Van Vlierberghe
    Polymer Chemistry & Biomaterials Group, Department of Organic and Macromolecular Chemistry, Ghent University, Ghent, Belgium
    Brussels Photonics, Department of Applied Physics and Photonics, Free University of Brussels, Brussels, Belgium
  • Nadia Zakaria
    Laboratory of Ophthalmology, Translational Neurosciences, University of Antwerp, Wilrijk, Belgium
    Department of Ophthalmology, Antwerp University Hospital, Edegem, Belgium
  • Footnotes
    Commercial Relationships   Bert Van den Bogerd, None; Jasper Van Hoorick, None; Jürgen Van Erps, None; Hugo Thienpont, None; Sorcha Ní Dhubhghaill, None; carina koppen, None; Peter Dubruel, None; Sandra Van Vlierberghe, None; Nadia Zakaria, Novartis Institute for Biomedical Research (E)
  • Footnotes
    Support  None
Investigative Ophthalmology & Visual Science July 2018, Vol.59, 1378. doi:
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    • Get Citation

      Bert Van den Bogerd, Jasper Van Hoorick, Jürgen Van Erps, Hugo Thienpont, Sorcha Ní Dhubhghaill, carina koppen, Peter Dubruel, Sandra Van Vlierberghe, Nadia Zakaria; Ultrathin Poly-DL-Lactic Membranes for Corneal Endothelial Transplantation. Invest. Ophthalmol. Vis. Sci. 2018;59(9):1378.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose : Corneal endothelial transplantation is limited by a donor shortage, making transplant lists long. As hypothesized in 1978, tissue-engineering could overcome this hurdle by growing corneal endothelial cells (CEnCs) on transplantable scaffolds. Despite the global interest, no tissue-engineered graft has made it yet to the clinic in part due to challenging properties scaffolds must fulfil. This study investigates the use of Poly-DL-lactic (PDLLA) membranes as scaffolds to aid in production of corneal endothelial grafts.

Methods : Membranes were synthesized by spincoating a 10 w/v% solution of gelatin type A on glass as a sacrificial layer (Fig 1A). PDLLA was spincoated on top and plasma treated for surface activation. Gelatin B derivatives, gelatine-methacrylamide (gel-MOD) and gel-MOD-2-aminoethyl methacrylate (gel-MOD-AEMA) were analyzed using Nuclear Magnetic Resonance (NMR) spectroscopy, spincoated on the PDLLA and UV crosslinked (Fig 1B). The membranes were characterized with x-ray photoemission spectroscopy (XPS), optical profilometry (OP) to determine thickness and glucose diffusion. Also, cytocompatibility was investigated using primary and immortalized CEnCs followed by immunocytochemistry.

Results : NMR spectra confirmed the modification of gel-MOD(-AEMA). The presence of stable gelatin layers during the synthesis was confirmed using XPS. The developed membranes exhibited thicknesses ranging from 1.1 - 2.6 µm as obtained via OP and the PDLLA showed a diffusion glucose coefficient of 0.0161cm/s. Finally, the membranes were seeded with primary and immortalized CEnCs and exhibited the characteristic hexagonal shape. Immunocytochemistry showed expression of Na+/K+ ATPase and ZO-1 along the cell membranes of primary and immortalized CEnCs cultured on both culture plastic and gel-MOD(-AEMA) membranes.

Conclusions : In this study, membranes were constructed using PDLLA for mechanical strength and functionalized gelatins to mimic the extracellular matrix. The membranes displayed glucose permeability and because of the ultrathin dimensions they qualify as scaffolds for endothelial keratoplasty. Cultured endothelial cells expressed hallmark proteins representing their pumping and barrier function. Therefore, these novel ultrathin membranes are potentially a suitable scaffold for corneal endothelial tissue engineering required to alleviate the donor shortage.

This is an abstract that was submitted for the 2018 ARVO Annual Meeting, held in Honolulu, Hawaii, April 29 - May 3, 2018.

 

(A) Principle of spincoating (B) A diagram of the PDLLA membranes

(A) Principle of spincoating (B) A diagram of the PDLLA membranes

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