June 2013
Volume 54, Issue 15
Free
ARVO Annual Meeting Abstract  |   June 2013
Engineering a biocompatible cell carrier with nanofeatured topography for retinal pigment epithelium transplantation
Author Affiliations & Notes
  • Zengping Liu
    Ophthalmology, University of Bonn, Bonn, Germany
  • Na Yu
    Department of Biomaterials, Radboud University Nijmegen Medical Center, Nijmegen, Netherlands
  • Frank Holz
    Ophthalmology, University of Bonn, Bonn, Germany
  • Fang Yang
    Department of Biomaterials, Radboud University Nijmegen Medical Center, Nijmegen, Netherlands
  • Boris Stanzel
    Ophthalmology, University of Bonn, Bonn, Germany
  • Footnotes
    Commercial Relationships Zengping Liu, None; Na Yu, None; Frank Holz, Acucela (C), Allergan (C), Genentech (F), Heidelberg Engineering (F), Zeiss (F), Novartis (F), Novartis (C), Optos (F), Merz (C), Bayer (F), Bayer (C), Boehringer Ingelheim (C); Fang Yang, None; Boris Stanzel, Geuder AG (F), Geuder AG (P)
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2013, Vol.54, 1386. doi:
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    • Get Citation

      Zengping Liu, Na Yu, Frank Holz, Fang Yang, Boris Stanzel; Engineering a biocompatible cell carrier with nanofeatured topography for retinal pigment epithelium transplantation. Invest. Ophthalmol. Vis. Sci. 2013;54(15):1386.

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

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Abstract

Purpose: An optimal carrier is crucial for retinal pigment epithelium (RPE) tissue engineering. We investigated the influence of fibrillar vs. smooth topography on RPE behavior and subretinal biocompatibility using biodegradable & biostable polymers.

Methods: Poly L-lactide/ε-caprolactone (PLCL) or poly ethylene terephthalate (PETP) fibrillar and smooth substrates were prepared by electrospinning or a heating-press method, respectively. Topography was characterized by scanning electron microscopy. Fetal hRPE growth curves were established from substrates seeded at 1×10E4 cells/cm2, while for differentiation cells were seeded at 6×10E4 cells/cm2. Morphology was monitored by phase contrast microscopy. Cultures were stained for tight junctions (ZO-1) at 6 weeks. Commercial membranes (Transwell®, Corning) coated with 200nm diameter PLCL or PETP fibers were implanted into the rabbit subretinal space. Repetitive in vivo images of the implants by SD-OCT and fundus photography were obtained after 4, 7 and 14days, followed by perfusion-fixed histology.

Results: Uniform PLCL fibers were achieved in 209±33, 568±187 & 1132±299nm diameter ranges, while PETP fibers were in wider ranges (175±86 & 993±596nm). RPE attached on all substrates at comparable densities (8958±2495 cells/cm2) at 24 hours. Cell amount on 200nm PLCL was 13.3% higher than on smooth, while increasing fiber diameters decreased cell counts at day 5. Cells on 200nm PETP showed decreased cell amount (33.6%) compared to smooth. In long term cultures, RPE monolayers partially/entirely detached from PLCL (13/15) and PETP (9/15) smooth films. Cell layers were maintained on all fibrillar scaffolds. 200nm PLCL fibers induced superior RPE pigmentation and uniformly hexagonal ZO-1 staining compared to other PLCL fibrillar or smooth surfaces. Intraoperative handling of all implants was safe. The retina overlying PET/PETP or PLCL subretinal implants appeared attached and transparent on ophthalmoscopy, with largely preserved and continuous layering on SD-OCT. On histology, outer nuclear layer thickness was reduced by half and photoreceptors inner segments were preserved in some regions above both implants.

Conclusions: RPE cells showed material and topography-depended proliferation behavior. Scaffolds created by 200nm fiber diameter topography induced superior RPE differentiation. Both substrates were well tolerated in subretinal space.

Keywords: 695 retinal degenerations: cell biology • 701 retinal pigment epithelium • 733 topography  
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