March 2012
Volume 53, Issue 14
ARVO Annual Meeting Abstract  |   March 2012
A New Model for In Vitro Testing of Vitreous Substitute Candidates
Author Affiliations & Notes
  • Henrik Barth
    Department of Ophthalmology, Lund University, Lund, Sweden
  • Sven W. Crafoord
    Ophthalmology, Orebro University Hospital, Orebro, Sweden
  • Cyrille Vinchon
    Anteis S.A., Plan Les Ouates, Switzerland
  • Timothy M. O’Shea
    Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts
  • Christopher D. Pritchard
    Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts
  • Fredrik K. Ghosh
    Department of Ophthalmology, Lund University, Lund, Sweden
  • Footnotes
    Commercial Relationships  Henrik Barth, None; Sven W. Crafoord, None; Cyrille Vinchon, Employee of Anteis S.A. (E); Timothy M. O’Shea, None; Christopher D. Pritchard, None; Fredrik K. Ghosh, None
  • Footnotes
    Support  The Swedish Research Council no90247201; The Wallenberg Foundation MMW 2011.0009; The Princess Margaretas Foundation for Blind Children; The Faculty of Medicine, University of Lund
Investigative Ophthalmology & Visual Science March 2012, Vol.53, 3765. doi:
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      Henrik Barth, Sven W. Crafoord, Cyrille Vinchon, Timothy M. O’Shea, Christopher D. Pritchard, Fredrik K. Ghosh; A New Model for In Vitro Testing of Vitreous Substitute Candidates. Invest. Ophthalmol. Vis. Sci. 2012;53(14):3765.

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

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Purpose: : To describe a new model for in vitro assessment of novel vitreous substitute candidates.

Methods: : The biological impact of three vitreous substitute candidates were explored in a retinal explant culture model; a polyalkyl-imide hydrogel (Bio-Alcamid®), a two component hydrogel of 20 wt.% poly(ethylene glycol) in phosphate buffered saline (PEG) and a cross-linked sodium hyaluronic acid hydrogel (Healaflow®). The gels where applied to explanted adult rat retinas and then kept in culture for 5 days. Gel exposed explants were compared with explants incubated under standard conditions (medium only). Cryosections of the specimens were stained with hematoxylin and eosin, immunohistochemical markers (GFAP, Rhodopsin), and TUNEL.

Results: : Explants kept under standard conditions as well as PEG exposed explants displayed disruption of retinal layers with moderate pyknosis of first, second and third order neurons. They also displayed moderate fragmentation of DNA (TUNEL). Bio-Alcamid® exposed explants displayed severe thinning and disruption of retinal layers with massive cell-death. Healaflow® treated explants displayed normal retinal lamination with significantly better preservation of retinal neurons compared with control specimens, and almost no DNA-fragmentation. Retinas exposed to Healaflow® and retinas kept under standard condition showed variable labeling of GFAP with generally low expression and some areas of upregulation, PEG-exposed retinas showed increased GFAP labeling, and Bio-Alcamid® exposed retinas showed sparse labeling of GFAP.

Conclusions: : Research into novel vitreous substitutes has important implications for both medical and surgical vitreoretinal disease. The in vitro model presented here provides a method of biocompatibility testing prior to more costly and cumbersome in vivo experiments. The explant culture system under standard conditions imposes reactions within the retina that can be used for comparison using vitreous substitute candidates. These reactions include disruption of layers, cell-death and GFAP upregulation. PEG gel imposes reactions similar to the control retinas whereas Healaflow® shows protection from culture induced trauma, indicating a favorable biocompatibility. Bio-Alcamid® adversely affects the retina strongly, which is consistent with the results of prior in vivo trials.

Keywords: vitreous substitutes • retina • vitreoretinal surgery 

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