May 2007
Volume 48, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2007
Targeting Activated Corneal Keratocytes Through CD90 Specific RNA Aptamers: Potential for Addressing Early Loss of Corneal Transparency
Author Affiliations & Notes
  • Y. M. Khan
    Ophthalmology, Rayne Institute , King's College London, London, United Kingdom
  • R. Angunawela
    Ophthalmology, Rayne Institute , King's College London, London, United Kingdom
  • D. Bunka
    Faculty of Biological Sciences, University of Leeds, Leeds, United Kingdom
  • P. Stockely
    Faculty of Biological Sciences, University of Leeds, Leeds, United Kingdom
  • J. Marshall
    Ophthalmology, Rayne Institute , King's College London, London, United Kingdom
  • Footnotes
    Commercial Relationships Y.M. Khan, None; R. Angunawela, None; D. Bunka, None; P. Stockely, None; J. Marshall, None.
  • Footnotes
    Support Royal College of Surgeons
Investigative Ophthalmology & Visual Science May 2007, Vol.48, 1477. doi:https://doi.org/
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      Y. M. Khan, R. Angunawela, D. Bunka, P. Stockely, J. Marshall; Targeting Activated Corneal Keratocytes Through CD90 Specific RNA Aptamers: Potential for Addressing Early Loss of Corneal Transparency. Invest. Ophthalmol. Vis. Sci. 2007;48(13):1477. doi: https://doi.org/.

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

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Abstract

Introduction:: Corneal keratocytes undergo injury induced cellular differentiation into activated keratocytes (fibroblasts). Early loss of corneal transparency is predominantly related to changes in number and density of activated keratocytes contributing to impairment of vision in post operative period. Specific targeting of differentiated cell types with therapeutic agents could minimize postoperative haze and scarring.

Purpose:: Based on our previous work we aimed to develop an aptamer specific to activated corneal keratocytes by selectively targeting the Thy-1 (CD-90) surface marker.

Methods:: A Thy-1 specific RNA aptamer was developed using a counter selective molecular strategy (SELEX).A defined stromal injury was created using a femtosecond laser platform. Tissue was cultured in a specifically designed artificial anterior chambers for seven days to allow keratocytes activation. Corneas were then fixed and sectioned prior to labelling with the RNA aptamer. Foreskin fibroblasts, passive keratocytes and a naïve aptamer pool were used as controls.

Results:: There was specific staining of activated keratocytes at the interface of tissue injury which labelled clearly with the CD-90 aptamer( p<0.01). Passive cells remained unlabelled.

Conclusions:: This is a significant step forward in specific cell targeting using an RNA aptamer. These developments bring us a step closer to in-vivo cellular targeting in the human cornea.

Keywords: cornea: stroma and keratocytes • cornea: basic science • cornea: epithelium 
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