May 2008
Volume 49, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2008
Development of an Artificial Corneal Bandage Using Primary Human Corneal Fibroblasts
Author Affiliations & Notes
  • T. Ng
    Schepens Eye Research Institute, Boston, Massachusetts
    Department of Ophthalmology, Harvard Medical School, Boston, Massachusetts
  • K. Bujold
    Wellman Center of Photomedicine, Boston, Massachusetts
  • R. Redmond
    Wellman Center of Photomedicine, Boston, Massachusetts
  • J. D. Zieske
    Schepens Eye Research Institute, Boston, Massachusetts
    Department of Ophthalmology, Harvard Medical School, Boston, Massachusetts
  • Footnotes
    Commercial Relationships  T. Ng, None; K. Bujold, None; R. Redmond, None; J.D. Zieske, None.
  • Footnotes
    Support  DOD W81XWH-07-2-0038
Investigative Ophthalmology & Visual Science May 2008, Vol.49, 2943. doi:https://doi.org/
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      T. Ng, K. Bujold, R. Redmond, J. D. Zieske; Development of an Artificial Corneal Bandage Using Primary Human Corneal Fibroblasts. Invest. Ophthalmol. Vis. Sci. 2008;49(13):2943. doi: https://doi.org/.

      Download citation file:


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

      ×
  • Supplements
Abstract

Purpose: : To make an artificial corneal bandage from corneal fibroblast secreted matrix in order to protect an injured cornea.

Methods: : Primary human corneal fibroblasts were isolated from human donors, cultured, propagated in vitro and characterized. The cells were seeded in transwell inserts and then allowed to grow for 4 weeks. During this time the cells stratified and produced an extra-cellular matrix to form a 3-D construct. A 2-mm superficial keratectomy was made in the central cornea of BALB/c mice to expose the stroma. Rose Bengal (0.05%) was applied to the exposed corneal stromal surface, and the 3-D construct or corneal bandage was laid down on the stroma and irradiated with a green laser (532nm) for 180 seconds. Prior to transplantation, the artificial corneal bandage was labeled with DTAF. The attachment of the corneal bandage was examined by the presence of the DTAF on days 1, 7 and 14. The mice were sacrificed and the eyes were processed and examined by immunofluorescence for the presence of infiltrating immune cells with anti-CD45.

Results: : The isolated corneal cells expressed keratan sulfate proteoglycan (KSPG), Ki67 and CD90; however, they did not express keratin AE1/3 nor HLA DR. After forming the 3-D construct, the KSPG positive cells laid down extra-cellular matrix components, including collagen VI, fibronectin and thrombospondin-1. The Rose Bengal and green laser irradiation made a tight bond between the bandage and the corneal stroma. The DTAF labeled collagen stayed on the corneal stromal surface up to day 14 and was covered by epithelium by day 3. CD45 positive immune cells were found on day 1 in the limbus and central cornea in eyes +/- the artificial bandage. However, by day 3, the number of immune cells decreased in the corneas with the bandage; whereas, the corneas without bandage still maintained numerous CD45 positive cells.

Conclusions: : The artificial corneal bandage provides a good barrier for protecting the injured corneal stroma.

Keywords: cornea: stroma and keratocytes • keratoprostheses • wound healing 
×
×

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.

×