June 2015
Volume 56, Issue 7
Free
ARVO Annual Meeting Abstract  |   June 2015
Perlecan and nidogen-2 up-regulation in stromal keratocytes after epithelial injury in human corneas
Author Affiliations & Notes
  • Andre A. M. Torricelli
    University Of Sao Paulo, Sao Paulo, Brazil
    Cole Eye Institute, Cleveland Clinic, Cleveland, OH
  • Jiahui Wu
    Cole Eye Institute, Cleveland Clinic, Cleveland, OH
  • Abirami Santhanam
    Cole Eye Institute, Cleveland Clinic, Cleveland, OH
  • Gustavo Kupper Kupper Marino
    Cole Eye Institute, Cleveland Clinic, Cleveland, OH
  • Arun Singh
    Cole Eye Institute, Cleveland Clinic, Cleveland, OH
  • Steven E. Wilson
    Cole Eye Institute, Cleveland Clinic, Cleveland, OH
  • Footnotes
    Commercial Relationships Andre Torricelli, None; Jiahui Wu, None; Abirami Santhanam, None; Gustavo Kupper Marino, None; Arun Singh, None; Steven Wilson, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2015, Vol.56, 710. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to Subscribers Only
      Sign In or Create an Account ×
    • Get Citation

      Andre A. M. Torricelli, Jiahui Wu, Abirami Santhanam, Gustavo Kupper Kupper Marino, Arun Singh, Steven E. Wilson; Perlecan and nidogen-2 up-regulation in stromal keratocytes after epithelial injury in human corneas. Invest. Ophthalmol. Vis. Sci. 2015;56(7 ):710.

      Download citation file:


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

      ×
  • Supplements
Abstract
 
Purpose
 

To investigate stromal production of corneal epithelial basement membrane (EBM) proteins in wounded human corneas.

 
Methods
 

A total of 5 unwounded control eyes and 4 wounded corneas were obtained from fresh corneoscleral buttons from human eyes enucleated due to choroidal melanoma with normal anterior segments. In the wounded corneas the central epithelium was scraped to remove an 8 mm patch of central epithelium and EBM. These corneas were collected 30 minutes after injury. Immunohistochemistry were performed to detect perlecan and nidogen-2 proteins that are important components of the EBM.

 
Results
 

Perlecan and nidogen-2 proteins were detected in the epithelium and BM zone in unwounded corneas. Little immunoreactivity was detected for either BM components in stromal cells of the unwounded corneas. However, after epithelial injury there was a strong signal for nidogen-2 in stromal keratocytes in all wounded corneas (Fig 1). Perlecan was also detected at higher levels in the stromal tissue and associated keratocytes in all wounded corneas (Fig 2).

 
Conclusions
 

After epithelial and EBM injury, stromal keratocytes produce perlecan and nidogen-2 components of the epithelial BM and likely contribute to regeneration of the EBM. We hypothesize that, after self-polymerizing laminin is secreted by epithelial cells to form the nascent EBM, some deeper components must be provided by keratocytes and/or corneal fibroblasts.  

 
Immunohistochemistry for nidogen-2. Column A. Nidogen-2 protein noted in the epithelial BM (arrows) and epithelium in unwounded corneas B. Negative control in unwounded cornea. C. At 30 minutes after epithelial scrape, nidogen-2 protein was detected in stromal keratocytes (arrowheads). Staining near the stromal surface (arrows) could represent BM not completely removed or expression in keratocytes D. Negative control cornea after epithelial scrape<br /> <br /> <br />
 
Immunohistochemistry for nidogen-2. Column A. Nidogen-2 protein noted in the epithelial BM (arrows) and epithelium in unwounded corneas B. Negative control in unwounded cornea. C. At 30 minutes after epithelial scrape, nidogen-2 protein was detected in stromal keratocytes (arrowheads). Staining near the stromal surface (arrows) could represent BM not completely removed or expression in keratocytes D. Negative control cornea after epithelial scrape<br /> <br /> <br />
 
 
Immunohistochemistry for perlecan. Column A. Perlecan protein noted in the epithelium and epithelial BM (arrows) in unwounded corneas B. Negative control in unwounded cornea C. At 30 minutes after epithelial scrape, strong perlecan immunoreactivity was noted in keratocytes (arrowheads). Staining near the stromal surface (arrows) could represent BM not completely removed or expression in keratocytes D. Negative control cornea after epithelial scrape<br />
 
Immunohistochemistry for perlecan. Column A. Perlecan protein noted in the epithelium and epithelial BM (arrows) in unwounded corneas B. Negative control in unwounded cornea C. At 30 minutes after epithelial scrape, strong perlecan immunoreactivity was noted in keratocytes (arrowheads). Staining near the stromal surface (arrows) could represent BM not completely removed or expression in keratocytes D. Negative control cornea after epithelial scrape<br />

 
×
×

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.

×