June 2013
Volume 54, Issue 15
Free
ARVO Annual Meeting Abstract  |   June 2013
Soluble collagen protects the corneal fibrils during riboflavin crosslinking
Author Affiliations & Notes
  • Marcos Garza-Madrid
    Translational Tissue Engineering Center, Wilmer Eye Institute, Johns Hopkins School of Medicine, Baltimore, MD
    Ophthalmology and Visual Sciences Research Chair, School of Medicine and Health Sciences, Tecnológico de Monterrey, Monterrey, Mexico
  • Jennifer Elisseeff
    Translational Tissue Engineering Center, Wilmer Eye Institute, Johns Hopkins School of Medicine, Baltimore, MD
Investigative Ophthalmology & Visual Science June 2013, Vol.54, 4074. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      Marcos Garza-Madrid, Jennifer Elisseeff; Soluble collagen protects the corneal fibrils during riboflavin crosslinking. Invest. Ophthalmol. Vis. Sci. 2013;54(15):4074.

      Download citation file:


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

      ×
  • Supplements
Abstract
 
Purpose
 

Cornea crosslinking with riboflavin and UV-A is a novel procedure able to stop the progression of keratoconus. Our aim is to characterize and prevent the fibrillar damage associated with this procedure.

 
Methods
 

Fresh bovine corneas were obtained from a local slaughterhouse. Six mm biopsies, devoid of epithelium and endothelium were treated with a Riboflavin isotonic solution with or without soluble collagen. The tissue was then crosslinked using UV-A light. Hydration was maintained by adding crosslinking solution periodically. Samples were then either processed for TEM or analyzed through DSC to identify the temperature at which the molecular structure breaks down. Fibril counting was performed manually on a minimum of 180 fibrils per group.

 
Results
 

After 30 minutes of crosslinking the denaturation temperature of the corneas increased from 66.77±0.92°C to 70.94±1.72°C (p<0.01). After 45 minutes, however, the temperature dropped to 67.99±1.15°C (p<0.01). When compared to an untreated control (Figure 1: A), cornea crosslinked for one hour (Figure 1: B) shows an accumulation of material in the space between fibers. This accumulation bears remarkable similarity to those formed when collagen is added to the cornea tissue without exposure to UV light (Figure 1: C). There was a significant increase in fibril width after crosslinking (25.91±4.1 nm vs 28.41±5.5 nm, p<0.01). The addition of collagen was able to prevent the change in fibril width (28.41±5.5 nm without collagen vs 24.66±4.2 nm with collagen, p<0.01). Adding collagen to the solution, without exposing the tissue to UV light, had no effect on fibril width (25.91±4.1 nm without collagen vs 25.79±4.1 nm with collagen, p=0.78).

 
Conclusions
 

Cornea collagen fibrils are damaged during riboflavin-UV crosslinking. This damage can be seen through fibril thickening, collagen release and decreased denaturation temperatures. Adding collagen to the crosslinking solution can prevent the damage to the cornea nanostructure during crosslinking.

 
 
Figure 1. TEM images of bovine cornea treated with crosslinking solution alone (A), crosslinking solution and UV (B) and crosslinking solution and collagen (C).
 
Figure 1. TEM images of bovine cornea treated with crosslinking solution alone (A), crosslinking solution and UV (B) and crosslinking solution and collagen (C).
 
Keywords: 484 cornea: stroma and keratocytes • 480 cornea: basic science  
×
×

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.

×