Biomechanical Changes After In Vivo Collagen Cross-Linking With Rose Bengal-Green Light and Riboflavin-UVA

Invest Ophthalmol Vis Sci. 2017 Mar 1;58(3):1612-1620. doi: 10.1167/iovs.17-21475.


Purpose: To compare corneal biomechanical properties after in vivo and ex vivo cross-linking (CXL) using rose bengal-green light (RGX) or riboflavin-UVA (UVX).

Methods: Corneas of 30 rabbits were treated in vivo by the two CXL modalities monolaterally (Group 1) or bilaterally (Group 2). Rabbits in Group 1 were euthanized 1 month after treatments and in Group 2 two months after treatment. Ex vivo CXL was also performed. Eyes were measured by Scheimpflug air puff corneal deformation imaging (Corvis ST) under constant IOP. Corneal deformation parameters were assessed. Inherent corneal biomechanical properties were estimated using inverse finite element modeling.

Results: Peak to peak distance decreased 16% 2 months after RGX, and 4% and 20% 1 and 2 months after UVX, respectively. The equivalent Young's modulus (Eeq) increased relative to the control during the post treatment period for both RGX and UVX. The Eeq increased by factors of 3.4 (RGX) and 1.7 (UVX) 1 month and by factors of 10.7 (RGX) and 7.3 (UVX) 2 months after treatment. However, the Eeq values for ex vivo CXL were much greater than produced in vivo. The ex vivo Eeq was greater than the 1-month in vivo values by factors of 8.1 (RGX) and 9.1 (UVX) and compared with 2 month by factors of 2.5 (RGX) and 2.1 (UVX).

Conclusions: These results indicate that corneal stiffness increases after CXL, and further increases as a function of time after both RGX and UVX. Also, while biomechanical properties determined after ex vivo CXL are indicative of corneal stiffening, they may not provide entirely accurate information about the responses to CXL in vivo.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Collagen / pharmacology*
  • Corneal Diseases / drug therapy*
  • Corneal Diseases / pathology
  • Corneal Diseases / physiopathology
  • Corneal Stroma / drug effects
  • Corneal Stroma / pathology
  • Corneal Stroma / physiopathology*
  • Cross-Linking Reagents / pharmacology*
  • Disease Models, Animal
  • Elasticity
  • Fluorescent Dyes / pharmacology
  • Photosensitizing Agents / pharmacology
  • Rabbits
  • Riboflavin / pharmacology*
  • Rose Bengal / pharmacology*
  • Ultraviolet Rays*


  • Cross-Linking Reagents
  • Fluorescent Dyes
  • Photosensitizing Agents
  • Rose Bengal
  • Collagen
  • Riboflavin