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Collagen Structural Changes in Rat Tarsus After Crosslinking.

Sruti S Akella1, Juan Liu1,2, Yuan Miao1,3

  • 1Department of Ophthalmology and Visual Sciences, Albert Einstein College of Medicine, Montefiore Medical Center, Bronx, NY, USA.

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Photo-activated crosslinking of tarsal collagen shows structural changes and increased stiffness in rat models. This offers a potential new treatment for floppy eyelid syndrome, moving beyond traditional surgery.

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Area of Science:

  • Ophthalmology
  • Biomaterials Science
  • Tissue Engineering

Background:

  • Floppy eyelid syndrome (FES) is typically treated with surgery.
  • Tarsal crosslinking (CXL) is an emerging alternative treatment for FES.
  • Understanding the effects of CXL on tarsal collagen is crucial for its clinical application.

Purpose of the Study:

  • To investigate the structural changes in tarsal collagen after photo-activated crosslinking using second-harmonic generation (SHG) microscopy.
  • To assess the impact of crosslinking on tissue stiffness indirectly using fluorescence recovery after photobleaching (FRAP).
  • To evaluate the efficacy of different riboflavin concentrations in the crosslinking process.

Main Methods:

  • Rat tarsal plates were subjected to crosslinking with varying concentrations of riboflavin (0.1%, 0.3%, 0.5%) and UVA irradiation.
  • Second-harmonic generation (SHG) microscopy was used to visualize collagen fiber organization before and after crosslinking.
  • Fluorescence recovery after photobleaching (FRAP) was employed to indirectly measure changes in tissue stiffness.

Main Results:

  • SHG imaging revealed significant structural alterations in tarsal collagen at a 0.5% riboflavin concentration, with fibers becoming more densely packed and wavy.
  • FRAP analysis demonstrated a significant increase in the half-time of fluorescence recovery (P < 0.05), indicating enhanced tissue stiffness.
  • Lower riboflavin concentrations (0.1% and 0.3%) did not induce observable structural changes or significant stiffness increases.

Conclusions:

  • This study is the first to utilize SHG microscopy for imaging tarsus collagen before and after crosslinking.
  • Photo-activated crosslinking, particularly at higher riboflavin concentrations, induces structural changes in tarsal collagen.
  • These findings support the potential of targeting tarsal collagen with crosslinking as a therapeutic strategy for floppy eyelid syndrome.