Comparative Two-Photon Fluorescence Microscopy Analysis of Riboflavin Penetration in Two Different Solutions: Dextran and Hydroxypropyl Methylcellulose

Thales Antonio Abra De Paula, Fernando Betty Cresta, Milton Ruiz Alves

University of São Paulo USP, Ophthalmology, Medical School, São Paulo, Brazil


Correspondence: Thales Antonio Abra De Paula 
 Rua Apeninos 800,apt 1004, São Paulo, SP 04104-020, Brasil
Email thalesaap@hotmail.com

Purpose: This study compared riboflavin penetration in porcine corneas using two different solutions: dextran and hydroxypropyl methylcellulose (HPMC).
Methods: Porcine corneas were de-swollen and after prepared in an artificial anterior chamber with balanced saline solution. The pressure was checked with a portable tonometer. We used 0.1% riboflavin in 20% dextran for 30 min or 0.1% riboflavin in 1% HPMC for 10 min. The fluorescence intensity in multiphoton microscopy was used to evaluate concentration into the cornea, and ultrasonic pachymetry was performed before and after using riboflavin.
Results: Riboflavin concentration up to 340 μm was higher in the HPMC group (p 0.146). From 340 microns to 500 μm, the concentration was higher in the dextran group (p 0.248). There was no statistically significant difference between the groups when considering all the corneal thickness studied (p 0.528). Pachymetric measurements in the group with dextran showed a 12.9% corneal thickness decrease after 30 min, while the HPMC group showed a 9.1% increase after 10 min.
Conclusion: We concluded that 1% HPMC riboflavin for 10 min had the same level of safety when compared to the Dresden protocol with dextran riboflavin for 30 min. Ultrasonic pachymetry during corneal crosslinking has fundamental importance in controlling corneal hydration due to changes in thickness caused by different types of riboflavin. This study used different methods than previous studies to better simulate corneal physiological conditions and used 1% HPMC concentration which differs from previous studies performed with 1.1% HPMC.

Keywords: crosslinking, fluorescence, riboflavin