Abstract
Abstract: :
Purpose: Human Amniotic Membrane (HAM) preparations are used for the reconstruction of the ocular surface in an expanding number of applications. Due to its transparency, thin structure, and adhesive quality the surgical use of HAM can be challenging. Staining of HAM has been proposed to facilitate its intraoperative visualization and manipulation. We evaluated the staining properties of de–epithelialized HAM using five commonly available dyes. Methods:De–epithelialized HAM was stained with different concentrations of five dyes, indocyanine green (1.0%, 0.5%, and 0.1%), flourescein (1.0%, 0.25%, 0.1%), lissamine green (0.5%, 0.1%), rose bengal (1.0%, 0.1%), and trypan blue (0.5%, 0.1%). The initial staining characteristics of each dyed sample was evaluated using an operating microscope by a single observer. Each positively staining membrane was then rinsed and resuspended with 3ml of balanced salt solution at thirty–minute intervals for a total of four hours and re–evaluated at each interval for positive staining. All remaining stained samples were then placed in 5ml balanced salt saline and then re–examined for staining at 24 hours. Results: All tested concentrations of indocyanine green, flourescein, lissamine green, rose bengal, and trypan blue stained de–epithelialized HAM initially. After 120 minutes had elapsed flourescein 0.1%, lissamine green B 0.5%, lissamine green B 0.1% no longer stained the membrane. Flourescein 0.5% no longer stained the de–epithelialized HAM at 210 minutes, and flourescein 1.0% no longer stained at 24 hours. Indocyanine green (1.0%, 0.5%, and 0.1%), rose Bengal (1.0%and 0.1%), and trypan blue (0.5% and 0.1%) all demonstrated positive staining at 24 hours. Conclusions: Indocyanine green, flourescein, lissamine green, rose bengal, and trypan blue stain de–epithelialized HAM. Flourescein (1.0%, 0.25%, and 0.1%) and lissamine green B (0.5%, 0.1%) may be superior to other tested dyes for intraoperative visualization of de–epithelialized HAM, because these dyes stain the membrane and then fade between 120 and 210 minutes after application. Rose Bengal (1.0%, 01%) and trypan blue (0.5%, 0.1%) may not be suitable dyes for de–epithelialized HAM as they demonstrate persistent staining at 24 hours after initial application.
Keywords: cornea: basic science • anterior segment