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Isolation, Cryopreservation and Culture of Human Amnion Epithelial Cells for Clinical Applications
Published on: December 21, 2014
Staining characteristics of preserved human amniotic membrane
Daniel J Hu1, Surendra Basti, Paul J Bryar
1Department of Ophthalmology, Northwestern University, Feinberg School of Medicine, Chicago, IL 60611, USA.
Purpose:
Amniotic membrane is an ultra-thin cellophane-like membrane that is used in ocular surface reconstruction. We evaluated the staining characteristics of commonly available dyes on preserved human amniotic membrane to aid in handling of amniotic membrane during transplantation.
Methods:
Five dyes, indocyanine green (2.5%, 1.0%, and 0.5%), fluorescein (0.25%), rose bengal (1%), lissamine green B (1%), and trypan blue (0.5%), were used to stain amniotic membrane. After staining, the specimens were observed under a dissecting microscope to evaluate for the uptake of the stains. Positively stained membranes were evaluated for the persistence of staining by placing them in 2 to 3 mL of balanced saline solution that was changed every 30 minutes over 6 hours.
Results:
Preserved human amniotic membrane is stained by indocyanine green, rose bengal, lissamine green B, and trypan blue. Of these four dyes, only the membrane stained with 1% lissamine green B was free of stain after 120 minutes. Indocyanine green, rose bengal, and trypan blue continued to strongly stain the membrane after 24 hours.
Conclusions:
Indocyanine green, rose bengal, trypan blue, and lissamine green B all stain amniotic membrane. Lissamine green B appears to have advantages over the other dyes in that it will stain the membrane well, and in our model, dissipate in 120 minutes. Intraoperative staining with lissamine green B may be a simple and effective way to assist surgeons in the proper handling of amniotic membrane.
Insights
Lissamine green B effectively stains amniotic membrane for surgical handling and dissipates within 120 minutes, unlike other tested dyes. This makes it a potentially useful tool for intraoperative use in amniotic membrane transplantation.
Area of Science:
- Ophthalmology
- Biomaterials Science
- Tissue Engineering
Background:
- Amniotic membrane is crucial for ocular surface reconstruction.
- Proper handling during transplantation is essential for successful outcomes.
- Evaluating dye characteristics aids surgical procedures.
Purpose of the Study:
- To assess the staining properties of common dyes on preserved human amniotic membrane.
- To determine dye uptake and persistence for surgical guidance.
- To identify optimal dyes for handling amniotic membrane during transplantation.
Main Methods:
- Five dyes (indocyanine green, fluorescein, rose bengal, lissamine green B, trypan blue) were applied to amniotic membrane specimens.
- Stain uptake was evaluated using a dissecting microscope.
- Stain persistence was assessed in balanced saline solution over 6 hours.
Main Results:
- Indocyanine green, rose bengal, lissamine green B, and trypan blue all stained the amniotic membrane.
- Only lissamine green B (1%) showed stain dissipation within 120 minutes.
- Indocyanine green, rose bengal, and trypan blue exhibited persistent staining even after 24 hours.
Conclusions:
- Lissamine green B, indocyanine green, rose bengal, and trypan blue all stain amniotic membrane.
- Lissamine green B offers an advantage due to its temporary staining, dissipating in 120 minutes.
- Intraoperative staining with lissamine green B may facilitate better handling of amniotic membrane during surgery.
