Optimizing Visualization of Descemet Membrane Endothelial Keratoplasty Tissue: Assessing the Impact of Trypan Blue Exposure on Stain Duration and Corneal Endothelial Cell Function
| Autor: | Mark A. Greiner, M. Bridget Zimmerman, Benjamin T. Aldrich, Chris Conwell, Cynthia R. Reed, Jennifer J. Ling, Jennifer Y. Li, Jessica M. Skeie, Tiffany Ramirez, Kimberlee A. Burckart, Gregory A. Schmidt, Ralph Kyrillos |
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| Rok vydání: | 2020 |
| Předmět: |
medicine.medical_specialty
Time Factors Descemet membrane Balanced salt solution Stain 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Ophthalmology Respiration medicine Humans Coloring Agents Staining and Labeling business.industry Endothelium Corneal Trypan Blue Corneal Endothelial Cell Loss Middle Aged Tissue Donors Staining Mitochondria chemistry 030221 ophthalmology & optometry Tissue and Organ Harvesting Trypan blue Corneal endothelial cell sense organs business 030217 neurology & neurosurgery Methylene blue Descemet Stripping Endothelial Keratoplasty |
| Zdroj: | Cornea. 40(3) |
| ISSN: | 1536-4798 |
| Popis: | Purpose To assess how trypan blue staining affects Descemet membrane endothelial keratoplasty (DMEK) graft visibility and corneal endothelial cell (CEC) mitochondrial respiration. Methods DMEK grafts (n = 20) were stained with trypan blue 0.06% for 1, 3, 5, or 10 minutes. Each graft was injected into an artificial anterior chamber. Surgery was simulated with tapping and sweeping motions on the corneal surface and injections of balanced salt solution (BSS). Graft visibility was assessed at 5, 10, 20, and 30 minutes. Effects of trypan blue on mitochondrial respiration were assessed using primary CECs cultured from donor corneas (n = 43). Treatment wells exposed to trypan blue 0.06% (1, 5, or 30 minutes) and donor-matched control wells to methylene blue 1% (1 minute) or BSS (1, 5, or 30 minutes) were assayed for key respiration parameters. Results After 5 minutes of surgical manipulation, grafts stained for 5 minutes were significantly more visible than grafts stained for 1 or 3 minutes; there was no added benefit of staining for 10 minutes. After 10 minutes of surgical manipulation, grafts stained for 3 minutes were more visible than grafts stained for 1 minute, without additional benefits of staining ≥5 minutes. No visibility differences were observed after ≥20 minutes of surgical manipulation. CEC mitochondrial respiration did not change significantly following trypan blue exposure for all intervals tested compared to BSS. Conclusions Staining DMEK grafts with trypan blue for 3 to 5 minutes optimizes visibility during surgical manipulation without mitochondrial impairment. Corneal surgeons learning DMEK will benefit from optimizing this critical step. |
| Databáze: | OpenAIRE |
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