Validation of preloaded DMEK donor tissues: a laboratory-based study on endothelial cell viability and comparison of two F-mark inks

• Published in: Canadian journal of ophthalmology 2024-12, Vol.59 (6), p.e673-e681
• Main Authors: Dhupar, Narisa, AlShaker, Sara, Chai, Jenny, Yan, Judy, Little, Ness, Humphreys, Christine, Chan, Clara C., Ong Tone, Stephan
• Format: Article
• Language: English
• Subjects: Aged; Aged, 80 and over; Cell Count; Cell Survival - physiology; Corneal Endothelial Cell Loss - diagnosis; Descemet Stripping Endothelial Keratoplasty - methods; Endothelium, Corneal - cytology; Endothelium, Corneal - pathology; Eye Banks - methods; Female; Humans; Ink; Male; Middle Aged; Tissue and Organ Harvesting - methods; Tissue Donors
• ISSN: 0008-4182; 1715-3360; 1715-3360
• DOI: 10.1016/j.jcjo.2024.02.012

To investigate endothelial cell loss (ECL) associated with Descemet membrane endothelial keratoplasty (DMEK) donor tissues preloaded in the DMEK RAPID transport system after 1 and 5 days and to compare prestamping with 2 different F-mark inks. DMEK donor tissues were stripped, marked with gentian violet dye applied as an F-mark, trephined, stained with trypan blue, and then preloaded into the DMEK RAPID transport system by an eye bank technician. Preloaded DMEK tissues were then unfolded and stained with calcein AM after 1 or 5 days of storage. Tissues were imaged, analyzed for total tissue ECL, and immunostained for corneal endothelium markers zonular occludens-1 and xCD166. Additionally, ECL and the intensity of an F-mark caused by 2 different inks were quantified. Preloaded DMEK tissues displayed an average ECL of 11.9% ± 4.5% (n = 8) at 1 day and 9.9% ± 4.2% (n = 9) at 5 days. No difference was found between the 2 groups. Zonular occludens-1 and activated leukocyte cell adhesion molecule (ALCAM; also know as CD166) staining showed that the corneal endothelial monolayer remained intact on preloaded tissues. On 5-day preloaded DMEK tissues, the average ECL and mean grayscale caused by the Keir Surgical ink F-mark and the Cardinal Health ink F-mark were 4.3% ± 0.8% and 158.5 ± 13.9% and 5.0% ± 1.1% and 142.9% ± 20.0%, respectively. No difference was found between the F-mark inks. Preloaded DMEK donor tissues resulted in an acceptable ECL range after 1 and 5 days of storage and were deemed suitable for transplantation. Both F-mark inks are acceptable for prestamping preloaded DMEK tissues prior to surgical transplantation with comparable ECL and intensities. Examiner la perte cellulaire endothéliale (PCE) après 1 jour et 5 jours dans des tissus de donneurs préchargés dans le système de transport DMEK RAPID en vue d'une kératoplastie endothéliale de la membrane de Descemet (DMEK) et comparer le marquage préalable d'un F au moyen de 2 encres différentes. Lestissus de donneurs ont été décapés, marqués d'un F au violet de gentiane, trépanés, colorés au bleu trypan et, enfin, préchargés dans le système de transport DMEK RAPID par un technicien de la banque d'yeux. Les tissus préchargés en vue de la DMEK ont ensuite été dépliés et colorés à la calcéine-AM après 1 jour ou 5 jours d'entreposage. Les tissus ont été imagés et analysés afin de mesurer la PCE tissulaire totale; ils ont ensuite subi une immunocoloration pour faire ressortir 2 marqueurs de l'endothélium