In vitro endothelial cell migration from limbal edge-modified Quarter-DMEK grafts


Autoři: Alina Miron aff001;  Daniele Spinozzi aff001;  Sorcha Ní Dhubhghaill aff001;  Jessica T. Lie aff001;  Silke Oellerich aff001;  Gerrit R. J. Melles aff001
Působiště autorů: Netherlands Institute for Innovative Ocular Surgery, Rotterdam, The Netherlands aff001;  Melles Cornea Clinic Rotterdam, Rotterdam, The Netherlands aff002;  Amnitrans EyeBank Rotterdam, Rotterdam, The Netherlands aff003
Vyšlo v časopise: PLoS ONE 14(11)
Kategorie: Research Article
doi: 10.1371/journal.pone.0225462

Souhrn

Endothelial cell migration plays a crucial role in achieving corneal clearance after corneal transplantation when using smaller-sized endothelial grafts to increase the donor pool. In this study we investigated how different strategies of Quarter-Descemet Membrane Endothelial Keratoplasty (Quarter-DMEK) limbal graft edge modification influence peripheral endothelial cell migration in an in vitro culture environment. For this study, 15 Quarter-DMEK grafts, prepared from 7 corneas deemed ineligible for transplantation but with intact and viable endothelial cells, were embedded in a cooled biocompatible, thermoresponsive matrix for culture. The limbal edge of ten Quarter-DMEK grafts were modified, either by using a small diameter punch or by peripheral radial cuts. All Quarter-DMEK grafts showed substantial collective endothelial cell migration from the radial cut graft edges, as observed by light microscopy at standardized time intervals. Grafts were retrieved from the polymer matrix after the two-week culture for immunohistochemistry analyses of the newly formed cell monolayers; this demonstrated the presence of tightly packed and viable cells that showed higher migratory ability at the leading edge. Peripheral endothelial cell migration, however, was not triggered by increasing cell exposure to free space through surgical modifications of the far periphery. Our data suggest that alterations in the far peripheral area of Quarter-DMEK grafts were insufficient to triggering cell migration from the limbal graft edge. This may be due to transient-amplifying cells that reside in the far periphery and which lack cytokinetic directional cues. Understanding the migration capacity of the peripheral endothelium could unlock cells’ therapeutic potential which are, at present, routinely discarded from transplantation. Encouraging peripheral cell migration may also improve clinical outcomes from Quarter-DMEK, but a more effective solution is required prior to clinical implementation of modified grafts.

Klíčová slova:

Cell migration – Cornea – Cytoskeletal proteins – Endothelial cells – Immunohistochemical analysis – Light microscopy – Vimentin – Endothelium


Zdroje

1. Baydoun L, Zygoura V, Hsien S, Birbal RS, Spinozzi D, Lie JT, et al. Clinical feasibility of using multiple grafts from a single donor for Quarter-DMEK. Acta Ophthalmol. 2018;96(5):e656–658. doi: 10.1111/aos.13720 29498213

2. Zygoura V, Baydoun L, Ham L, Bourgonje VJA, van Dijk K, Lie JT, et al. Quarter-Descemet membrane endothelial keratoplasty (Quarter-DMEK) for Fuchs endothelial corneal dystrophy: 6 months clinical outcome. Br J Ophthalmol. 2018;102(10):1425–1430. doi: 10.1136/bjophthalmol-2017-311398 29343529

3. Muller TM, Lavy I, Baydoun L, Lie JT, Dapena I, Melles GR. Case report of Quarter-Descemet membrane endothelial keratoplasty for Fuchs endothelial dystrophy. Cornea. 2017;36(1):104–107. doi: 10.1097/ICO.0000000000001008 27583798

4. Miron A, Spinozzi D, Bruinsma M, Lie JT, Birbal RS, Baydoun L, et al. Asymmetrical endothelial cell migration from in vitro Quarter-Descemet membrane endothelial keratoplasty grafts. Acta Ophthalmol. 2018;96(8):828–833. doi: 10.1111/aos.13841 30171674

5. Lie JT, Lam FC, Groeneveld-van Beek EA, van der Wees J, Melles GR. Graft preparation for hemi-Descemet membrane endothelial keratoplasty (hemi-DMEK). Br J Ophthalmol. 2016;100(3):420–424. doi: 10.1136/bjophthalmol-2015-307335 26508780

6. He Z, Campolmi N, Gain P, Ha Thi BM, Dumollard JM, Duband S, et al. Revisited microanatomy of the corneal endothelial periphery: new evidence for continuous centripetal migration of endothelial cells in humans. Stem Cells. 2012;30911):2523–2534. doi: 10.1002/stem.1212 22949402

7. Scarpa E, Mayor R. Collective cell migration in development. J Cell Biol. 2016;212(2):143–155. doi: 10.1083/jcb.201508047 26783298

8. Curchoe CL, Maurer J, McKeown SJ, Cattarossi G, Cimadamore F, Nilbratt M, et al. Early acquisition of neural crest competence during hESCs neuralization. PLoS One. 2010;5(11):e13890. doi: 10.1371/journal.pone.0013890 21085480

9. Adzic M, Nedeljkovic N. Unveiling the role of Ecto-5'-Nucleotidase/CD73 in astrocyte migration by using pharmacological tools. Front Pharmacol. 2018;9:153. doi: 10.3389/fphar.2018.00153 29545748

10. Du D, Xu F, Yu L, Zhang C, Lu X, Yuan H, et al. The tight junction protein, occludin, regulates the directional migration of epithelial cells. Dev Cell. 2010;18(1):52–63. doi: 10.1016/j.devcel.2009.12.008 20152177

11. Schwab A, Fabian A, Hanley PJ, Stock C. Role of ion channels and transporters in cell migration. Physiol Rev. 2012;92(4):1865–1913. doi: 10.1152/physrev.00018.2011 23073633

12. Trepat X, Chen Z, Jacobson K. Cell migration. Compr Physiol. 2012;2(4):2369–2392. doi: 10.1002/cphy.c110012 23720251

13. Liu Y, Sun H, Guo P, Hu M, Zhang Y, Tighe S, et al. Characterization and prospective of human corneal endothelial progenitors. Int J Med Sci. 2017;14(8):705–710. doi: 10.7150/ijms.19018 28824304


Článek vyšel v časopise

PLOS One


2019 Číslo 11