Asymmetrical endothelial cell migration from in vitro Quarter-Descemet membrane endothelial keratoplasty grafts

Alina Miron; Daniele Spinozzi; Marieke Bruinsma; Jessica T. Lie; Rénuka S. Birbal; Lamis Baydoun; Silke Oellerich; Gerrit R.J. Melles

doi:10.1111/aos.13841

Asymmetrical endothelial cell migration from in vitro Quarter-Descemet membrane endothelial keratoplasty grafts

Alina Miron, Daniele Spinozzi, Marieke Bruinsma, Jessica T. Lie, Rénuka S. Birbal, Lamis Baydoun, Silke Oellerich, Gerrit R.J. Melles

Ophthalmology - Eye surgery

Research output: Contribution to journal › Article › peer-review

14 Citations (Scopus)

Abstract

Purpose: To investigate in vitro central and peripheral corneal endothelial cell (EC) migration from Quarter–Descemet membrane endothelial keratoplasty (Quarter-DMEK) grafts. Methods: Quarter-DMEK grafts were obtained from 10 corneas ineligible for transplantation but with intact and viable ECs. Ten Quarter-DMEK grafts were ‘sandwiched’ between two glass slides and cultured over 1 week in a humidified atmosphere at 37 °C and 5% CO₂. Cell migration was evaluated by light microscopy at standardized time intervals. In addition, immunohistochemistry analyses were performed to assess the detailed structural organization of ECs in the corneal centre and far periphery. Results: Endothelial cell (EC) migration occurred from the radial cut graft edges, but not from the far peripheral area. Cell migration followed three different migration patterns: (1) individual cell migration, (2) uncoordinated cell migration of cell clusters and (3) collective migration in which ECs moved as a sheet. Immunostaining showed the presence of ECs up to the far periphery but with different expression patterns of phenotypical markers ZO-1, Na⁺/K⁺ -ATPase and vimentin compared to central ECs. Conclusion: In vitro EC migration from Quarter-DMEK grafts occurs along the radial cut edges with a decrease in migration activity towards the corneal far periphery. No migration occurred along the outer peripheral corneal edge possibly due to a different anatomical matrix in the far periphery. Hence, ECs from the far periphery may not contribute to corneal clearance of the adjacent bare area after Quarter-DMEK surgery, but these cells may constitute a valuable cellular reserve on the graft.

Original language	English
Pages (from-to)	828-833
Number of pages	6
Journal	Acta Ophthalmologica
Volume	96
Issue number	8
DOIs	https://doi.org/10.1111/aos.13841
Publication status	Published - Dec 2018

Bibliographical note

Funding Information:
Key words: corneal clearance – Descemet membrane endothelial keratoplasty – endothelial cell migration – immunohistochemistry – peripheral endothelial cells – Quarter-DMEK Part of the project leading to this study has received funding from the European Union’s Horizon 2020 Research and Innovation Programme under grant agreement number 667400 (ARREST BLINDNESS Consortium).

Publisher Copyright:
© 2018 The Authors. Acta Ophthalmologica published by John Wiley & Sons Ltd on behalf of Acta Ophthalmologica Scandinavica Foundation.

Keywords

corneal clearance
Descemet membrane endothelial keratoplasty
endothelial cell migration
immunohistochemistry
peripheral endothelial cells
Quarter-DMEK

Access to Document

10.1111/aos.13841

Cite this

@article{bff741eca0ea4efb8f88e6e1684b633d,

title = "Asymmetrical endothelial cell migration from in vitro Quarter-Descemet membrane endothelial keratoplasty grafts",

abstract = "Purpose: To investigate in vitro central and peripheral corneal endothelial cell (EC) migration from Quarter–Descemet membrane endothelial keratoplasty (Quarter-DMEK) grafts. Methods: Quarter-DMEK grafts were obtained from 10 corneas ineligible for transplantation but with intact and viable ECs. Ten Quarter-DMEK grafts were {\textquoteleft}sandwiched{\textquoteright} between two glass slides and cultured over 1 week in a humidified atmosphere at 37 °C and 5% CO2. Cell migration was evaluated by light microscopy at standardized time intervals. In addition, immunohistochemistry analyses were performed to assess the detailed structural organization of ECs in the corneal centre and far periphery. Results: Endothelial cell (EC) migration occurred from the radial cut graft edges, but not from the far peripheral area. Cell migration followed three different migration patterns: (1) individual cell migration, (2) uncoordinated cell migration of cell clusters and (3) collective migration in which ECs moved as a sheet. Immunostaining showed the presence of ECs up to the far periphery but with different expression patterns of phenotypical markers ZO-1, Na+/K+ -ATPase and vimentin compared to central ECs. Conclusion: In vitro EC migration from Quarter-DMEK grafts occurs along the radial cut edges with a decrease in migration activity towards the corneal far periphery. No migration occurred along the outer peripheral corneal edge possibly due to a different anatomical matrix in the far periphery. Hence, ECs from the far periphery may not contribute to corneal clearance of the adjacent bare area after Quarter-DMEK surgery, but these cells may constitute a valuable cellular reserve on the graft.",

keywords = "corneal clearance, Descemet membrane endothelial keratoplasty, endothelial cell migration, immunohistochemistry, peripheral endothelial cells, Quarter-DMEK",

author = "Alina Miron and Daniele Spinozzi and Marieke Bruinsma and Lie, {Jessica T.} and Birbal, {R{\'e}nuka S.} and Lamis Baydoun and Silke Oellerich and Melles, {Gerrit R.J.}",

note = "Funding Information: Key words: corneal clearance – Descemet membrane endothelial keratoplasty – endothelial cell migration – immunohistochemistry – peripheral endothelial cells – Quarter-DMEK Part of the project leading to this study has received funding from the European Union{\textquoteright}s Horizon 2020 Research and Innovation Programme under grant agreement number 667400 (ARREST BLINDNESS Consortium). Publisher Copyright: {\textcopyright} 2018 The Authors. Acta Ophthalmologica published by John Wiley & Sons Ltd on behalf of Acta Ophthalmologica Scandinavica Foundation.",

year = "2018",

month = dec,

doi = "10.1111/aos.13841",

language = "English",

volume = "96",

pages = "828--833",

journal = "Acta Ophthalmologica",

issn = "1755-375X",

publisher = "Wiley-Blackwell",

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}

Asymmetrical endothelial cell migration from in vitro Quarter-Descemet membrane endothelial keratoplasty grafts. / Miron, Alina; Spinozzi, Daniele; Bruinsma, Marieke; Lie, Jessica T.; Birbal, Rénuka S.; Baydoun, Lamis; Oellerich, Silke; Melles, Gerrit R.J.

In: Acta Ophthalmologica, Vol. 96, No. 8, 12.2018, p. 828-833.

Research output: Contribution to journal › Article › peer-review

TY - JOUR

T1 - Asymmetrical endothelial cell migration from in vitro Quarter-Descemet membrane endothelial keratoplasty grafts

AU - Miron, Alina

AU - Spinozzi, Daniele

AU - Bruinsma, Marieke

AU - Lie, Jessica T.

AU - Birbal, Rénuka S.

AU - Baydoun, Lamis

AU - Oellerich, Silke

AU - Melles, Gerrit R.J.

N1 - Funding Information: Key words: corneal clearance – Descemet membrane endothelial keratoplasty – endothelial cell migration – immunohistochemistry – peripheral endothelial cells – Quarter-DMEK Part of the project leading to this study has received funding from the European Union’s Horizon 2020 Research and Innovation Programme under grant agreement number 667400 (ARREST BLINDNESS Consortium). Publisher Copyright: © 2018 The Authors. Acta Ophthalmologica published by John Wiley & Sons Ltd on behalf of Acta Ophthalmologica Scandinavica Foundation.

PY - 2018/12

Y1 - 2018/12

N2 - Purpose: To investigate in vitro central and peripheral corneal endothelial cell (EC) migration from Quarter–Descemet membrane endothelial keratoplasty (Quarter-DMEK) grafts. Methods: Quarter-DMEK grafts were obtained from 10 corneas ineligible for transplantation but with intact and viable ECs. Ten Quarter-DMEK grafts were ‘sandwiched’ between two glass slides and cultured over 1 week in a humidified atmosphere at 37 °C and 5% CO2. Cell migration was evaluated by light microscopy at standardized time intervals. In addition, immunohistochemistry analyses were performed to assess the detailed structural organization of ECs in the corneal centre and far periphery. Results: Endothelial cell (EC) migration occurred from the radial cut graft edges, but not from the far peripheral area. Cell migration followed three different migration patterns: (1) individual cell migration, (2) uncoordinated cell migration of cell clusters and (3) collective migration in which ECs moved as a sheet. Immunostaining showed the presence of ECs up to the far periphery but with different expression patterns of phenotypical markers ZO-1, Na+/K+ -ATPase and vimentin compared to central ECs. Conclusion: In vitro EC migration from Quarter-DMEK grafts occurs along the radial cut edges with a decrease in migration activity towards the corneal far periphery. No migration occurred along the outer peripheral corneal edge possibly due to a different anatomical matrix in the far periphery. Hence, ECs from the far periphery may not contribute to corneal clearance of the adjacent bare area after Quarter-DMEK surgery, but these cells may constitute a valuable cellular reserve on the graft.

AB - Purpose: To investigate in vitro central and peripheral corneal endothelial cell (EC) migration from Quarter–Descemet membrane endothelial keratoplasty (Quarter-DMEK) grafts. Methods: Quarter-DMEK grafts were obtained from 10 corneas ineligible for transplantation but with intact and viable ECs. Ten Quarter-DMEK grafts were ‘sandwiched’ between two glass slides and cultured over 1 week in a humidified atmosphere at 37 °C and 5% CO2. Cell migration was evaluated by light microscopy at standardized time intervals. In addition, immunohistochemistry analyses were performed to assess the detailed structural organization of ECs in the corneal centre and far periphery. Results: Endothelial cell (EC) migration occurred from the radial cut graft edges, but not from the far peripheral area. Cell migration followed three different migration patterns: (1) individual cell migration, (2) uncoordinated cell migration of cell clusters and (3) collective migration in which ECs moved as a sheet. Immunostaining showed the presence of ECs up to the far periphery but with different expression patterns of phenotypical markers ZO-1, Na+/K+ -ATPase and vimentin compared to central ECs. Conclusion: In vitro EC migration from Quarter-DMEK grafts occurs along the radial cut edges with a decrease in migration activity towards the corneal far periphery. No migration occurred along the outer peripheral corneal edge possibly due to a different anatomical matrix in the far periphery. Hence, ECs from the far periphery may not contribute to corneal clearance of the adjacent bare area after Quarter-DMEK surgery, but these cells may constitute a valuable cellular reserve on the graft.

KW - corneal clearance

KW - Descemet membrane endothelial keratoplasty

KW - endothelial cell migration

KW - immunohistochemistry

KW - peripheral endothelial cells

KW - Quarter-DMEK

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U2 - 10.1111/aos.13841

DO - 10.1111/aos.13841

M3 - Article

C2 - 30171674

AN - SCOPUS:85052828169

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EP - 833

JO - Acta Ophthalmologica

JF - Acta Ophthalmologica

SN - 1755-375X

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