ROCK inhibitor enhances adhesion and wound healing of human corneal endothelial cells.
Maintenance of corneal transparency is crucial for vision and depends mainly on the endothelium, a non-proliferative monolayer of cells covering the inner part of the cornea. When endothelial cell density falls below a critical threshold, the barrier and "pump" functions of the endothelium...
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doaj-b7f2d8476f134625b916c44c859d5b172020-11-24T22:06:49ZengPublic Library of Science (PLoS)PLoS ONE1932-62032013-01-0184e6209510.1371/journal.pone.0062095ROCK inhibitor enhances adhesion and wound healing of human corneal endothelial cells.Aurélien PipparelliYvan ArsenijevicGilles ThuretPhilippe GainMichael NicolasFrançois MajoMaintenance of corneal transparency is crucial for vision and depends mainly on the endothelium, a non-proliferative monolayer of cells covering the inner part of the cornea. When endothelial cell density falls below a critical threshold, the barrier and "pump" functions of the endothelium are compromised which results in corneal oedema and loss of visual acuity. The conventional treatment for such severe disorder is corneal graft. Unfortunately, there is a worldwide shortage of donor corneas, necessitating amelioration of tissue survival and storage after harvesting. Recently it was reported that the ROCK inhibitor Y-27632 promotes adhesion, inhibits apoptosis, increases the number of proliferating monkey corneal endothelial cells in vitro and enhance corneal endothelial wound healing both in vitro and in vivo in animal models. Using organ culture human cornea (N = 34), the effect of ROCK inhibitor was evaluated in vitro and ex vivo. Toxicity, corneal endothelial cell density, cell proliferation, apoptosis, cell morphometry, adhesion and wound healing process were evaluated by live/dead assay standard cell counting method, EdU labelling, Ki67, Caspase3, Zo-1 and Actin immunostaining. We demonstrated for the first time in human corneal endothelial cells ex vivo and in vitro, that ROCK inhibitor did not induce any toxicity effect and did not alter cell viability. ROCK inhibitor treatment did not induce human corneal endothelial cells proliferation. However, ROCK inhibitor significantly enhanced adhesion and wound healing. The present study shows that the selective ROCK inhibitor Y-27632 has no effect on human corneal endothelial cells proliferative capacities, but alters cellular behaviours. It induces changes in cell shape, increases cell adhesion and enhances wound healing ex vivo and in vitro. Its absence of toxicity, as demonstrated herein, is relevant for its use in human therapy.http://europepmc.org/articles/PMC3633902?pdf=render |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Aurélien Pipparelli Yvan Arsenijevic Gilles Thuret Philippe Gain Michael Nicolas François Majo |
spellingShingle |
Aurélien Pipparelli Yvan Arsenijevic Gilles Thuret Philippe Gain Michael Nicolas François Majo ROCK inhibitor enhances adhesion and wound healing of human corneal endothelial cells. PLoS ONE |
author_facet |
Aurélien Pipparelli Yvan Arsenijevic Gilles Thuret Philippe Gain Michael Nicolas François Majo |
author_sort |
Aurélien Pipparelli |
title |
ROCK inhibitor enhances adhesion and wound healing of human corneal endothelial cells. |
title_short |
ROCK inhibitor enhances adhesion and wound healing of human corneal endothelial cells. |
title_full |
ROCK inhibitor enhances adhesion and wound healing of human corneal endothelial cells. |
title_fullStr |
ROCK inhibitor enhances adhesion and wound healing of human corneal endothelial cells. |
title_full_unstemmed |
ROCK inhibitor enhances adhesion and wound healing of human corneal endothelial cells. |
title_sort |
rock inhibitor enhances adhesion and wound healing of human corneal endothelial cells. |
publisher |
Public Library of Science (PLoS) |
series |
PLoS ONE |
issn |
1932-6203 |
publishDate |
2013-01-01 |
description |
Maintenance of corneal transparency is crucial for vision and depends mainly on the endothelium, a non-proliferative monolayer of cells covering the inner part of the cornea. When endothelial cell density falls below a critical threshold, the barrier and "pump" functions of the endothelium are compromised which results in corneal oedema and loss of visual acuity. The conventional treatment for such severe disorder is corneal graft. Unfortunately, there is a worldwide shortage of donor corneas, necessitating amelioration of tissue survival and storage after harvesting. Recently it was reported that the ROCK inhibitor Y-27632 promotes adhesion, inhibits apoptosis, increases the number of proliferating monkey corneal endothelial cells in vitro and enhance corneal endothelial wound healing both in vitro and in vivo in animal models. Using organ culture human cornea (N = 34), the effect of ROCK inhibitor was evaluated in vitro and ex vivo. Toxicity, corneal endothelial cell density, cell proliferation, apoptosis, cell morphometry, adhesion and wound healing process were evaluated by live/dead assay standard cell counting method, EdU labelling, Ki67, Caspase3, Zo-1 and Actin immunostaining. We demonstrated for the first time in human corneal endothelial cells ex vivo and in vitro, that ROCK inhibitor did not induce any toxicity effect and did not alter cell viability. ROCK inhibitor treatment did not induce human corneal endothelial cells proliferation. However, ROCK inhibitor significantly enhanced adhesion and wound healing. The present study shows that the selective ROCK inhibitor Y-27632 has no effect on human corneal endothelial cells proliferative capacities, but alters cellular behaviours. It induces changes in cell shape, increases cell adhesion and enhances wound healing ex vivo and in vitro. Its absence of toxicity, as demonstrated herein, is relevant for its use in human therapy. |
url |
http://europepmc.org/articles/PMC3633902?pdf=render |
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