ROCK Inhibitor-Induced Promotion of Retinal Pigment Epithelial Cell Motility during Wound Healing
Purpose. No standard therapy for RPE tear, a complication of neovascular age-related macular degeneration, exists even though RPE tears cause severe vision loss, and promotion of cell proliferation and/or migration could be a candidate RPE tear therapy. The aim of this study is to evaluate the effec...
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| Format: | Article |
| Language: | English |
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Wiley
2019-01-01
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| Series: | Journal of Ophthalmology |
| Online Access: | http://dx.doi.org/10.1155/2019/9428738 |
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| _version_ | 1832550970407518208 |
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| author | Hiroyuki Kamao Atsushi Miki Junichi Kiryu |
| author_facet | Hiroyuki Kamao Atsushi Miki Junichi Kiryu |
| author_sort | Hiroyuki Kamao |
| collection | DOAJ |
| description | Purpose. No standard therapy for RPE tear, a complication of neovascular age-related macular degeneration, exists even though RPE tears cause severe vision loss, and promotion of cell proliferation and/or migration could be a candidate RPE tear therapy. The aim of this study is to evaluate the effect of Rho-associated coiled-coil containing kinase (ROCK) inhibitor Y27632 on retinal pigment epithelial (RPE) cell motility during wound healing. Methods. Human RPE cells were cultured in media with and without 10 μM Y27632. A luminescent cell viability assay and vinculin immunocytochemistry were used to test the Y27632 effect on RPE cell adhesion. The mean size of vinculin puncta was quantified from immunofluorescence images. RPE cell motility during wound healing was evaluated using time-lapse imaging and measuring cell migration distances and cell coverage rate in wound fields. Results. The number of adhered RPE and mean size of vinculin puncta were, respectively, 20519 cells and 3.65 μm2 under nontreatment and 23569 cells and 0.66 μm2 under Y27632 treatment. Cell migration distance and cell coverage percentage for untreated and Y27632-treated cells were 98.9 and 59.4% and 203.4 and 92.5%, respectively. Conclusions. Inhibition of ROCK signaling by using 10 μM Y27632 promoted RPE cell motility during wound healing by reducing RPE cell adhesion strength. |
| format | Article |
| id | doaj-art-4ebe876740a4400ba1899383a8253611 |
| institution | Kabale University |
| issn | 2090-004X 2090-0058 |
| language | English |
| publishDate | 2019-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Journal of Ophthalmology |
| spelling | doaj-art-4ebe876740a4400ba1899383a82536112025-02-03T06:05:21ZengWileyJournal of Ophthalmology2090-004X2090-00582019-01-01201910.1155/2019/94287389428738ROCK Inhibitor-Induced Promotion of Retinal Pigment Epithelial Cell Motility during Wound HealingHiroyuki Kamao0Atsushi Miki1Junichi Kiryu2Department of Ophthalmology, Kawasaki Medical School, 577 Matsushima, Kurashiki, Okayama 701-0114, JapanDepartment of Ophthalmology, Kawasaki Medical School, 577 Matsushima, Kurashiki, Okayama 701-0114, JapanDepartment of Ophthalmology, Kawasaki Medical School, 577 Matsushima, Kurashiki, Okayama 701-0114, JapanPurpose. No standard therapy for RPE tear, a complication of neovascular age-related macular degeneration, exists even though RPE tears cause severe vision loss, and promotion of cell proliferation and/or migration could be a candidate RPE tear therapy. The aim of this study is to evaluate the effect of Rho-associated coiled-coil containing kinase (ROCK) inhibitor Y27632 on retinal pigment epithelial (RPE) cell motility during wound healing. Methods. Human RPE cells were cultured in media with and without 10 μM Y27632. A luminescent cell viability assay and vinculin immunocytochemistry were used to test the Y27632 effect on RPE cell adhesion. The mean size of vinculin puncta was quantified from immunofluorescence images. RPE cell motility during wound healing was evaluated using time-lapse imaging and measuring cell migration distances and cell coverage rate in wound fields. Results. The number of adhered RPE and mean size of vinculin puncta were, respectively, 20519 cells and 3.65 μm2 under nontreatment and 23569 cells and 0.66 μm2 under Y27632 treatment. Cell migration distance and cell coverage percentage for untreated and Y27632-treated cells were 98.9 and 59.4% and 203.4 and 92.5%, respectively. Conclusions. Inhibition of ROCK signaling by using 10 μM Y27632 promoted RPE cell motility during wound healing by reducing RPE cell adhesion strength.http://dx.doi.org/10.1155/2019/9428738 |
| spellingShingle | Hiroyuki Kamao Atsushi Miki Junichi Kiryu ROCK Inhibitor-Induced Promotion of Retinal Pigment Epithelial Cell Motility during Wound Healing Journal of Ophthalmology |
| title | ROCK Inhibitor-Induced Promotion of Retinal Pigment Epithelial Cell Motility during Wound Healing |
| title_full | ROCK Inhibitor-Induced Promotion of Retinal Pigment Epithelial Cell Motility during Wound Healing |
| title_fullStr | ROCK Inhibitor-Induced Promotion of Retinal Pigment Epithelial Cell Motility during Wound Healing |
| title_full_unstemmed | ROCK Inhibitor-Induced Promotion of Retinal Pigment Epithelial Cell Motility during Wound Healing |
| title_short | ROCK Inhibitor-Induced Promotion of Retinal Pigment Epithelial Cell Motility during Wound Healing |
| title_sort | rock inhibitor induced promotion of retinal pigment epithelial cell motility during wound healing |
| url | http://dx.doi.org/10.1155/2019/9428738 |
| work_keys_str_mv | AT hiroyukikamao rockinhibitorinducedpromotionofretinalpigmentepithelialcellmotilityduringwoundhealing AT atsushimiki rockinhibitorinducedpromotionofretinalpigmentepithelialcellmotilityduringwoundhealing AT junichikiryu rockinhibitorinducedpromotionofretinalpigmentepithelialcellmotilityduringwoundhealing |