title: Y-27632, a ROCK Inhibitor, Promoted Limbal Epithelial Cell Proliferation and Corneal Wound Healing.
abstract: Transplantation of ex vivo cultured limbal epithelial cells is proven effective in restoring limbal stem cell deficiency. The present study aimed to investigate the promoting effect of Y-27632 on limbal epithelial cell proliferation. Limbal explants isolated from human donor eyes were expanded three weeks on culture dishes and outgrowth of epithelial cells was subsequently subcultured for in vitro experiments. In the presence of Y-27632, the ex vivo limbal outgrowth was accelerated, particularly the cells with epithelial cell-like morphology. Y-27632 dose-dependently promoted the proliferation of in vitro cultured human limbal epithelial cells as examined by phase contrast microscopy and luminescent cell-viability assay 30 hours after the treatment. The colony forming efficacy determined 7 days after the treatment was enhanced by Y-27632 also in a dose-dependent manner. The number of p63- or Ki67-positive cells was dose-dependently increased in Y-27632-treated cultures as detected by immunofluorescent staining and western blotanalysis. Cell cycle analysis by flow cytometric method revealed an increase in S-phase proliferating cells. The epithelial woundclosure rate was shown to be faster in experimental group received topical treatment withY-27632 than the sham control using a rat corneal wounding model. These resultsdemonstrate that Y-27632 can promote both the ex vivo and in vitro proliferation oflimbal epithelial cell proliferation. The in vivo enhanced epithelial wound healingfurther implies that the Y-27632 may act as a new strategy for treating limbal stem cell deficiency.
authors.affiliation: Department of Ophthalmology, Chang Gung Memorial Hospital, Keelung, Taiwan.|Department of Chinese Medicine, College of Medicine, Chang Gung University, Kwei-shan, Taoyuan, Taiwan.
authors.forename: Chi-Chin
authors.initials: CC
authors.lastname: Sun
authors.affiliation: Department of Ophthalmology, Chang Gung Memorial Hospital, Keelung, Taiwan.
authors.forename: Hsiao-Ting
authors.initials: HT
authors.lastname: Chiu
authors.affiliation: Department of Ophthalmology, Chang Gung Memorial Hospital, Keelung, Taiwan.
authors.forename: Yi-Fang
authors.initials: YF
authors.lastname: Lin
authors.affiliation: Department of Ophthalmology, Chang Gung Memorial Hospital, Keelung, Taiwan.
authors.forename: Kuo-Ying
authors.initials: KY
authors.lastname: Lee
authors.affiliation: Graduate Institute of Clinical Medical Sciences, College of Medicine, Chang Gung University, Kwei-shan, Taoyuan, Taiwan.
authors.forename: Jong-Hwei Su
authors.initials: JH
authors.lastname: Pang
mesh_terms: D000577:Amides / Q000494:pharmacology*; D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D015415:Biomarkers / Q000378:metabolism; D002453:Cell Cycle / Q000187:drug effects; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D003114:Colony-Forming Units Assay; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism / Q000473:pathology*; D006801:Humans; D053551:Keratin-12 / Q000378:metabolism; D019394:Ki-67 Antigen / Q000378:metabolism; D016850:Limbus Corneae / Q000473:pathology*; D047428:Protein Kinase Inhibitors / Q000494:pharmacology*; D011725:Pyridines / Q000494:pharmacology*; D017207:Rats, Sprague-Dawley; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism; D014945:Wound Healing / Q000187:drug effects*; D054460:rho-Associated Kinases / Q000037:antagonists & inhibitors* / Q000378:metabolism
chemical_list: D000577:Amides; D015415:Biomarkers; D053551:Keratin-12; D019394:Ki-67 Antigen; D047428:Protein Kinase Inhibitors; D011725:Pyridines; D016159:Tumor Suppressor Protein p53; C108830:Y 27632; D054460:rho-Associated Kinases