June 2022
Volume 63, Issue 7
Open Access
ARVO Annual Meeting Abstract  |   June 2022
Novel ROCK Inhibitors on the Regeneration of Corneal Endothelial Cells
Author Affiliations & Notes
  • Young Joo Shin
    Department of Ophthalmology, College of Medicine, Hallym University Medical Center, Hallym University, Seoul, Other, Korea (the Republic of)
  • Yoochang Kim
    pH Pharma Co., LTD., Seoul, Other, Korea (the Republic of)
  • Sammy Chi Sam Lee
    pH Pharma Co., LTD., Seoul, Other, Korea (the Republic of)
  • Footnotes
    Commercial Relationships   Young Joo Shin pH Pharma Co., LTD, Code C (Consultant/Contractor); Yoochang Kim pH Pharma Co., LTD, Code E (Employment); Sammy Lee pH Pharma Co., LTD, Code E (Employment)
  • Footnotes
    Support  None
Investigative Ophthalmology & Visual Science June 2022, Vol.63, 2750 – A0239. doi:
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      Young Joo Shin, Yoochang Kim, Sammy Chi Sam Lee; Novel ROCK Inhibitors on the Regeneration of Corneal Endothelial Cells. Invest. Ophthalmol. Vis. Sci. 2022;63(7):2750 – A0239.

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      © ARVO (1962-2015); The Authors (2016-present)

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Purpose : The cornea may lose transparency and cause blindness when Human corneal endothelial cells (HCEnCs) are damaged or diseased. Replacing diseased cells with a healthy donor endothelium is the only currently available treatment. There is increasing interest in the development of cultured graft substitutes and artificial corneas due to the global shortage of donor corneas. The aim of the study is to investigate whether novel Rho-kinase (ROCK) inhibitors can aid in the cultivation and regeneration of HCEnCs.

Methods : Human CEnCs were cultured and treated with Y-27632 (Y0503, Sigma-Aldrich), sovesudil (also known as PHP-201; pH Pharma), or PHP-0961 (pH Pharma) for 24h. We observed cellular responses, including cell viability, cytotoxicity, proliferation and Ki67 expression with ROCK inhibitors. We also assessed wound healing and cell adhesion assays. Porcine corneas were used ex vivo to evaluate wound healing and regeneration effects of Y-27632, sovesudil, and PHP-0961. We performed live/dead cell assays and immunofluorescence staining (for SOX2, b-catenin, and ZO-1) on porcine corneas treated with ROCK inhibitors.

Results : Cell viability, BrdU proliferation assay, and number of Ki67-positive cells was higher in Y-27632, sovesudil and PHP-0961 treated compared with control (p<0.05). There was no difference in LDH cytotoxicity test between any groups. Cells treated with Y-27632, sovesudil and PHP-0961 showed faster migration, wound healing, and cell adhesion. In the porcine ex vivo experiments, wound healing, the number of live cells, and SOX-positive cells were higher in Y-27632, sovesudil and PHP-0961 treated corneas (p<0.05). Across all experiments, the results of sovesudil and PHP-0961 were equal or superior to the results of the gold standard ROCK inhibitor Y-27632.

Conclusions : Novel ROCK inhibitors have a capacity to regenerate HCEnCs through enhancing the cell proliferation and adhesion between cells.

This abstract was presented at the 2022 ARVO Annual Meeting, held in Denver, CO, May 1-4, 2022, and virtually.


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