Effect of Rho-kinase Inhibitor, Y27632, on Porcine Corneal Endothelial Cell Culture, Inflammation and Immune Regulation

Abstract

Purpose: To investigate the effect of the Rho-kinase inhibitor, Y27632, on pig corneal endothelial cell (pCEC) culture, and on inflammation and immune regulation of the responses of human cells to pCECs.

Methods: pCECs were cultured with/without Y27632 to assess cell proliferation and in vitro wound healing assay. The level of MCP-1 and VEGF in pCECs stimulated with human TNF-α were measured. Proliferation of human PBMCs stimulated with pCECs, and cytokine production in human T cells, and monocyte migration after stimulation were investigated.

Results: Y27632 promoted pCEC proliferation, prevented pCEC death, and enhanced in vitro wound healing. After stimulation, there were significantly lower levels of MCP-1 and VEGF measured in pCECs cultured with Y27632, and significantly reduced human PBMC proliferation, cytokine production, and monocyte migration.

Conclusions: The application of the Rho-kinase inhibitor will be beneficial when culturing pCECs, and may provide a novel therapy to reduce inflammation after corneal xenotransplantation.

Keywords:

• Corneal endothelial cell
• immune response
• inflammation
• pig
• Rho-kinase
• xenotransplantation

Author contributions

WL participated in the performance of the research, interpretation of the results, and in writing of the manuscript. YM, CL and MZ participated in the performance of the research, and in review of the manuscript. DKCC participated in research design, in interpretation of the results, and in writing of the manuscript. HH participated in research design, in the performance of the research, interpretation of the results, and in writing of the manuscript.

Declaration of interest

The authors report no conflicts of interest. The authors alone are responsible for the content and writing of the paper.

Funding

Research on xenotransplantation at the University of Pittsburgh is funded in part by NIH Grant Numbers: 1RO3A1096296-01 (HH), IU19A1090959-01 (DKCC), U01A1066331 (DKCC), and 5P01 HL107152-02 (DKCC), by an Ocular Tissue Engineering and Regenerative Ophthalmology (OTERO) Postdoctoral Fellowship (WL), and by Sponsored Research Agreements between the University of Pittsburgh and Revivicor, Blacksburg, VA.