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Potential Functional Restoration of Corneal Endothelial Cells in Fuchs Endothelial Corneal Dystrophy by ROCK Inhibitor (Ripasudil)
- Source :
- American Journal of Ophthalmology. 224:185-199
- Publication Year :
- 2021
- Publisher :
- Elsevier BV, 2021.
-
Abstract
- Purpose Rho-associated kinase (ROCK) inhibitors have been successfully used as a rescue strategy in eyes that failed to clear after descemetorhexis without endothelial graft for treatment of Fuchs endothelial corneal dystrophy (FECD). The functional mechanisms by which ROCK inhibitors modulate corneal endothelial cell regeneration in FECD patients have, however, not been clarified. Here, we analyzed the effect of the ROCK inhibitor ripasudil on corneal endothelial cells of FECD patients and normal donors using ex vivo tissue and in vitro cellular models. Design Experimental study: laboratory investigation. Methods This institutional study used endothelial cell–Descemet membrane lamellae from FECD patients (n = 450) undergoing Descemet membrane endothelial keratoplasty (FECD ex vivo model), normal research-grade donor corneas (n = 30) after scraping off central endothelial cells (ex vivo wound healing model), normal donor corneas (n = 20) without endothelial injury, and immortalized cell lines (n = 3) generated from FECD patients (FECD in vitro model). Descemet membrane lamellae were dissected into halves and incubated for 24-72 hours in storage medium with or without a single dose of 30 μM ripasudil. The effects of ripasudil on expression of genes and proteins related to endothelial cell proliferation, migration, functionality, and endothelial-to-mesenchymal transition were analyzed and complemented by functional assays on FECD cell lines. Results A single dose of ripasudil induced significant upregulation of genes and proteins related to cell cycle progression, cell-matrix adhesion and migration, as well as endothelial barrier and pump function up to 72 hours, whereas classical markers of endothelial-to-mesenchymal transition were downregulated in both FECD and normal specimens compared to unstimulated controls ex vivo. In addition to stimulation of proliferation and migration, ripasudil-induced changes in expression of functional signature genes could be also verified in FECD cell lines in vitro. Conclusions These data support the concept that inhibition of ROCK signaling represents a potent tool in regenerative therapies in FECD patients through reactivation of cell proliferation and migration as well as restoration of endothelial pump and barrier function without inducing adverse phenotypic changes.
- Subjects :
- Male
Endothelium
Cell Cycle Proteins
Cell-Matrix Junctions
03 medical and health sciences
chemistry.chemical_compound
0302 clinical medicine
Cell Movement
medicine
Humans
Cells, Cultured
Barrier function
Aged
Cell Proliferation
030304 developmental biology
Sulfonamides
rho-Associated Kinases
0303 health sciences
Dose-Response Relationship, Drug
Chemistry
Cell growth
Cell Cycle
Endothelium, Corneal
Fuchs' Endothelial Dystrophy
Middle Aged
Isoquinolines
Endothelial stem cell
Ophthalmology
medicine.anatomical_structure
Descemet Stripping Endothelial Keratoplasty
030221 ophthalmology & optometry
Cancer research
Female
Ripasudil
Wound healing
Immortalised cell line
Subjects
Details
- ISSN :
- 00029394
- Volume :
- 224
- Database :
- OpenAIRE
- Journal :
- American Journal of Ophthalmology
- Accession number :
- edsair.doi.dedup.....eada6a9c91b1990c9b64f32d5e5d3309