Predicative Factors for Corneal Endothelial Cell Migration

Abstract

PURPOSE.To characterize the effects of Descemet's stripping, Rho-associated protein kinase inhibitor Y-27632, and donor age on endothelial migration in human corneas maintained in ex vivo culture.METHODS. Twenty-eight cadaveric human corneas underwent ex vivo culture in either standard or Y-27632-supplemented culture medium for 14 days. The posterior surface of each cornea was manipulated to create two types of wounds: scratched wound-corneal endothelial cells (CECs) were denuded from the Descemet's membrane (DM) to leave behind a bare but intact DM; and peeled wound-both the DM and overlying CECs were stripped to leave behind bare corneal stroma. Endothelial migration was assessed via Trypan blue staining. Morphologic traits of CECs were assessed via Alizarin red microscopy and scanning electron microscopy.RESULTS. The CECs migrated preferentially over scratched wounds compared with peeled wounds. Y-27632 supplementation accelerated endothelial migration over scratched wounds. Endothelial migration decreased with advanced donor age for both wound types, regardless of exposure to Y-27632. Y-27632 supplementation resulted in a less rapid decline in endothelial migration for donors older than 50 years of age for scratched surfaces. Greater cell density and hexagonality was observed over scratched wounds compared with peeled wounds, regardless of Y-27632 supplementation.CONCLUSIONS. The presence of an intact DM, Y-27632 supplementation, and young donor age are factors that promote endothelial migration in an ex vivo human cornea culture model. The negative effect of age on endothelial migration can be mitigated by the presence of an intact DM and Y-27632 supplementation.

ROCK inhibitors in ocular disease

“…The first two patients had spontaneous detachment of Descemet's membrane during surgery, which could have further impaired the healing of the wound. However, in contrast with the in vitro findings [32], in these cases the corneal endothelium was reconstructed on bare corneal stroma and the corneal clarity recovered. When the first patient was referred to the cornea clinic she was only able to count fingers with her right eye.…”

“…In a rabbit organ culture model (treated with 10 µM Y27632 for 48 hours) there was no difference between the presence and absence of Descemet's membrane, and the ROCK inhibitor increased CEC proliferation to approximately the same level in both samples [31]. In contrast, in human CEC culture (treated with 10 µM Y27632 for 14 days), results suggest that the presence of an intact Descemet's membrane does affect corneal endothelial wound healing [32]. Soh et al [32] observed that the maximum endothelial recovery in culture without Descemet's membrane was around 20 %, whereas with intact Descemet's membrane recovery was approximately 80 % due to increased endothelial migration.…”

“…In contrast, in human CEC culture (treated with 10 µM Y27632 for 14 days), results suggest that the presence of an intact Descemet's membrane does affect corneal endothelial wound healing [32]. Soh et al [32] observed that the maximum endothelial recovery in culture without Descemet's membrane was around 20 %, whereas with intact Descemet's membrane recovery was approximately 80 % due to increased endothelial migration. These differing results may be due to species differences or differences in the experimental models.…”

“…It can detach spontaneously, or be removed during cataract surgery, or other injuries. The importance of an intact Descemet's membrane in corneal endothelial recovery has been investigated in in vitro studies [31,32]. In a rabbit organ culture model (treated with 10 µM Y27632 for 48 hours) there was no difference between the presence and absence of Descemet's membrane, and the ROCK inhibitor increased CEC proliferation to approximately the same level in both samples [31].…”

Rho‐kinase inhibitor Y‐27632 facilitates the proliferation, migration and pluripotency of human periodontal ligament stem cells

“…In earlier studies, dental pulp cells (DPCs) migration was increased by inhibition of ROCK , and the findings differed from other studies in which ROCK inhibition reduced cell motility . In our study, Y‐27632 supplementation promoted PDLSCs migration and accelerated cell migration over scratched wounds, which was in accordance with another study that Y‐27632 enhanced endothelial migration in a wound‐healing assay .…”