Construction of a tissue-engineered human corneal endothelium and its transplantation in rabbit models

Abstract

Human corneal endothelium (HCE) is vital in maintaining corneal transparency and thickness, and damages to it may result in endotheliopathy and even corneal blindness, which can only be cured by keratoplasty. Due to the shortage of donor corneas, tissueengineered HCE (TE-HCE) has become an equivalent to HCE. This study constructs a high endothelial cell density (ECD) TE-HCE and evaluates its functions by corneal transplantation in rabbits. A TE-HCE was constructed by culturing DiI-labeled nontransfected HCE cells on modified denuded amniotic membrane (mdAM) using collagen IV, fibronectin, and laminin, and rabbit corneas were characterized both in vivo and ex vivo after TE-HCE transplantation. Our results indicated that the constructed TE-HCE with a high ECD of 3611 ± 56.66 cells/mm2 had similar morphology and structure to a native HCE. In vivo postsurgery detections showed that the TE-HCE-transplanted cornea gained transparency and recovered its thickness gradually during a monitoring period of 358 days, while the mdAM-transplanted cornea remained opaque and edematous. Ex vivo examinations revealed that a native-like corneal endothelium with an ECD of 2703 ± 70.37 cells/mm2 was reconstructed by the transplanted TE-HCE. Our findings suggested that the TE-HCE might be used as a HCE equivalent and has promising applications in therapy of corneal endotheliopathy.

Natural Biomaterials for Corneal Tissue Engineering, Repair, and Regeneration

“…Rabbits transplanted with the modified AM without cells were used as a control group. Those eyes transplanted with the tissue engineered corneal endothelium cleared and remained transparent over the almost yearlong follow‐up period, whereas those with just the modified AM remained opaque over the entire period . These results show that even though the AM itself was not transparent, when endothelial cells were present, the cornea was able to clear and remain transparent.…”

“…There are two reports in recent literature of groups working to engineer a corneal endothelium using the AM. In one study, after removing the epithelial layer from the AM, the remaining portion was coated with a mixture of fibronectin and collagen IV and used as a substrate to engineer a high density (≈3600 cells mm −2 ) corneal endothelial cell monolayer . The endothelium was then transplanted into rabbit corneas by removing a full thickness corneal button, stripping the existing endothelium and Descemet's membrane from the button and replacing it with the tissue engineered endothelium before re‐transplantation into the rabbit eye .…”

Corneal endothelium tissue engineering: An evolution of signaling molecules, cells, and scaffolds toward 3D bioprinting and cell sheets

“…Various types of synthetic and natural scaffolds have been investigated for the CETE with a set of advantages and disadvantages. Nonetheless, only a limited number of expected clinical outcomes have been reported (Cen & Feng, 2018; Kennedy et al, 2019; D. K. Kim et al, 2016; Kimoto et al, 2014; J. Zhao et al, 2016).…”

“…To overcome these disadvantages, the hAM properties are improved by functionalization, chemical crosslinking, and coating with further materials (Palchesko et al, 2018). J. Zhao et al (2016) modified the denuded amniotic membrane by coating with collagen IV, fibronectin, and laminin to improve cell adhesion. HCECs are expanded with high density and natural morphology under these modifications, which reduce the corneal opacity and edema (J. Zhao et al, 2016).…”

“…J. Zhao et al (2016) modified the denuded amniotic membrane by coating with collagen IV, fibronectin, and laminin to improve cell adhesion. HCECs are expanded with high density and natural morphology under these modifications, which reduce the corneal opacity and edema (J. Zhao et al, 2016).…”