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Development of a tissue engineered corneal endothelium

Tsai, Meng-Chen; (2023) Development of a tissue engineered corneal endothelium. Doctoral thesis (Ph.D), UCL (University College London).

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Normal corneal endothelial cells are pivotal in maintaining the cornea's transparency. Limited in vivo cell proliferative capacity results in a corneal endothelium that cannot regenerate to compensate for natural or acquired cell density loss. Therefore, corneal opacity due to endothelial cell failure is a leading cause of blindness. Transplantation of corneal tissue is the standard treatment for irreversible corneal endothelium decompensation by replacing the malfunctioning corneal endothelial cells. However, this surgery depends on limited supplied donor tissue. Thus, developing suitable alternatives for donor grafting material through a tissue engineering approach is needed. Here, we proposed using a plastic compressed collagen 3D matrix called Real Architecture For 3D Tissues (RAFT) as a scaffold for cell culture and a novel tissue equivalent for corneal endothelial transplantation. This study aimed to establish the isolation and expansion of primary porcine corneal endothelial cells (PCECs), to optimise the biomechanical properties of RAFT to construct a suitable environment for endothelial cell expansion, and to establish an ex vivo anterior chamber infusion model. Finally, the cellseeded-RAFT graft was transplanted onto a human cornea and maintained in the ex vivo organ culture model. These findings suggested that PCECs can be successfully expanded in vitro as a reliable primary cell source for endothelial research. RAFT was a suitable material for corneal endothelial tissue engineering as it shared similar mechanical properties to native corneal tissue. PCECs formed a high-density monolayer on RAFT expressing endothelial cell markers, ZO-1 and Na/K ATPase. An organ culture model was successfully established, in which cornea tissue can be maintained, and the endothelial cell function can be evaluated by measuring the thickness changes to the cornea. More importantly, the cell-seeded-RAFT transplantation successfully restored corneal endothelium function, drawing the thickness of endotheliumwounded cornea back to normal in two weeks of the ex vivo organ culture model.

Type: Thesis (Doctoral)
Qualification: Ph.D
Title: Development of a tissue engineered corneal endothelium
Language: English
Additional information: Copyright © The Author 2023. Original content in this thesis is licensed under the terms of the Creative Commons Attribution-NonCommercial 4.0 International (CC BY-NC 4.0) Licence (https://creativecommons.org/licenses/by-nc/4.0/). Any third-party copyright material present remains the property of its respective owner(s) and is licensed under its existing terms. Access may initially be restricted at the author’s request.
UCL classification: UCL
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Brain Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Brain Sciences > Institute of Ophthalmology
URI: https://discovery.ucl.ac.uk/id/eprint/10184335
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