Plastic Compressed Collagen as a Novel Carrier for Expanded Human Corneal Endothelial Cells for Transplantation

Abstract

Current treatments for reversible blindness caused by corneal endothelial cell failure involve replacing the failed endothelium with donor tissue using a one donor-one recipient strategy. Due to the increasing pressure of a worldwide donor cornea shortage there has been considerable interest in developing alternative strategies to treat endothelial disorders using expanded cell replacement therapy. Protocols have been developed which allow successful expansion of endothelial cells in vitro but this approach requires a supporting material that would allow easy transfer of cells to the recipient. We describe the first use of plastic compressed collagen as a highly effective, novel carrier for human corneal endothelial cells. A human corneal endothelial cell line and primary human corneal endothelial cells retained their characteristic cobblestone morphology and expression of tight junction protein ZO-1 and pump protein Na+/K+ ATPase α1 after culture on collagen constructs for up to 14 days. Additionally, ultrastructural analysis suggested a well-integrated endothelial layer with tightly opposed cells and apical microvilli. Plastic compressed collagen is a superior biomaterial in terms of its speed and ease of production and its ability to be manipulated in a clinically relevant manner without breakage. This method provides expanded endothelial cells with a substrate that could be suitable for transplantation allowing one donor cornea to potentially treat multiple patients.

Propagation of Human Corneal Endothelial Cells: A Novel Dual Media Approach

“…These results suggested that propagated hCECs were amenable to the dual media approach presented in this study and could be plated at a higher density to achieve a denser monolayer of CECs with compact cellular pattern. Indeed, when seeded at a higher density onto a compressed collagen-based hydrogel (a potential transplantable corneal graft substitute), a cell density of approximately 1,941 cells per mm 2 was achieved (33).…”

Biomaterials for Regenerative Medicine Approaches for the Anterior Segment of the Eye

“…Compressed Type 1 collagen using the RAFT technology, as described above, has been investigated as a potential substrate for the expansion and delivery of corneal endothelial cells. [45] It was demonstrated that the mechanical properties of the samples were sufficient to be clinically manipulated into an eye using the standard transplant instruments. Furthermore, primary human corneal endothelial cells were demonstrated to attach and grow as a monolayer on the RAFT substrates and showed evidence of tight junctions and functional behavior after only 4 d of culture while maintaining their endothelial phenotypic morphology in longer term cultures.…”

Corneal Tissue Engineering: Recent Advances and Future Perspectives

“…60,[69][70][71][72][73][74] Decellularized aminiotic membrane was clinically evaluated in combination with human corneal endothelial cells in a lamellar keratoplasty model, where the removal of the endothelium and part of the Descemet's membrane was performed and able to function as a corneal endothelium equivalent. 73 …”