1. Evaluation of the Suitability of Biocompatible Carriers as Artificial Transplants Using Cultured Porcine Corneal Endothelial Cells
- Author
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Gerrit R. J. Melles, Marieke Bruinsma, Alina Miron, Silke Oellerich, Daniele Spinozzi, Perry S. Binder, Itay Lavy, Mehrdad Rafat, and Isabel Dapena
- Subjects
Biocompatibility ,Cell Survival ,Swine ,Cell- och molekylärbiologi ,Lens Capsule, Crystalline ,Biocompatible Materials ,Porcine endothelial cells ,cell culture ,donor material ,endothelial cell transplantation ,cell carriers ,03 medical and health sciences ,Cellular and Molecular Neuroscience ,0302 clinical medicine ,Stroma ,Animals ,Humans ,Viability assay ,Fluorescent Antibody Technique, Indirect ,Cells, Cultured ,Cell Proliferation ,Tissue Engineering ,Tissue Scaffolds ,Chemistry ,Endothelium, Corneal ,Adhesion ,Sensory Systems ,In vitro ,Ophthalmology ,Ki-67 Antigen ,Cell culture ,Descemet Stripping Endothelial Keratoplasty ,Zonula Occludens-1 Protein ,030221 ophthalmology & optometry ,Immunohistochemistry ,Collagen ,Sodium-Potassium-Exchanging ATPase ,Cell and Molecular Biology ,030217 neurology & neurosurgery ,Biomedical engineering - Abstract
Purpose/Aim: Evaluating the suitability of bioengineered collagen sheets and human anterior lens capsules (HALCs) as carriers for cultivated porcine corneal endothelial cells (pCECs) and in vitro assessment of the cell-carrier sheets as tissue-engineered grafts for Descemet membrane endothelial keratoplasty (DMEK). Materials and Methods: pCECs were isolated, cultured up to P2 and seeded onto LinkCell (TM) bioengineered matrices of 20 mu m (LK20) or 100 mu m (LK100) thickness, and on HALC. During expansion, pCEC viability and morphology were assessed by light microscopy. ZO-1 and Na+/K+-ATPase expression was investigated by immunohistochemistry. Biomechanical properties of pCEC-carrier constructs were evaluated by simulating DMEK surgery in vitro using an artificial anterior chamber (AC) and a human donor cornea without Descemet membrane (DM). Results: During in vitro expansion, cultured pCECs retained their proliferative capacity, as shown by the positive staining for proliferative marker Ki67, and a high cell viability rate (96 +/- 5%). pCECs seeded on all carriers formed a monolayer of hexagonal, tightly packed cells that expressed ZO-1 and Na+/K+-ATPase. During in vitro surgery, pCEC-LK20 and pCEC-LK100 constructs were handled like Descemet stripping endothelial keratoplasty (DSEK) grafts, i.e. folded like a "taco" for insertion because of challenges related to rolling and sticking of the grafts in the injector. pCEC-HALC constructs behaved similar to the DMEK reference model during implantation and unfolding in the artificial AC, showing good adhesion to the bare stroma. Conclusions: In vitro DMEK surgery showed HALC as the most suitable carrier for cultivated pCECs with good intraoperative graft handling. LK20 carrier showed good biocompatibility, but required a DSEK-adapted surgical protocol. Both carriers might be notional candidates for potential future clinical applications. Funding Agencies|European Unions Horizon 2020 research and innovation programme [667400]
- Published
- 2018