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N. Kopsachilis, U. Welge - Luessen, F. E. Kruse; Human Anterior Lens Capsule as an Artificial Descemet Membrane Substrate in Corneal Surgery. Invest. Ophthalmol. Vis. Sci. 2010;51(13):2835. doi: https://doi.org/.
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© ARVO (1962-2015); The Authors (2016-present)
To study the potential use of human anterior lens capsule as a scaffold for human endothelial cell transplantation in treatment of endothelial cell deficiency.
Anterior lens capsules were recovered from 15 Cornea donors 24 to 72 hours post mortem. Human corneal endothelial cells were recovered from the remaining corneal sclera rims of donor corneas used for penetrating keratoplasty. Capsules were treated in distilled water for 2 hours to eliminate the crystalline epithelium and stored at 4ºC. After initial processing, primary cultured human corneal endothelial (HCE) cells were added on the anterior capsule and allowed to attach in culture at 37°C. Samples were sorted into 3 groups. Group 1 consisted of 5 samples in which the endothelial cells were allowed to grow on corresponding capsules. Group 2 consisted of human endothelial cells that were allowed to grow on a collagen membrane (Resorba, Nuremberg). Group 3 consisted of 5 samples in which human endothelial cells were allowed to grow on polystyrene culture plates (Control). All specimens were incubated for 2 weeks at 37°C and 5% CO2. Cell density, viability, morphology, and adherence of the cell-capsule complex were evaluated at 1, 4, 7, and 14 days with a phase-contrast microscope and an electron microscope. Expression of zonula occludens-1 and Na/K-adenosine triphosphatase were investigated by immunohistochemistry
A mean diameter of 8 mm anterior capsule was obtained as a substrate for cell culture. The rate of cell growth and density in group 1 was comparable to the control group whereas in group 2 there was a slight lower growth rate. Cell viability was 95% or superior in all groups, and tight junctions developed between growing cells. Immunohistochemical analysis demonstrated strongly positive staining for zonula occludens-1 and Na/K-adenosine triphosphatase. Electron microscopy confirmed the adherence and monolayer growth of the endothelial cells.
Phenotypical properties of the cell-capsule complex imply that the HCE sheets are capable of maintaining intact barrier and ionic pump functions in vitro. A mean diameter of 8 mm of the cell-capsule complex is comparable to the size of the Descemet membrane transplant used in Descemet membrane endothelial keratoplasty. Human anterior lens capsule might therefore be a potential scaffold for ex vivo expansion of human corneal endothelial cells and can be used as an artificial Descemet membrane substrate in corneal surgery
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