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huixiang ma, Lingli Li, Xiaojuan Hao, Timothy Hughes, Hao Chen; Cross-linking Bovine lens capsule as a potential corneal endothelium carrier. Invest. Ophthalmol. Vis. Sci. 2014;55(13):2061.
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The objective of our study is to examine the possibility of using cross-linked bovine lens capsule as a carrier for BCECs in vitro and acting as a tissue engineered membrane for use in future transplant surgery.
BLCs were prepared from 30 bovine eyes obtained from local abattoir. BLCs were cross-linked by riboflavin and UVA light with a wavelength of 370 nm and a dosage of 5.4 J/cm2. Compare the mechanical, glucose and protein permeability, and optical transparency properties between cross-linked and fresh BLC samples. Then bovine corneal endothelial cells (BCECs) which were were harvested from fresh bovine eyes obtained from the same source were seeded on BLCs. Cell morphology was assessed with a phase-contrast microscope after 6 hours of incubation and at days 1,3,5, and 7. The density of BCECs was expressed as cells per square millimeter after 7 days in culture. Live/Dead viability/cytotoxicity assay was also performed and analyzed by fluorescence microscopy after 7 days of incubation during confluence. Expression of zonula occludens-1, Na+/K+-adenosine triphosphatase were investigated by immunohistochemistry.
Cross-linking BLCs had similar mechanical, glucose and protein permeability, and optical transparency properties to fresh samples. And they are hard to crimp and more easily to handle and control during culturing and transplantation. Moreover, they could support the growth of BCECs and creat a continuous viable monolayer of cells which resembled the healthy corneal endothelium with a mean cell density of 3230±215cells per square millimeter. Immunohistochemistry analysis demonstrated strongly positive staining for zonula occludens-1 and Na+/k+-adenosine triphosphatase.
In the present study, we have demonstrated the suitability of cross-linked bovine lens capsule to support the bovine corneal endothelial cells growth (as a carrier), indicating that the cross-linked BLC has a great potential in tissue engineered corneal endothelium.
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