Abstract
Purpose:
Purpose A tissue engineered corneal endothelium could overcome potential problems associated with donor tissues. We have developed a thin (50µm) synthetic hydrogel film suitable for corneal endothelial cell (CEC) culture and implantation. An ovine model was used to develop and evaluate the film in vitro and in vivo.
Methods:
Methods Synthetic hydrogel films (SHF) with superior properties were developed using a novel approach. Ovine CEC (OCEC) density in vivo was determined by specular microscopy and compared to cell density achieved on tissue culture plastic and the hydrogel film as determined using Computer Assisted Steriological Toolbox (Leica). Films were implanted into an ovine anterior chamber for 30d before histological evaluation.
Results:
OCEC density in vivo was determined to be 3150cells/mm2 in 14 month old merino sheep (n=3). Cell density at 28d was 3150 (SEM 459) on SHF, 2530 (SEM 526) on collagen I coated SHF and 3770 (SEM 201) tissue culture plastic (n=4). The films had sufficient tensile strength for implantation and readily un-scrolled within the anterior chamber. Histology determined that PHF did not produce and inflammatory response in vivo (n=4).
Conclusions:
Hydrogel films suitable for implantation that support CEC cultivation have been produced. A combination of a SFH with a cultured CEC monolayer may be able to replace the donor tissue lenticule used in Descemet’s Stripping Automated Endothelial Keratoplasty.
Keywords: 481 cornea: endothelium