Abstract
Corneal keratoplasty (penetrating or lamellar) using cadaveric human tissue, is nowadays the main treatment for corneal endotelial dysfunctions. However, there is a worldwide shortage of donor corneas available for transplantation and about 53% of the world’s population have no access to corneal transplantation. Generating a complete cornea by tissue engineering is still a tough goal, but an endothelial lamellar graft might be an easier task. In this study, we developed a tissue engineered corneal endothelium by culturing human corneal endothelial cells on a human purified type I collagen membrane. Human corneal endothelial cells were cultured from corneal rims after corneal penetrating keratoplasty and type I collagen was isolated from remnant cancellous bone chips. Isolated type I collagen was analyzed by western blot, liquid chromatography -mass spectrometry and quantified using the exponentially modified protein abundance index. Later on, collagen solution was casted at room temperature obtaining an optically transparent and mechanically manageable membrane that supports the growth of human and rabbit corneal endothelial cells which expressed characteristic markers of corneal endothelium: zonula ocluddens-1 and Na+/K+ ATPase. To evaluate the therapeutic efficiency of our artificial endothelial grafts, human purified type I collagen membranes cultured with rabbit corneal endothelial cells were transplanted in New Zealand white rabbits that were kept under a minimal immunosuppression regimen. Transplanted corneas maintained transparency for as long as 6 weeks without obvious edema or immune rejection and maintaining the same endothelial markers that in a healthy cornea. In conclusion, it is possible to develop an artificial human corneal endothelial graft using remnant tissues that are not employed in transplant procedures. This artificial endothelial graft can restore the integrality of corneal endothelium in an experimental model of endothelial dysfunction. This strategy could supply extra endothelial tissue and compensate the deficit of cadaveric grafts for corneal endothelial transplantation.
Highlights
Corneal transplantation is the main treatment for patients suffering corneal endothelial dysfunctions
human purified type I collagen membranes (HPCM), in rabbits transplanted with cultured cells, was attached tightly to the corneal stroma and rabbit corneal endothelial cells (CECs) formed a continuous monolayer with the same morphology and phenotypical markers, zonula ocluddens-1 (ZO-1) and Na+/K+ ATPase, as a healthy control eye (Fig 11)
Human corneal endothelial cellular loss or damage leads to stromal edema, loss of transparency, and will eventually lead to blindness, requiring a healthy endothelial layer to reverse the edema [44]
Summary
Corneal transplantation is the main treatment for patients suffering corneal endothelial dysfunctions. There are several drawbacks with regard to PK such as graft rejection, suture-related problems, infection and astigmatism New surgical options, such as Descemet’s membrane endothelial keratoplasty (DMEK) or Descemet’s striping automated endothelial keratoplasty (DSAEK), have become increasingly popular since they tend to optimize corneal resources by replacing only the damaged part of the cornea [4,5,6,7]. These new techniques require even better endothelial quality in order to perform the corneal graft (cell density, hexagonality ratio, etc), so only 30–35% of the corneas are suitable for lamellar endothelial keratoplasty [8]
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