Abstract
The aim of this study was to construct a rabbit anterior corneal replacement for transplantation using acellular porcine corneal matrix (APCM) and rabbit epithelial or stromal cells. APCM was prepared from fresh porcine cornea treated with 0.5% (wt./vol.) sodium dodecyl sulfate (SDS) solution. The expanded stromal cells were first injected into APCM parallel to its surface and were cultured in a shaking culture system for 7 days to obtain the stromal construct. Next, corneal epithelial cells were cultured on the stromal construct surface for another 7 days to obtain rabbit anterior corneal lamella. The construct had a phenotype similar to that of normal cornea, with high expression of cytokeratin 3 in the epithelial cell layer and vimentin in the stromal cells. More importantly, the construct integrated well with the implanted host corneal tissue, and the implant cornea maintained transparency in the 6-month follow-up, although there was a slight haze in the central corneal area. The endothelium in the surgery cornea had a similar cell density and mosaic pattern with normal cornea as shown by confocal laser corneal microscopy, and the regenerated corneal epithelial cells on the implant surface showed a similar morphology to that of natural epithelial cells. These results demonstrate that the constructed anterior corneal replacement exhibits an excellent biological property for lamellar keratoplasty and might be a possible alternative to human corneal tissue in the future.
Highlights
The cornea is a transparent tissue outside the eye and serves as a barrier for the protection of the eye inner tissues and is a major refractive element of the visual system [1]
acellular porcine corneal matrix (APCM) was transparent after being soaked in sterile glycerol for 30 min (Fig 2B)
Two or three layers of epithelial cells with high expression of cytokeratin 3 (CK3) formed on top of the stromal substitutes after 7 days of submerged culture (Fig 3A, 3B, and 3C)
Summary
The cornea is a transparent tissue outside the eye and serves as a barrier for the protection of the eye inner tissues and is a major refractive element of the visual system [1]. Irreversible damage to the cornea can result in permanent visual loss or blindness. Corneal disease is the largest cause of vision loss, especially in developing countries [1]. There are more than 50 million people in the world with visual loss caused by corneal opacification resulting from disease or trauma [2]. The most successful and widely used treatment for corneal blindness is transplantation with human donor corneal tissue in several procedures such as penetrating keratoplasty [3], PLOS ONE | DOI:10.1371/journal.pone.0168084. The most successful and widely used treatment for corneal blindness is transplantation with human donor corneal tissue in several procedures such as penetrating keratoplasty [3], PLOS ONE | DOI:10.1371/journal.pone.0168084 December 8, 2016
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