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(Investigative Ophthalmology and Visual Science. 2007;48:3537-3544.)
© 2007 by The Association for Research in Vision and Ophthalmology, Inc.
DOI:  10.1167/iovs.06-1483
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Innervation of Tissue-Engineered Corneal Implants in a Porcine Model: A 1-Year In Vivo Confocal Microscopy Study

Neil S. Lagali,1 May Griffith,1,2 Naoshi Shinozaki,3 Per Fagerholm,4 and Rejean Munger1

1From the University of Ottawa Eye Institute and the 2Department of Cellular and Molecular Medicine, University of Ottawa, Ottawa, ON, Canada; the 3TDC Eye Bank and Cornea Centre, Ichikawa General Hospital, Chiba, Japan; and the 4Department of Ophthalmology, Linköping University Hospital, Linköping, Sweden.

PURPOSE. To examine the pattern of nerve regeneration within tissue-engineered corneal substitutes grafted into host porcine corneas over a 1-year postoperative period.

METHODS. Biodegradable corneal substitutes from cross-linked collagen were implanted into the left eyes of 12 pigs by deep lamellar keratoplasty. Regeneration of severed nerves into the central implant region was investigated with in vivo confocal microscopy. Both implant-recipient and control (right) eyes were examined before surgery and 2, 6, 10, and 12 months after surgery, to quantify the number, density, diameter, and branching of nerve fiber bundles at various corneal depths. Transmission electron microscopy was used to confirm the presence of nerve bundles.

RESULTS. Two months after surgery, corneal nerve ingrowth was observed within the deep anterior stroma, with a number and density of regenerated nerves significantly higher than in nonsurgical control eyes (P < 0.01). Nerves within the superficial anterior stroma regenerated by 6 to 10 months after surgery, and the first subbasal epithelial nerves were seen 10 months after surgery. After 1 year, subbasal nerve density recovered to preoperative levels. Nerve fibers in the deep anterior stroma remained significantly thinner relative to control eyes after 1 year (P < 0.001), where both superficial anterior and subbasal nerve diameter did not change relative to control eyes.

CONCLUSIONS. The pattern of reinnervation within tissue-engineered corneal substitutes has been quantified in vivo. Innervation proceeded rapidly in the deep anterior stroma, followed by repopulation of more superficial regions. One year after surgery, nerve density within the tissue-engineered cornea increased or remained unchanged relative to controls in all corneal regions examined.






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Copyright © 2007 by the Association for Research in Vision and Ophthalmology