Survival of corneal nerve/sheath structures in organ-cultured donor corneas.

Abstract

To study the morphology of human corneal nerves in eye bank organ-cultured corneas and in corneal grafts post-transplantation. Thirty-seven organ-cultured corneas were divided into: Group-A, anterior 300-400μm of 20 corneas used for Descemets stripping endothelial keratoplasty, and Group-B, 17 full-thickness corneas unsuitable for transplantation. Corneas whole mounts were stained for nerves using acetylcholinesterase technique and examined by NanoZoomer digital pathology microscope. Central and sub-Bowman's stromal nerves and the sub-basal nerve plexus including perforation sites and terminal bulbs were studied. Ten eyes were imaged following penetrating keratoplasty using in-vivo confocal microscopy (IVCM) for the presence of sub-basal and stromal nerves at 1, 4-5 and 7-8weeks postoperatively (five eyes) and in all the other five eyes, the final follow-up was at 12weeks. Fifteen of twenty (75%) corneas had stromal nerves in Group-A and 15 of 17 (88.2%) in Group-B. Average number of stromal nerves entering peripherally were 9.1 (range: 1-36). 7.5 in Group-A and 10.8 in Group-B. Central stromal nerves were seen in eight samples in Group-A and nine in Group-B. Many stromal nerves terminated abruptly without demonstrable continuity through Bowman's membrane. No terminal bulbs or sub-basal nerves were detected. In-vivo confocal microscopy (IVCM) showed 4 of 5 in 9 of 10 (90%) donor corneas had stromal nerves 1week postoperatively, which remained present in 8 of 10 (80%) corneas at 4-5weeks and in 9 of 10 (90%) at 7-8weeks postoperatively. All 5 corneas analysed at 12weeks showed the same stromal nerves from 1 to 12weeks postoperatively. Sub-basal nerves were absent in all corneas over the 12-week study period. This study provides further insight into the behaviour of corneal nerves in transplanted corneas. Corneal stromal nerves/nerve-sheaths are preserved in organ-cultured eye bank eyes and persist post-transplantation up to 3months. These could provide directional guidance to regenerating nerves from host stroma.