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Dong Won Hyun, Ah Young Koh, Yun Hee Kim, Eun Jung Shin, Saewha Jeon, Won Ryang Wee, Mee Kum Kim; Effect of NecroX-5, a novel necrosis inhibitor, on the survival of cultivated oral mucosal epithelial sheets. Invest. Ophthalmol. Vis. Sci. 2014;55(13):5518.
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Cell sheets are exposed to ischemic damages during the interval from procurement to attachment after transplantation. We intended to investigate the effect of NecroX-5,a novel necrosis inhibitor that shows scavenger activity against mitochondrial reactive oxygen species, on the survival of cultivated oral mucosal epithelial sheets after procurement.
Human oral mucosal epithelial cells were cultivated to make sheets for 2 weeks. Sheets were harvested and divided into 5 groups. One was stained with trypanblue(TB) immediately after detachment from the bottom of the culture well as a negative control. Another one was kept in the media at room temperature for 3.5 hours after procurement as a positive control. The other sheets were immersed in 10µm, 20µm, and 30µm of NecroX-5 for 30 minutes after detachment, and were re-suspended in culture media for 3 hours. In vivo, six limbal deficient rabbits received cultivated oral mucosal epithelial sheet transplant(COMET) after the sheet had been immersed in 10 µm NecroX-5 for 30 minutes (group 1), and the other six limbal deficient rabbits received COMET as control (group 2). 7 days after transplantation, periodic-acid-Schiff (PAS) staining and immunohistochemistry for P63 was performed.
Negative control showed a live cell percentage of 80.5%, whereas positive control showed 100% of cell death. Live to dead cell ratio was 51.5% vs 48.5% in the 10 µm NecroX-5 treated sheet. There were no goblet cells on the transplanted corneal epithelial cells in bothgroups. The expression of P63 showed no significant difference between groups.
Our results suggest that immersion in 10µm NecroX-5 may increase the cell survival of cultivated oral mucosal cell sheets.
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