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Rashed Alhabshan, David Belyea, Mary Ann Stepp, Jeffrey Barratt, Sanjeev Grewal, Alexey Shashurin, Michael Keidar; Effects of In-vivo Application of Cold Atmospheric Plasma on Corneal Wound Healing in New Zealand White Rabbits. Invest. Ophthalmol. Vis. Sci. 2013;54(15):3866.
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Cold Atmospheric Plasma (CAP) has been shown to reduce corneal infection but little is known about the impact of CAP on healthy corneal tissues and their ability to respond to injuries. To examine the effect of Cold Atmospheric Plasma (CAP) on wound healing after corneal epithelial and basement membrane ablation in New Zealand white rabbits.
Twelve New Zealand white rabbits were assigned into two groups, Group A (7 rabbits) and Group B (5 rabbits). Five rabbits from each group underwent surgical intervention to the right eye, a 6 mm central corneal ablation to the epithelium and stroma using an Alger brush with 0.5 mm burr. After ablation, all rabbits in group A received 120s application of CAP. Two rabbits in Group A received CAP application without ablation. Eyes monitored for haze, inflammation, and reepithelialization with slit lamp examination. 24 hours after ablation, two corneas were harvested from each group. The 20th day, the remaining corneas were harvested, fixed in formalin and stained with H&E. In addition, immunofluorescence microscopy was performed to assess scar formation using antibodies against fibronectin and αsmooth muscle actin.
Corneal reepithelialization on day 1 showed that rabbits in group A (treated) had an average epithelial defect of 9.25 mm2 and group B (untreated) had a defect of 12.05 mm2, not a statistically significant difference. H & E stained sections showed the expected responses of stromal fibroblasts to debridement injury in both groups. Epithelial thickness and stromal cell counts on both groups 20 days after injury showed no significant statistical differences. At 24 hours and 20 days after injury, analyses show that CAP-treatment increased fibronectin deposition in the anterior stroma and αSMA localization within stromal fibroblasts; quantitative analysis of immunofluorescence data will be needed to determine whether these differences are significant.
CAP treatment following corneal injury does not interfere with rate of wound closure or induce increased inflammation. CAP treatment did affect corneal tissues since αSMA and fibronectin localization within stromal fibroblasts appears to increase slightly after CAP treatment regardless of whether corneas have been wounded. More studies are needed to evaluate the potential effects of CAP on the cornea and its possible applications in the field of Ophthalmology.
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