Abstract
Purpose :
To evaluate the topical use of crystalline alpha B minipeptides for ameliorating inflammation, rendering cytoprotection and neuroprotection to the cornea following lamellar flap surgery.
Methods :
Internalization of the FICT tagged minipeptides was assessed by treatment of the cultured corneal cells with the minipeptides. Cultured corneal cells pretreated with or without minipeptide were exposed to H2O2 and cell viability was examined by MTT assay, similarly, neuroprotection to trigeminal neurons was evaluated by beading and neurite elongation quantification.
Rabbit model of corneal nerve ablation were treated topically with crystalline alpha B- peptide at a concentration of 0.5mg/ml thrice daily for 14days while the control eye received sterile PBS. Corneal healing was evaluated by slit lamp biomicroscopy, Corneas were harvested for gene expression of inflammatory cytokines and oxidative stress protein. Histopathology immunostaining were performed to evaluate healing and expression of CRYAB protein in injured, treated and normal cornea.
Results :
Internalization of the tagged minipeptides was evidenced in the cytosol and nucleus of cultured corneal cells. The MTT assay recorded significant number of live cells in stressed corneal cells cotreated with the minipeptides versus the cells without treatment p<0.05. The minipeptides did not influence the elongation of trigeminal neurites, they significantly (p<0.05) reduced beading on the stressed neurites. The treated cornea showed reduced hyper-reflective fibrotic zones (p<0.05) and suppression in the expressions of inflammatory cytokines and oxidative stress genes. Histopathology and immunostaining supported clinical changes.
Conclusions :
The cytoprotective, neuroprotective role of crystalline alpha B minipeptides on corneal cells and trigeminal ganglion neurons enables this natural protein emerge as a promising agent for corneal healing.
This abstract was presented at the 2023 ARVO Annual Meeting, held in New Orleans, LA, April 23-27, 2023.