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F. Luengo Gimeno, V. Lavigne, S. Gatto, J. Ferro, J. O. Croxatto, L. Correa, J. E. Gallo; Preclinical Study Results: Six Months Follow Up of Autograft Epithelial Corneal Cell Layers Implantation in Rabbits. Invest. Ophthalmol. Vis. Sci. 2008;49(13):5737.
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To evaluate the efficacy of autologous corneal epithelial sheet implantation in restoring limbal deficiency at six months due to severely alkaline injured corneas in rabbits
Limbal stem cell deficiency (LSCD) was induced in 24 rabbits by alkali burns. Limbal cell biopsy was done in contralateral eye and cells were cultured on a fibroblast feeder layer grown on autologous clotted platelet poor plasma (PPP). Deep anterior keratectomy followed by suturing of corneal cell sheet to the sclera over the stroma was performed in 16 animals. The remaining animals constituted the control group and were treated only with a PPP membrane. Ciprofloxacin and dexamethasone drops were postoperatively used and argon laser photocoagulation was applied if corneal neovascularization was seen. Clinical signs were recorded according to Bagley scoring system for LSCD. The corneas were processed for histopathology and inmunohistochemistry (antiK3/12, antiK19 and antiK4)
No signs of limbal stem cells deficiency were observed in the majority of treated animals that showed a notorious improvement in clinical parameters. A full thickness continuous layer of stratified epithelial cells resembling corneal epithelium was seen in grafted corneas together with several clear cytoplasm epithelial-like cells in the deepest part of the epithelium. Neither clinical nor histological evidence of immune reaction was detected. Control animals showed LCSD signs with a thin regenerative epithelium and a pronounced fibroblastic proliferation. Expression of cytokeratins 4, 19 and 3/12 on the corneas of 16 animals treated with stem cells transplantation was similar to that found in normal corneas.
Autografts of bioengineered limbal epithelium promoted normal corneal reepithelialization restoring the LSCD at six months. Autologous corneal epithelial cell transplantation may be soon used in a pilot study of Argentinean patients.
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