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Babyla G. Monteiro, Gustavo B. Melo, Fernando Malerbi, Nelson F. Lizier, Bruna P. Morais, Rui C. Serafim, Marcelo C. Silva, Ricardo L. Smith, Irina Kerkis, Jose A. Gomes; Human Immature Dental Pulp Stem Cells in the Reconstruction of Cornea with Mild and Severe Injury. Invest. Ophthalmol. Vis. Sci. 2011;52(14):5142.
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The successful regeneration of cornea depends of the severity and cell environment in the site of injury. Herein we compared the corneal regeneration after transplantation of IDPSC in to mild and severe eye surface chemical injury in rabbits.
The injury was induced by 0.5 M NaOH for 25 seconds (mild chemical burn -MCB) or 45 seconds (severe chemical burn - SCB. After 1 month, a superficial keratectomy was performed and a tissue-engineered by IDPSC was transplanted onto the corneal bed and then covered with deepithelialized human amniotic membrane (AM). The denuded cornea of control groups were covered with AM only. After 3 months, a detailed analysis of the rabbit eyes has been done with histology, transmission electron microscopy (TEM), and immunofluorescence (IF) analysis and also clinical aspect was analyzed.
The progressive transparence of corneal surface was observed in all animals that received IDPSC. Histological analysis and TEM revealed that MCB group presented organized structure similar to normal cornea, in SCB group, the corneal epithelium showed disorganized, but the stroma showed a cell density similar to normal eye. In SCB group the presence of IDPSC in stromal was observed by IF and also presented expression of cornea-specific proteins in the epithelial layer. These results suggest that second surgery in SCB group would improve the reconstruction of cornea. MCB group presented expression of cornea-specific protein, and few expression of IDPSC antibody in stromal. Control group none of these antibodies were detected. We also started the analysis of limbal region, since we detected the presence of expression of p63 antibody in corneal epithelium, indicating that IDPSC were capable to differentiate in cornea and part of these cells still maintain undifferentiated.
Our data showed that transplantation of hIDPSC was successful for the reconstruction of cornea in an animal model with mild and severe injury.
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