Bilateral limbal stem cell deficiency (LSCD) can be effectively treated with cultivated oral mucosa epithelial cell transplantation (COMET). However, COMET is associated with greater superficial neovascularization than limbal stem cell (LESC) transplantation, the gold standard for unilateral LSCD. To investigate the intrinsic molecular features of cells intended for grafting, we assessed the in vitro expression of genes involved in vascularization and inflammation using real-time quantitative PCR and multifactorial linear models. Oral mucosal epithelial cells (OMECs) and limbal epithelial cells (LECs) were cultured in either conventional (COM) or xenobiotic-free (XF) media on fibrin substrates. Gene expression profiling revealed distinct transcriptional signatures. The pro-angiogenic genes AGR2, ANGPTL2, CRYAB, EREG, JAM3, and S100A4 were significantly higher in LECs (adjusted p < 0.01), whereas FGF2 was higher in OMECs (adjusted p < 0.001). The anti-angiogenic genes TIMP3 and SERPINF1 were higher in LECs (adjusted p < 0.01), while COL18A1 was higher in OMECs (adjusted p < 0.01). OMECs also showed significantly greater expression of the immunoregulatory genes IL1B, IL6, TNF, CXCL10, and IL1RN (adjusted p < 0.01). Cultivation induced phenotypic changes in OMECs, with COM and XF media exerting comparable effects. These results highlight the contribution of inflammatory mediators to neovascularization following COMET.

Department of Pathology of the 3rd Medical Faculty University Hospital Kralovske Vinohrady 100 34 Prague Czech Republic

Institute of Biology and Medical Genetics 1st Faculty of Medicine Charles University and General University Hospital Prague 128 00 Prague Czech Republic

Ophthalmology Department of the 3rd Medical Faculty University Hospital Kralovske Vinohrady 100 34 Prague Czech Republic

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