Voukali Eleni, Cabral Joao Victor, Smorodinova Natalia, Kolin Vojtech, Netukova Magdalena, Vacík Tomáš, Jirsova Katerina
Institute of Biology and Medical Genetics, First Faculty of Medicine, Charles University and General University Hospital in Prague, 128 00 Prague, Czech Republic.
Department of Pathology of the Third Medical Faculty, University Hospital Kralovske Vinohrady, 100 34 Prague, Czech Republic.
Bioengineering (Basel). 2025 Jun 24;12(7):688. doi: 10.3390/bioengineering12070688.
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 , , , , , and were significantly higher in LECs (adjusted < 0.01), whereas was higher in OMECs (adjusted < 0.001). The anti-angiogenic genes and were higher in LECs (adjusted < 0.01), while was higher in OMECs (adjusted < 0.01). OMECs also showed significantly greater expression of the immunoregulatory genes IL1B, IL6, TNF, CXCL10, and IL1RN (adjusted < 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.
双侧角膜缘干细胞缺乏症（LSCD）可通过培养的口腔黏膜上皮细胞移植（COMET）得到有效治疗。然而，与单侧LSCD的金标准——角膜缘干细胞（LESC）移植相比，COMET与更严重的浅层新生血管形成相关。为了研究用于移植的细胞的内在分子特征，我们使用实时定量PCR和多因素线性模型评估了参与血管生成和炎症的基因的体外表达。口腔黏膜上皮细胞（OMECs）和角膜缘上皮细胞（LECs）在纤维蛋白基质上的常规（COM）或无外源物（XF）培养基中培养。基因表达谱揭示了不同的转录特征。促血管生成基因 、 、 、 、 和 在LECs中显著更高（校正 < 0.01），而 在OMECs中更高（校正 < 0.001）。抗血管生成基因 和 在LECs中更高（校正 < 0.01），而 在OMECs中更高（校正 < 0.01）。OMECs还显示出免疫调节基因IL1B、IL6、TNF、CXCL10和IL1RN的表达显著更高（校正 < 0.01）。培养诱导了OMECs的表型变化，COM和XF培养基发挥了类似的作用。这些结果突出了炎症介质对COMET后新生血管形成的作用。
