Rat limbal niche cells (LNCs) have been proven to induce transdifferentiation of oral mucosal epithelial cells (OMECs) into corneal epithelial-like cells termed transdifferentiated oral mucosal epithelial cells (T-OMECs). This investigation aimed to evaluate the effect of subconjunctival T-OMEC injections on alkali-induced limbal stem cell deficiency (LSCD) in rats. LNCs were cocultured with OMECs in the Transwell system to obtain T-OMECs, with NIH-3T3 cells serving as a control. Subconjunctival injection of single T-OMEC or OMEC suspension was performed immediately after corneal alkali injury. T-OMECs were prelabeled with the fluorescent dye CM-DiI in vitro and tracked in vivo. Corneal epithelial defect, opacity, and neovascularization were quantitatively analyzed. The degree of corneal epithelial defect (from day 1 onward), opacity (from day 5 onward), and neovascularization (from day 2 onward) was significantly less in the T-OMEC group than in the OMEC group. Cytokeratin 12 (CK12), pigment epithelium–derived factor, and soluble fms-like tyrosine kinase-1 were expressed at a higher rate following T-OMEC injection. Some CM-DiI-labeled cells were found to be coexpressed with CK12, Pax6, and ΔNp63α in the corneal epithelium after subconjunctival injection. Subconjunctival injection of T-OMECs prevents conjunctival invasion and maintains a normal corneal phenotype, which might be a novel strategy in the treatment of LSCD:

大鼠角膜缘小生境细胞 (LNC) 已被证明可诱导口腔黏膜上皮细胞 (OMEC) 转分化为角膜上皮样细胞，称为转分化口腔黏膜上皮细胞 (T-OMEC)。本研究旨在评估结膜下 T-OMEC 注射对大鼠碱诱导的角膜缘干细胞缺乏症 (LSCD) 的影响。在 Transwell 系统中将 LNC 与 OMEC 共培养以获得 T-OMEC，以 NIH-3T3 细胞作为对照。角膜碱损伤后立即进行单次 T-OMEC 或 OMEC 悬液的结膜下注射。T-OMEC 在体外用荧光染料 CM-DiI 进行预标记并在体内进行跟踪。定量分析角膜上皮缺损、混浊和新生血管。角膜上皮缺损的程度（从第 1 天开始），T-OMEC 组的混浊（从第 5 天开始）和新生血管（从第 2 天开始）明显少于 OMEC 组。细胞角蛋白 12 (CK12)、色素上皮衍生因子和可溶性 fms 样酪氨酸激酶-1 在 T-OMEC 注射后以更高的速率表达。结膜下注射后，发现一些 CM-DiI 标记的细胞在角膜上皮中与 CK12、Pax6 和 ΔNp63α 共表达。结膜下注射 T-OMEC 可防止结膜侵袭并维持正常的角膜表型，这可能是治疗 LSCD 的新策略：T-OMEC 注射后，可溶性 fms 样酪氨酸激酶 1 的表达率更高。结膜下注射后，发现一些 CM-DiI 标记的细胞在角膜上皮中与 CK12、Pax6 和 ΔNp63α 共表达。结膜下注射 T-OMEC 可防止结膜侵袭并维持正常的角膜表型，这可能是治疗 LSCD 的新策略：T-OMEC 注射后，可溶性 fms 样酪氨酸激酶 1 的表达率更高。结膜下注射后，发现一些 CM-DiI 标记的细胞在角膜上皮中与 CK12、Pax6 和 ΔNp63α 共表达。结膜下注射 T-OMEC 可防止结膜侵袭并维持正常的角膜表型，这可能是治疗 LSCD 的新策略：