Department of Ophthalmology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
Department of Ophthalmology, Renmin Hospital of Wuhan University, Wuhan, China.
J Histochem Cytochem. 2021 Mar;69(3):177-190. doi: 10.1369/0022155420980071. Epub 2020 Dec 21.
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 niche 细胞（LNCs）已被证明可诱导口腔黏膜上皮细胞（OMECs）向角膜上皮样细胞转分化，称为转分化的口腔黏膜上皮细胞（T-OMECs）。本研究旨在评估结膜下注射 T-OMEC 对大鼠碱诱导的角膜缘干细胞缺乏症（LSCD）的影响。将 LNCs 与 OMECs 在 Transwell 系统中共培养，以获得 T-OMECs，NIH-3T3 细胞作为对照。角膜碱烧伤后立即进行结膜下注射单一 T-OMEC 或 OMEC 悬液。T-OMECs 在体外预先用荧光染料 CM-DiI 标记，并在体内进行跟踪。定量分析角膜上皮缺损、混浊和新生血管形成。T-OMEC 组的角膜上皮缺损程度（从第 1 天开始）、混浊程度（从第 5 天开始）和新生血管形成程度（从第 2 天开始）均明显低于 OMEC 组。T-OMEC 注射后，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 的表达率更高。在结膜下注射后，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 可防止结膜侵犯并维持正常的角膜表型，这可能是治疗 LSCD 的一种新策略。
