Liang Wentao, Huang Li, Clayton Joseph M, Nicholas Sarah E, Hefley Brenna S, Ma Jian-Xing, Karamichos Dimitrios
Department of Biochemistry, Wake Forest University School of Medicine, Winston-Salem, North Carolina, United States.
Aier Eye Hospital, Jinan University, Guangzhou, Guangdong Province, China.
Invest Ophthalmol Vis Sci. 2025 Aug 1;66(11):8. doi: 10.1167/iovs.66.11.8.
This study aimed to evaluate the therapeutic potential of salivary exosomes (SEs) in corneal epithelial wound healing by assessing their effects on wound closure, cellular function, and molecular mechanisms in both in vivo and in vitro models.
Corneal epithelial wounds were induced in C57BL/6J mice and treated with topical SEs (10 µg/eye) twice daily. Wound closure was monitored using fluorescein staining. In vitro, primary human corneal epithelial cells (HCECs) and human limbal epithelial cells (HLECs) were treated with SEs (0, 5, and 25 µg/mL) to assess migration, proliferation, and mitochondrial function. Western blot and immunohistochemistry were used to evaluate key molecular markers, including integrin α6, integrin β4, thrombospondin-1 (TSP1), and transforming growth factor-β1 (TGF-β1).
SE treatment significantly accelerated corneal wound closure in vivo. In vitro, SEs enhanced HCEC and HLEC migration, proliferation, and mitochondrial function. SEs upregulated integrin α6, integrin β4, and TSP1 expression in both wounded corneas and cultured HCECs. TGF-β1 levels were transiently increased in exosome-treated corneas but returned to baseline as healing progressed. Mitochondrial stress assays revealed that SEs enhanced oxidative phosphorylation in HCECs and HLECs.
SEs promote corneal epithelial wound healing by enhancing cellular migration, proliferation, and mitochondrial function while modulating key molecular pathways. These findings suggest that SEs represent a novel therapeutic strategy for corneal injury, warranting further investigation into their mechanisms and clinical applications.
本研究旨在通过评估唾液外泌体（SEs）在体内和体外模型中对伤口闭合、细胞功能及分子机制的影响，来评价其在角膜上皮伤口愈合中的治疗潜力。
在C57BL/6J小鼠中诱导角膜上皮伤口，并每天两次局部给予SEs（10μg/眼）进行治疗。使用荧光素染色监测伤口闭合情况。在体外，用SEs（0、5和25μg/mL）处理原代人角膜上皮细胞（HCECs）和人角膜缘上皮细胞（HLECs），以评估细胞迁移、增殖和线粒体功能。采用蛋白质免疫印迹法和免疫组织化学法评估关键分子标志物，包括整合素α6、整合素β4、血小板反应蛋白-1（TSP1）和转化生长因子-β1（TGF-β1）。
SE治疗在体内显著加速了角膜伤口的闭合。在体外，SEs增强了HCEC和HLEC的迁移、增殖及线粒体功能。SEs上调了受伤角膜和培养的HCECs中整合素α6、整合素β4和TSP1的表达。在经外泌体处理的角膜中，TGF-β1水平短暂升高，但随着愈合进展恢复至基线水平。线粒体应激试验表明，SEs增强了HCECs和HLECs中的氧化磷酸化。
SEs通过增强细胞迁移、增殖和线粒体功能，同时调节关键分子途径来促进角膜上皮伤口愈合。这些发现表明，SEs代表了一种治疗角膜损伤的新策略，值得对其机制和临床应用进行进一步研究。
