Li Xiaoxue, Li Kuanshu, Gu Jianing, Sun Xihao, Liang Yuqin, Ding Chengcheng, Chen Yuexi, Chen Hang, Chen Jiansu, Cui Zekai
Aier Academy of Ophthalmology, Central South University, Changsha, Hunan, China.
Changsha Aier Eye Hospital, Changsha, Hunan, China.
Mater Today Bio. 2025 Jul 23;34:102130. doi: 10.1016/j.mtbio.2025.102130. eCollection 2025 Oct.
Keratoconus (KC) is a prevalent ectatic corneal disease influenced by corneal biomechanics, though its pathogenesis remains unclear. Constructing an in vitro corneal model with adjustable biomechanics is essential for studying keratoconus. In this study, a KC disease group (L group) was created using a low matrix stiffness collagen hydrogel containing human corneal stromal cells (hCSCs), while a high matrix stiffness group (H group) was established using a plastic compression method. Additionally, a cross-linking treatment model was applied to the L group using corneal collagen cross-linking (CXL). Results showed that the matrix stiffness of the L group was significantly lower compared to the H group. Both the L group and clinical KC samples exhibited lower collagen fibrils density. Transcriptomic and proteomic analyses revealed decreased antioxidant capacity and increased levels of inflammatory factors and reactive oxygen species (ROS) in the L group. The NRF2 pathway activity was downregulated. In the L group cells, inflammation-related genes were upregulated, antioxidant-related genes were downregulated, and ROS levels were elevated, accompanied by a reduction in mitochondrial membrane potential. The concentrations of IL-6 and TNF-α in the culture medium were increased. Histological results indicated that the L group, similar to clinical KC samples, exhibited high expression of inflammatory and oxidative stress factors, while signals of antioxidant-related factors were decreased. After cross-linking treatment, the L group demonstrated increased matrix stiffness, higher collagen fibrils density, upregulated expression of antioxidant factors, and decreased expression of inflammatory and immune factors. This study successfully established an in vitro corneal stromal model with adjustable matrix stiffness, providing a platform for investigating the pathogenesis of KC.
圆锥角膜（KC）是一种受角膜生物力学影响的常见扩张性角膜疾病，但其发病机制尚不清楚。构建具有可调节生物力学的体外角膜模型对于研究圆锥角膜至关重要。在本研究中，使用含有人类角膜基质细胞（hCSCs）的低基质硬度胶原水凝胶创建了一个圆锥角膜疾病组（L组），而通过塑料压缩法建立了一个高基质硬度组（H组）。此外，对L组采用角膜胶原交联（CXL）进行交联处理模型。结果表明，L组的基质硬度明显低于H组。L组和临床圆锥角膜样本的胶原纤维密度均较低。转录组学和蛋白质组学分析显示，L组的抗氧化能力下降，炎症因子和活性氧（ROS）水平升高。NRF2信号通路活性下调。在L组细胞中，炎症相关基因上调，抗氧化相关基因下调，ROS水平升高，同时线粒体膜电位降低。培养基中IL-6和TNF-α的浓度增加。组织学结果表明，L组与临床圆锥角膜样本相似，炎症和氧化应激因子表达较高，而抗氧化相关因子的信号降低。交联处理后，L组的基质硬度增加，胶原纤维密度更高，抗氧化因子表达上调，炎症和免疫因子表达降低。本研究成功建立了一个具有可调节基质硬度的体外角膜基质模型，为研究圆锥角膜的发病机制提供了一个平台。
