Kalha Solja, Kuony Alison, Michon Frederic
Helsinki Institute of Life Science, Institute of Biotechnology, University of Helsinki.
Helsinki Institute of Life Science, Institute of Biotechnology, University of Helsinki; School of Medicine and Institute for Science and Technology in Medicine, Keele University;
J Vis Exp. 2018 Jul 10(137):58071. doi: 10.3791/58071.
The murine cornea provides an excellent model to study wound healing. The cornea is the outermost layer of the eye, and thus is the first defense to injury. In fact, the most common type of eye injury found in clinic is a corneal abrasion. Here, we utilize an ocular burr to induce an abrasion resulting in removal of the corneal epithelium in vivo on anesthetized mice. This method allows for targeted and reproducible epithelial disruption, leaving other areas intact. In addition, we describe the visualization of the abraded epithelium with fluorescein staining and provide concrete advice on how to visualize the abraded cornea. Then, we follow the timeline of wound healing 0, 18, and 72 h after abrasion, until the wound is re-epithelialized. The epithelial abrasion model of corneal injury is ideal for studies on epithelial cell proliferation, migration and re-epithelialization of the corneal layers. However, this method is not optimal to study stromal activation during wound healing, because the ocular burr does not penetrate to the stromal cell layers. This method is also suitable for clinical applications, for example, pre-clinical test of drug effectiveness.
小鼠角膜为研究伤口愈合提供了一个极佳的模型。角膜是眼睛的最外层,因此是抵御损伤的第一道防线。事实上,临床上发现的最常见的眼外伤类型是角膜擦伤。在此,我们利用眼科磨钻在麻醉的小鼠体内造成擦伤,从而去除角膜上皮。这种方法能够实现有针对性的、可重复的上皮破坏,而其他区域保持完整。此外,我们描述了用荧光素染色观察擦伤上皮的方法,并就如何观察擦伤角膜提供了具体建议。然后,我们追踪擦伤后0、18和72小时的伤口愈合时间线,直至伤口重新上皮化。角膜损伤的上皮擦伤模型对于研究角膜层上皮细胞的增殖、迁移和重新上皮化非常理想。然而,这种方法对于研究伤口愈合过程中的基质激活并非最佳选择,因为眼科磨钻不会穿透到基质细胞层。这种方法也适用于临床应用,例如药物有效性的临床前测试。