Huang Stephanie U, Yoon Jinny J, Ismail Salim, McGhee Jennifer J, Sherwin Trevor
Department of Ophthalmology, Faculty of Medical and Health Sciences, University of Auckland, Auckland, New Zealand.
Cell Biol Int. 2015 Nov;39(11):1274-87. doi: 10.1002/cbin.10501. Epub 2015 Jul 29.
The cornea is the initial refractive interface of the eye. Its transparency is critical for clear vision and is maintained by stem cells which also act to repair injury inflicted by external insults, such as chemical and thermal burns. Damage to the epithelium compromises its clarity and can reduce or eliminate the stem cell population, diminishing the ability for self-repair. This condition has been termed "limbal stem cell deficiency"; severe cases can lead to corneal blindness. Sphere-forming cells isolated from peripheral cornea are a potential source of stem and progenitor cells for corneal repair. When provided with appropriate substrate, these spheres have the ability to adhere and for cells to migrate outwards akin to that of their natural environment. Direct compression injury and remote scratch injury experiments were conducted on the sphere cells to gauge their wound healing capacity. Measures of proliferation, differentiation, and migration were assessed by immunohistochemical detection of EdU incorporation, α-smooth muscle actin expression and confocal image analysis, respectively. Both modes of injury were observed to draw responses from the spheres indicating wound healing processes. Direct wounding induced a rapid, but transient increase in expression of α-SMA, a marker of corneal myofibroblasts, followed by a proliferative and increasing migratory response. The spheres were observed to respond to remote injury as entire units, with no directional response seen for targeted repair over the scratch injury area. These results give strength to the future use of these peripheral corneal spheres as transplantable units for the regeneration of corneal tissue.
角膜是眼睛的初始屈光界面。其透明度对于清晰视力至关重要,由干细胞维持,这些干细胞还能修复外部损伤(如化学和热烧伤)造成的伤害。上皮损伤会损害其透明度,并可能减少或消除干细胞群体,削弱自我修复能力。这种情况被称为“角膜缘干细胞缺乏症”;严重病例可导致角膜失明。从周边角膜分离出的成球细胞是角膜修复的干细胞和祖细胞的潜在来源。当提供合适的基质时,这些球体有能力黏附,细胞能够向外迁移,类似于它们在自然环境中的情况。对球体细胞进行直接压迫损伤和远程划痕损伤实验,以评估它们的伤口愈合能力。分别通过免疫组化检测EdU掺入、α-平滑肌肌动蛋白表达和共聚焦图像分析来评估增殖、分化和迁移情况。观察到两种损伤模式均能引起球体的反应,表明存在伤口愈合过程。直接损伤诱导角膜肌成纤维细胞标志物α-SMA的表达迅速但短暂增加,随后出现增殖和迁移增加的反应。观察到球体作为一个整体对远程损伤做出反应,在划痕损伤区域未观察到针对靶向修复的定向反应。这些结果为未来将这些周边角膜球体用作角膜组织再生的可移植单元提供了依据。