Department of Ophthalmology, Tokyo Dental College, Chiba, Japan.
Invest Ophthalmol Vis Sci. 2011 Oct 17;52(11):8132-7. doi: 10.1167/iovs.11-7984.
To characterize corneal epithelial cells separated from limbus in vivo by transplantation of a stainless steel ring with or without creating a defect inside the ring.
A stainless steel ring (diameter, 8 mm; width, 300 μm; depth, 250 μm) was transplanted into rabbit corneal stroma using 10-0 nylon interrupted suture after cutting to a 250 μm depth by corneal vacuum trephine (diameter, 8.0 mm). Epithelial cells were removed inside the ring, and re-epithelization was evaluated after 1 week. Hematoxylin staining and immunostaining against p63, Ki67, and cytokeratin 3 were performed for phenotypic analysis of corneal epithelia. A corneal epithelial defect was centrally created inside the ring (4, 5, and 6 mm diameter) after transplantation. When re-epithelization was achieved, a central epithelial defect was continuously created until cells were exhausted within the ring. The number of created defects was also analyzed to assess the potential of re-epithelialization.
Ring-transplanted corneal stroma showed few signs of inflammation, and when epithelium was totally removed from inside the ring, complete epithelial defects were persistent for ≥ 1 month. Corneal sensation was significantly decreased in corneas with the ring (P < 0.05). Immunostaining demonstrated similar expression patterns for p63, Ki67, and cytokeratin3 as the controls. When rings were transplanted into the intact cornea, inside epithelia prevented epithelial defects in vivo for ≤ 6 months.
Transient-amplifying cells might maintain homeostasis for >1 month when separated from their limbus in vivo. This model will be useful for future stem cell research or wound healing models.
通过移植不锈钢环（环内有或无缺损），对活体分离的角膜缘上皮细胞进行特征描述。
使用 10-0 尼龙间断缝线，将直径为 8mm、宽 300μm、深 250μm 的不锈钢环移植到经角膜真空环钻切割至 250μm 深度的兔角膜基质中。在环内去除上皮细胞，1 周后评估再上皮化情况。进行苏木精染色和针对 p63、Ki67 和细胞角蛋白 3 的免疫染色，以对角膜上皮进行表型分析。在移植后，在环内中央（直径 4、5 和 6mm）创建角膜上皮缺损。当再上皮化完成时，在环内持续创建中央上皮缺损，直至细胞耗尽。还分析了创建的缺损数量，以评估再上皮化的潜力。
环移植的角膜基质炎症反应较少，当环内上皮完全去除时，完全性上皮缺损可持续≥1 个月。角膜环移植后角膜感觉明显下降（P<0.05）。免疫染色显示 p63、Ki67 和细胞角蛋白 3 的表达模式与对照组相似。当将环移植到完整的角膜中时，内侧上皮在体内可防止≤6 个月的上皮缺损。
当活体分离的角膜缘上皮细胞从其缘部分离时，短暂扩增细胞可能会维持 1 个月以上的内稳态。该模型将有助于未来的干细胞研究或伤口愈合模型。
