Kanayama S, Nishida K, Yamato M, Hayashi R, Maeda N, Okano T, Tano Y
Department of Ophthalmology, University of Washington Medical Center, 1959 NE Pacific Street, Seattle, WA 98195, USA.
Br J Ophthalmol. 2009 Feb;93(2):263-7. doi: 10.1136/bjo.2008.141580.
In clinical trials, eyes transplanted with cultured oral mucosal epithelial cell sheets have shown increased neovascularisation compared with eyes treated with cultured corneal epithelial cell sheets. As reported recently, soluble vascular endothelial growth factor receptor-1 (soluble VEGFr-1) is a main factor to maintain a corneal avascularity.
To investigate soluble VEGFr-1 of cultured corneal epithelial cells (CCE) and cultured oral mucosal epithelial cells (COE) in vitro.
Rabbit corneal and oral mucosal epithelial cells were co-cultured with mitomycin C-treated NIH/3T3 cells on culture plates. After CCE and COE were multilayered, culture medium was replaced by basal medium and incubated. Protein secretion of soluble VEGFr-1 was assessed in conditioned medium from CCE and COE by ELISA. Angiogenic potential was examined by invasion, migration assays with human umbilical vein endothelial cells (HUVECs) in addition to recombinant soluble VEGFr-1.
CCE secreted a significantly higher amount of soluble VEGFr-1 than did COE. Recombinant soluble VEGFr-1 significantly suppressed HUVEC migration induced by COE, without suppression in CCE. In conclusion, these findings suggest that low protein levels of soluble VEGFr-1 may lead to corneal neovascularisation after COE sheet transplantation.
在临床试验中，与接受培养的角膜上皮细胞片治疗的眼睛相比，接受培养的口腔黏膜上皮细胞片移植的眼睛显示出新生血管形成增加。最近有报道称，可溶性血管内皮生长因子受体-1（可溶性VEGFr-1）是维持角膜无血管状态的主要因素。
在体外研究培养的角膜上皮细胞（CCE）和培养的口腔黏膜上皮细胞（COE）中的可溶性VEGFr-1。
将兔角膜和口腔黏膜上皮细胞与丝裂霉素C处理的NIH/3T3细胞在培养板上共培养。在CCE和COE形成多层后，用基础培养基替换培养基并进行孵育。通过ELISA评估CCE和COE条件培养基中可溶性VEGFr-1的蛋白分泌。除重组可溶性VEGFr-1外，还通过用人脐静脉内皮细胞（HUVEC）进行侵袭和迁移试验来检测血管生成潜力。
CCE分泌的可溶性VEGFr-1量明显高于COE。重组可溶性VEGFr-1显著抑制了由COE诱导的HUVEC迁移，而对CCE无抑制作用。总之，这些发现表明，可溶性VEGFr-1的低蛋白水平可能导致COE片移植后角膜新生血管形成。
