[关键词]
[摘要]
目的:通过体外细胞实验和在体动物实验初步评价MIL60在眼局部应用的安全性。
方法:常规培养人角膜上皮细胞,CCK8法体外检测MIL60抗体对角膜上皮细胞毒副作用。流式细胞仪检测MIL60对角膜上皮细胞的凋亡影响。正常SD大鼠结膜下注射MIL60抗体,观察眼部反应情况,通过裂隙灯显微镜检查、Draize的评分系统和病理切片等,分析结膜和角膜病理改变。建立SD大鼠角膜上皮缺损模型,结膜下注射MIL60抗体,观察角膜上皮愈合情况。
结果:MIL60不影响角膜上皮细胞增殖,不促进角膜上皮细胞的凋亡。结膜下注射MIL60抗体后,大鼠角结膜和眼部其他各组织形态正常,组织学检查显示结构正常,未见炎症细胞浸润。结膜下注射MIL60不影响角膜上皮愈合。
结论:MIL60结膜下应用安全性较好,不影响角膜上皮细胞正常功能。
[Key word]
[Abstract]
AIM:To verify the safety application of MIL60 in the treatment of corneal neovascularization both
in vivo and
in vitro.
METHODS: We observed the biological characteristics of human corneal epithelial cells. The cell proliferation was analyzed using CCK-8 assay, which also used to test the toxicity of MIL60 and the solvent on cultured human corneal epithelial(HCE). FACs was used to analyze the apoptosis of HCE after treated with MIL60. Also we evaluated the effect of subconjunctival injection of MIL60 on corneal epithelial healing model in normal rat and rats with epithelium defect through slit lamp-microscopy, Draize scores and histopathology way.
RESULTS: The proliferation speed of HCE in three groups was the same. MIL60 did no harm on the proliferation of HCE and the apoptosis of HCE, and has no effect on corneal epithelial healing and other parts of the ocular in rats without inflammation cells infiltration.
CONCLUSION: When given subconjunctival injection, Mil60 does no harm to the proliferation and apoptosis of HCE, and is safe with ocular application.
[中图分类号]
[基金项目]
武警总医院院内课题(No.WZ2015011)