Ozgurtas Taner, Tekin Sercan, Yesildal Fatih, Karaca Umut, Aydin Fevzi Nuri, Ugurlu Muhammed Talha, Ozler Mehmet, Durukan Hakan
Department of Medical Biochemistry, Gulhane Military Medical Academy, Ankara 06010, Turkey.
School of Medicine, Gulhane Military Medical Academy, Ankara 06010, Turkey.
Int J Ophthalmol. 2016 Sep 18;9(9):1265-70. doi: 10.18240/ijo.2016.09.05. eCollection 2016.
To examine changes in retinal vasculature after treatment with different oxygen concentrations from common retinopathy of prematurity (ROP) models and to determine a novel and practical ROP model.
A sample of 14 newborn Sprague-Dawley rats was used. The study group (n=7) was exposed to 95% oxygen for 4h per day followed by normoxic laboratory conditions for 20h. This cycle was repeated for 14d. The control group (n=7) was subjected to normobaric normoxic conditions. On postnatal day 14 (P14), the two groups were placed in room air for 7d. On P21, the two groups were examined using indirect ophthalmoscopy. All eyes were enucleated for immunofluorescence (IF) staining of the vasculature of retinas and analysis of vascular endothelial growth factor (VEGF), hypoxia inducible factor-1 alpha (HIF-1α), placental growth factor (PLGF) in vitreous humor, and then the rats were sacrificed by decapitation. All procedures were repeated using another litter of 14 pups.
In the study group and under normobaric hyperoxic conditions, retinal neovascularization and peripheral avascular retina were determined in 85% of the rats through indirect ophthalmoscopic examination. Also IF staining of retina of the study group showed retarded peripheral vascular growth. The difference between the two groups for VEGF, HIF-1α and PLGF concentrations of vitreous humor was statistically significant (P=0.003, 0.007, 0.027 respectively).
Fluctuating oxygen concentrations are primarily responsible for retinal neovascularization. Our new ROP model is practical and applicable for all retinal neovascularization studies, considering the laboratory procedures.
研究不同氧浓度处理常见早产儿视网膜病变(ROP)模型后视网膜血管系统的变化,并确定一种新颖实用的ROP模型。
使用14只新生Sprague-Dawley大鼠作为样本。研究组(n = 7)每天暴露于95%氧气中4小时,随后置于常氧实验室条件下20小时。此循环重复14天。对照组(n = 7)置于常压常氧条件下。出生后第14天(P14),两组置于室内空气中7天。在P21时,使用间接检眼镜检查两组。所有眼睛摘除后进行视网膜血管系统的免疫荧光(IF)染色,并分析玻璃体液中的血管内皮生长因子(VEGF)、缺氧诱导因子-1α(HIF-1α)、胎盘生长因子(PLGF),然后通过断头处死大鼠。使用另一窝14只幼崽重复所有程序。
在研究组和常压高氧条件下,通过间接检眼镜检查在85%的大鼠中确定有视网膜新生血管形成和周边无血管视网膜。研究组视网膜的IF染色也显示周边血管生长迟缓。两组玻璃体液中VEGF、HIF-1α和PLGF浓度的差异具有统计学意义(分别为P = 0.003、0.007、0.027)。
氧浓度波动是视网膜新生血管形成的主要原因。考虑到实验室操作,我们的新ROP模型实用且适用于所有视网膜新生血管形成研究。