期刊
MICROCIRCULATION
卷 27, 期 6, 页码 -出版社
WILEY
DOI: 10.1111/micc.12623
关键词
Fick's Law; fundus fluorescein angiography; non-toxic; permeability; quantitative; retina; sensitive; vessel
资金
- Medical Research Council [MR/P003214/1, MR/L01985X/1]
- Masonic Charitable Foundation
- National Eye Research Centre
- Medical Research Council [MR/P003214/1] Funding Source: researchfish
- MRC [MR/L01985X/1, MR/P003214/1] Funding Source: UKRI
Objective The gold standard for measuring blood-retinal barrier permeability is the Evans blue assay. However, this technique has limitations in vivo, including non-specific tissue binding and toxicity. This study describes a non-toxic, high-throughput, and cost-effective alternative technique that minimizes animal usage. Methods Sodium fluorescein fundus angiography was performed in non-diabetic and diabetic Brown Norway rats on days 0, 7, 14, 21, and 28. Sodium fluorescein intensity in the retinal interstitium and a main retinal vessel were measured over time. The intensity gradients were used to quantify retinal vascular permeability. Post-study eyes were fixed, dissected, and stained (isolectin B4) to measure required parameters for permeability quantification including total vessel length per retinal volume, radius, and thickness. Results In the non-diabetic cohort retinal permeability remained constant over the 28-day study period. However, in the diabetic cohort there was a significant and progressive increase in retinal permeability from days 14-28 (P < .01, P < .001, P < .0001). Conclusions This novel imaging methodology in combination with mathematical quantification allows retinal permeability to be non-invasively and accurately measured at multiple time points in the same animal. In addition, this technique is a non-toxic, rapid, sensitive, and cost-effective alternative to the Evans blue assay.
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