Please use this identifier to cite or link to this item: https://doi.org/10.1159/000353869
Title: Effect of Novel Vital Dyes on Outer Blood-Retina Barrier Function in Cultured Human Retinal Pigment Epithelium
Authors: LIU ZENGPING 
Meyer, Carsten H
Stanzel, Boris V
Keywords: Science & Technology
Life Sciences & Biomedicine
Ophthalmology
Blood-retina barrier
Cell culture
Chromovitrectomy
Retinal pigment epithelium
Tight junctions
Transepithelial electrical resistance
INTERNAL LIMITING MEMBRANE
MACULAR HOLE SURGERY
INDOCYANINE GREEN
TIGHT JUNCTIONS
IN-VITRO
CELLS
PERMEABILITY
MONOLAYERS
MECHANISMS
SAFETY
Issue Date: 1-Jan-2013
Publisher: KARGER
Citation: LIU ZENGPING, Meyer, Carsten H, Stanzel, Boris V (2013-01-01). Effect of Novel Vital Dyes on Outer Blood-Retina Barrier Function in Cultured Human Retinal Pigment Epithelium. OPHTHALMOLOGICA 230 (s2) : 33-40. ScholarBank@NUS Repository. https://doi.org/10.1159/000353869
Abstract: PURPOSE: To assess tight junction (TJ) integrity in cultured human fetal retinal pigment epithelium (HFRPE) after exposure to clinically relevant novel vital dyes. METHODS: HFRPE floater cells were harvested from RPE primary cultures of 4 donor eyes and seeded on polyester Transwell® for 4-6 weeks. The apical compartments of well-differentiated cultures were exposed to 0.005 mg/ml Coomassie violet R200 (CVR), methyl 2B (M2B) or Orange II. Periods of 30-300 s were chosen to mimic surgical exposure times, while 3 h was used for toxicity assays, with subsequent washout. Cell-cell junctions were studied by immunofluorescence (zonula occludens-1, ZO-1). Transepithelial electrical resistance (TER) was measured regarding blood-retina barrier (BRB) function. RESULTS: At 4-6 weeks after confluence, HFRPE had grown into pigmented hexagonal monolayers with stable TER values (451-1,520 Ω·cm(2)). After 300-second dye treatments, a continuous ZO-1 signal was detected in all vital dye-treated groups 1.5 h after exposure, whereas trypsin controls showed patchy loss of the TJ stain. TER of CVR-, M2B- and Orange-II-treated groups had dropped 1.5 h after exposure to 148 ± 58.4, 162 ± 23.7 and 164 ± 18.5 Ω·cm(2), respectively, compared to 73 ± 44.9 Ω·cm(2) in positive controls. After 3 h of exposure to 0.005 mg/ml vital dyes in thick drops, TER maintained similar levels to those prior to exposure (90.8 ± 4.7% of the original values, 93.8 ± 6.5 and 91.9 ± 3.6%, respectively), together with no difference from the vehicle controls (94.8 ± 6.6%). TER values recovered in all groups to prior levels within 3 days. CONCLUSION: Novel vital dyes (CVR, M2B and Orange II) caused no outer BRB function alteration.
Source Title: OPHTHALMOLOGICA
URI: https://scholarbank.nus.edu.sg/handle/10635/206752
ISSN: 0030-3755
1423-0267
DOI: 10.1159/000353869
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