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. 2015 Jul;7(7):517-24.
doi: 10.14740/jocmr2085e. Epub 2015 May 8.

Effect of distance and duration of illumination on retinal ganglion cells exposed to varying concentrations of brilliant blue green

Kakarla V Chalam  1 , Wenhua Li  1 , Keyvan Koushan  1 , Sandeep Grover  1 , Sankarathi Balaiya  1
Affiliations

Effect of distance and duration of illumination on retinal ganglion cells exposed to varying concentrations of brilliant blue green

Kakarla V Chalam et al. J Clin Med Res. 2015 Jul.

Abstract

Background: The objective of the study was to determine the safety parameters of using brilliant blue green (BBG) for chromovitrectomy by assessing the cytotoxicity of BBG on cultured retinal ganglion cells (RGCs) exposed to illumination.

Methods: RGCs were exposed to two concentrations of BBG (0.25 and 0.5 mg/mL) under metal halide illumination at varying distances (1 and 2.5 cm), intensities (990 and 2,000 Fc), and durations (1, 5 and 15 minutes). Cell viability was assessed using the WST-1 and CellTiter 96(®) AQueous One solution cell proliferation assays.

Results: Using the WST-1 assay, with high-intensity illumination, viability of RGCs ranged from 97.5±16.4% of controls with minimum BBG and light exposure (0.25 mg/mL BBG and illuminated for 1 minute at 2.5 cm distance) to 53.1±11.3% of controls with maximum BBG and light exposure (0.50 mg/mL and illuminated for 15 minutes at 1 cm distance; P < 0.01). With medium-intensity illumination, RGCs showed better viability, ranging from 95.1±7.2% of controls with minimum BBG and light exposure to 72.3±12.8% of controls with maximum BBG and light exposure. CellTiter 96(®) AQueous One assay showed similar results.

Conclusion: RGCs seem to safely tolerate up to 5 minutes of exposure to 0.5 mg/mL BBG under diffuse medium-intensity illumination (990 Fc).

Keywords: Brilliant blue green; Chromovitrectomy; Cytotoxicity; Endoillumination; Retinal ganglion cells.

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Figures

Figure 1
Figure 1
Schematic representation explaining the exposure of RGCs to BBG during intraocular surgery as well as cell culture setup. (a) Diffuse light illumination. (b) Focal illumination. LS: light source; LP: light pipe; D: distance; RGC: retinal ganglion cell; BBG: brilliant blue green.
Figure 2
Figure 2
Experimental illustration. (a) Measurement of metal halide light illumination using a light meter at a certain distance from the light source. (b) Measurement of metal halide light illumination using a light meter directly from the light source. (c) Exposure of cultured RGCs to metal halide illumination. LS: light source; LP: light pipe; LM: light meter; dist: distance; RGC: retinal ganglion cell; BBG: brilliant blue green.
Figure 3
Figure 3
Evaluating the cytotoxic effect of brilliant blue green at 1 cm metal halide distance illumination on retinal ganglion cells using WST-1 assay at high (a) and medium illumination (b). X-axis represents the time dependent exposure in minutes; Y-axis represents number of viable cells expressed as percent of control (N = 4); (c) represents the trend line.
Figure 4
Figure 4
Evaluating the cytotoxic effect of brilliant blue green at 2.5 cm metal halide distance illumination on retinal ganglion cells using WST-1 assay at high (a) and medium illumination (b). X-axis represents the time dependent exposure in minutes; Y-axis represents number of viable cells expressed as percent of control (N = 4); (c) represents the trend line.
Figure 5
Figure 5
Evaluating the cytotoxic effect of brilliant blue green at 1 cm metal halide distance illumination on retinal ganglion cells using CellTiter 96® AQueous One proliferation assay at high (a) and medium illumination (b). X-axis represents the time dependent exposure in minutes; Y-axis represents number of viable cells expressed as percent of control (N = 4); (c) represents the trend line.
Figure 6
Figure 6
Evaluating the cytotoxic effect of brilliant blue green at 2.5 cm metal halide distance illumination on retinal ganglion cells using CellTiter 96® AQueous One proliferation assay at high (a) and medium illumination (b). X-axis represents the time dependent exposure in minutes; Y-axis represents number of viable cells expressed as percent of control (N = 4); (c) represents the trend line.
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