Staining Properties of Brilliant Blue Depending on Different Incubation Times and Solvents in Humans

Abstract

Purpose: Intravitreous injection of vital dyes, e.g. brilliant blue (BBG), promotes better visualization of the internal limiting membrane (ILM). This paper investigates the staining properties of BBG depending on different incubation times and 2 types of solvents - 5% glucose (GL) or saline solution - in a prospective study in patients. Methods: This paper investigates various aspects of BBG in various methods. An interventional prospective study was conducted in patients to examine the binding properties of the blue dye diluted in either saline or 5% GL to epiretinal membranes (ERMs) and ILMs. Forty-nine consecutive patients older than 18 years scheduled for macular ERM, vitreomacular traction and macular hole surgeries were prospectively recruited. The primary outcomes of this study were the degree of ILM and ERM staining. The secondary outcomes of the study were the need of reinjection of BBG or any other dye, the ability of BBG to stain the vitreous, and frequency of complications. The staining of the ILM and ERM were graded as no staining, little, moderate or strong staining. Results: There was no correlation between age (p = 0.32) or gender (p = 0.33) in the staining affinity of BBG to either the ILM or ERM. BBG may be an appropriate staining agent for the ILM in the majority (82%) of surgeries; however, in approximately half of the cases (45%) surgeons considered BBG not enough for ERM coloring and visualization. There is a tendency of BBG to stain the ILM better when saline solution is used compared to GL 5%; however, this was not statistically significant (p = 0.64). There was no difference in the staining efficacy of BBG to the ERMs by either solution (p = 0.70), despite the low staining affinity. Conclusion: BBG became the state-of-the-art dye for ILM identification. Differences in staining properties may imply that BBG should not be considered as first-line stain for ERM surgery. BBG is effective in ILM staining in over 80% of macular hole surgeries.

An evaluation of two heavier-than-water internal limiting membrane-specific dyes during macular hole surgery

“…We standardised the technique of staining between the two dyes and importantly used the same contact time between the dyes and retina, before washout. Both dyes showed selective ILM staining at this duration with low affinity for epiretinal membrane (ERM) as previously described for BBG, [22] and allowing the presence of epiretinal tissue on the ILM to be detected by negative staining as we have previously described for BBG [11]. This is a useful tool during surgery, allowing areas of bare ILM to be identified, but we were also careful to avoid areas of negative staining when making the contrast measurements by using areas of maximum stain around the peeled area as others have done [23].…”

“…Importantly the two groups were well matched in terms of the amount of vitreous debris present on the ILM, which, if unequal between the two groups, could have confounded contrast measurements. It is possible that AV17 may require a longer time for optimum ILM staining than BBG although contact time was found to have a limited effect on staining intensity with BBG in previous studies [22].…”

Foetal bovine serum can reduce toxicity of indocyanine green, brilliant blue G and trypan blue in ARPE‐19 cellular model that suggests new surgical staining protocols for internal limiting membrane peeling procedure

“…Rodrigues et al 24 and Hernandez et al 25 However, this study was done on the in vitro porcine retinas and the results may not be comparable to the situation in the human retina clinically. In other words, the dyes staining capability may differ from species to species.…”

Assessment of Macular Function Following Internal Limiting Membrane Peeling With ILM Blue

“…BBG performs a selective ILM staining with a low affinity for ERM [4,22]. This is important because the visualization and removal of the ILM is challenging as the structure is about 1 µm thick and is formed by the end feet of Müller cells and astrocytes.…”

“…A possible explanation of Januschowski et al for the improved biocompatibility of ILM Blue could be that the dye solutions sold by a manufacturer place special emphasis on the purity of the ingredients and quality control [3]. It could also be argued that the presence of 4% PEG has a potential neuroprotective effect [22]. In particular, potential toxicity is a decisive factor in the choice of the vital dye and is part of the current research, as the currently used vital dyes have shown retinal toxic reactions [5,[9][10][11]18,19].…”