Purchase this article with an account.
C.–C. Lai, L.–H. Chuang, N.–K. Wang, L. Yeung, Y.–P. Chen, K.–J. Chen, W.–C. Wu, T.–L. Chen; Selective Staining of Membrane Adjuvant With Whole Blood in Trypan Blue–Assisted Membrane Peeling . Invest. Ophthalmol. Vis. Sci. 2006;47(13):4074.
Download citation file:
© ARVO (1962-2015); The Authors (2016-present)
To reduce the toxic effect of trypan blue (TB) on retinal pigment epithelium (RPE) and the retina by selectively staining the internal limiting membrane (ILM) or epiretinal membrane with aid of whole blood (WB) during TB–assisted membrane peeling.
In the in vitro study, cultured human RPE cells were covered with balanced saline solution (BSS), heparinized WB and then exposed to various concentrations of TB solution. One cohort was incubated in the dark, the other cohort was exposed to light, both for 20 minutes. The toxic effect of TB in cultured human RPE cells was evaluated by mitochondrial dehydrogenase assay. In the clinical study, a total of 30 patients who underwent TB–assisted membrane peeling were enrolled. Autologous heparinized WB was applied to cover the macular area, then selectively removed to expose the central macula (except the macular hole area) for staining. TB solution (0.75 mg/ml) diluted with 5 % glucose was used to stain the macular area. The stained membrane was removed completely.
Cultured human RPE cells covered by WB showed no decrease of mitochondrial dehydrogenase even in 3.0mg/ml concentration of TB for 20 minutes either with or without light exposure. Conversely, those exposed to TB and covered with BSS only demonstrated a significant decrease of mitochondrial dehydrogenase after incubation in 3 mg/ml for 20 minutes in the dark and in 3 to 0.75 mg/ml with light exposure. Clinically, we studied 24 patients with macular holes and 6 with macular pucker. The ILM or epiretinal membrane was selectively stained by TB adjuvant with WB and the stained ILM could be removed completely. The TB solution diluted with 5% glucose showed minimal dispersion during application.
WB can be used to reduce the toxic effect of TB in PRE cell culture, as well as selective staining of ILM and epiretinal membrane during TB–assisted membrane peeling. TB diluted with 5% glucose can decrease dispersion during surgical application. Complete removal of the stained membrane could prevent TB retention, and may further reduce its toxic effect on RPE and retina.
This PDF is available to Subscribers Only