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KMID : 0350519940470041777
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Journal of Catholic Medical College
1994 Volume.47 No. 4 p.1777 ~ p.1791
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In Vitro Study for Staining and Toxicity of Rose Bengal on Cultured Bovine Corneal Endothelial Cells
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Abstract
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It has been believed that rose bengal does not stain normal healthy cells but rather stains degenerated or dead cells and mucous strands in vivo. And that rose bengal does not have an intrinsic toxiceffect. For therapeutical purposes, rose bengal
has
been applied in the treatment of viral infections and neovascularization as a photosensitizing dye.
But it has been recently reported that rose bengal is not actually a vital dye and that it has an intrinsic toxic effect. Therefore we conducted a study to determine whether rose bengal is actually a vital dye and whether it has an intrinsic
toxic
effect and we also examined the effects influencing the proliferation of cultured bovine corneal endothelial cells.
We observed staining patterns and morphological changes by means of fluorescein photomicroscope and invers microscope, measured the intrinsic toxic effect and additional phototoxic effect by a Cr51 labelling study, and counted the number of cell
by
a
coulter counter in cel proliferation study.
@ES The results were as follows:
@EN 1. Cells were stained even after a short exposure(one second) to rose bengal in a fluorescein photomicroscope. The cell nucleus was initially stained, and followed by the staining of cytoplasm. The longer the exposure time, the more
cytoplasmic
staning was observed. At a concentration of 0.001%, the entire cell layer was not stained and at a concentration above 0.01%, the cell layer was diffusely stained, but at a concentration of 5% some areas of cellular detachment and lysis were
noted.
2. The higher the concentration of rose bengal and the longer the exposure time, the more cellular changes were observed in the invert microscope. Cellular damage included cell swelling, intracytoplasmic vacuole formation, cell detachment and
lysis.
3. An intrinsic toxic effect of rose bengal was observed in proportion to concentration and exposure time, and the light induced additional toxic effect was higher than the intrinsic toxic effect without light, which was statistically
significant.
4. After exposure to rose bengal, cell proliferation was inhibited and concentration-dependent, which was statistically significant above a concentration of 0.01%.
These results suggest that rose bengal may not actually be a vital dye on cultured bovine corneal endothelial cells, and that rose bengal has intrinsic toxic effect with additional phototoxic effect. Furthermore, after exposure to rose bengal,
cell
proliferation was markedly inhibited.
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