In vitro toxicity of gentamicin to corneal epithelial cells.
We report morphologic and viability changes in the rabbit corneal epithelium exposed in vitro to gentamicin sulfate. Rabbit corneal epithelial cells (3 x 10(5)) were seeded in replicate 24-well plates at their first in vitro passage, and gentamicin in concentrations of 0, 50, 250, 500, 1,000, or 5,000 micrograms/ml was added to the tissue culture medium beginning 7 days after subculture. By phase contrast microscopy, changes in cell morphologic appearance, particularly increased cytoplasmic granularity, were observed in the 5,000-micrograms/ml groups as early as 24 h after introduction of the drug. At 48 h, similar findings were observed in the 250-micrograms/ml group and at all higher concentrations. The cytoplasmic granularity was not noted in the 0- or 50-micrograms/ml groups. By electron microscopy, these observations correlated with ultrastructural findings of increased accumulations of intralysosomal bodies beginning in the 250-micrograms/ml group after 48 h of exposure to gentamicin. Furthermore, a statistically significant difference (p = 0.001) was demonstrated between the total number of viable cells in the low-dose group (50-micrograms/ml) and the high-dose groups (greater than or equal to 250-micrograms/ml) for exposure periods of 48 h or more. These findings demonstrate aminoglycoside toxicity to corneal epithelial cells in vitro similar to that seen in the human kidney and conjunctiva.
Alfonso
E C
EC
Howe Laboratory, Massachusetts Eye and Ear Infirmary, Boston.
Albert
D M
DM
Kenyon
K R
KR
Robinson
N L
NL
Hanninen
L
L
D'Amico
D J
DJ
D000818:Animals; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D003315:Cornea / Q000187:drug effects* / Q000648:ultrastructure; D004848:Epithelium / Q000187:drug effects / Q000648:ultrastructure; D005839:Gentamicins / Q000633:toxicity*; D008858:Microscopy, Phase-Contrast; D011817:Rabbits; D013997:Time Factors
D005839:Gentamicins