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In vitro toxicity of gentamicin to corneal epithelial cells
E C Alfonso1, D M Albert, K R Kenyon
1Howe Laboratory, Massachusetts Eye and Ear Infirmary, Boston.
Abstract:
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.
Insights
Gentamicin sulfate causes cell damage in rabbit corneal epithelial cells at high concentrations. This aminoglycoside toxicity was observed in vitro, affecting cell morphology and viability.
Area of Science:
- Ophthalmology
- Cell Biology
- Toxicology
Background:
- Gentamicin sulfate is an aminoglycoside antibiotic.
- Aminoglycosides are known to cause toxicity in human kidney and conjunctiva.
- The in vitro effects of gentamicin on corneal epithelium are not well-documented.
Purpose of the Study:
- To investigate the morphologic and viability changes in rabbit corneal epithelial cells exposed in vitro to gentamicin sulfate.
- To determine the concentration-dependent toxicity of gentamicin sulfate on corneal epithelial cells.
- To correlate light and electron microscopy findings with cell viability.
Main Methods:
- Rabbit corneal epithelial cells were cultured in vitro.
- Cells were exposed to varying concentrations of gentamicin sulfate (0-5,000 µg/ml).
- Morphologic changes were assessed using phase contrast and electron microscopy.
- Cell viability was quantified and statistically analyzed.
Main Results:
- Increased cytoplasmic granularity and ultrastructural intralysosomal bodies were observed at gentamicin concentrations ≥250 µg/ml after 48 hours.
- No significant changes were noted at 0 or 50 µg/ml.
- A statistically significant reduction in viable cells was observed at concentrations ≥250 µg/ml compared to 50 µg/ml for exposure periods of 48 hours or more.
Conclusions:
- Gentamicin sulfate exhibits dose-dependent toxicity to rabbit corneal epithelial cells in vitro.
- The observed toxicity is characterized by morphologic alterations and reduced cell viability.
- These findings suggest a potential risk of aminoglycoside-induced corneal toxicity.