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Published on: November 16, 2016
In vitro toxicity of antibiotics to LLC-RK(1) rabbit kidney cells
1Cellular and Experimental Toxicology, Safety of Medicines, ICI Pharmaceuticals, Mereside, Alderley Park, Macclesfield, Cheshire SK10 4TJ, England.
Abstract:
The renal toxicity of cephalosporin and aminoglycoside antibiotics has been studied in an in vitro system using the rabbit kidney cell line LLC-RK(1). The effect of a rabbit kidney S-9 metabolizing system on the toxicity the antibiotics was also studied. Cultures of the LLC-RK(1) cells were established in microtitre plates and exposed to test compounds in the concentration range 0 to 2000 mug/ml, either in the presence or absence of an S-9 metabolizing system. The compounds were in contact with the cultures for a period of 48 hr before viability of the cells was determined. Assay of cell viability was based on the mitochondrial conversion of 3-(4,5-dimethyl thiazol-2-yl)-2,5-diphenyl tetrazolium bromide to a coloured formazan product (MTT Assay). Duplicate cultures were also tested with a neutral red staining assay. Cephalosporins tested were cephaloridine, cephalothin, cephapirin, cefotaxime, ceftazidime, cephaloglycin, cefoperazone and cefazolin. The aminoglycoside antibiotics tested were gentamicin, neomycin and kanamycin. A good correlation was seen between in vitro toxicity and in vivo effects for these compounds. Further techniques were investigated to assess sub-cytolethal impairment of renal function in the LLC-RK(1) cells. Both in vivo clearance studies and in vitro kidney slice methods have used transport of the organic ions tetraethyl ammonium and p-aminohippurate as indices of renal function. Therefore the ability of LLC-RK(1) cells to transport these ions was investigated. Where ion transport was demonstrated the effect of nephrotoxic antibiotics on this kidney function was studied.
Insights
This study evaluated the in vitro renal toxicity of cephalosporin and aminoglycoside antibiotics using rabbit kidney cells. The findings show a good correlation between in vitro toxicity and known in vivo effects, aiding in predicting drug nephrotoxicity.
Area of Science:
- Pharmacology
- Toxicology
- Cell Biology
Background:
- Renal toxicity is a significant concern for cephalosporin and aminoglycoside antibiotics.
- Existing methods for assessing nephrotoxicity can be limited.
- In vitro models offer a promising avenue for predicting in vivo drug effects.
Purpose of the Study:
- To establish and validate an in vitro system for assessing the renal toxicity of cephalosporin and aminoglycoside antibiotics.
- To investigate the influence of a metabolic activation system (S-9) on antibiotic-induced cytotoxicity.
- To explore the potential of LLC-RK(1) cells to model sub-lethal renal function impairment.
Main Methods:
- Utilized the rabbit kidney cell line LLC-RK(1) cultured in microtitre plates.
- Exposed cells to a range of cephalosporin and aminoglycoside antibiotics (0-2000 µg/ml) with and without a rabbit kidney S-9 metabolizing system for 48 hours.
- Assessed cell viability using MTT assay and neutral red staining; investigated organic ion transport (tetraethyl ammonium and p-aminohippurate) as a marker of renal function.
Main Results:
- Demonstrated a good correlation between the in vitro toxicity observed in LLC-RK(1) cells and established in vivo nephrotoxic effects of the tested antibiotics.
- The S-9 metabolizing system's effect on antibiotic toxicity was evaluated.
- Investigated the capacity of LLC-RK(1) cells to transport organic ions, a key indicator of renal function.
Conclusions:
- The LLC-RK(1) cell line, in conjunction with MTT and neutral red assays, provides a reliable in vitro model for predicting the nephrotoxicity of cephalosporins and aminoglycosides.
- The study supports the utility of this in vitro system for drug safety screening.
- Further research into ion transport in LLC-RK(1) cells can enhance the assessment of sub-lethal nephrotoxicity.
