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A comparison of BGM and LLC-PK1 cells for the evaluation of nephrotoxicity
Carlos A Tagliati1, Diego Romero, Elisabete C R Dutra
1Laboratório de Toxicologia Experimental, Departamento de Análises Clínicas e Toxicológicas da Faculdade de Farmácia da Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais, Brasil. carlostagliati@ufmg.br
Abstract:
Nephrotoxicity is one of the most frequent effects observed after the use of medicine. Such situations have been tardily discovered because of existing methods to determine toxicity. The validation of sensitive, alternative methods for the early identification of toxic effects is as important as restrictions on the use of animals. In this light, the present study evaluated the effects of gentamicin on BGM and LLC-PK1 cells, using MTT and Neutral Red (NR). Although the LLC-PK1 cell line is used for toxicological studies, the BGM cell line is relatively new for this purpose. MTT (BGM: EC(50) = 6.29 mM; LLC-PK1: EC(50) = 8.01 mM) was found to be more sensitive than NR (EC(50) was greater than 10 mM for both cells). By using MTT, both cells demonstrated the involvement of mitochondria in a manner that was dose dependent, with an apoptotic process occurring at the concentrations of 1 and 3 mM and necrosis at concentrations above 4 mM. It could, therefore, be concluded that 1) BGM appears to be useful in the study of the mechanism of nephrotoxicity caused by gentamicin and 2) because of its sensitivity to MTT, in addition to its ease of manipulation, it is believed that the BGM cell line can also be used as an alternative method to evaluate nephrotoxicity.
Insights
The BGM cell line shows promise as a new method for detecting gentamicin nephrotoxicity early. Using MTT assays, it proved more sensitive than NR, identifying apoptosis and necrosis dose-dependently.
Area of Science:
- Toxicology
- Cell Biology
- Pharmacology
Background:
- Nephrotoxicity is a common adverse drug effect, often detected late.
- Current toxicity testing methods need improvement for early detection and reduced animal use.
Purpose of the Study:
- To evaluate gentamicin's nephrotoxic effects on BGM and LLC-PK1 cells.
- To compare the sensitivity of MTT and Neutral Red (NR) assays for toxicity detection.
- To explore the utility of the BGM cell line as an alternative toxicity testing model.
Main Methods:
- Gentamicin exposure to BGM and LLC-PK1 cell lines.
- Cell viability assessed using MTT and Neutral Red (NR) assays.
- Dose-dependent effects and cell death mechanisms (apoptosis, necrosis) analyzed.
Main Results:
- MTT assay was more sensitive (EC50: BGM=6.29 mM, LLC-PK1=8.01 mM) than NR (EC50 >10 mM for both).
- Gentamicin induced mitochondrial involvement, apoptosis at 1-3 mM, and necrosis above 4 mM.
- BGM cells exhibited clear dose-dependent responses.
Conclusions:
- The BGM cell line is suitable for studying gentamicin-induced nephrotoxicity mechanisms.
- BGM cells, with MTT assay sensitivity, offer a viable alternative for nephrotoxicity evaluation, reducing animal testing.
