FENTANYL GENOTOXICITY EVALUATION VIA COMET ASSAY
Abstract
DNA is our main genetic structure that can be affected by various molecules we come into contact with. Therefore, there is a high probability of DNA damage. Genotoxicity tests involve systematic methods designed to assess the safety of drugs and chemicals. Comet assay evaluates the possible genotoxicity resulting from DNA single-strand breaks.
Fentanyl is a powerful painkiller from the group called opioid analgesics administered intravenously. In this research, we evaluated the possible DNA damage in human lymphocytes after fentanyl exposure via single-cell gel electrophoresis (Comet Assay).
DNA damage in healthy human peripheral lymphocytes treated with fentanyl was investigated via comet assay. Lymphocytes were treated with 5, 10, 20, and 40 µg/mL doses of fentanyl for 1 hour. After the incubation period, the cells' DNA tail length, tail intensity, and tail moment values were evaluated by comparing them with the spontaneous control and positive control data. EtBr stained slides were visualized under the fluorescent microscope. Fentanyl induced the comet parameters such as tail length, tail intensity, and tail moment but dose-dependent increase was not obtained after fentanyl administration. Fentanyl showed the highest tail length and tail moment value at the dose of 10 µg/mL. The highest tail intensity value was obtained at the dose of 40 µg/mL fentanyl administration.
This study aimed to reveal previously undiscovered genotoxicity of fentanyl on healthy human lymphocytes in vitro via comet assay. Fentanyl exposure induces DNA damage in healthy human lymphocytes, as shown via comet parameters (tail length, tail intensity, and tail moment). Genotoxic effect does not display a consistent dose-dependent increase.
Keywords
Comet assay, Single-cell gel electrophoresis, DNA damage, Lymphocyte culture, Genotoxicity, Fentanyl
References
- [1] Waskell L. A study of the mutagenicity of anesthetics and their metabolites. Mutation Research 1978; 57(2): 141–153.
- [2] Kant I, van Rijssen-Moll M, Borm PJ. Simulation of nitrous oxide concentrations in operating and recovery rooms. Annals of Occupational Hygiene 1990; 34(6): 575–583.
- [3] Stanley TH, Philbin DM, Coggins CH. Fentanyl-oxygen anaesthesia for coronary artery surgery: cardiovascular and antidiuretic hormone responses. Canadian Anaesthetists' Society Journal 1979; 26(3): 168–172.
- [4] Stanley TH. The fentanyl story. The Journal of Pain 2014; 15(12): 1215–1226.
- [5] Gasperini S, et al. The genotoxicity of acrylfentanyl, ocfentanyl and furanylfentanyl raises the concern of long-term consequences. International Journal of Molecular Sciences 2022; 23(22): 14406.
- [6] Collins AR. The comet assay for DNA damage and repair. Mol Biotechnol. 2004;26(3):249-261.
- [7] Singh NP, McCoy MT, Tice RR, Schneider EL. A simple technique for quantitation of low levels of DNA damage in individual cells. Experimental Cell Research 1988; 175(1): 184–191.
- [8] Olive PL, Banáth JP, Durand RE. Heterogeneity in radiation-induced DNA damage and repair in tumor and normal cells measured using the "comet" assay. Radiation Research 1990; 122(1): 86–94.
- [9] Yeager MP, et al. Intravenous fentanyl increases natural killer cell cytotoxicity and circulating CD16(+) lymphocytes in humans. Anesthesia and Analgesia 2002; 94(1): 113–119.
- [10] Sardaş S, et al. DNA damage evaluated by the alkaline comet assay in lymphocytes of humans anaesthetized with isoflurane. Mutation Research 1998; 418(1): 1–6.