Determination of DNA damage by alkaline halo and comet assay in patients under sevoflurane anesthesia

Abstract

In this study, we report data on the possible genotoxic effect of inhalation anesthetic sevoflurane (SVF) by comparing two techniques, comet and alkaline halo assay, in peripheral blood lymphocytes (PBL) of patients before, during, and after anesthesia and in controls. DNA single strand breaks were detected in PBL of malignant breast cancer diagnosed patients (stage II-III), who were undergoing mastectomy. Blood samples were taken before the induction of anesthesia, at 120 min of SVF anesthesia, and on the postoperative fifth day. The nuclear spreading factor (NSF) for each cell was assessed by alkaline halo assay, and the total comet score (TCS) was evaluated by comet assay. A statistically significant increase (P < 0.0001) was observed in the mean NSF at 120 min of anesthesia (38.24 +/- 14.14) as compared with samples before anesthesia (12.33 +/- 6.14), and the mean NSF was significantly decreased after the postoperative fifth day (17.89 +/- 9.44). Similar results were obtained by the comet assay with significant increase (P < 0.0001) in DNA damage at 120 min of anesthesia (79.66 +/- 15.28) as compared with samples before anesthesia (36.30 +/- 11.39). The DNA damage was almost with the preoperative damage rates after the fifth day of anesthesia (43.40 +/- 12.19). In conclusion, the study points out a reversible genotoxic effect of SVF and the similar DNA damage levels obtained by comet and alkaline halo assay indicate that although halo assay has a completely different principle, it can conveniently be utilized for the assessment of DNA single strand breakage in individual mammalian cells with its experimental advantages.

DNA damage in patients who underwent minimally invasive surgery under inhalation or intravenous anesthesia

“…Because of its sweet smell, SVF is used for anesthesia induction in both adults and children. Conflicting data have been presented concerning the genotoxic and mutagenic potential of SVF in vitro, in animals and in patients [10][11][12][13][14].…”

Does Neuraxial Anesthesia as General Anesthesia Damage DNA? A Pilot Study in Patients Undergoing Orthopedic Traumatological Surgery

“…The authors of the cited studies followed the state of the DNA on the several postoperative days. They agree that full DNA repair will occur between the 3rd and 5th postoperative day [5,23,34].…”

“…They used two methods-the comet assay and the alkaline halo assay. They described statistically significant damage to DNA by both methods after two-hour surgery [34]. Nogueira and et al described statistically significant damage the day after minor surgery under general anesthesia maintained with desflurane [35].…”

Assessment of Individual Susceptibility to Baseline DNA and Cytogenetic Damage in a Healthy Turkish Population: Evaluation with Lifestyle Factors

“…It has been modified by using lesion-specific enzymes (endonuclease III [Endo III] and formamidopyrimidine glycosylase [Fpg]) to be a more specific and a reliable marker for oxidative DNA damage (Collins et al, 1993). Besides the intensive use of these cytogenetic biomarkers in occupational settings (Karahalil et al, 1998;Burgaz et al, 2002;Karakaya et al, 2005;Cakmak et al, 2010), clinical studies (Sardas et al, 2001;Karahalil et al, 2005;Kadioglu et al, 2009;Burgaz et al, 2011), and environmental exposures (Rossner et al, 2011), they are also used in healthy population studies to determine background cytogenetic damage levels (Barale et al, 1998).…”