ABSTRACT
Leukocytes have been shown to play an important role in the development of tissue injury after ischemia and reperfusion (I/R). In the present study, the effects of tourniquet-ischemia on induction of DNA damage in peripheral leukocytes and on respiratory burst of neutrophils in humans were examined. The DNA damage was measured as increased migration of DNA using the single-cell gel-electrophoresis technique (comet assay). Intracellular production of reactive oxygen species by neutrophils was measured flow-cytometrically using dihydrorhodamine 123 as indicator. Postischemic, significantly increased migration of DNA was found in leukocytes of 20 patients (tourniquet-ischemia of the lower limb for 65-130 min, anterior-cruciate-ligament-reconstruction) and in 10 experiments (1 volunteer, repeated tourniquet-ischemia of the upper limb for 60 min, no operation). DNA effects were most pronounced 5-30 min after tourniquet release, and then declined over a 2 h period, but did not return to preischemic baseline values. A similar time course showed the oxidative status of unstimulated granulocytes during reperfusion. Simultaneously, opposing changes were measured in formyl peptide (f-MLP)- or phorbol ester (PMA)-stimulated granulocytes, which showed a significantly declined respiratory burst reaction after tourniquet-release indicating preactivation of neutrophils by IR. Our data suggest that IR induces genotoxic effects in human leukocytes presumably in response to oxidative stress during reperfusion.
Subject(s)
DNA Damage , Ischemia/pathology , Leukocytes/chemistry , Reperfusion Injury/pathology , Adult , Arm/blood supply , Chemotaxis, Leukocyte/drug effects , Humans , Ischemia/genetics , Lactates/blood , Leg/blood supply , Leukocyte Count , Leukocytes/pathology , Male , Middle Aged , N-Formylmethionine Leucyl-Phenylalanine/pharmacology , Neutrophils/pathology , Oxidative Stress , Reperfusion Injury/genetics , Respiratory Burst , Tetradecanoylphorbol Acetate/pharmacology , TourniquetsABSTRACT
Laser therapy has gained wide acceptance and application in many medical disciplines. Nevertheless, during surgical procedures, the thermal destruction of tissue creates a smoke plume. Recent research data indicate that pyrolysates liberated during vaporisation of tissue induce DNA damage. However, assessing potential health hazards during medical laser treatment requires comprehensive insight into the cytotoxic, genotoxic, clastogenic and mutagenic capacity of laser pyrolysis products (LPP). Therefore, the aim of this study was to evaluate the cytotoxic, genotoxic, clastogenic and mutagenic potential of substances resulting from laser irradiation. Four different types of porcine tissues were irradiated with a surgical CO2 laser, the aerosols were sampled under defined conditions and subjected to the SCE test, micronucleus test and the HPRT test. The results showed that the pyrolysis products are strong inducers of cytotoxic effects. The pyrolysis products induced positive effects in the SCE test, micronucleus test and the HPRT test. The ability and extent to induce genotoxic and mutagenic effects turned out to be dependent on the type of tissue that had been irradiated. In general, the effects were most pronounced with liver pyrolysate. In all test systems, a clear dose relationship could be established. In conclusion, we were able to prove that the particulate fraction of laser pyrolysis aerosols originating from biological tissues undoubtedly have to be classified as cytotoxic, genotoxic, clastogenic and mutagenic. Therefore, they could be potential health hazards for humans.
Subject(s)
Adipose Tissue/radiation effects , Lasers/adverse effects , Liver/radiation effects , Muscle, Skeletal/radiation effects , Mutagenicity Tests , Adult , Aerosols/toxicity , Animals , Cell Line , Cell Survival/radiation effects , Cricetinae , Cricetulus , DNA Damage , Humans , Hypoxanthine Phosphoribosyltransferase/genetics , Hypoxanthine Phosphoribosyltransferase/radiation effects , Lymphocytes/radiation effects , Male , Micronucleus Tests , Sister Chromatid Exchange , SwineABSTRACT
The purpose of this study was to examine whether changes in the repair capacity of blood cells could be used as a valuable biomarker for radiation exposure. To characterize the repair kinetics in nonirradiated and irradiated cells we first performed in vitro split dose experiments. DNA damage and DNA repair capacity were analysed using the comet assay. Our results showed that the first in vitro irradiation affects the repair system of the cells, resulting in a decreased repair capacity after the second irradiation. Furthermore, the second irradiation results in a large amount of DNA damage in the blood cells. To test whether the analysis of the DNA repair capacity after in vitro irradiation is also a valuable method for in vivo studies of donors exposed to radiation, we analysed the repair capacity of blood cells of two exposed groups: patients subjected to a radioiodine therapy and chronically irradiated volunteers from the Chernobyl region. Both groups also showed a significantly impaired repair capacity indicating a stress on the hematopoietic system. In addition, in the group of the Ukrainians DNA damage after in vitro irradiation was significantly higher than in a control group. These results lead to the presumption that the repair capacity and the DNA damage after in vitro irradiation might be a very useful biological marker for radiation exposure in population monitoring.
