[Comparison of the effects of laser and light-emitting diodes on lipid peroxidation in rat wound exudate].

The effect of laser and light-emitting diode radiation on lipid peroxidation in rat wound exudate was studied with the aim to compare the efficiency of coherent laser and incoherent light-emitting diode radiations. A model of aseptic wound in rat suggested by L.I. Slutskii was used. A He-Ne laser (632 nm) and a U-332B light-emitting diode were used in this study. The intensity of lipid peroxidation was estimated by the TBA assay. The antioxidative capacity of rat wound fluid was evaluated by means of chemiluminescent assays in two model systems: a) aqueous system with ABAP and luminol and b) in phospholipid liposome suspension with Fe2+ and cumarin. It was shown that exposure of rat wounds to both laser and light-emitting diode radiation decreased the concentration of TBA products and increased the antioxidative capacity of wound exudates, compared with the control group (without irradiation). The results obtained show that exposure of wounds to both laser and light-emitting diode irradiation causes a decrease in the oxidative stress in the rat wound fluid. No significant quantitative difference between the effects of laser and light-emitting diode irradiation was found.

[A comparison of the effects of laser and light-emitting diodes on superoxide dismutase activity and nitric oxide production in rat wound fluid].

The action of laser and light-emitting diode radiation in the visible region on the content of reactive nitrogen species and activity of superoxide dismutase in rat wound fluid was studied, and efficiency of action of coherent laser and incoherent light emitting diode radiations in the red region of the spectrum on the parameters under study was compared. A model of incised aseptic wounds in rats proposed by L.I. Slutskiy was used. A He-Ne laser (632 nm) and a Y-332B light emitting diode served as radiation sources. It was shown that (1) exposure of wounds to the visible light of both laser and light-emitting diodes causes dose-dependent changes in superoxide dismutase activity and production of nitrites and (2) the radiation coherence does not play any significant role in the changes of superoxide dismutase activity or nitrogen oxide formation by wound fluid phagocytes.

[A comparative study of the effects of laser and light-emitting diode irradiation on the wound healing and functional activity of wound exudate leukocytes].

The effects of coherent He-Ne laser and non-coherent light-emitting diode radiation on rat skin wound healing and functional activity of wound excudate leukocytes were compared. A comparative pathomorphological analysis showed that the He-Ne laser and light-emitting diode irradiation stimulated the transition of the inflammatory phase of the wound healing into the reparative (proliferative) and scarring phases sequentially. It was also detected that the functional activity of leucocytes changed in a dose-dependent manner. The leukocyte activity was found to be similar in the groups with laser and light-emitting diode irradiation. Thus, we can conclude that coherent laser and non-coherent light-emitting diode radiation have very close effects on wound healing and activity of wound exudate leukocytes, and coherence is not required for this activity.

[The role of lipid peroxidation products in the effect of He-Ne laser on human blood leukocytes].

The role of lipid peroxidation products formed in membranes of human blood leukocytes after irradiation with He-Ne laser was studied. It was found that low-intensity laser irradiation (0.3-1.6 J/cm2) leads to both cell activation and an increase in the content of lipid peroxidation products. The intensity of lipid peroxidation was analyzed by estimating the amount of TBA reactive products and lipid diene conjugates. Irradiation in the presence of an exogenous photosensitizer (protoporphyrin IX) enhanced the phenomena observed. The use of antioxidants (tocopherol and ionol) completely eliminated the laser-induced effects (changes in leukocyte activity and accumulation of lipid peroxidation products). These results can be explained by the fact that laser irradiation leads to the activation of lipid peroxidation in leukocyte membranes, which in turn enhances the response of cells to the stimulus (priming).

[The role of blood plasma porphyrins in the effects of He-Ne-laser on human leukocytes].

The He-Ne-laser induced effects in human blood leukocytes in the presence of autologic plasma were investigated. Experiments were performed in two ways: (1) He-Ne-laser irradiation of cells in the presence of autologic plasma or (2) laser irradiation and subsequent addition of autologic plasma to the cell suspension. The concentration dependencies of plasma additions were evaluated. To obtain different concentrations of porphyrins in plasma samples, we either diluted the samples with PBS or selected patients with different porphyrin plasma content. The effects of He-Ne-laser irradiation were characterized by the maximum effect dose (Dmax) of irradiation and the degree of maximum cell activation (Amax, priming index). In the first series of experiments, we irradiated leukocytes in the presence of autologic plasma taken from patients with pneumonia and bronchial asthma. It was found that Dmax decreased with increasing porphyrin concentration in plasma. It was observed that, at low porphyrin concentrations, Amax increased severalfold with increasing photosensitizer concentration. At a porphyrin concentration of 0.46 pmol a decrease in Amax was detected as the porhyrin concentration increased. The same effects were revealed at high doses of laser irradiation. Very similar effects were found in experiments with the addition of irradiated plasma to cells. However, the Amax value was considerably less compared to that after irradiation in the presence of plasma (160% vs. 230 - 270% upon combined irradiation of cells and plasma). The Dmax value was higher in the series of experiments in which plasma was irradiated separately. The results suggest that laser-induced leukocyte activation can be mediated by blood plasma porphyrins and the products of lipid peroxidation formed as a result of porphyrin-photosensitized lipid oxidation.

Changes in superoxide dismutase activity and peroxynitrite content in rat peritoneal macrophages exposed to He-Ne laser radiation.

The formation of reactive oxygen and nitrogen species by rat peritoneal macrophages induced by a low-intensity He-Ne laser radiation (LR) was studied in this work. It was found that the formation of reactive oxygen species, nitric oxide, and peroxynitrite as well as changes in the activity of superoxide dismutase (SOD) depended to a large extent on the LR dose. In particular, it was found that activation of SOD at low LR doses was accompanied by nitric oxide level increase, while the level of peroxynitrite showed no significant changes. On the other hand, an enhanced LR dose inhibited the enzyme, and this was accompanied by peroxynitrite accumulation. All the measurements were carried out the day after LR treatment. The revealed regularities consequently demonstrate the existence of a deferred LR action on macrophages associated with the production of reactive oxygen and nitrogen species.

[Free radical mechanisms of low-intensive laser radiation]. / Svobodnoradikal'nye mekhanizmy stimuliruiushchego deistviia nizkointensivnogo lazernogo izlucheniia.

A hypothesis of free radical mechanisms of stimulating action of low-intensive laser radiation (LILR) used for therapy of a variety of inflammatory diseases is formulated. The main points of the above hypothesis are as follows. Endogenous porphins are a LILR chromofor in the red band (lambda = 632.8 nm). Light absorption induces the production of initiating radicals that are involved in subsequent free radical reactions, in lipid peroxidation in particular. Modified lipid peroxidation in the cell membranes causes an increase in ion permeability, including that for Ca2+. The higher levels of Ca2+ in the leukocytic cytosol result in Ca(2+)-dependent cellular priming, which appeared as the increased cell functional potential and which is seen in subsequent leukocytic stimulation of the greater production of prooxidants and other biologically active products. These products include nitric oxide and a number of cytokines involved in the regulation of microcirculation. The paper presents experimental findings that can be regarded as evidence for some points of the above hypothesis which are used to provide a chain of events underlying the free radical mechanisms of stimulating action of low-intensive laser radiation.

Hematoporphyrin derivative or phthalocyanine photosensitized hemolysis of erythrocytes under laser irradiation.

The effect of laser irradiation on the photosensitized hemolysis of rabbit erythrocytes was investigated. Two photosensitizers of domestic production were used: Photohem (a hematoporphyrin derivative) and Photosense (sulfonated phthalocyanine aluminium). The hemolysis was activated by laser irradiation in the presence of Photohem or Photosense. The process depended on the irradiation dose, the concentration of the photosensitizers and on the time during which the erythrocytes were incubated with the photosensitizers. Three lasers with wavelengths 520, 632.8 and 850 nm were used. The photosensitized hemolysis of rabbit erythrocytes was found to be inhibited by antioxidants.

Effects of endogenous photosensitizers on the laser-induced priming of leucocytes.

The influence of He-Ne (lambda = 632.8 nm) laser irradiation on the functional activity of leucocytes was investigated. The functional activity of leucocytes irradiated in the presence of plasma with phthalocyanine raised. Irradiation of leucocytes without plasma had no influence on the chemiluminescent response of the blood cells. An increase in the concentration of phthalocyanine in plasma first led to an increase and then a decrease in the functional activity of the leucocytes. Similar results were obtained for the case of endogenous porphyrins in plasma. These results make it possible to consider the influence of laser irradiation on the leucocytes' functional activity as the priming. The basic concepts of the free-radical mechanism of laser therapy were formulated.

Low-power laser irradiation induces leukocyte priming.

Laser radiation (LR) of various spectral composition has been broadly used in clinical practice. However, the mechanism of the stimulating effects of LR remains obscure. The effect of He-Ne LR (633 nm) on human blood leukocytes was investigated both in the absence and presence of 8.65 nmol/l phthalocyanine (PhC). Irradiation of non-stimulated leukocytes with 0.025 to 0.5 J/cm2 did not lead to any activation of their luminol-dependent chemiluminescence (LCL). On the other hand, LR increased in most cases the subsequent CL response of the cells to opsonized zymosan (priming action of He-Ne-laser light). The effect of LR on the leukocytes was not standard. In irradiated leukocytes isolated from patients with severe acute or chronic pneumonia, or chronic bronchitis, the maximal LCL exceeded that for non-irradiated cells by 80% (0.05 J/cm2), 20-25% (0.15 J/cm2), and 0%, respectively (doses are shown in parentheses). Further increase of the exposure brought about a dose-dependent inhibition of LCL in cells from patients with severe acute and chronic pneumonia. There was an intriguing relationship between maximal CL responses of leukocytes subjected to laser irradiation in the presence and without PhC. When the priming effect of LR on isolated cells was small, it increased in the presence of exogenous photosensitizer, phthalocyanine; in cells of severely ill patients where the initial effect of LR was strong, Pc inhibited the priming action of LR. Apparently, different cells contained different amounts of endogenous photosensitizer(s); the addition of exogenous sensitizer increased the priming action of LR at low concentrations and decreased it at higher concentrations of the endogenous photosensitizer.