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Radiation damage of hematopoietic organs is tremendously significant in the growth of radiation sickness disease in human beings and animals. The blood system damage is distinctly revealed in changes of the cell number in peripheral blood [1]. A decrease of the amount of leukocytes (radiation leucopenia) and bone marrow karyocytes is the most characteristic feature. In this connection, the search of opportunities to improve the hematopoiesis recovery after the ionizing radiation action on the organism is an important task for radiation protection.
It is well-known that at present lasers are widely used in biology and medicine. The application of the low level laser radiation (LLLR) has been especially used in therapy [2-5]. One of the most accepted methods of therapeutic action of the low level radiation on the human organism is the intravenous laser irradiation of blood (ILIB) that is successfully applied in various fields of medicine. It is shown [6] that after ILIB changes occur in the regular blood elements, blood properties in general (plasma composition, rheological properties, etc.), as well as in the system response on the level of different organs and tissues. Laser irradiation of blood brings a stimulating effect on hematopoiesis in the form of an increase of the amount of hemoglobin, erythrocytes and leukocytes. The non-specific protection system is stimulated, the functional and phagocytal activity of lymphocytes increases. The totality of changes in blood observed at ILIB is to a great extent regarded as a response of the homeostasis management system to pathological processes in separate organs and tissues, without allocating any link in principle as a leading one.
Fig. 1 shows the spectra of arterial and venous blood
absorption [7].
One can see that blood absorbs in a wide spectral region (in different spectral regions different photoreceptors, blood components, absorb). It can be assumed that it would have been most efficient to use laser radiation at ILIB with the wavelength about 410 nm, where the absorption is maximal. However, such laser diodes are still too expensive and cannot be purchased by a wide range of consumers [8]. That is why ultraviolet and red spectral ranges have still been most common (at least, until now) in the methodology of the intravenous laser irradiation of blood.
We showed earlier [9] (invention patent RU 2 330 695 C2) that the mice fibroblasts cells survival increases under the action of 633 nm laser radiation applied before and after both ?-rays and 150 MeV protons. The simultaneous action of laser and ionizing radiation on these cells also caused an increase of cell survival. The maximal radioprotection effect was observed when the energy density of the laser radiation was about 1 mJ/cm2. Further investigation of the mechanism of radioprotective effect of the 633 nm optical radiation on fibroblast cells shows [10] that the radioprotective action of the laser is transferred to the fibroblast cells according to the mechanism of the“bystander” effect [11] via direct intercellular communication through gap junctions, as well as via medium transfer from the irradiated with laser and ionizing radiation ones to irradiated just with ionizing radiation cell population. The radioprotective action of laser radiation was established also when irradiated with ionizing radiation cells were co-cultured as a mixture with laser and ionizing radiation irradiated cells [10].
As the search for the effective radioprotectors, especially those that are effective after the action of ionizing radiation, is still one of the important problems in radiation biology, space biology and medicine, studies of opportunities to apply radioprotective effect of the laser radiation conducted on laboratory mice in in vivo experiments are of great interest. In this connection, research was done on the action of various doses of laser radiation of the red spectrum region, alone and combined with ?-rays, on
2. Methods
The experiments were performed on young male C57ВL/6 mice with the mass of 11-15 g. The conditions of managing and experiments complied with “The regulations of working with animals for experiments” (Addendum to Order of the USSR ministry of health service 755 of 12.08.1977). The animals were fed with the standard briquetted feeding staff and drinking water from drinking cups. Before killing the mice were weighed; then some blood was taken from the tails of the experimental animals to count the amount of leukocytes of peripheral blood and to determine the amount of hemoglobin in blood. To analyze the number of karyocytes, a buttock of the animal, with muscles cleaned off, was put into a chemical cup of 50 mL and ground up in 6 mL of the 3% solution of acetic acid.
To determine the content of hemoglobin in blood we used the measuring device “Gemoglobinometr Mini-Gem 540” (CJSC Scientific-Industrial Enterprise TEKHNOMEDIKA, Russia). The number of leukocytes and karyocytes was calculated with the Goryaev chamber according to the conventional methods.
Each experimental group included 10 mice. The intact group contained 12 mice.
(Table 2) that in such exposure after 24 h the number of leukocytes and karyocytes truly increases at both doses of the laser (in comparison with the mice exposed only to gamma-rays). After 72 h a true decrease of the number of leukocytes was observed at both doses and an increase of hemoglobin at the laser dose 1 mJ/cm2, in comparison with the parameters of the mice irradiated only with gamma rays after a 72-hour exposure.
The obtained data testify that distant irradiation of the body may lead to changes in blood, and stimulated blood cell formation. Furthermore, stimulating effect is observed not only in the case of irradiation with laser,
but also after the action of ionizing radiation.
15 days after the irradiation of the mice with gamma-rays the quantitative amount of hemoglobin in blood was repaired, but the values of the quantity of leukocytes and karyocytes of the bone marrow were still lower, compared to those of the intact mice. However, the mice that had combined irradiation with?-rays and laser radiation showed the quantity of karyocytes of the bone marrow on the 15th day after the exposure on the level of the intact mice already.
It can be assumed (as an increase of the bone marrow karyocytes was observed) that laser radiation stimulates hematogenesis.
Distant laser irradiation of the body may lead to changes in blood, and stimulated blood cell formation.
Laser radiation stimulates blood cell formation not only in the case of irradiation with laser, but also after the action of ionizing radiation.
The red spectral range laser radiation can be applied to improve the recovery of hematogenesis after the action of ionizing radiation on biological objects.
It is well-known that at present lasers are widely used in biology and medicine. The application of the low level laser radiation (LLLR) has been especially used in therapy [2-5]. One of the most accepted methods of therapeutic action of the low level radiation on the human organism is the intravenous laser irradiation of blood (ILIB) that is successfully applied in various fields of medicine. It is shown [6] that after ILIB changes occur in the regular blood elements, blood properties in general (plasma composition, rheological properties, etc.), as well as in the system response on the level of different organs and tissues. Laser irradiation of blood brings a stimulating effect on hematopoiesis in the form of an increase of the amount of hemoglobin, erythrocytes and leukocytes. The non-specific protection system is stimulated, the functional and phagocytal activity of lymphocytes increases. The totality of changes in blood observed at ILIB is to a great extent regarded as a response of the homeostasis management system to pathological processes in separate organs and tissues, without allocating any link in principle as a leading one.
Fig. 1 shows the spectra of arterial and venous blood
absorption [7].
One can see that blood absorbs in a wide spectral region (in different spectral regions different photoreceptors, blood components, absorb). It can be assumed that it would have been most efficient to use laser radiation at ILIB with the wavelength about 410 nm, where the absorption is maximal. However, such laser diodes are still too expensive and cannot be purchased by a wide range of consumers [8]. That is why ultraviolet and red spectral ranges have still been most common (at least, until now) in the methodology of the intravenous laser irradiation of blood.
We showed earlier [9] (invention patent RU 2 330 695 C2) that the mice fibroblasts cells survival increases under the action of 633 nm laser radiation applied before and after both ?-rays and 150 MeV protons. The simultaneous action of laser and ionizing radiation on these cells also caused an increase of cell survival. The maximal radioprotection effect was observed when the energy density of the laser radiation was about 1 mJ/cm2. Further investigation of the mechanism of radioprotective effect of the 633 nm optical radiation on fibroblast cells shows [10] that the radioprotective action of the laser is transferred to the fibroblast cells according to the mechanism of the“bystander” effect [11] via direct intercellular communication through gap junctions, as well as via medium transfer from the irradiated with laser and ionizing radiation ones to irradiated just with ionizing radiation cell population. The radioprotective action of laser radiation was established also when irradiated with ionizing radiation cells were co-cultured as a mixture with laser and ionizing radiation irradiated cells [10].
As the search for the effective radioprotectors, especially those that are effective after the action of ionizing radiation, is still one of the important problems in radiation biology, space biology and medicine, studies of opportunities to apply radioprotective effect of the laser radiation conducted on laboratory mice in in vivo experiments are of great interest. In this connection, research was done on the action of various doses of laser radiation of the red spectrum region, alone and combined with ?-rays, on
2. Methods
The experiments were performed on young male C57ВL/6 mice with the mass of 11-15 g. The conditions of managing and experiments complied with “The regulations of working with animals for experiments” (Addendum to Order of the USSR ministry of health service 755 of 12.08.1977). The animals were fed with the standard briquetted feeding staff and drinking water from drinking cups. Before killing the mice were weighed; then some blood was taken from the tails of the experimental animals to count the amount of leukocytes of peripheral blood and to determine the amount of hemoglobin in blood. To analyze the number of karyocytes, a buttock of the animal, with muscles cleaned off, was put into a chemical cup of 50 mL and ground up in 6 mL of the 3% solution of acetic acid.
To determine the content of hemoglobin in blood we used the measuring device “Gemoglobinometr Mini-Gem 540” (CJSC Scientific-Industrial Enterprise TEKHNOMEDIKA, Russia). The number of leukocytes and karyocytes was calculated with the Goryaev chamber according to the conventional methods.
Each experimental group included 10 mice. The intact group contained 12 mice.
(Table 2) that in such exposure after 24 h the number of leukocytes and karyocytes truly increases at both doses of the laser (in comparison with the mice exposed only to gamma-rays). After 72 h a true decrease of the number of leukocytes was observed at both doses and an increase of hemoglobin at the laser dose 1 mJ/cm2, in comparison with the parameters of the mice irradiated only with gamma rays after a 72-hour exposure.
The obtained data testify that distant irradiation of the body may lead to changes in blood, and stimulated blood cell formation. Furthermore, stimulating effect is observed not only in the case of irradiation with laser,
but also after the action of ionizing radiation.
15 days after the irradiation of the mice with gamma-rays the quantitative amount of hemoglobin in blood was repaired, but the values of the quantity of leukocytes and karyocytes of the bone marrow were still lower, compared to those of the intact mice. However, the mice that had combined irradiation with?-rays and laser radiation showed the quantity of karyocytes of the bone marrow on the 15th day after the exposure on the level of the intact mice already.
It can be assumed (as an increase of the bone marrow karyocytes was observed) that laser radiation stimulates hematogenesis.
Distant laser irradiation of the body may lead to changes in blood, and stimulated blood cell formation.
Laser radiation stimulates blood cell formation not only in the case of irradiation with laser, but also after the action of ionizing radiation.
The red spectral range laser radiation can be applied to improve the recovery of hematogenesis after the action of ionizing radiation on biological objects.