RU2376043C1 - Method for suppression of functions and damage of malignant tumor cells - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention is related to medicine and veterinary and is intended for suppression of functions and damage of malignant tumor cells. Malignant tumor is radiated with controlled local sign-alternating infralow frequency pulse electromagnet field (SIP EMF), with induction of magnetic field B=(120÷220) mTl, with time of relaxation of anterior front of pulse 0.01 sec., duration of electromagnetic impulse t_imp ⁼0.75 sec and pause of t_pause ⁼0.5 sec, with overall time of radiation of 30 min. for 30 days. Additionally surgical operation is executed to remove malignant tumor at 3-5 day on completion of course of SIP EMF radiation.

EFFECT: suggested method makes it possible to suppress functions and to destroy cells of malignant tumors.

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Description

The invention relates to medicine and to veterinary medicine, more specifically to oncology, and can be used in the treatment of humans and animals with malignant tumors.

In practical oncology, ionizing radiation is widely used for the treatment of malignant neoplasms (see Lindenbraten L.D. Medical Radiology. - M .: Medicine, 2000, 672 p.).

The ionizing radiation used in radiation therapy destroys the tumor cells, but it also infects healthy tissues and cells that fall into the radiation beam. This leads to radiation burns, radiation fibrosis of healthy organs and tissues, as well as to complications from the hematopoiesis system and the immune system. Radiation therapy has a limit and is carried out taking into account the accumulated dose. It should be noted that radiation therapy requires expensive equipment and special facilities.

The use of non-ionizing electromagnetic radiation (microwave fields) in oncology is also known, which is based on thermal effects and is used for antitumor hyperthermia with heating of the tumor to 43.5 ° (see A. Dunchenko, Microwave hyperthermia in oncology. // Abstracts reports "Medical Physics" - 95. - M., 1995. - P.113). In this case, there is a real danger of provoking denaturation processes that cause endotoxemia and irreversible damage to healthy tissues.

Studies are also being conducted in the field of antistress and antitumor effects of weak infra-low-frequency electromagnetic fields (EMFs) for the slow growth of tumors. It was found that EMF, preventing the onset of oxidative stress and the functional integrity of brain cell membranes, indirectly influenced the dynamics of the development of neoplasms (reduced their growth rate) (see AS No. 826587, A61K 1/42, publ. 30.08.90, Bull No. 32).

Studies are also being conducted on the use of a low-frequency periodic EMF with an induction amplitude of 3 mT in the treatment of a generalized form of melanoma to stimulate the neuroendocrine system, which was achieved by simultaneous exposure to the whole body of the patient (see Bakhmutsky N.G. Treatment of the genitalized form of skin melanoma with a vortex magnetic field. / / Russian Oncological Journal. - 2000, No. 6, p. 32-35). The author of the study concludes that there is a significant decrease in postoperative relapse of the disease under the condition of exposure to EMF compared with the control.

The positive results of the indirect effect of infra-low-frequency electromagnetic fields indicate the prospects of its use in oncology.

A method for the treatment of malignant neoplasms by direct exposure to intermittent sinusoidal EMF (the induction amplitude was 40 mT) was claimed (see A.S. No. 5222688, A61H 1/42, publ. 05.03.77., Bull. No. 9). The results of the studies revealed softening of the tumor tissue. However, the effectiveness of such exposure was low compared to radiation and chemotherapy used. As a result, the method has not received clinical use.

Work was carried out using the influence of intense infra-frequency electromagnetic fields directly on malignant neoplasms. A short-term, ~ 5 min, pulsed EMF effect was applied with an magnetic induction amplitude of up to 1.8 T, the intervals between pulses were 60 ms. Studies were conducted on outbred rats with neoplasms of the soft tissues of the thigh (sarcomas). The effects of magnetotherapy were manifested in necrotic processes of varying intensity in tumors and surrounding healthy tissues. This result allows us to search for optimal parameters and modes of influence of the low-frequency electromagnetic field on neoplasms (see Kaplan MA Prospects for the use of high-intensity pulsed magnetic fields in the treatment of malignant neoplasms. // Russian Oncological Journal. - 1998, No. 8, p. 34-37) .

The objective of the invention is to develop a method of suppressing functions and the subsequent destruction of cancer cells by local exposure to neoplasms with intense infra-low-frequency electromagnetic fields. The parameters of the electromagnetic field, the method and modes of irradiation obtained in the course of research should allow for the selective action of malignant neoplasms on cells. The consequences of such an effect on the tumor tissue should be comparable with the consequences obtained after courses of radiation and chemotherapy (carried out according to standard schemes), without damage to adjacent healthy tissues.

The problem is solved in that after verification of the tumor (diagnosis), the detected malignant tumor is irradiated with a local alternating infra-low-frequency pulsed electromagnetic field (ZII EMF) with proven and justified parameters: with an induction amplitude of B = (120 ÷ 220) mT, electromagnetic pulse time t _imp = 0.75 s, pause time t _pauses = 0.5 s, with a duration of a session of electromagnetic radiation of 30 minutes for 30 days daily. Surgical removal of the irradiated malignant tumor is carried out 3-5 days after the end of the irradiation course. When irradiating, two coils are used, which represent the electromagnetic field generator, while the axis of the ZII EMF coils is located in the center of the irradiated neoplasm, and the coils are located above and below the tumor at a distance of 2-5 cm from it with a working irradiation area of S≈5 ÷ 35 cm ² . Using the developed and created experimental setup, for the first time it was possible to obtain a ZII EMF with an induction amplitude of B = (120 ÷ 220) mT, which made it possible to achieve the claimed effect.

The method is characterized in that in order to suppress the functions and division of malignant tumor cells, they are exposed to bipolar ZII EMF without overheating of the affected object under conditions of natural convection at B = (120 ÷ 220) mT, formed in the space between two solenoids directed in the opposite direction, and with a relaxation time of the leading edge of the impulse of 0.01 s, which provides selective, in relation to healthy, morphologically reversible and irreversible changes in the tissues of malignant neoplasms.

To implement the method, an experimental magnetotherapeutic stationary installation was designed and manufactured, the main elements of which are a generator of ZII EMF, a power supply, a pulse forming unit and an electronic control unit.

The main characteristics of the used experimental magnetotherapy unit are shown in table 1.

Table 1 Main characteristics of the magnetotherapy unit Characteristics Values Magnetic field amplitude (B) ≤220 mT Relaxation time of the leading edge of the pulse 0.01 s Coil Current (I) up to 27 A Coil spacing (h) (10-60) cm Sectional area of the working window (S) ~ 5 ÷ 35 cm ² Pulse Duration (t _and ) from 0.02 s The duration of the pause (t _p ) from 0.05 s Power consumption (W) up to 1.5 kW

During the experimental work, the parameters created by the ZII EMF generators were controlled.

Material for conducting studies to substantiate the proposed method were selected malignant mammary tumors in outbred dogs. The time of tumor development (from the moment of detection to the start of treatment) ranged from 3 to 24 months. The size of the tumor at the beginning of treatment (exposure) ranged from 2.5 to 9 cm in diameter. The age of the dogs is from 4 to 13 years. The weight of animals is from 7 to 45 kg.

Before the experiment, 215 animals (dogs) were examined. Of these, 70 animals were selected for the experiment. The object of the study was outbred dogs with a spontaneous malignant tumor - according to the histological structure - highly and moderately differentiated ductal carcinoma of one or two mammary glands. Dogs were divided into groups. Animals in the amount of 10 dogs made up the control group, they were not irradiated with ZII EMF, and only surgical removal of neoplasms was carried out, followed by histological examination of the material obtained. The remaining 60 animals were divided into 3 groups of 20 animals each. The groups were distinguished by the applied magnitude of the induction amplitude of ZII EMF:

- group A - control group (10 animals);

- group B - working group - the impact of ZII EMF with an amplitude of B≤80 mT;

- group C - working group - the impact of ZII EMF with an amplitude of B = (80 ÷ 120) mT;

- group D - working group - the impact of ZII EMF with an amplitude of B = (120 ÷ 220) mT;

In the process of irradiation, the animal was placed between the coils on a special table. The axis of the coils corresponded to the center of the irradiated neoplasm.

Prior to the course of exposure, all animals underwent a puncture biopsy of the tumor formation, followed by histological and histostereometric examination of the obtained material. For (histostereometry) estimates of the volumetric ratios of the structural components of the studied tissues and their conditions, a grid containing 100 points was used. Separately, tumor cells in a stable state, as well as in a state of dystrophic changes and necrosis, lymphoid elements and stroma were counted. When calculating, the number of points and grids randomly coinciding with each of the above types (states) of cells was taken into account. Counting results were expressed as a percentage. For each drug, to ensure the reliability of the result according to the specified methodology, the calculation was carried out ten times.

On 3-5 days after the end of the course of exposure to ZII EMF, the animals underwent surgical removal of the irradiated mammary glands affected by the tumor. The resulting surgical material was subjected to histological and histostereometric studies, and the studies were conducted in different areas of the tumor.

The effectiveness of the impact of ZII EMF on neoplasms was determined by comparing the results of histological studies and histostereometry of irradiated and non-irradiated tumor tissues. A comparison was also made of the effects on tumor and healthy tissues. Additionally, a qualitative comparison was made with the morphological consequences after chemotherapy.

The reliability of clinical, laboratory and histometric studies was evaluated by standard methods of mathematical statistics for each determined indicator (sign) with a confidence probability of P <0.05.

In histological preparations of spontaneous malignant neoplasms of the mammary glands without the influence of ZII EMF (the diagnosis is highly and moderately differentiated ductal carcinoma), glandular and glandular-cribrotic structures from polymorphic atypical cells with hyperchromic nuclei, single sections of dystrophies and foci of necrosis are determined among the stroma. There is a predominance of the epithelial tumor component over the stroma.

In a comparative study of histological preparations of spontaneous malignant neoplasms of the mammary glands (the diagnosis is moderate and highly differentiated ductal carcinoma) without exposure and after a course of exposure to ZII EMF with parameters and modes: B≤80 mT; t _and = 0.75 s; t _p = 0.75 s;

T _exposure = (30 min × 30 sessions), the following features are noted:

- dystrophic changes in epithelial cells in the form of focal diffuse karyopiknosis and karyorexis, vacuolization of the cytoplasm up to (40 ÷ 70)%;

- increase in the area of necrosis to (25 ÷ 35)%;

- change in the ratio of the stroma - epithelial tumor component in the direction of the stroma;

- moderate focal lymphoid infiltration of the stroma in the form of clusters of reactive lymphocytes;

- the appearance of areas of hyalinosis and fibrosis;

- the presence of hemosiderosis in the stroma.

The listed changes are more pronounced in the histological preparations of those neoplasms that were subjected to the course of exposure to ZII EMF with an induction amplitude of B = (80 ÷ 120) mT and exposure T _exposure = (30 min × 30 sessions), while therapeutic pathomorphosis reaches II-III degree . Dystrophic changes and necrosis increase and make up (60 ÷ 90)% and (35 ÷ 55)%, respectively. Stroma significantly prevails over other structural elements.

In the study of histological preparations of tumors exposed to ZII EMF with an induction amplitude of B = (12 ÷ 220) mT and exposure

T _exposure = (30 min × 30 sessions), almost complete replacement of other cellular components with the stroma was revealed. Defined in a small amount of cellular components are in a dystrophic state or in a state of necrosis.

At the same time, during histological examination of healthy mammary gland tissues with all the used parameters of exposure to ZII EMF, dystrophic changes are (3 ÷ 7)%, necrosis is not detected.

The histostereomic treatment of histological preparations obtained after surgical removal of malignant neoplasms of the mammary glands in dogs was performed.

The effects of the course effect of ZII EMF on the tumor were evaluated according to histostereometry of histological preparations. The statistical reliability of the results was provided by:

- averaging the relative number of structural components and their conditions for each section of the tumor over its different sections;

- averaging the relative number of structural components in the aggregate of the same type of tumor under the same exposure conditions.

As a result of irradiation with ZII EMF, the following structural changes in the tissues of the mammary glands were revealed, which are presented in table 2.

Table 2 shows that when exposed to an increase in the induction of MP in the tissues of malignant neoplasms, the following changes are determined:

- a significant decrease in tumor cells (in%) with (28.49 ± 2.08) - no effect, up to (18.58 ± 1.09); (7.23 ± 1.87) and (0.61 ± 0.43) upon induction of MP B ≤80 mT; B = (80 ÷ 120) mT and B = (120 ± 220) mT after the course of exposure, respectively;

- increase in the number of necrosis (in%) with (0.80 ± 0.35) - without exposure, from (1.46 ± 0.29) mT to (1.96 ± 0.43) - MP induction B≤80 mT and B = (80 ÷ 20) mT after the course of exposure, respectively;

table 2 A quantitative characteristic of the structural components of spontaneous malignant neoplasms of the mammary glands after a course of exposure at various amplitudes of magnetic induction (t _and = 0.75 s; t _n = 0.5 s; T _exposure = (30 min × 30 sessions)); (M ± m) Structural components and their condition No effect (in%) After a course of exposure with MP induction ≤80 mT (in%) After a course of exposure with induction of MP (80-120) mTl (in%) After a course of exposure with induction of MP (120 ÷ 220) mTl (in%) one Tumor cells 28.49 ± 2.08 * 18.58 ± 1.09 * 7.23 ± 1.87 * 0.61 ± 0.43 * 2 Dystrophically altered cells 2.71 ± 0.67 2.80 ± 0.59 2.97 ± 0.58 * 0.85 ± 0.21 3 Necrosis 0.80 ± 0.35 * 1.46 ± 0.29 * 1.96 ± 0.43 * 1.28 ± 0.50 * four Lymphoid elements 1.51 ± 0.42 1.56 ± 0.31 1.80 ± 1.05 2.12 ± 0.78 5 Stroma 66.49 ± 2.16 * 75.60 ± 3.27 * 87.04 ± 3.27 * 95.14 ± 3.46 * * - p <0.05

- a significant increase in the stroma up to the almost complete replacement of other cellular components (in%) with (66.49 ± 2.16) - no effect, with (75.60 ± 3.27); (87.04 ± 3.27) to (95.14 ± 3.46) upon induction of MP B ≤80 mT; B = (80 ÷ 120) mT and B = (120 ÷ 220) mT after the course of exposure, respectively;

The results obtained during numerous studies of the effects of ZII EMF with induction B≤ (80 ÷ 220) mT, allow us to draw the following conclusions:

1. The selectivity and morphological irreversibility of the effects of exposure to tumor tissues are established in comparison with similar healthy ones. The revealed therapeutic pathomorphism of malignant neoplasms according to their structural characteristics proves the prospects and novelty of using ZII EMF in oncological practice.

2. The proposed regimes of local ZII EMF exposure reliably lead to therapeutic pathomorphosis of the II-III degree of severity of spontaneous malignant neoplasms in experimental animals, which corresponds to the consequences after a cycle of chemotherapy courses according to standard methods.

3. Structural changes in the cells of malignant tumors when exposed to ZII EMF are phase in nature and depend on its magnitude of the amplitude of induction: first, dystrophic changes, then cessation of division, an increase in necrosis and, in the final phase, the replacement of all cellular components with the stroma.

The proposed method, parameters and tested modes of exposure to alternating infra-low-frequency pulsed EMFs are promising for the selective suppression of functions and further death of cancer cells, which can be considered for possible use in medical and veterinary oncological practice, either alone or in combination with other treatment methods.

Claims (1)

A method of suppressing the functions and destruction of malignant tumor cells, including irradiating a tumor with an electromagnetic field before the removal operation, characterized in that the malignant tumor is irradiated with an adjustable local alternating infra-low-frequency pulsed electromagnetic field (ZII EMF) with magnetic field induction B = (120 ÷ 220) mT, if the relaxation time of the front pulse edge of 0.01 s, the pulse duration of the electromagnetic _pulses t = 0.75 s and t pause _pauses = 0.5 s, with a total irradiation time of 30 minutes for 30 days second, the surgery to remove a malignant tumor was performed on day 3-5 after the end of the FIL EMF exposure.