Method of combination treatment of abscesses in experiment

FIELD: medicine.

SUBSTANCE: invention refers to medicine, namely surgery, and may be used for treating abscesses, festered residual cavities and ganglions. The cavity contents are daily aspirated through a catheter in laboratory animals (male rabbits) suffering simulated hepatic abscesses of the diameter of 1.5 cm. Then the abscess cavity is filled with copper nanoparticle suspension in 0.9% physiologic saline of the concentration of 1 mcg/ml. A laser light guide provided with a diffuser is placed in the cavity centre to expose to laser light at wave length 630 nm in a continuous mode, output power 35 mWt for 3 minutes.

EFFECT: method enables intensifying antimicrobial action of copper nanoparticles under laser light, starting treating without specifying an agent, suppressing pathogenic microogranisms over a relatively short time, stimulating repair processes successfully, reducing toxic action of nanoparticles on macroorganism, considerably reducing a length of treating festered cavities.

2 dwg, 1 tbl, 1 ex

 

The invention relates to medicine, in particular to surgery, and can be used to treat abscesses, festering residual cavities and cystic formations.

There are many ways to treat abscesses of the liver: restoration of purulent, antibiotic therapy, anti-inflammatory, desensibiliziruyuschee therapy (Akhaladze GG, Tsereteli YOU Colagiovanni abscesses in the liver // Consilium medicum. - 2005. - V.7, №2. Surgery (ADJ.). - P.1-9. Parhisenko Y.A., deaf A.A. et al. Diagnosis and treatment of liver abscesses // Surgery. - 2000. No. 8. - S. - 35-37. The shochet Y.N., Fokeev S.D. et al. The influence of local continuous intraarterial antimicrobial therapy in the postoperative period in patients with liver abscesses // Annals of surgery. - 2008. No. 1. - S. - 50-53), and how puncture and aspiration and drainage treatment of the abscess under ultrasound control (Afernoon, Adetimehin, Vueling, Wiesco. "Puncture and aspiration and drainage treatments in abdominal surgery", Annals of surgery. 1997, No. 2, pp.42-47), lies in the puncture and drainage of the purulent cavity by one or more drains, depending on its volume, with lavage with antibiotics and antiseptics. The presence of a large number of different tools and techniques of rehabilitation of purulent foci testifies to their low efficiency.

Present methods do not allow the quality of the public treatment of the walls of the cavity, as dense fibrin overlay covering the inside of the abscess cavity, prevent the action of drugs, at the same time because of the formation of antibiotic-resistant forms of bacteria have to change the dosage and the drug group.

In complex treatment of purulent surgical infection recently widely used phage therapy. A method for the treatment of postoperative intra-abdominal abscesses with local injection of bacteriophage (Kurmangaliev S.M. Purulent infection in surgery. - M.: Medicine, 1985, p.106-109). "For the treatment of inflammatory diseases (patent RF №2105544), "drug for the treatment of inflammatory diseases (patent RF №2144368), "a Method for the treatment of postoperative abscesses of the abdominal cavity (patent RF №2243780).

However, despite some increase the effectiveness of drug therapy scope of these drugs in the treatment of abscesses is limited, and treatment is prolonged, the bacteriophage is possible only after determining the type of pathogen - minimum research time 5-7 days.

The tools and techniques used to combat intraoral infections, not as effective as required. Existing tools are not able to stimulate sufficient regeneration processes, so the AK now changed the structure of the agents of purulent complications, and commonly used drugs are ineffective, also, due to the formation of polyantibiotic-resistant strains of bacteria have to constantly change the drug.

Bactericidal properties of copper and its ions known to man since ancient times. In 2008, the Federal Agency for Environmental Protection (US EPA) officially assigned the copper status of substances with antibacterial surface. It has been proven bactericidal and bacteriostatic effect of copper on the most toxic species of bacteria, fungi and viruses: Acinetobacter Baumann, Black mold, Campylobacter, Aerobacter, Helicobacter pylori, Legionella, MRCS (including E-MRS), Pseudomonas aeruginosa, Staphylococcus aureus, Enterococcus resistant to vancomycin, an Adenovirus, the Fungus Candida, Clostridium difficile, E. coli, influenza Virus type a (H1N1), Listeria monocytogenes, Polio, Salmonella (Bacillus Gartner), the Tuberculosis Bacillus.

In literature there are reports of the use of copper nanoparticles in vitro closest to this paper (I.V. Babushkin study of the antibacterial action of nanoparticles of copper and iron on clinical strains of Staphylococcus aureus / Ivebeen, Vbboolean, Gwisoon, Dampaknya // Saratov scientific medical journal. - 2010. - V.6. No. 1. - S-014.). In the study of antimicrobial effects of copper nanoparticles on gram-negative microorganisms found that most who possesses suspension of copper nanoparticles at a concentration of 1 mg/ml (Vasariano et al., 2010). In several experimental studies demonstrated that the introduction of copper nanoparticles in the wound stimulates the mechanisms of regulation of microelement composition and activity of antioxidant enzymes observed wound healing effect (Pea and others, 2010), which is the prospect of creating a new class of antibacterial drugs. Copper nanoparticles through multilateral action on bacterial cell unlike antibiotics do not cause the selection of resistant strains, allowing further be recommended for use in the treatment of purulent diseases caused by strains resistant to currently used antibiotics.

Known negative changes of biochemical parameters of blood serum after oral administration of copper nanoparticles. When evaluating the biological effect of nanopowder of copper on the activity of transaminases (ASAT and Alat), used as a test of the change in membrane permeability and the expression of cytolytic syndrome hepatocytes, an increase in their activity, increasing in proportion to the increase in the concentration of 0.05-5 mg/kg). The increase in the concentration of copper nanoparticles causes an increase in the activity of LDH, the most important biochemical indicator of carbohydrate metabolism, reflecting the severity of interstitial hypoxia, and CPK, which assetsaamounting-cytoplasmic enzyme (the Study of biological activity of copper nanopowder. / Uschakov, Tasedunova. // Materials 70 scientific-practical conference of students and young scientists points out: "Young scientists for the health of the region." - 2009. - P.76-77).

In the literature there is information on the series of Toxicological studies of copper nanoparticles. Most animals treated with copper nanoparticles, observed symptoms of the gastrointestinal tract, decreased appetite, diarrhoea, vomiting. In animals treated with copper nanoparticles, observed lethargy, oligopol, tremor, opistotonus. In experimental animals at necropsy marked color change of renal tissue in bronze, as well as the death of the cells of the proximal tubules, glomerulonephritis, massive necrobiosis, spleen atrophy and changes color. In addition to the above changes marked changes of biochemical parameters blood urea nitrogen, creatinine, total bile acids and alkaline phosphatase, indicating renal and hepatic dysfunction (Chen z, Meng h, Xing G. et al. Acute toxicological affects of copper nanoparticles in vivo // The journal of physical chemistry. Toxicology letters, 2006. - 163. - 109-120).

Target organs for toxic effects of copper nanoparticles were liver, spleen, kidneys (GG Onishchenko, Begbick, V.I. Pokrovsky, Aiyetoro. "The concept of Toxicological studies, methodology of risk assessment methods to identify and measure the military definition of nanomaterials", 2007).

The value of the maximum tolerated dose (MTD) (Studying the security of the introduction of copper nanoparticles with different physico-chemical characteristics in the organism of animals / Oaasa, Eaazyshv, Westosha etc. // Vestnik OSU. - 2009. No. 2. - P.124-127) differences lie in the range of 2.5-10 mg/kg weight of the animal, LD50 - 7-15 mg/kg weight of the animal and LD100 - 20-30 mg/kg weight of the animal.

Widespread in the treatment of purulent diseases of the received laser technology. The basis of the mechanism of interaction of low-intensity laser (LLLT) with the bioobject are photophysical and photochemical reactions associated with resonant absorption of light fabrics and a violation of the weak intermolecular bonds, as well as the perception and the transfer effect of laser irradiation bodily uids (Illarionov VE fundamentals of laser therapy. - M.: Publishing house "RESPECT" Association INOTECH-Progress, 1992, pp.42-43).

The task of the invention is to develop an effective combined treatment of abscesses, to reduce the treatment time, reduce the toxic effects of nanoparticles.

The technical result is the complete suppression of pathogenic microorganisms and successful stimulation of reparative processes. The technical result is achieved by the combination of the processing cavity abscess the s suspension of nanoparticles of copper and irradiation NEELY.

The severity of antimicrobial action of copper nanoparticles and laser radiation against a clinical strain of Staphylococcus aureus was evaluated in vitro bacteriological methods in several stages:

1) evaluation of the influence of laser radiation on the growth of colonies of microorganisms;

2) evaluation of the effect of different concentrations of copper nanoparticles on the growth of colonies of microorganisms;

3) the study of the enhancement of the antimicrobial activity of copper nanoparticles in combination with the effect of LLLT.

Used a standardized optical standard turbidity of Mcfarland suspension of Staphylococcus aureus. Microbial load was 3×104microbial cells/ml In the first series of experiments, the culture of microorganisms were irradiated three minutes a laser therapeutic apparatus "matrix vlok", type MLOCR, the wavelength is 630 nm, the radiation power of 35 mW in continuous mode with the scattering nozzle (spherical dispersion). In the second series in the culture of microorganisms contributed 0.2 ml suspension of nanopowders of copper (average particle size of copper 60-80 nm) with final concentrations of 1000, 100, 10, 1 μg/ml In the third combined laser irradiation (wavelength is 630 nm, the radiation power of 35 mW in continuous mode with the scattering nozzle) and the introduction of copper nanoparticles. Control was a culture of microorganisms without laser irradiation and d is bauleni nanoparticles. Immediately and after 1, 2, 3 hours of cultivation produced dimensional seeding (0.1 ml) cups with meat-peptone agar and after 24 hours incubation at 37°C under a constant stirring speed of 50 rpm was calculated the number of grown colonies. When carrying out statistical processing was guided by the method of determining the standard deviation found in the experience of values, the proposed Ipodmini and Age.

The number of colonies of a strain of S. aureus, depending on the time of seeding and culturing conditions.
Stages
work
Conditions
cultivation
The time of sowing, h
0123
To*3153±5,2lawnlawnlawn
ILaser irradiation3086±2,099±2,31212±4,25416±8,
IINanoparticles of copper 1 mg/ml3123±2,2lawnlawnlawn
Copper nanoparticles 10 mg/ml3163±3,13550±4,61094±5,20
Copper nanoparticles 100 µg/ml3140±4,342±1,100
Nanoparticles of copper 1000 mcg/ml3128±3,7000
IIILaser + nanomed 1 mg/ml152±3,6000
Laser + nanomed 10 mg/ml141±4,5000
Laser + nanomed 100 µg/ml103±5,0000
Laser + nanomed 1000 µg/ml98±6,8000
*To control variant strain (physiological solution of sodium chloride without ultrafine copper powder and laser irradiation)

The study established that the first and second stages of the determined number of grown colonies immediately after sowing did not differ significantly from the control (see table).

At 1 and 2 hours of cultivation after laser irradiation, there was a decrease in the number of colonies experienced strain to 99±2.3 and 1212±4,2 respectively, however, at the third hour the growth of Staphylococcus aureus was resumed. Antibacterial properties NEELY is not effective enough.

The copper nanopowder (second stage) at a concentration of 1 μg/ml had no effect on the growth experienced culture. The marked decrease in the number of colonies of microorganisms by increasing the concentration of ultra-fine copper powder and time of cultivation in comparison with the control, where there was an increase in the lawn. The concentration of copper nanopowder 10 μg/ml causes a reduction in the number of colonies of Staphylococcus aureus in the early hours of contact with the culture in the complete suppression of growth after 3 hours of exposure. The concentration of nanomate 100 and 1000 MK who/ml inhibited the growth experienced culture for 2 and 1 h, respectively.

At the third stage immediately after exposure there has been a marked growth inhibition of S. aureus. The concentration of copper nanopowder 1, 10, 100 and 1000 μg/ml in combination with NEELY completely inhibited the activity of Staphylococcus aureus at 1, 2, and 3 hours of cultivation.

For the first time revealed the effect of enhancement of antimicrobial activity of copper nanoparticles in combination with the effect of LLLT on the culture of Staphylococcus aureus, which allows to obtain the required antibacterial effect at lower concentrations of copper nanoparticles, thereby reducing the toxic effect of the substance on the organism in vivo.

To create abscess of the liver of laboratory animals (rabbits male) was modeled fibrous cyst with a diameter of 1.5 cm according to the method described in the patent of the Russian Federation 2394278 (performed percutaneous puncture of the liver with laser support under ultrasound guidance, the needle was introduced through a two-channel catheter with a balloon which was inflated to the desired size of the simulated cavity in the liver tissue through the second channel of the catheter was introduced into the cyst cavity sclerosing solutions, on the 5th day the tank was drained, was carried out by x-ray and ultrasonic testing cysts)on the 6th day after sanitizing the channel of a catheter formed in the cyst was injected 2×109SOME in 1.0 ml of a clinical strain of Staphylococcus aureus. Analysis of the obtained data showed that the and the day 3 after infection by a strain of S. aureus the average number of bacterial cells in asperrimum detachable from the cavity of the cyst animals were 5,12×105CFU/ml detected for liver abscess.

Laboratory animals with the simulated abscess version of local treatment were divided into 4 groups: 1 - standard sanitation (furatsilin, dioxidine), 2 - the use of laser therapy, 3 - applying the suspension of nanoparticles of copper, 4 - copper nanoparticles in combination with the effect of LLLT.

In all groups, treatment was started with the aspiration of the abscess contents. In the first group performed daily oral hygiene with furatsilina 1:5000, followed by administration of 0.5 ml of 1% solution dioksidina. The second group of laser treatment is carried out by the apparatus ALT "matrix", type MLOCR (wavelength is 630 nm, continuous mode, the radiation power is 35 mW, scattering the head with a spherical dispersion; the power flux-density (MRP) for a cavity with a diameter of 1.5 cm was 5 mW/cm2). The sessions were carried out daily for 3 minutes within 14 days. In the third group once a day, the cavity was filled with a suspension of copper nanoparticles in physiological solution of sodium chloride concentration of 100 µg/ml In the fourth daily for ten days filled abscess cavity suspension of copper nanoparticles in physiological solution of sodium chloride concentration of 1 μg/ml, were placed the center cavity of the fiber laser, equipped with a diffuser, through the sanitizing channel of the catheter and perform laser irradiation (a mode of action similar to the second group experience).

For a comprehensive assessment of the inflammatory process used ultrasound investigation (USI), which was carried out on the 3rd, 5th, 7th, 10th, 14th, 30th day, every day spent microbiological research content abscesses and assessed the overall condition of the animals. Using ultrasound was determined by the diameter of the cavity and the thickness of the capsule. Bacteriological examination of the content of liver abscesses included a review of the qualitative composition and quantitative accounting of microorganisms.

The treatment results

Figure 1 - photograph of the control of sowing detachable from the cavity of the abscess on the seventh day of treatment, where the animals of the fourth group, the growth of colonies of the pathogen was absent microbiological study, the animals of the third group had more pronounced growth inhibition of S. aureus (104±2,6) compared with the first group (1936±4,2), in animals of the second group of inhibiting the growth of colonies of microorganisms was not (lawn).

Purification of cavities, abscesses of the pathogen in animals of the first group celebrated the fourteenth day. When treating isolated NEELY (the second group) of the cavities, abscesses animals continued pus throughout the observations In animals of the third group of the cavity was cleared for the twelfth day of treatment.

Figure 2 - photo ultrasound on the 14th day of treatment, where the animals of the first group of the diameter of the cavity was reduced to 1.1 cm, the second remained the same (1.5 cm), the animals of the third group - 0.9 cm, the fourth - 0.5, see

The application of the standard rehabilitation of liver abscesses in the experiment (animals of the first group) cavity was cleansed of the pathogen only to the fourteenth day, the amount was reduced to 1.1 cm, and by the thirtieth day, the size of the cavities was 0.7, see Treatment NEELY (the second group) did not lead to the purification of the abscess cavity even to the thirtieth day of the experiment, when ultrasound diameter cavity remained the same throughout the observation. In the processing of nanoparticles of copper (third group) to the fourteenth day, the size of the residual cavity was decreased to 0.9 cm from the pathogen cavity was cleared only for the twelfth day, and thirtieth days of the diameter of the abscesses was 0.5 cm

For the first time revealed the effect of enhancement of antimicrobial activity of copper nanoparticles under the action of LLLT allows you to start treatment without determining the type of pathogen, significantly reduce the concentration of the nanoparticles, completely inhibiting the growth of pathogenic microorganisms, and reduce its toxic effects on the macro-organism.

The combination of nanomed and NEELY promptly cleaned the infected cavity from which Sudetes, as evidenced by the lack of growth of colonies of microorganisms already by the seventh day. According to the U.S. by the fourteenth day of combined treatment of liver abscesses the size of the cavities were reduced three times, to the thirties was not determined.

The method is illustrated by the following example.

EXAMPLE

Rabbit-male breed chinchilla age 1 year weight 3.5 kg with the simulated liver abscess with a diameter of 1.5 cm daily aspirated the contents from the cavity through the catheter, then filled the cavity of the abscess suspension of copper nanoparticles in 0.9% physiological solution of sodium chloride concentration of 1 μg/ml, was placed in the center cavity of the fiber laser, equipped with a diffuser, through the sanitizing channel of the catheter and perform laser irradiation with a wavelength of 630 nm in continuous mode output power of 35 mW for 3 minutes. On the seventh day noted the lack of growth of colonies of Staphylococcus aureus (reference sowing of the abscess cavity. On the ninth day of treatment had removed the catheter. According to the U.S. by the fourteenth day of the diameter of the residual cavity was 0.5, see the Control ultrasound was performed on the thirtieth day, and the abscess cavity was not determined.

Thus, the effect of enhancement of antimicrobial activity of copper nanoparticles under the action of LLLT allows you to start treatment without determining the type of pathogen, in the shortest possible time is fully suppress pathogenic microorganisms, successfully stimulate reparative processes, to reduce the toxic effect of nanoparticles on macroorganism, significantly reduce the duration of treatment of purulent cavities.

The method of combined treatment of abscesses in the experiment, including the aspiration of the cavity through the catheter and the impact of the suspension of copper nanoparticles in physiological solution of sodium chloride, characterized in that the abscess cavity is filled with a suspension of copper nanoparticles in physiological solution of sodium chloride concentration of 1 μg/ml, while conducting intracavitary laser irradiation with a wavelength of 630 nm in continuous mode output power of 35 mW daily for 3 minutes within ten days.



 

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3 ex

FIELD: medicine.

SUBSTANCE: invention relates to field of medicine, namely to ophthalmology, and is intended for transpupillary thermotherapy of amelanotic melanoma of choroid of medium size. Transpupillary thermotherapy by infrared diode laser 810 nm is performed to tumour tissue temperature 50-60°C, irradiation power being selected depending on degree of tumour pigmentation and on its size in central zone. At the first stage of treatment laser with wavelength 561 nm is used to perform laser barrage around neoplasm with power parameters - 200 mW, exposure - 0.05 msec, coagulant diameter 300 mcm, coagulants being laid in tiled way with formation of coagulant strip of III degree, up to 1 mm wide. After that, coagulation of intra-tumour vessels is performed with parameters of radiation power 1200 mW, exposure 0.5 sec, spot diameter 1000 mcm, in tiled way for coagulant of II degree. The second stage is carried out after 2 weeks by laser 810 nm, power of radiation is selected depending on rate of manifestation of calorimetric signs of laser impact. Coagulants are applied by method of "daisy", starting with the edge of tumour, allowing their small overlay on each other, central the highest part of tumour being processed last.

EFFECT: method makes it possible to reduce the number of intra- and post-operative complications.

FIELD: medicine.

SUBSTANCE: invention relates to medicine and can be used for photodynamic therapy of intraocular neoplasms. For this purpose photosensitiser chlorine e6 monoethylene diamine monoamide adipinate is introduced intravenously in dose 0.4 mg/kg during 10 minutes. 60 minutes after finishing intravenous introduction of PS, fluorescent diagnostics of PS accumulation in neoplasm is started. Intensity of PS fluorescence in intraocular neoplasm is controlled. When maximal fluorescence of neoplasm in comparison with surrounding tissue is achieved, neoplasm is irradiated transpupillarily with laser radiation. Wavelength corresponds to maximum of luminous radiation absorption by photosensitiser.

EFFECT: method makes it possible to perform treatment of intraocular neoplasms of small and medium size, reduce dose of introduced preparation and density of laser radiation energy, accelerate complete tumour regress due to higher PS tropism by tumour cells.

3 cl, 3 dwg, 2 ex

FIELD: medicine.

SUBSTANCE: invention relates to medicine, namely to pulmonology, and can be used for treatment of bronchopulmonary diseases. For this purpose traditional drug treatment of pathology is carried out, additionally performed is impact by infrared laser radiation (IRLR) and successive inhalation of mineral water aerosols. Impact by infrared laser radiation is performed on projection of bronchi and lungs with total superficial energy density 0.06-0.65 J/cm2 for 4-8 minutes per 1 procedure. For inhalation used are aerosols of mineral water in dose 6-15 ml with temperature 40-42°C. Inhalation is performed for 4-10 minutes per 1 procedure. Course of therapy constitutes 7-12 daily performed procedures.

EFFECT: method ensures increase of treatment efficiency due to effect of synergistic effect on cell elements and walls of bronchi as a result of photoionophoresis, namely quantum modification of membrane of epithelium cells, activation of state of ion channels, and therefore selective permeability of ion channels in the following inhalation, which ensures enhancement of ion diffusion into tissues of bronchi and lungs, intracellular and intercellular exchange.

8 tbl, 7 ex

FIELD: medicine.

SUBSTANCE: invention refers to medicine, namely to otorhinolaryngology, and can find application in engraftment of a neotympanic flap following tympanoplasty. A method involves application of the flap to borders of a tympanic perforation and physical effect on the flap. The flap is exposed to a gas flow containing 0.1-0.3% nitrogen monoxide at temperature 18-20°C with exposition of 1 min. 30 sec. A manipulator tip is introduced in an external auditory canal for max. 1 cm with its output placed at 45°C to a plane of a posterior wall of the external auditory canal. The treatment is conducted once a day for 5-10 days.

EFFECT: use of the given invention enables providing effective engraftment of the neotympanic membrane due to stimulation of repair processes, as well as avoiding attachment of a secondary infection.

2 ex

FIELD: medicine.

SUBSTANCE: invention refers to veterinary science. The preparation metallosuccinate 10 ml is introduced intramuscularly three times before calving and two times after every 10 days.

EFFECT: method is effective for correction of the immunobiochemical status in ante- and postpartum cows, preventing reproduction pathologies and producing high-resistant newborn calves.

2 tbl, 1 ex

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