RU2466720C2 - Method of treating infected wounds in experiment - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: during phases of regeneration and epithelisation, a wound surface is coated with a dressing impregnated with an ointment of the following composition, wt %: organic-silicon glycerohyrogel - 95.25, ciprofloxacin - 0.2, oxytocin - 4.55.

EFFECT: method provides faster regenerative processes at the II and III phases of the wound process, prevented the onset of persistent infection and purulent-septic complications, reduced length of treatment.

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Description

The invention relates to medicine, in particular to medical microbiology, and may find application in various fields of practical medicine for the treatment of purulent wounds.

It is known that the wound process is a complex set of reactions developing in the body in response to tissue damage, which can be divided into three successive phases: purulent-necrotic phase (phase I), characterized by the presence of necrotic tissue and purulent contents in the wound; phase of regeneration (granulation) (II phase) - cleansing the wound from purulent-necrotic contents and the formation of granulation tissue in it, gradually filling the wound cavity; epithelization phase of the wound surface and reorganization of the scar (phase III) [Wounds and wound infection. Ed. M.I. Kuzina, B.M. Kostyuchenko. M., Medicine, ed. 2nd rev. and add., 1991. - 608 p.].

A correctly selected treatment algorithm, the effectiveness of which depends on the optimal selection of therapeutic agents taking into account the phase of the wound process, allows you to quickly eliminate the acute purulent process, suppress microorganisms remaining in the wound or prevent reinfection of the wound surface, accelerate epithelization and, as a result, reduce the length of hospital stay of patients .

Currently, modern methods of treating purulent wounds involve the use of drugs, depending on the objectives of therapy, taking into account the phase of the wound process.

However, despite the successes of clinical medicine and pharmacological science, the problem of treatment of purulent wounds does not cease to be urgent, since the issues of effective treatment taking into account the phase of the wound process are not always solved successfully, and the use of antimicrobials in relation to antibiotic-resistant persistent microflora is ineffective, which leads to the development of microbial biofilms resistant to therapeutic effects and recurrence of the inflammatory process in the wound with the development of purulent-septic complications.

Known methods for treating the wound surface in the purulent-necrotic phase (phase I of the wound process) are that after surgical treatment of the wound, dressings with a drug having a wide spectrum of antimicrobial action, osmotic properties and exhibiting anti-inflammatory and analgesic effects are applied to its surface (" Levomekol ”,“ Levosil ”,“ Dioxol ”, etc.) [Blatun L.A., Svetukhin AM, Paltsin A.A., Lyapunov N.A., Agafonov V.A. Clinical and laboratory effectiveness of modern polyethylene glycol-based ointments in the treatment of purulent wounds. // Antibiotics and chemotherapy, 1999. - No. 7, - S.25-31].

Known methods traditionally used in clinical practice for treating a wound surface in the regeneration phase (phase II) are applying a dressing with ointment on a hydrophilic or hydrophobic emulsion base to the wound surface, which has a weak osmotic effect, protects granulation tissue from mechanical damage and other negative factors, suppresses the remaining in a small number of microorganisms in the wound, prevents secondary infection and stimulates regenerative processes wound (methyluracyl ointment, "Betadine", "Eksalb", "Metildoksitsillin" et al.) [Blatun LA Phlegmon and abscesses - modern treatment options. // Attending physician, 2002. - No. 2. - S.30-40].

The methods for treating purulent wounds in phase III coincide with the methods for treating purulent wounds in the regeneration phase (phase II of the wound healing process) [Bagirova VL, Demina NB, Kulinichenko NA Ointments. A modern look at the dosage form. // Pharmacy. - 2002. - No. 2. - S.24-26].

A known method of treating purulent wounds using multicomponent ointments based on a hydrophilic base (Levomekol, Levosin, Dioxol, etc.), where a mixture of polyethylene oxides of different molecular weights (PEO-400 and PEO-1500 in the ratio 4: 1) [Datsenko B.M., Shalimov O.O., Bezuglaya E.P. Theory and practice of local treatment of purulent wounds. Kiev, 1995. - S.238-260].

However, the known method of treatment is carried out without taking into account the phases of the wound process, and therefore is not effective in the phases of regeneration and epithelization due to the fact that the use of an ointment based on a mixture of polyethylene oxides overdries the wound surface (due to the strong osmotic effect) and does not affect the processes regeneration in the wound. Treatment with this method does not give the desired result also because the development of the chronic inflammatory process impedes the process of regeneration of the wound surface, since antimicrobial agents, traditionally included in the ointment, do not have an antipersistent effect on microorganisms and do not inhibit their formation of biofilms, which leads to the formation of a focus of chronic persistent infection and the occurrence of purulent-septic complications in patients.

A known method of treating purulent wounds taking into account the phases of the wound process, including surgical treatment of the wound surface and subsequent application of a dressing with drugs on the wound surface: ointments with high osmotic properties are used in the purulent-necrotic phase (Levomekol, Levosin, Dioxikol ”And others), and in the phases of regeneration and epithelization - multicomponent ointments that stimulate regeneration processes in the wound (methyluracil ointment, Betadin, Exalb, Methyldoxycillin, etc.) [Blatun L.A. Phlegmon and abscesses - modern treatment options. // Attending physician, 2002. - No. 2. - S.30-40].

The disadvantages of this method include the fact that the antimicrobial agents that make up the ointments, traditionally used to treat purulent wounds in the first, second and third phases of the wound healing process, do not have an antipersistent effect on microorganisms, and therefore a focus of chronic persistent infection is formed , interfering with the regeneration process in the wound due to a sluggish inflammatory process.

Closest to the claimed method in terms of essential features is a method of treating acute purulent-inflammatory diseases of soft tissues of microbial etiology, including surgical treatment of a wound (opening a purulent foci, pus evacuation, necrectomy, washing the wound with an antiseptic solution) and applying a dressing soaked in antibacterial hydrophilic ointment based on polyethylene oxide. The composition of the ointment includes 0.2 g of ciprofloxacin, 5 ME oxytocin, 19.05 g of polyethylene oxide-1500, 76.2 g of polyethylene oxide-400 (RF patent No. 2306947, IPC A61K 38/11, A61K 31/47, A61K 31/765 , A61P 31/04, A61B 17/00, 2007).

The disadvantages of this method is the limitation of the use of ointments based on a mixture of polyethylene oxides in the phases of regeneration and epithelization of wounds (II and III phases), since they dry the wound surface due to high osmotic properties, worsening the regeneration processes, and do not have regenerative activity, which increases the healing time of wounds due to the longer course of the II and III phases of the wound process.

The objective of the invention is to increase the effectiveness of treatment of purulent wounds by enhancing regenerative processes in the wound in the II and III phases of the wound process and preventing the occurrence of persistent infection.

The problem is achieved due to the fact that in the method of treating purulent wounds in an experiment by surgical treatment and subsequent application of a dressing with drugs on the wound surface, in the phases of regeneration and epithelization on the wound surface, a dressing impregnated with an ointment of the following composition, wt.%:

organosilicon glycerohydrogel - 95.25

ciprofloxacin - 0.2

oxytocin - 4.55

The technical result achieved during the implementation of the invention is that the claimed method, by potentiating the regenerative and antipersistent action of the preparations, prevents the occurrence of persistent infection, accelerates the regenerative processes in phases II and III of the wound healing process and, therefore, reduces the patient’s hospital stay.

The application of the proposed method expands the arsenal of tools and treatment methods used in the treatment of purulent wounds.

A distinctive feature of the proposed method is the application in the phases of regeneration and epithelialization (II and III phases) of a dressing soaked in ointment of the following composition, wt.%:

organosilicon glycerohydrogel - 95.25

ciprofloxacin - 0.2

oxytocin - 4.55

Organosilicon glycerohydrogel (Silativit) is a substance with the chemical formula Si (C ₃ H ₇ O ₃ ) ₄ · 6C ₃ H ₈ O ₃ · 24H ₂ O (RF Patent 2255939, MKI C07F 7/04, 2005 Reg. No. 1075114) (patent holder - I.Y.Postovsky Institute of Organic Synthesis, Ural Branch of the Russian Academy of Sciences, Ekaterinburg), which is a gel-like translucent mass of white color, odorless. It has wound healing, regenerative, anti-inflammatory, decongestant and antioxidant activity, high transcutaneous conductivity of various medicinal substances, the ability to protect tissues from drying out and edema, increase their oxygenation, structural compatibility with the lipid component of cell membranes. At the same time, it does not have any negative side effects, it is stable, convenient for storage and is well washed off, it is highly soluble in 95% alcohol and water and easily transfers the medicinal substances included in it to the skin.

Ciprofloxacin (1-cyclopropyl-6-fluoro-1,4-dihydro-4-oxo-7- (1-piperazinyl) -3-quinoline carboxylic acid hydrochloride monohydrate) refers to monofluoroquinolones with a wide antibacterial spectrum, including gram-positive and gram-negative pathogens processes. Manufacturers: Tsiprolet (Dr. Reddy s, India), tsiprobay, tsifran, etc. Available in tablet (250 mg, 500 mg) and injectable (2% solutions) forms.

Oxytocin is a peptide hormone of the posterior pituitary gland (nonapeptide), obtained synthetically, molecular weight 1007.2. Manufacturers: Gideon Richter Chemical Plant (Hungary), Endocrine Drug Plant (Moscow). Available in 1 ml ampoules containing 5 ME oxytocin.

The use of oxytocin in combination with antibiotics is known in the treatment of purulent diseases of soft tissues [Kurlaev P.P. Experimental and clinical justification for the use of oxytocin in the complex antibacterial therapy of purulent diseases of soft tissues: Abstract. Cand. honey. sciences. - Perm, 1986 - 24 pp.], To stimulate reparative processes in the wound [Gavrilenko V.G. Clinical rationale for the use of oxytocin in the complex treatment of diabetic purulent-necrotic lesions of the feet: Abstract. Cand. honey. sciences. - Orenburg, 1999 - 25 p.].

It is known that oxytocin potentiates the inhibitory effect of ciprofloxacin on the persistent potential of bacterial pathogens and increases the antibacterial activity of the antimicrobial agent against pathogens of acute purulent-inflammatory diseases of soft tissues [Bukharin OV, Skorobogatykh Yu.I., Kurlaev P.P. Experimental substantiation of the effectiveness of the combined use of ciprofloxacin with oxytocin. // Journal. microbiol. 2007. No. 5, p. 70-73].

In purulent-inflammatory diseases, the most frequent chronization of the infectious process contributes to an increase in the antilysocyme trait of microorganisms of the pathogens of purulent wound healing, and due to the formation of biofilms, the resistance of pathogens to the action of antibiotics, antiseptics and natural defense factors of the human body [Bukharin OV The persistence of pathogenic bacteria. M .: - Medicine; Yekaterinburg: Uro RAS, 1999. S.195-231; Lewis K. Persister Cells and the Riddle of Biofilm survival. // Biochemistry, 2005. V.70, No.2. P.267-274].

The authors experimentally established the potentiation of the inhibitory effect on biofilm formation and antilysocyme activity of the microorganism population of the pathogenic purulent pathogen pathogens and stimulation of regenerative processes in the tissue using a combination of ciprofloxacin with oxytocin and organosilicon glycerohydrogel.

At the first stage of the study, the antimicrobial activity of ciprofloxacin, oxytocin, organosilicon glycerohydrogel and the combined effect of ciprofloxacin with oxytocin and organosilicon glycerohydrogel were evaluated using the method of holes in the thickness of the agar [Egorov N.S. The basics of the doctrine of antibiotics. Textbook for students a biologist. specialist. high boots. 4th ed., Rev. and add. M .: Higher. shk., 1986. P.160] on the strains of bacteria that are most often encountered in a purulent wound process: Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosae (40 strains of each species).

The antimicrobial effect of ciprofloxacin (200 mg), oxytocin (5 ME), organosilicon glycerohydrogel, and their combination, which was the following composition, ointment of the following composition, wt.%: Organosilicon glycerohydrogel — 95.25, ciprofloxacin — 0.2 and oxytocin — 4.55 were evaluated. The ointment was prepared as follows: organosilicon glycerohydrogel in an amount of 95.25 g was heated in a water bath at 50 ° C until a homogeneous transparent mass was formed for 20 minutes, cooled to 37 ° C, and then 4 ml of 5% ciprofloxacin (0.2 g) was added ) and 1 ml oxytocin (5 IU / ml). The final concentration of ciprofloxacin in the ointment (200 mg / g) corresponded to the recommended single dose of the drug [Vidal 2011. Reference Vidal. Medicines in Russia // Publisher: AstraFarmServis, 2011. 1728 p.] And oxytocin - the minimum average therapeutic dose of 5 ME [Zak V.I., Kurlaev P.P. Topical treatment of post-injection abscesses with a combination of oxytocin and antibiotics // Surgery. - 1985. - No. 5. - C.112-115].

It was found that the most pronounced antimicrobial effect (inhibition of the growth of 100% of the studied bacterial strains) was observed when the effects of ciprofloxacin with oxytocin and organosilicon glycerohydrogel were combined. Ciprofloxacin had antimicrobial activity against 75% of the studied microorganism strains, organosilicon glycerohydrogel - 30% of bacterial strains, and oxytocin did not show antimicrobial activity.

At the next stage, to determine the effects of ciprofloxacin, oxytocin, organosilicon glycerogel hydrogel, and the combined effect of ciprofloxacin with oxytocin and organosilicon glycerogel hydrogel on antilysocyme activity and biofilm formation of microorganisms, in vitro experiments were also performed on bacterial strains of S. aureus, E. coli bacteria, E. coli (40 strains of each species).

Ciprofloxacin, oxytocin, organosilicon glycerohydrogel and the prepared ointment were introduced into the nutrient broth at a ratio of 1:10 (Nutrient Medium NPO, Makhachkala), inoculated with the studied bacterial strains and cultured for 9-10 hours (until the transition of the microorganism strains from logarithmic to stationary population growth phase). A mixture of nutrient broth with isotonic sodium chloride and placebo oxytocin was used as a control.

Then, the resulting bacterial broth cultures were centrifuged at 3000 rpm for 15 minutes, the cells were washed twice with a 0.85% sodium chloride solution and scattered on solid nutrient agar (NPO Nutrient Sredy, Makhachkala).

Antilysocyme activity (ALA) and the ability to form bacterial biofilms (BPO) were determined by the photometric method [O. Bukharin. The persistence of pathogenic bacteria. - M .: Medicine, 1999. - P.88-89; O'Toole G.F., Kaplan H. B., Kolter R. Biofilm formation as micribial development. Ann Rev Microbiol, 2000; 54: p. 49-79].

The effect of ciprofloxacin, oxytocin, organosilicon glycerogel hydrogel and the combined effect of ciprofloxacin with oxytocin and organosilicon glycerogel hydrogel on the antilysocyme activity and biofilm formation of bacterial strains S. aureus, E. coli and P.aeraginosae was judged by a change in the ratio of bacterial strains to high, low activity bacteria and biofilm forming ability. To identify statistically significant differences in the compared groups used nonparametric Wilcoxon test [Lakin G.F. Biometrics. - M .: Higher School, 1990. - S.128-134].

The results of the effect of ciprofloxacin, oxytocin, organosilicon glycerogel hydrogel and the combined effect of ciprofloxacin with oxytocin and organosilicon glycerogel hydrogel on the antilysocyme activity and biofilm formation of bacterial strains S. aureus, E. coli and P. aeruginosae are presented in the diagrams (Figs. 1 and 2).

Figure 1 shows the effect of oxytocin-placebo, organosilicon glycerogel hydrogel, oxytocin, ciprofloxacin and the combination of ciprofloxacin with oxytocin and organosilicon glycerogel hydrogel on bacterial antilysimic activity, where horizontally indicated in the numerator: 1 - S. aureus, 2 - E. coli P. aeruginosae, and in the denominator: 1 - control (ALA of bacteria before drug exposure), 2 - ALA after exposure to oxytocin-placebo, 3 - ALA after exposure to organosilicon glycerogel hydrogel, 4 - ALA after exposure to oxytocin, 5 - ALA after exposure effects of ciprofloxacin, 6 - ALA after exposure to a combination of ciprofloxacin with oxytocin and organosilicon glycerohydrogel.

Vertical indicates% bacteria. In the columns, white indicates low ALA (0-0.8 μg / ml * OD), oblique shading indicates average ALA (0.81-1.5 μg / ml * OD), black indicates high ALA (more 1.5 μg / ml * OD).

As can be seen from the presented diagrams, in the ALA control of the studied bacterial groups varied: 10% of S. aureus cultures had low ALA values, 55% had medium and 35% high; 15% of E. coli cultures had low ALA, 65% had medium, 20% had high; 10% of P. aeruginosae cultures had low ALA values, 65% had medium ALA and 25% had high ALA.

Analysis of the data showed that placebo-oxytocin did not affect the ALA of the studied bacteria.

Among all the drugs, the most pronounced decrease in ALA of microorganisms was caused by a combination of ciprofloxacin with oxytocin and organosilicon glycerohydrogel (columns 1/6, 2/6 and 3/6) (p <0.05), under the influence of which the majority (from 90% to 100% bacteria) of S. aureus, E. coli and P. aeruginosae strains showed low ALA values, and high ALA values in bacteria were absent.

Figure 2 shows the effect of oxytocin-placebo, organosilicon glycerogel hydrogel, oxytocin, ciprofloxacin and the combination of ciprofloxacin with oxytocin and organosilicon glycerogel hydrogel on bacterial biofilm formation, where in the numerator are indicated horizontally in the numerator: 1 - control (after BPO of bacteria - before drug exposure) exposure to oxytocin-placebo, 3 - BPO after exposure to organosilicon glycerohydrogel, 4 - BPO after exposure to oxytocin, 5 - BPO after exposure to ciprofloxacin, 6 - BPO after exposure combining of ciprofloxacin with oxytocin and organosilicon glitserogidrogelem.

Vertical indicates% bacteria. In the columns, white values indicate low BPO (0.2 OD units or less), oblique shading indicate average BPO values (0.21-0.4 OD units), black indicate high BPO values (more than 0.4 units) . OD).

The ability to form biofilms among the studied microorganisms in the control also varied: in S. aureus, 20% of the strains had low BPO, 45% had medium, and 35% had high BPO; in E. coli, 10% of the strains showed low BPO, 60% medium, and 30% high BPO; in P. aeruginosae, 20% of the strains had low BPO, 60% had medium, and 20% had high BPO.

Analysis of the data showed that placebo-oxytocin did not affect the BPO of the studied bacteria.

Among all the acting drugs, the most pronounced decrease in the BPO of the studied microorganisms was caused by the combination of ciprofloxacin with oxytocin and organosilicon glycerohydrogel (columns 1/6, 2/6 and 3/6) (p <0.05), under the influence of which the majority (from 85 ± 9, 0% to 95 ± 5.0% bacteria) of S. aureus, E. coli and P. aeruginosae strains showed low BPO values, and high BPO values in bacteria were absent.

In vivo experiments were carried out on 48 white mongrel male rats weighing 150 g - 200 g, in which a purulent wound was simulated. The conditions for keeping and feeding the animals were the same.

A model of an infected wound in rats was created as follows: after excision of a skin flap in the scapular region 2 × 2 cm (400 mm ² ) to the muscle layer, crushing of the edges of the wound and muscle with a Kocher clamp was performed. The wounds of the experimental animals were infected with a strain of S.aureus 25923 ATCC isolated from a patient with a purulent process of soft tissues in the amount of 10 ⁹ CFU / ml. The operation was performed under canned anesthesia in non-sterile conditions.

For the experiment, animals were divided into three groups.

In all three groups, wound treatment was started in the presence of signs of acute purulent inflammation (hyperemia of the wound edges, presence of inflammatory edema, purulent plaque, purulent wound secretion) and it was carried out in the first phase of the wound process by traditional methods by surgical treatment of the wound (opening of the purulent focus, pus evacuation , necrectomy, washing the wound with an antiseptic solution) and applying a bandage to the wound surface with Levomekol ointment once a day until there are signs of the beginning of the regeneration phase, the criteria of which were Is: the absence of purulent-necrotic discharge from the wound, the formation of granulations and the appearance of marginal epithelization of the wound.

In the phases of regeneration and epithelization, a bandage was applied to the wound surface once a day:

1) group 1 - with ointment "Methyldoxycillin" at a dose of 1 mg per wound;

2) group 2 - with a combination of ciprofloxacin with oxytocin based on a mixture of polyethylene oxides (RF patent No. 2306947, IPC A61K 38/11, A61K 31/47, A61K 31/765, A61P 31/04, A61B 17/00, 2007), c dose of 1 mg per wound;

3) group 3 - with an ointment based on organosilicon glycerohydrogel of the following composition, wt.%: Organosilicon glycerohydrogel - 95.25, ciprofloxacin - 0.2 and oxytocin - 4.55, at a dose of 1 mg per wound.

An ointment based on organosilicon glycerohydrogel was prepared as follows: organosilicon glycerohydrogel in the amount of 95.25 g was heated in a water bath at 50 ° C until a homogeneous transparent mass was formed for 20 minutes, cooled to 37 ° C, and then 4 ml of 5% ciprofloxacin was added ( 0.2 g) and 1 ml oxytocin (5 IU / ml).

The composition of the ointment (wt.%): Organosilicon glycerohydrogel - 95.25; ciprofloxacin - 0.2; oxytocin - 4.55.

The course of treatment lasted depending on the speed of wound healing.

The decapitation of animals followed by histological examination of the regenerate tissue was carried out at least three times in each of the phases of the wound healing process, starting from 3 days after the wound was applied.

The molten scab along with the wound was removed, the remaining regenerate tissues together with the surrounding intact skin were excised sectorally, fixed in 10% neutral formalin and embedded in paraffin. Sections 5-1 μm thick were stained with hematoxylin-eosin. The percentage reduction in the area of the wound was found by the formula

where S is the area of the previous measurement (in mm ² );

S _n is the area of this measurement (in mm ² );

S _t is the number of days between measurements.

Statistical processing was performed with the calculation of student criterion.

Comparative results of the treatment of three groups of rats with purulent soft tissue wounds are presented in the table.

As can be seen from the data obtained, the timing of detection of signs of the onset of phase II of the wound process did not differ in all compared groups.

In the phases of regeneration and epithelialization of purulent wounds in animals, signs of secondary infection of the wound were detected in 58% of rats in group I and only 8.3% of cases in groups II and III.

The number of macrophages and fibroblasts reflecting the regenerative processes occurring in the wound was significantly higher in rats of group III, compared with group I and group II. As a result of the treatment of purulent wounds in animals of group III, complete healing of the wound was faster (16.4 ± 0.7 days from the start of treatment) than in rats I and II of the compared groups (20.4 ± 2.5 and 21, respectively) , 4 ± 2.5 days from the start of treatment).

Thus, the treatment of purulent wounds with the use of a new ointment based on organosilicon glycerohydrogel in the phases of regeneration and epithelialization contributed to the stimulation of regenerative processes in the tissue and led to faster healing of the wound: 4 days earlier than with the traditional treatment with methyldoxicillin ointment and 5 days earlier than during treatment using a combination of ciprofloxacin with oxytocin based on a mixture of polyethylene oxides.

The method is as follows:

1. Under general ether anesthesia in white outbred rats in the scapular region, a skin wound is applied to a muscle 2 × 2 cm (400 mm ² ) in size and crush the edges of the wound and muscle with a Kocher clamp. 0.2 ml of S. aureus suspension diluted to 10 ⁹ CFU / ml are applied to the wound with a pipette;

2. In the purulent-necrotic phase of the wound healing process in the presence of signs of acute purulent inflammation (hyperemia of the edges of the wound, the presence of inflammatory edema, purulent plaque, purulent wound secretion), wounds are treated by traditional methods consisting in surgical treatment (opening of the purulent focus, pus evacuation, necrectomy washing the wound with an antiseptic solution) and then applying a bandage to the wound surface with a medicinal product with a wide spectrum of antimicrobial action, osmotic properties and showing anti-inflammatory and analgesic effects (“Levomekol”, “Levosil”, “Dioxol”, etc.). The dressing is applied once a day until there are signs of the beginning of the regeneration phase. The onset of the regeneration phase is determined by the criteria: the absence of purulent-necrotic discharge from the wound, the formation of granulations and the appearance of marginal epithelization of the wound;

3. Prepare an ointment of the following composition (wt.%): Organosilicon glycerohydrogel - 95.25; ciprofloxacin - 0.2; oxytocin - 4.55. To do this, organosilicon glycerohydrogel in an amount of 95.25 g is heated in a water bath at 50 ° C until a homogeneous transparent mass is formed for 20 minutes, cooled to 37 ° C, and then 4 ml of 5% ciprofloxacin (0.2 g) and 1 are added ml oxytocin (5 IU / ml);

4. At the onset of the regeneration phase, a dressing with the obtained ointment is applied to the wound for a day;

5. Dressings are carried out daily until the wound surface heals, which is judged by the presence of complete epithelization of the wound surface.

Examples of specific performance of the method.

Example 1

Laboratory animal No. 15.

Under general ether anesthesia in a white outbred rat in the scapular region, a skin wound was applied to a muscle 2 × 2 cm (400 mm ² ) in size and crushed the wound edges and muscles with a Kocher clamp. 0.2 ml of S. aureus suspension diluted to 10 ⁹ CFU / ml was applied to the wound with a pipette.

In the first phase of the wound process, when the wound took signs of acute purulent inflammation (hyperemia of the edges of the wound, the presence of inflammatory edema, purulent plaque, purulent secretion of the wound), the treatment of the wound was started in the traditional way, consisting in surgical treatment and subsequent application of a dressing with ointment on the wound surface water-soluble base "Levomekol" once a day.

A bacteriological study of the purulent-necrotic phase of the wound process was performed on day 3 from the start of treatment. The microbial contamination rate of the wound (PMO), antilysocyme activity (ALA), and biofilm formation (BP) of the S. aureus strain were determined.

On the 3rd day, the indicator of microbial dissemination of staphylococcus was 5 * 10 ⁷ CFU / ml, antilysocyme activity of S. aureus was 1.53 μg / ml * OD, the ability to form biofilms was 0.58 units. OP.

Treatment was performed until signs of the onset of the regeneration phase appeared. The onset of the regeneration phase was determined by the criteria: the absence of purulent-necrotic discharge from the wound, the formation of granulations and the appearance of marginal epithelization of the wound.

The regeneration phase occurred on the 7th day from the start of treatment. At this time, slight swelling of the tissues was noted, there was a small amount of purulent contents under the edges of the wound, and marginal epithelization of the wound was observed.

Bacteriological examination of the wound on day 7 showed the presence of staphylococcus in the wound, the number of which decreased compared to the initial data (5 * 10 ⁵ CFU / ml), but persistence factors and the ability to form biofilms increased and amounted to 1.0 μg / ml * OD, respectively and 1.1 units OP.

With the onset of the regeneration phase (7 days after the start of treatment), a dressing impregnated with an ointment of the following composition (wt.%) Is applied to the wound surface: organosilicon glycerohydrogel - 95.25, ciprofloxacin - 0.2 and oxytocin - 4.55.

The ointment was prepared as follows: organosilicon glycerohydrogel in an amount of 95.25 g was heated in a water bath at 50 ° C until a homogeneous transparent mass was formed for 20 minutes, cooled to 37 ° C, and then 4 ml of 5% ciprofloxacin (0.2 g) was added ) and 1 ml oxytocin (5 IU / ml).

The dressing was changed daily, 1 time per day.

On day 10, with a wound area of 65%, a control bacteriological culture was performed. Growth of S. aureus in the wound was absent.

The completion of epithelialization of the wound surface was noted at 16 days.

Example 2

Laboratory animal No. 24.

Under general ether anesthesia in a white outbred rat in the scapular region, a skin wound was applied to a muscle 2 × 2 cm (400 mm ² ) in size and crushed the wound edges and muscles with a Kocher clamp. 0.2 ml of S. aureus suspension diluted to 10 ⁹ CFU / ml was applied to the wound with a pipette.

In the first phase of the wound process, when the wound took signs of acute purulent inflammation (hyperemia of the wound edges, the presence of inflammatory edema, purulent plaque, purulent wound secretion), the wound treatment was started in the traditional way, consisting in surgical treatment and subsequent application of a dressing with dioxicol ointment on the wound surface "Once a day.

During bacteriological examination of the wound in the purulent-necrotic phase on the 3rd day, the PMA of staphylococcus was 1 * 10 ⁵ CFU / ml, ALA - 1.57 μg / ml * OD, BPO - 0.53 units. OP.

The regeneration phase began on the 6th day: there was a slight swelling of the tissues, there was a small amount of purulent contents under the edges of the wound, and marginal epithelization of the wound was observed.

Bacteriological examination of the wound on the 6th day showed a slight decrease in the PMO of S. aureus (1 * 10 ⁴ CFU / ml) and an increase in ALA and BP of S. aureus (2 μg / ml * OD, - 1.05 units, respectively) .

With the onset of the regeneration phase (6 days after the start of treatment), a dressing impregnated with an ointment of the following composition (wt.%) Was applied to the wound surface: organosilicon glycerohydrogel - 95.25, ciprofloxacin - 0.2 and oxytocin - 4.55.

The ointment was prepared as follows: organosilicon glycerohydrogel in an amount of 95.25 g was heated in a water bath at 50 ° C until a homogeneous transparent mass was formed for 20 minutes, cooled to 37 ° C, and then 4 ml of 5% ciprofloxacin (0.2 g) was added ) and 1 ml oxytocin (5 IU / ml).

The dressing was changed daily, 1 time per day.

On day 10, with a wound area of 65%, a control bacteriological culture was performed. Growth of S. aureus in the wound was absent.

The completion of epithelialization of the wound surface was noted on the 15th day from the start of treatment.

Example 3

Laboratory animal No. 30.

Under general ether anesthesia in a white outbred rat in the scapular region, a skin wound was applied to a muscle 2 × 2 cm (400 mm ² ) in size and crushed the wound edges and muscles with a Kocher clamp. 0.2 ml of S. aureus suspension diluted to 10 ⁹ CFU / ml was applied to the wound with a pipette.

In the first phase of the wound process, when the wound took signs of acute purulent inflammation (hyperemia of the edges of the wound, the presence of inflammatory edema, purulent plaque, purulent secretion of the wound), the treatment of the wound was started in the traditional way, consisting in surgical treatment and subsequent application of a dressing soaked in ointment “ Syntomycin "once a day.

During bacteriological examination of the wound on the 3rd day, the PMA of staphylococcus was 1 * 10 ⁷ CFU / ml, ALA - 1.71 μg / ml * OD, BPO - 0.67 units. OP.

The regeneration phase began on the 9th day: there was a slight swelling of the tissues, there was a small amount of purulent contents under the edges of the wound, and marginal epithelization of the wound was observed.

Bacteriological examination on the 9th day showed a decrease in S. aureus PMO (1 * 10 ⁵ CFU / ml) and an increase in ALA and BPO of staphylococcus (2.1 μg / ml * OD, 1.3 units OD, respectively).

With the onset of the regeneration phase (9 days from the start of treatment), a dressing impregnated with an ointment of the following composition (wt.%) Was applied to the wound surface: organosilicon glycerohydrogel — 95.25, ciprofloxacin — 0.2, and oxytocin — 4.55.

The ointment was prepared as follows: 95.25 g organosilicon glycerohydrogel was heated in a water bath at 50 ° C until a homogeneous transparent mass was formed for 20 minutes, cooled 40 minutes to 37 ° C, and then 4 ml of 5% ciprofloxacin was added (0, 2 g) and 1 ml oxytocin (5 IU / ml).

The dressing was changed daily, 1 time per day.

Treatment has begun by applying dressings with a new ointment based on a combination of ciprofloxacin with oxytocin and organosilicon glycerohydrogel.

On day 13, with a wound area of 70%, a control bacteriological culture was performed. Growth of S. aureus in the wound was absent.

The completion of epithelialization of the wound surface was obtained on the 18th day from the start of treatment.

The proposed method for the treatment of purulent wounds in the experiment allows you to accelerate the regenerative processes in the II and III phases of the wound process, prevent the occurrence of persistent infections and purulent-septic complications and, therefore, reduce the treatment time. The application of the proposed method expands the arsenal of tools and treatment methods used in the treatment of purulent wounds.

Table Comparative results of treatment of rats with purulent soft tissue wounds. Phrases of the wound healing process Rat groups
Indicators group 1 (n-12) group 2 (n = 12) group 3 (n = 12) I phase (purulent-necrotic) Inoculation of microflora on 3-5 days of treatment (number of rats) 3 3 3 PMO bacteria 1 * 10 ⁵ -1 * 10 ⁷ CFU / ml 1 * 10 ⁵ -1 * 10 ⁷ CFU / ml 1 * 10 ⁵ -1 * 10 ⁷ CFU / ml ALA bacteria 1.54 mcg / ml * OD 1.43 mcg / ml * OD 1.49 mcg / ml * OD Ability to form bacteria biofilms 0.6 units OP 0.59 units OP 0.6 units OP Duration of wound cleansing from the start of treatment (24 hours) 7.8 ± 0.8 7.8 ± 0.8 7.8 ± 0.8 Dates of normalization of peripheral blood counts (days) 4.2 ± 0.5 4.2 ± 0.5 4.2 ± 0.5 Terms of the appearance of regional epithelization (days) 8.9 ± 0.8 8.9 ± 0.8 8.9 ± 0.8 II and III phases (regeneration and epithelization) Joining secondary infection (% rats) 58.3% 8.3% * 8.3% * PMO bacteria 1 * 10 ⁵ CFU / ml 1 * 10 ¹ -1 * 10 ² CFU / ml 1 * 10 ¹ -1 * 10 ² CFU / ml ALA bacteria 2.02 mcg / ml * OD 0.61 mcg / ml * OD 0.57 mcg / ml * OD Ability to form bacteria biofilms 0.69 units OP 0.27 units OP 0.23 units OP Macrophages (abs.) In conditional fields of view 3.7 ± 0.8 10.8 ± 1.9 * 16.6 ± 4.1 * Fibroblasts (poorly differentiated),% 1.7 ± 0.2 2.7 ± 0.1 * 3.9 ± 0.2 * Proliferating Epidermocytes unit cells + + Duration of complete wound healing (days) 21.4 ± 2.5 20.4 ± 2.5 16.4 ± 0.7 * Note: * - p <0.01 compared with the main group

Claims (1)

A method of treating purulent wounds in an experiment by surgical treatment and subsequent application of a dressing with drugs on a wound surface, characterized in that in the phases of regeneration and epithelialization, a dressing impregnated with an ointment of the following composition is applied to the wound surface, wt.%:
organosilicon glycerohydrogel 95.25 ciprofloxacin 0.2 oxytocin 4,55

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