RU2308960C1 - Wound-healing sponge - Google Patents

Abstract

FIELD: medicine, pharmaceutical industry, pharmacy.

SUBSTANCE: invention relates to wound-healing collagen sponge. Wound-healing collagen sponge with the definite content of Bergenia pacifica black leaves extract is prepared by sublimation of mixture of 2% collagen solution, purified water and Bergenia pacifica black leaves dry extract for a definite time followed by additional drying of prepared plates and conditioning under the definite conditions. Sponge possesses the expressed wound-healing activity in wound injuries.

EFFECT: valuable medicinal properties of sponge.

7 tbl, 3 ex

Description

The invention relates to medicine and the pharmaceutical industry, in particular to the creation of drugs used as external agents in the treatment of wounds.

The problem of wound healing is one of the most important branches of medicine that have not lost their significance even now. Comprehensive treatment of wound injuries includes, mainly, surgical and pharmacotherapeutic methods. For topical treatment of wound injuries, the use of complex preparations with antimicrobial, anti-inflammatory, immunomodulatory, wound healing effects is relevant. Of the large number of available products, biologically active substances of plant origin possess a combined effect. The problem of developing external medicines based on substances of plant origin is relevant.

In practical surgery, wound dressings in the form of films, sponges based on polymers, which are convenient to use, easily release active substances and are easily removed when the dressing is removed, are widespread. In this regard, the collagen biopolymer, which has good form-forming properties, is absorbed by biofluids and body cells [Collagenoplasty in medicine. / Ed. Kovanova V.V., Sychenikova I.A. - M .: Med., 1978. - 256 p .; Collagen sponge “Cytotimacol” - a stimulator of wound healing. / Malinin V.V., Burakov M.A., Sidorova N.D. and others // Chem. pharmacist. Journal. - 1998. - No. 11. - S. 54-56].

The problem to which the invention is directed, is to create an application collagen sponge with a wound healing effect. The problem is solved in that the inventive wound healing sponge as a medicine contains the previously obtained dry extract from the black leaves of frankincense (conventional name - "badaton"). [Lubsandorzhieva P. B., Tsyrenzhapova O. D., Nikolaev S. M., Nikolaev G. G. The method of obtaining the sum of extractive substances with adaptogenic activity: RF Patent 2076727. Application No. 93028222/14. Claim 06/02/93. publ. 04/10/97. Bull. No. 10].

The technical result obtained by carrying out the invention is expressed in expanding the range of highly effective and harmless drugs for the treatment of wound injuries.

To achieve the specified technical result, badaton is mixed with diluted water (1: 1) with a 2% collagen solution in a homogenizer to a homogeneous mass, dried in a freeze-drying apparatus at a temperature of minus 20-25 ° С under a vacuum of 16-17 Pa for 30 hours, dried for 10 hours, then incubated for conditioning at room temperature for 24 hours. The obtained plates of a highly porous collagen sponge, conventionally called “badanoplast”, are cut into even rectangular plates 30 × 30 mm in size. In the obtained collagen sponges, the moisture and pH of the aqueous solution were determined. The determination of the pH of the badanoplast plates was carried out by the potentiometric method according to the Global Fund XI ed. [State Pharmacopoeia of the USSR. General methods of analysis. - 11th ed. - Issue 1. M.: Med., 1989. (GF XI ed.)], Moisture weight method [GF XI ed. issue 1., p.176].

To determine the pH, the plates were crushed with scissors, 0.5 g samples were taken, 50 ml of distilled water was poured, kept for 15 min with periodic stirring. The pH of the solution is measured on a universal ionomer I-160.

Determination of moisture content of badanoplast plates: about 0.5 g (exact weight) of badanoplast plates is placed in a bottle and dried in a vacuum oven for 3 hours at a temperature of 60 ± 1 ° C and a residual pressure not exceeding 0.667 kPa ( 5 mmHg). The calculation of the mass fraction of moisture in percent is carried out by the mass difference before and after drying. The results of the analyzes are given in table 1.

In the above-described way, 7 compositions were obtained that differed from each other in the quantitative content of badaton (minimum, average, maximum). The resulting collagen sponge is conditionally called "badanoplast." The data are shown in table 1.

Table 1
Physico-chemical data of the obtained badanoplast plates Composition number Appearance of plates The content of the extract of incense in the plate,% pH plates Moisture content, % one Light gray plates 0.3 4.32 10.48 2 Light gray plates 0.5 4.21 10.50 3 Light gray plates 0.6 4.43 9.19 four Light gray plates 1,5 4.15 9.95 5 Gray plates 4.0 4.13 11.0 6 Gray plates 5,0 4.38 10.70 7 Unbreakable, gray colored plate 10.0 4,54 10.80

When preparing a collagen sponge with a badaton content above 5% (composition No. 7, Table 1), brittle, brittle plates are obtained. When conditioning a collagen sponge with a 10% badatone content, badatone powder spills out of the collagen pores. Thus, the high content of badaton in the collagen sponge "badanoplast" (composition No. 7, table 1) does not allow to obtain a means of constant composition.

To assess the pharmacological activity, “badanoplast” plates were selected — compositions No. 1, 2 (Table 1) with a minimum content of badatone (0.5%; 1.5%) and composition No. 5 with an average content of badatone (5%). A pharmacological study of their effect on the healing of flat muscular skin wounds showed the greatest effectiveness of composition No. 5 with a 5% badaton content. Below are 3 examples of obtaining "badanoplast" with a 5% content of badaton.

Example 1. In a homogenizer was loaded with 250 ml of water for injection, 250 g of a viscous colorless 2% collagen solution, 0.25 g of dry extract of black leaves of frankincense. The resulting mixture was stirred until homogeneous in an Erweka apparatus (made in Germany) at a stirrer speed of 336 rpm at a temperature of 30 ° C for 5 minutes. The obtained homogeneous composition of collagen with frankincense extract in a special container was transferred for freeze-drying to a TG-5 apparatus (made in Germany). The mass was poured into a stainless steel cassette measuring 250 × 250 mm, the height of the liquid column did not exceed 7 mm.

The spilled cartridge was mounted on sublimator shelves cooled to minus 20-25 ° С. After freezing the product to minus 20 ° С, the chamber was evacuated to an equilibrium working pressure of 16 Pa (minus 40 ° С). Then the standard course of the process, the period of sublimation itself, was established. After 30 hours of drying, the temperature of the material coincided with the temperature of the shelf at which the preparation was dried for another 9 hours. The vacuum readings ranged from 16.0-17.7 Pa. The drying temperature did not exceed 28 ° C.

After unloading from the sublimation machine, the dried sheets of badanoplast plates were kept for 24 hours at a relative humidity of 70% and a temperature of 18-24 ° C in a clean room for conditioning. After conditioning, the plates are dry, smooth, highly porous, light gray in color with a pearl tint. The humidity of the obtained plates is 10.70%, pH is 4.38. Sheets of “badanoplast” were cut into 30 × 30 mm plates, hermetically packed in plastic bags on a PDM-1 bag making machine.

We studied the effect of "badanoplast" on the healing of planar muscular skin wounds. The experiments were carried out on rats of the Wistar strain of both sexes weighing 160-180 g. The model of planar muscular skin wounds was reproduced under aseptic conditions according to the generally accepted method [Ponomareva-Astrakhantseva L.Z. A method of experimental receiving wounds and ulcers. // Reproduction of diseases in animals for experimental and therapeutic research. - M., 1954. - S. 66-73]. The animals of the experimental group immediately after the operation produced an application of 0.5; 1.5 and 5% "badanoplast" and fixed it with adhesive plaster. The test agent was replaced daily in the morning during the entire experiment. In rats of the control group, the wound surface remained open during the entire observation period. As an additional control, a collagen sponge (combutec) was used, as a comparison preparation, a porous algipor preparation was used, which were applied in a similar manner. Collagen sponge (kombutek) is a sponge coating for wounds obtained from soluble collagen of animal origin. The drug is non-toxic, does not cause tissue irritation, does not have antigenic activity, has antiseptic properties due to the addition of antiseptics (Khilkin A.M., Schreter A.B., Istranov L.P., Lemenev V.L. Collagen and its use in medicine. - M .: Medicine. - 1976. - 208 p .; Saschikova V.G., Vasiliev M.P., Pukhova Z.I. Use of a fibrous soluble collagen preparation in the treatment of trophic ulcers of the lower extremities // Herald of Surgery. - 1992. - No. 4. - S.71-75). Algipor is a porous preparation created on the basis of alginic acid. Algipor has a stimulating effect on reparative processes and, primarily, on the development of connective tissue; also has antibacterial properties due to additives of furatsilin and furagin (Sedlarik K.M. Alginates for the treatment of wounds // Surgery. - 1993. - No. 1. - P.62-65). In order to assess the dynamics of wound healing on days 3, 7, 14, 21 and 28, the area of the wound surface was determined by the planimetric method (Table 2). On the 3rd, 7th and 14th day, a wound cytological examination was performed using the “wound imprints” method (Table 3) and on the 21st day, a histological examination of wounds was performed.

As follows from the data given in table 2, the use of "badanoplast" in all the studied concentrations had a wound healing effect. At the same time, 5% “badanoplast” had the most pronounced wound healing activity. So, on the 3rd day of the experiment, the size of the skin defect in animals of the experimental groups on the background of the use of 0.5 and 1.5% “badanoplast”, as well as collagen sponge and algipore did not statistically differ from that in animals of the control group, whereas when using 5% “Badanoplast” by this time the area of damage was on average 22% less compared to the control.

On the 7th, 14th and 21st day of the study in animals of the experimental group, with the use of 5% badanoplast, a decrease in the area of the wound defect was noted by 30, 49 and 82%, respectively, compared with similar indicators in animals of the control group. At the same time, 0.5 and 1.5% "badanoplast", like collagen sponge and algipore, had a less pronounced wound healing effect in the indicated periods. In particular, it was shown that the statistically significant wound healing effect in the algipore comparison drug appears on the 7th day, and in the collagen sponge on the 14th day of the experiment. On the 28th day of the experiment, in animals with 5% badanoplast, complete closure of the wound skin defect was observed, while in rats of the control group, as well as using a collagen sponge, 0.5 and 1.5% of badanoplast and algipore remained, respectively an average of 9.0; 4.7; 5.0; 4.3; and 4.5% of an unhealed wound surface.

table 2
The influence of “badanoplast” on the dynamics of changes in the area of planar musculocutaneous wounds in white rats Duration of experience, day The area of the wound, mm ² Control group Collagen sponge Badanoplast 0.5% Badanoplast 1.5% Badanoplast 5.0% Algipor one 342.8 ± 17.2 341.3 ± 20.2 340.6 ± 15.4 342.7 ± 12.6 340.5 ± 14.6 341.3 ± 18.2 3 315.0 ± 15.4 302.4 ± 15.5 294.2 ± 14.2 293.3 ± 16.6 245.7 ± 12.7 * 296.3 ± 10.3 7 168.2 ± 11.1 144.6 ± 10.3 143.3 ± 12.1 131.2 ± 11.2 117.6 ± 9.7 * 134.2 ± 10.3 * fourteen 100.5 ± 9.2 70.5 ± 6.4 * 70.9 ± 5.7 * 65.7 ± 3.6 * 51.5 ± 4.3 * 64.6 ± 4.5 * 21 64.3 ± 3.3 43.5 ± 3.6 * 42.1 ± 2.2 * 24.7 ± 1.9 * 11.5 ± 1.8 * 21.4 ± 2.7 * 28 31.1 ± 1.8 16.2 ± 1.7 * 17.1 ± 1.3 * 14.8 ± 1.1 * - 15.6 ± 0.9 * Note: * - hereinafter, the values significantly different from the data of the control group at P≤0.05.

Table 3
The influence of 5% "badanoplast" on the cellular composition of the "wound prints" of muscular skin wounds in white rats. Indicators Groups of animals Duration of experience, day 3 7 fourteen White blood cells / of them destroyed Control ++++ / +++ +++ / ++ ++ / + Badanoplast ++++ / ++ ++ / + ++ / - Algipor ++++ / ++ +++ / + - ++ / + Macrophages / phagocytic Control - / - + / + ++ / + Badanoplast + / + ++ / + ++ / ++ Algipor - / - + / + ++ / ++ Fibroblasts Control - + Badanoplast - ++ +++ Algipor - + ++ Note: (-) lack of bacterial growth, (+) pronounced bacteriostatic effect, (++) bacteriostatic effect, (+++) weak bacteriostatic effect, (++++) lack of antibacterial action.

The data of a cytological study, shown in table 3, indicate that the use of 5% "badanoplast" has a positive effect on the cellular structure of the wound surface. In particular, on the 3rd day of the experiment, a large number of neutrophilic leukocytes was noted in the “wound prints” of animals from the control and experimental groups. Moreover, in the group of animals against the background of the use of 5% "badanoplast" and algipore, the number of destructive forms of leukocytes was on average 20% less than in the control. The appearance of single phagocytic macrophages was also noted. On the 7th day of the experiment in “wound prints” in rats of the control and experimental groups, a decrease in the number of neutrophilic leukocytes and the appearance of macrophages were noted. At the same time, against the background of using 5% “badanoplast”, the number of macrophages was on average 2 times higher than in the control, and the appearance of fibroblasts was also noted. On day 14, in animals of the experimental group using 5% "badanoplast" in the "wound imprint" fibroblasts and fibrocytes prevailed; phagocytosis was more pronounced than in control; free-lying microbes were not detected.

The results of a histological study conducted on the 21st day of the experiment showed that in the animals of the control group there was an increase in the epithelium from the basal epithelial layer of cells in the form of finger-like growths deep into. The epithelium was thickened, the wound surface is not completely covered by the epithelial layer. In all cases, the skin defect in the rats of the control group is filled with granulation tissue, consisting of fibroblasts of varying degrees of maturity and collagen fibers arranged randomly. In animals, against the background of using 5% “badanoplast”, filling of the skin defect with mature granulation tissue was noted, collagen fibers are located mainly in parallel bundles, among which are mature fibrocytes. The wound surface is completely epithelized; in the upper layers of the dermis, the beginnings of hair follicles are noted. A histological examination of wounds with the use of an algipore comparison drug revealed the filling of a skin defect with dense young granulation tissue containing randomly located collagen fibers, young fibroblasts. Epithelialization marginal, wound defect is not completely epithelized. Inflammatory changes around the wound are minor.

Thus, "badanoplast" with a 5% content of badaton has the most pronounced wound healing effect.

Example 2. In a homogenizer was loaded with 250 ml of water for injection, 250 g of a viscous colorless 2% solution of collagen, 0.25 g of dry extract of black leaves of frankincense. The resulting mixture was stirred until homogeneous in an Erweka apparatus (made in Germany) at a stirrer speed of 336 rpm at a temperature of 30 ° C for 5 minutes. The obtained homogeneous composition of collagen with frankincense extract in a special container was transferred for freeze-drying to a TG-5 apparatus (made in Germany). The mass was poured into a 250 × 250 mm stainless steel cassette, the height of the liquid column was 9 mm.

The spilled cartridge was mounted on sublimator shelves cooled to minus 20-25 ° С. After freezing the product to minus 20 ° С, the chamber was evacuated to an equilibrium working pressure of 16 pa (minus 40 ° С). Then the standard course of the process, the period of sublimation itself, was established. Sublimation was carried out for 32 hours, dried for 8 hours at a temperature of 25 ± 3 ° C under a vacuum of 16-17 Pa. After unloading from the sublimation machine, the dried sheets of badanoplast plates were kept for 24 hours at a relative humidity of 70% and a temperature of 18-24 ° C in a clean room for conditioning. After conditioning, dry, smooth, highly porous plates were obtained. The humidity of the obtained plates is 10.20%, pH 4.42. Sheets of “badanoplast” were cut into 30 × 30 mm plates, hermetically packed in plastic bags on a PDM-1 bag making machine.

The influence of "badanoplast" on the healing of infected skin and muscle wounds.

The experiments were carried out on Wistar rats of both sexes weighing 160-180 g. The model of infected wounds was reproduced by irrigation of a flat muscular skin wound with 1 ml of a suspension of Staphylococcus aureus 209 P strain with a titer of 1 billion / ml [Methodical recommendations for experimental (preclinical) study of drugs for local treatment of purulent wounds. - M., 1989. - 45 p.]. 5% Badanoplast wound application was performed starting from the first day and then daily throughout the experiment. Algipore was used as a comparison drug according to a similar scheme. Animals with "natural healing" of the wound served as control. In order to assess the dynamics of healing of the wound process on days 3, 7, 14, 21 and 28, the area of the wounds was determined by the planimetric method, and bacteriological, cytological and histological studies of the wound were performed. The data obtained are shown in tables 4, 5.

Table 4
The effect of 5% "badanoplast" on the area of infected musculocutaneous wounds in white rats. Duration of experience, day The area of the wound, mm ² Control group Badanoplast Algipor 3 376.2 ± 7.29 368.7 ± 7.05 365.8 ± 17.15 7 317.0 ± 9.02 291.6 ± 15.92 304.0 ± 21.60 fourteen 166.6 ± 11.91 114.9 ± 8.78 * 140.8 ± 16.60 21 57.6 ± 4.65 10.4 ± 8.78 * 30.0 ± 3.82 * 28 24.5 ± 2.10 - 11.4 ± 1.1 *

As follows from the data given in table 4, the use of 5% "badanoplast" contributed to the acceleration of healing of infected skin and muscle wounds, as evidenced by a decrease in the area of the wound defect on days 3, 7, 14 and 21 of the experiment, respectively, on 2, 8, 31, 82% compared with similar rats in the control group. On the 28th day of the experiment, against the background of the use of "badanoplast", complete wound epithelization was noted, while in the rats of the control group, a skin defect remained. In this case, the comparison drug algipor had a less pronounced wound healing effect.

A bacteriological study showed that in the biopsy of the wounds of rats of the experimental group on the 3rd day of the study, the bacterial contamination of the wound was 6.50 × 10 ⁵ , while in rats of the control group by the indicated time the level of bacterial contamination of the wound was 8.04 × 10 ⁷ . On the 7th day of the experiment, against the background of the use of "badanoplast", the number of bacteria decreased to 1.30 × 10 ³ , while in animals of the control group the infection level by the indicated observation period averaged 3.50 × 10 ⁶ . On the 14th day of the study, the level of bacterial contamination of the wounds of animals of the experimental and control groups was 9.60 × 10 ¹ and 9.28 × 10 ^5, respectively. On day 21, the bacterial contamination of the wounds of animals of the control group was 8.50 × 10 ³ ; in animals of the experimental group, bacterial growth was not determined. It was found that the antibacterial activity of "badanoplast" exceeds that of the comparison drug. Thus, the level of bacterial contamination of animal wounds during the use of algipore on days 3, 7, 14, 21 was 1.81 × 10 ⁶ , 5.58 × 10 ⁵ , 6.20 × 10 ⁴ , 4.04 × 10 ^2, respectively.

Table 5
The influence of 5% "badanoplast" on the cellular composition of the "wound prints" of infected wounds in white rats Indicators Groups of animals Duration of experience, day 3 7 fourteen White blood cells / of them destroyed Control ++++ / +++ +++ / +++ +++ / ++ Badanoplast +++ / ++ +++ / + ++ / - Algipor ++++ / +++ +++ / ++ ++ / + Macrophages / phagocytic Control - / - +/- ++ / + Badanoplast + / + ++ / ++ ++ / + Algipor - / - + / + ++ / ++ Fibroblasts Control - - + Badanoplast - + +++ Algipor - - ++ Note: (-) lack of bacterial growth, (+) pronounced bacteriostatic effect, (++) bacteriostatic effect, (+++) weak bacteriostatic effect, (++++) lack of antibacterial action.

It was found that on the 3rd day of the experiment in the "wound prints" of rats of the control and experimental groups, a significant number of leukocytes was noted. Moreover, in the experimental group, there were fewer destroyed white blood cells than in the control, and the appearance of single macrophages was also noted. A similar trend was observed on the 7th day of the experiment: in the “wound imprints” of the rats of the experimental group, the destroyed leukocytes were less than in the control, as well as a greater number of phagocytic macrophages and the appearance of fibroblasts. On day 14, against the background of the use of "badanoplast", destroyed leukocytes were not observed; the number of macrophages decreased; along with this in the "wound prints" the predominance of fibroblasts was noted.

Histological examination showed that the use of 5% "badanoplast" led to the formation of mature granulation tissue and complete wound epithelization in animals of the experimental group on days 21-28. Moreover, in the control group on day 28 complete epithelization was not observed in any case, the skin defect was filled with granulation tissue, the collagen fiber strands were loose.

Example 3. In a homogenizer was loaded with 250 ml of water for injection, 250 g of a viscous colorless 2% solution of collagen, 0.25 g of dry extract of black leaves of frankincense. The resulting mixture was stirred until homogeneous in an Erweka apparatus (manufactured in Germany) at a stirrer speed of 336 rpm at a temperature of 30 ° C for 10 minutes. The obtained homogeneous composition of collagen with frankincense extract in a special container was transferred for freeze-drying to a TG-5 apparatus (made in Germany). The mass was poured into a stainless steel cassette measuring 250 × 250 mm, the height of the liquid column did not exceed 9 mm.

The spilled cartridge was mounted on sublimator shelves cooled to minus 20-25 ° С. After freezing the product to minus 20 ° C, the chamber was evacuated to an equilibrium working pressure of 16 Pa (minus 40 ° C), then the standard course of the process was established, the period of sublimation itself. After 28 hours of drying, the plates were dried for another 10 hours. The vacuum readings ranged from 16-17 Pa. The drying temperature did not exceed 25 ± 3 ° C.

After unloading from the sublimation machine, the dried sheets of badanoplast plates were kept for 24 hours at a relative humidity of 70% and a temperature of 18-24 ° C in a clean room for conditioning. After conditioning, dry, smooth, highly porous plates were obtained. The humidity of the obtained plates 16.50%, pH 4.10.

Sheets of “badanoplast” were cut into 30 × 30 mm plates, hermetically packed in plastic bags on a PDM-1 bag making machine.

A clinical study of "badanoplast" in combination with phlebosclerotherapy in the local treatment of trophic ulcers of the lower extremities was conducted on the basis of the surgical department of the Republican Clinical Hospital named after N.A.Semashko of the Ministry of Health of the Republic of Buryatia (Ulan-Ude). 89 patients with trophic ulcers were examined, including 72 (80.9%) women and 17 (19.1%) men. In 51 (57.3%) patients, the cause of trophic ulcers was post-thrombophlebic disease, after acute thrombophlebitis of deep veins of the lower extremities. In 38 (42.7%) trophic ulcers formed on the background of varicose veins of the lower extremities. In a history of 15 (16.7) patients, phlebectomy was performed for varicose veins of the lower extremities. In 67 (75.3%) patients, the wound surface area reached 40 cm ² ; in 9 (10.1%) patients - up to 100 cm ² and ulcers with an area of more than 100 cm ² were found in 13 patients. In 21 (23.4%) patients, trophic ulcers were on the left lower limb, in 16 (17.9%) - on the right and in 52 (58.7%) on both lower limbs. Concomitant diseases were detected in 76.4% of patients; including: coronary heart disease - 33.7%; hypertension - 23.5%; obesity - 44.1%; diabetes mellitus - 10.2%; chronic diseases of the respiratory system - 5.7%; obliterating diseases of the vessels of the lower extremities - 4.4%, kidney disease - 2.8% of patients.

The duration of the disease ranged from 3 months to 30 years or more. 56.2% of patients were of working age (up to 60 years). All patients were divided into 3 groups:

Group 1 - control (15 patients), the treatment was carried out by generally accepted methods with the use of drugs and dressings with furatsilinom. The ulcer surface was treated with a 3% hydrogen peroxide solution. In the presence of purulent-necrotic masses, iruxol, an aqueous solution of chlorhexidine, boric acid were used, and then a bandage with furatsilin, including the Unna dressing, was applied. When granulations appeared (the beginning of focal epithelization), the latter was replaced by ointment dressings - with solcoseryl, sea buckthorn or rosehip oil.

Group 2 - comparisons (29 patients), treatment was carried out with kuriosin without sclerotherapy. The wound was treated with saline and antiseptic, after which a 0.2% solution of curiosin was applied at the rate of 1 drop per 1 cm ^{2 of the} wound. After a two-minute exposure, parchment paper was applied over a gauze napkin and bandage of the limb with an elastic bandage was performed. Bandaging was performed 1 time per day.

Group 3 - the main group (45 patients), received treatment in the form of a combination of phlebosclerotherapy with an application of 5% "badanoplast". The wound was treated with saline. Fragments were cut from the badanoplast plate for the calculation of wound coverage in two-thirds of its area. After that, fragments of "badanoplast" were placed on a wound in a checkerboard pattern, an absorption bandage was applied from above.

For phlebosclerotherapy, a 3% thrombovar solution was used at a rate of 2.0 ml per session (per leg). Sclerotherapy began with veins located in the trophic ulcer area or located in trophically altered tissues, which were determined visually and by palpation. For injection, insulin syringes were used, when blood appeared in the syringe, the tip of the needle was tightly pressed against the wall of the vein and a sclerosant was slowly introduced. After removing the needle from the vein, the puncture site was crushed with a gauze ball and tightly bandaged with an elastic bandage. For 1 session, 3-4 injections were performed in one shin. Then the bandage was removed and an elastic compression bandage was applied (the limb was bandaged from the foot to the knee joint). If the patient had tissue edema, induction, hyperpigmentation, varicose veins were absent due to edema or lymphostasis, then blockade of 2% lidocaine in a volume of 5.0 ml was performed next to the trophic ulcer. As a rule, after 5-6 blocks, the edema significantly decreased or completely disappeared, while the configuration of the veins passing around the trophic disorders of the lower leg clearly emerged, which made it possible to perform phlebosclerotherapy. If blood appeared in the needle during the blockade, then 3% thrombovar was injected in the same place in a volume of 0.5 ml and elastic shin compression was performed. When technically correctly performed sclerotherapy, necrosis of subcutaneous fat in the area of sclerosant administration was not observed. The number of phlebosclerotherapy sessions depended on the degree of trophic changes in tissues, the size of ulcers, the number of varicose veins, the nature of varicose veins (loose or main) and corresponded from 3 to 9. Puncture therapy was completed after obliteration of all varicose veins located in trophically altered tissues was achieved. When evaluating the effectiveness of treatment, the clinical course of the disease was primarily taken into account: reduction or disappearance of pain and swelling in the lower extremity, rate of cleansing of the ulcer lesion, the timing of the appearance of granulations and ulcer epithelization. The data obtained are shown in table 6.

As follows from the data given in Table 6, in patients receiving basic therapy (control group), a decrease in the severity of disease symptoms (pain, hyperemia, edema, etc.) was noted on days 13-16, cleansing of the ulcer surface and the appearance of granulations - on day 19, and partial epithelization (an average of 70-80% of ulcerative defect) was observed on day 39-40. In patients of the comparison group, during the treatment of ulcerative defects with kuriosin, the appearance of granulation symptoms was observed on the 10th day, partial epithelization (70-80%) occurred on the 18-20th day of treatment.

Table 6
The effect of the integrated use of phlebosclerotherapy and "badanoplast" on the clinical parameters of patients with trophic ulcers Indicators Patient groups Control (standard therapy) Comparisons (curiosin) The main (phlebosclerotherapy + "badanoplast") The terms of the disappearance of pain, day 13.0 ± 1.6 8.1 ± 0.8 4.1 ± 0.9 * Duration of edema, days 16.0 ± 3.0 6.0 ± 0.7 4.1 ± 1.8 * The timing of the appearance of granulation, day 19.1 ± 0.8 9.3 ± 1.4 7.5 ± 1.3 * Duration of ulcer healing
day 39.4 ± 1.1 18.2 ± 1.2 15.6 ± 1.3 *

In patients of the 3rd group who received complex therapy in the form of a combination of phlebosclerotherapy and “badanoplast”, 4–5 days after the start of treatment, the state of health improved, pain sensations in the area of the pathological focus decreased, there was a tendency to decrease swelling, hyperemia and inflammatory infiltration along the periphery of the ulcer lesion . By the end of 5-12 days from the start of treatment in patients of this group, the wound surface was gradually cleared of necrotic tissues, healthy juicy granulations appeared, inflammatory phenomena around the ulcers subsided, and a pale pink corolla of young epithelial tissue appeared on the periphery of the ulcer surface. After a two-week treatment period, the surface of the ulcer was completely covered with epithelium.

One of the important criteria for the effectiveness of any treatment method is the percentage of healing of trophic ulcers at the hospital stage. With the combined treatment of trophic ulcers with phlebosclerotherapy and “badanoplast”, the decrease in the area of ulcer defects after 3, 5, and 7 days decreased by 19, 61, and 89%, respectively, compared with that before treatment. Moreover, in patients of the comparison group who received treatment with kuriosin, the area of ulcers in the indicated periods was respectively 7, 26 and 45% less. In the control group of patients, a significant decrease in the area of ulcerative defects was not observed during these periods.

A cytological study of “prints” from the surface of trophic ulcers and a histological study of biopsy samples from trophic ulcers before treatment, as well as on days 7, 14 and 21 from the start of treatment, were also performed.

The cytogram of smears of patients before treatment was characterized by neutrophils (89.3%), and most of them (91.2%) were in a state of decay. A morphological study revealed that necrotic masses and fibrous-purulent exudate predominate in the bottom and edges of trophic ulcers. Granulation tissue was with severe structural changes, characterized by the presence of areas of granulation tissue with scar inclusions. Significant microcirculatory disorders were noted in the form of stasis, erythrocyte sludge, microthrombosis, infiltrative-productive endovasculitis, fibrinous necrosis of the vascular walls, erythrocyte diapedesis, and leukodiapedesis. Marked edema and tissue lymphostasis, neutrophilic infiltration were also noted. The response of macrophages and mast cells was sharply inhibited.

In the control group of patients receiving standard therapy, a cytological study of smears of the wound discharge showed a slight increase in macrophages and fibroblasts only on days 15-21, although the inflammatory reaction remained pronounced (81.7 ± 3.09 neutrophils in the smear). Only by 21 days a focal typical granulation tissue appeared, necrosis, edema and inflammatory tissue infiltration decreased, and a proliferative reaction appeared. At the same time, the wound healing process was slowed down, microcirculatory disorders and the inflammatory process remained pronounced.

A cytological study of patients in the comparison group showed that under the influence of kuriosin, the exudate was much faster cleared of dystrophically altered and necrotic neutrophils, it quickly, starting from 5 days, there was an increase in the number of immature mononuclear elements and macrophages, increased phagocytosis of microbes and detritus, an increase in fibroplastic elements, and from 14 days and epithelial cells. Morphological studies showed that the activation of reparative processes occurred on the 7th day of treatment, as evidenced by the proliferation of fibroblasts, the appearance of granulation tissue and traces of epithelization. At the same time, a decrease in the severity of the inflammatory reaction was noted. By the 14th day of treatment, granulation tissue filled the tissue defect, and by the 21st day rapid growth of epithelial cells was noted.

In patients of the third group who received combined therapy in the form of phlebosclerotherapy and applications of "badanoplast", a significant acceleration of reparative processes was noted. In particular, in patients of this group, there was a rapid cleansing of the ulcer surface of necrotic masses and fibrous-purulent exudate, the early appearance of granulation tissue of a typical structure and its maturation, scarring, epithelization and closure of a tissue defect. A cytological study showed that in patients of this group, on the 7th day from the start of treatment, a significant activation of phagocytosis of bacteria and detritus by neutrophils and macrophages is noted; normalization of microcirculation, a decrease in the severity of tissue edema and other inflammatory processes.

Given the great danger of the development of purulent complications in patients with trophic ulcers, on the 7th day of treatment in 30 patients of different groups samples of discharge from ulcers were taken for bacteriological examination (Table 7).

Table 7
The frequency of isolation of microorganisms from trophic ulcers before treatment and after 7 days from the start of treatment Type of microorganism The frequency of isolation of microorganisms,% Before treatment After 7 days Control (standard therapy) Comparisons (curiosin) The main (phlebosclerotherapy + "badanoplast") Staphylococcus aureus 44,2 33.6 21.1 17,2 Pseudomonas aerugenosae 31.8 20,2 10.1 5,6 Proteus vulgaris 26.3 10,4 5,6 - Streptococcus faecalis 19.2 2.2 1.9 - Escherichia coli 13.3 12.3 3.4 1.3 Other 6.1 5,6 2.9 1,1

As follows from the data given in table 7, in smears of trophic ulcers obtained before the start of treatment, a pronounced bacterial infection was observed, consisting mainly of coccal and rod microflora. During standard therapy (control group) on day 7, a slight decrease in microbial contamination was noted, the frequency of isolation of these microorganism strains remained quite high. Bacteriological studies of smears of the comparison group showed that treatment with curiosin helps to reduce the infection of the ulcer surface, however, as can be seen from the table, all studied pathogenic microorganisms were seeded in patients of this group. A more pronounced antibacterial effect was exerted by the combined use of sclerotherapy and "badanoplast", as evidenced by a significant change in the microbial background in patients of this group. In particular, a microbiological study of the “fingerprints” of trophic ulcers showed that 7 days after the start of treatment, the ulcer surface was completely cleansed of bacteria such as Proteus vulgaris and Streptococcus faecalis. The sowing frequency of other studied microorganisms was also significantly lower than in the control group and the comparison group.

Thus, the use of 5% "badanoplast" in the treatment of patients with trophic ulcers of the lower extremities in combination with phlebosclerotherapy can improve hemodynamics, reduce microbial contamination of ulcers, which helps to reduce the time of the first phase of the wound process and stimulates regeneration processes.

Claims (1)

A wound healing collagen sponge with a 5% content of blackberry leaf extract, obtained by sublimation for 28-32 hours of a mixture of a 2% solution of collagen, purified water and a dry blackberry leaf extract, by drying the plates for 8-10 hours and conditioning for 24 hours at room temperature.