RU2733137C1 - Method of producing biocomposite with regenerative properties based on hydrogel of bacterial cellulose - Google Patents

Abstract

FIELD: biotechnologies.

SUBSTANCE: invention relates to the biotechnology. Disclosed is a method of producing a biocomposite based on a hydrogel of bacterial cellulose with regenerative properties. Method involves cultivation of bacteria Gluconacetobacter sucrofermentans VKPM V-11267 on molasses in static conditions, separation of the obtained gel film of bacterial cellulose, its purification and mechanical grinding. Obtained hydrogel of bacterial cellulose with hydromodule 1:3 is mixed with 2 % chitosan solution in ratio of 50:50 or 2 % chitosan solution, 10 % gelatin solution, 20 % transglutaminase solution in ratio 5:5:15:5, respectively, then adding sodium fusidine and physiologically active compounds of polyphenolic nature, enzymes, low-molecular peptides.

EFFECT: invention widens the range of wound coatings based on bacterial cellulose hydrogel with antibacterial and regenerative properties.

1 cl, 13 ex

Description

The invention relates to the field of biotechnology and medicine, and in particular to a method for producing a biocomposite based on hydrogels of bacterial cellulose, used as a wound covering with antibacterial and regenerative properties.

The purpose of the biocomposite, obtained on the basis of bacterial cellulose hydrogels, is to create a material with high biocompatibility, low toxicity, adhesion, developed specific surface area, high moisture absorption capacity, the possibility of delivering biologically active substances to body tissues, the presence of regenerative properties and high antimicrobial activity.

In modern medicine, the role of wound coverings is reduced not only to protection from the external environment and mechanical influences, but also to an active effect on the wound process by creating a favorable microclimate and including biologically active substances in the composition of the coatings.

Quite a lot of various coatings are known for the treatment of wounds, in which drugs are applied to a base of natural origin or to a synthetic material. These drugs have a different composition of biologically active components: drugs with antimicrobial, antienzyme, reparative action, antibiotics, etc.

Known wound covering, consisting of bacterial cellulose. Moisture is retained under this coating, the wound is isolated from the air, and the bacterial cellulose itself is easily separated from the wound due to the fact that it is not biodegradable (CA 2632767, IPC A61K 37/715, publ. 07.06.2007).

The composition of this coating contains medicinal preparations of various directions of action for the treatment of wounds and a growth factor that contributes to the restoration of damaged tissues, including soft tissues.

Known biologically active wound dressing based on hydrated microbial cellulose with collagen gel layered on it using human cellular material. The gel contains a ten-fold concentrate of the M199 growth medium in an amount of 3-5% of the total volume, broad-spectrum antibiotics (for example, a mixture of antibiotics penicillin from 50 to 100 units / ml and streptomycin from 0.05 to 0.1 mg / ml) (RU 2430743, IPC A61L 15/28, A61F 13/02, A61L 15/44, publ. 10.10.2011).

The disadvantage of the known solution is the use of antibiotics penicillin and streptomycin, to which the resistance of microorganisms is rapidly developing

Known thermosetting neutralized composition of chitosan, forming a hydrogel, its lyophilisate and methods of production. Aqueous thermosetting neutralized chitosan composition, which forms a phosphate-free transparent hydrogel at temperatures above 5 ° C, which includes, in terms of the total weight of the composition, 0.1-5.0% reacetylated chitosan with a molecular weight of at least 100 kDa and a degree of deacetylation from 40 to 70%, neutralized with a hydroxylated base, and 1-30% of a complexing agent selected from polyoses (polysaccharides) and polyols derived from them (RU 2008130389, publ. 27.01.2010, IPC A61K 9/00, A61K 9/19, publ. 27.01.2010).

The resulting gel, as well as the film, cannot be used for deep burn wounds of III-A and III-B degrees due to rapid biodegradation in the wound environment and requires frequent repeated treatments and additional treatment of a purulent wound with antiseptic solutions.

Known pharmaceutical composition for the treatment of wounds and burns, which includes the following components: colloidal silver, colloidal zinc oxide, glycerin, methylcellulose and distilled water. The composite can be used in therapy and surgery for the healing of burn wounds, as a reparative, anti-inflammatory and microcirculation-improving agent, for the treatment of thermal, solar and chemical burns (RU 2636530, IPC A61K 33/30, A61K 33/38, A61P 17/02 , publ. 23.11.2016).

This composition has a highly effective regenerating, wound healing effect, but the disadvantage is that the composite is made in the form of a pharmaceutical hydrophilic ointment composition, and not a hydrogel, which is a more convenient form.

Known bioactive hydrogel for skin regeneration, obtained by chemical cross-linking of gelatin with glutaric dialdehyde. This hydrogel is characterized by a reduced content and consumption of a crosslinking agent, a reduced residual toxicity of a crosslinking agent, increased biocompatibility, and increased regenerative activity. The bioactive hydrogel additionally contains an anionic polymer and a bivalent metal in a final concentration of not more than 50 mM (RU 2659383, IPC A61K 38/17, publ. 06/29/2018).

The closest solution, taken as a prototype, is a wound dressing with a therapeutic effect, in which a single complex of perforated cellulose Acetobacter xylinum and biologically active ingredients with a healing effect are used. The wound covering includes a complex of fullerene C ₆₀ / Tween-80 (antioxidant), an antimicrobial component, an antienzyme and hemostatic component, a necrolytic component (RU 2437681, IPC A61L 15/18, A61L 15/44, A61L 15/28, publ. 27.12. 2011).

The technical result of the claimed invention is to expand the range of wound dressings based on a hydrogel of bacterial cellulose with regenerative properties and enhanced antimicrobial action.

The essence of the invention lies in the fact that the method of obtaining a biocomposite with regenerative properties, based on a hydrogel of bacterial cellulose, consists in obtaining a gel film of bacterial cellulose by cultivating a bacterial strain Gluconacetobacter sucrofermentans under static conditions on a medium with molasses, separating the obtained gel film of bacterial cellulose from culture medium and its purification, mechanical grinding of the purified gel film of bacterial cellulose for 10 min to obtain a hydromodule with a ratio of 1: 3, obtaining a hydrogel bacterial cellulose-chitosan in a 50:50 ratio by mixing a 2% solution of chitosan in 1% - acetic acid, bacterial cellulose hydrogel, 25% glutaraldehyde, as well as obtaining a hydrogel bacterial cellulose: chitosan: gelatin: transglutaminase in a ratio of 5: 5: 15: 5, respectively, and the addition of sodium fusidin, effective for external use, physiologically active compounds polyphenolic pr hydrodes in the form of dehydroquartzin or resveratrol, enzymes in the form of trypsin, chymotrypsin or lysozyme, low molecular weight peptides.

The bacterial strain Gluconacetobacter sucrofermentans H-110 was isolated at the Department of Biotechnology of the Mordovian State University from Kombucha with subsequent selection based on natural selection. The culture was identified to species by analysis of genes encoding 16S rRNA at FGUP GosNIIGenetics. The bacterial strain Gluconacetobacter sucrofermentans H-110 was deposited in the All-Russian Collection of Industrial Microorganisms (VKPM) under registration number: B-11267. The bacterial strain is not zoopathogenic, phytopathogenic and is not dangerous for other reasons.

The method for producing bacterial cellulose is described in the applicant's inventions (RU 2536973, IPC C12N 1/20, publ. 27.12.2014, RU 2536257, IPC C12N 1/20, publ. 20.12.2014).

Bacterial cellulose forms a matrix of a thin network of nanofibers, which has a high moisture-binding and water-holding capacity, selective permeability to gases and steam, and impermeable to bacteria.

The claimed invention uses the antibiotic fusidin sodium, the effectiveness of which has been proven for external use, in particular in the treatment of systemic and local staphylococcal infections of the skin and soft tissues. Fusidin sodium is effective against staphylococci resistant to penicillins, streptomycin, chloramphenicol, erythromycin. The drug is effective in the treatment of systemic and local staphylococcal infections of the skin and soft tissues, bones and joints, blood, endocardium, eyes, including those caused by methicillin-resistant strains. When applied externally, it penetrates deeply into the skin of the affected areas. Fusidic acid is well tolerated by patients and has low levels of toxicity, resistance and allergic reactions. There is no cross-resistance with other antibiotics. In addition, in addition to the antibacterial, fusidic acid has a weak immunomodulatory effect, which is associated with the suppression of the production and secretion of cytokines, especially interleukins and tumor necrosis factor. The foregoing makes it possible to include fusidic acid on a par with modern antibacterial drugs and recommend it for wider use, especially in infections caused by methicillin-resistant strains of Staphylococcus aureus.

By its chemical properties, dihydroquercetin is an extremely active antioxidant, that is, a substance that binds free radicals and deprives them of their harmful activity. It has the ability to penetrate into the lipid membrane. Due to its high antioxidant activity, it is able to regulate metabolic processes at the level of cell membranes and has a strong antiallergic and anti-inflammatory effect. It has a pronounced cytomembrane-protective effect, improves the state of the microvasculature in the lesions.

Resveratrol is a stilbene polyphenol of the phytoalexin family. It has a wide spectrum of action, including its antioxidant effect, which is manifested in a decrease in the level of malondialdehyde in mononuclear blood cells, in platelets damaged by peroxynitrite, and oxidative stress caused by cisplatin in platelets, plasma and lymphocytes. Resveratrol activates genes that help fight diseases associated with aging. Neutralizes free radicals produced during daily activities, increases the level of the coenzyme NAD ⁺ and sirtuins.

The main task of treating patients with wounds and burns is the fastest cleansing of the wound from necrotic tissues and purulent-fibrinous exudate, which contributes to the early appearance of granulations. For this purpose, proteolytic enzymes are relatively widely used. The most widespread are enzymes of the digestive tract of animals (trypsin, chymotrypsin, chymopsin); enzymes that depolarize DNA and RNA (deoxyribonuclease and ribonuclease), as well as bacterial enzymes with a similar effect. As well as lysozyme, these enzymes, due to their antimicrobial activity, contribute to the destruction of the cell walls of gram-positive bacteria. They have a powerful anti-inflammatory effect. Promote accelerated regeneration and complete wound healing. Proteolytic enzymes dissolve necrotic masses and cleanse the wound surface. Promotes the acceleration of the formation of fresh granulations. The use of enzymes helps to speed up wound healing. In the process of anti-inflammatory action, enzymes destroy damaged cells and necrotic material and, by inactivating mediators and toxic products, limit swelling and pain.

It is known that soluble forms of enzymes have some disadvantages, such as rapid inactivation, hydrolysis by the body's own proteases and bacteria. One of the ways to increase the stability of enzymes is to include them in composites, which increases the time of enzyme activity, simplifies delivery and makes it possible to quickly remove the enzyme.

Peptides attract the attention of researchers due to their antioxidant, anticarcinogenic, immunomodulatory, and other biologically active properties. Low molecular weight peptides cause a powerful regenerating effect associated with the presence of valuable biologically active components: growth factors, cytokines, cytamines and other substances that promote the regeneration and proliferation of skin cells. The use of a new cell preparation will enhance the regenerative effect: accelerate the renewal of skin cells by dividing collagen, elastin and keratinocytes by influencing the signaling mechanisms of intercellular interaction, which contributes to the activation of the regeneration process in the deeper tissues of the body.

Tinrostim is a product containing a complex of polypeptides isolated from nerve tissue (ganglia) of squid, and can exhibit a wide range of biological activities. Bioactive polypeptides are capable of exerting a physiological effect at low concentrations and for a short period of time, since they are quickly degraded by the enzyme systems of the body. In most cases, polypeptides are activators, initiators of certain functions of a living organism. These peptides have a positive effect on the quantitative and qualitative composition of peripheral blood, stimulate the functional activity and absorption capacity of phagocytic blood cells.

Tinrostim helps to reduce the duration of the first (inflammatory) phase of the wound process by 1.5 times. It can be noted that the disadvantage of the drug is a liquid helium base, which creates inconvenience when used in a purulent wound.

Chitosan is a naturally occurring cationic aminopolysaccharide, a copolymer of glucosamine and N-acetylglucosamine, which is obtained by partial deacetylation of chitin. Chitosan can be used as a promising matrix for immobilization due to the fact that it has a good ability to form a gel with low immunogenicity and toxicity, high mechanical strength and stability, and antibacterial properties.

The gel quickly and well adheres to the wound, isolating it from the penetration of a secondary infection. The intrinsic antibacterial nature of chitosan gives it an additional advantage over other polymers. The non-woven base creates conditions under which, after being removed from the wound, fibers and particles of matter do not remain in it, as with a gauze bandage, which is also very important for the prevention and reduction of the risk of additional infection.

The method of obtaining a biocomposite with regenerative properties, based on a hydrogel of bacterial cellulose, is carried out as follows.

Example 1. Get a gel film of bacterial cellulose by culturing the bacterial strain Gluconacetobacter sucrofermentans VKPM B-11267 on a medium with molasses. To remove cells and components of the culture medium, the resulting bacterial cellulose gel film is separated from the culture medium and purified. Sterilize bacterial cellulose gel film at 120 ° C for 20 minutes. Mechanical grinding of the purified gel-film of bacterial cellulose is carried out to obtain a hydromodule with a ratio of 1: 3.

To obtain a hydrogel "bacterial cellulose-chitosan" in a 50:50 ratio, mix a 2% solution of chitosan in 1% acetic acid, a hydrogel of bacterial cellulose, 25% glutaraldehyde (2%, v / v) and make 0.06% sodium fusidin and 0.1% resveratrol.

Example 2. Get a gel film of bacterial cellulose by culturing the bacterial strain Gluconacetobacter sucrofermentans VKPM B-11267 under static conditions on a medium with molasses. To remove cells and components of the culture medium, the resulting bacterial cellulose gel film is separated from the culture medium and purified. Sterilize bacterial cellulose gel film at 120 ° C for 20 minutes. Mechanical grinding of the purified gel-film of bacterial cellulose is carried out to obtain a hydromodule with a ratio of 1: 3.

To obtain a hydrogel "bacterial cellulose-chitosan" in a 50:50 ratio, mix a 2% solution of chitosan in 1% acetic acid, a hydrogel of bacterial cellulose, 25% glutaraldehyde (2%, v / v) and make 0.06% sodium fusidin and 2% dehydroquartsetin.

Example 3. Get a gel film of bacterial cellulose by culturing the bacterial strain Gluconacetobacter sucrofermentans VKPM B-11267 under static conditions on a medium with molasses. To remove cells and components of the culture medium, the resulting bacterial cellulose gel film is separated from the culture medium and purified. Sterilize bacterial cellulose gel film at 120 ° C for 20 minutes. Mechanical grinding of the purified gel-film of bacterial cellulose is carried out to obtain a hydromodule with a ratio of 1: 3.

To obtain a hydrogel "bacterial cellulose-chitosan" in a 50:50 ratio, mix a 2% solution of chitosan in 1% acetic acid, a hydrogel of bacterial cellulose, 25% glutaraldehyde (2%, v / v) and make 0.06% sodium fusidin and 0.14% trypsin.

Example 4. Get a gel film of bacterial cellulose by cultivating the bacterial strain Gluconacetobacter sucrofermentans VKPM B-11267 under static conditions on a medium with molasses. To remove cells and components of the culture medium, the resulting bacterial cellulose gel film is separated from the culture medium and purified. Sterilize bacterial cellulose gel film at 120 ° C for 20 minutes. Mechanical grinding of the purified gel-film of bacterial cellulose is carried out to obtain a hydromodule with a ratio of 1: 3.

To obtain a hydrogel "bacterial cellulose-chitosan" in a 50:50 ratio, mix a 2% solution of chitosan in 1% acetic acid, a hydrogel of bacterial cellulose, 25% glutaraldehyde (2%, v / v) and make 0.06% sodium fusidin and 0.15% chemotrypsin.

Example 5. Get a gel film of bacterial cellulose by culturing the bacterial strain Gluconacetobacter sucrofermentans VKPM B-11267 under static conditions on a medium with molasses. To remove cells and components of the culture medium, the resulting bacterial cellulose gel film is separated from the culture medium and purified. Sterilize bacterial cellulose gel film at 120 ° C for 20 minutes. Mechanical grinding of the purified gel-film of bacterial cellulose is carried out to obtain a hydromodule with a ratio of 1: 3.

To obtain a hydrogel "bacterial cellulose-chitosan" in a 50:50 ratio, mix a 2% solution of chitosan in 1% acetic acid, a hydrogel of bacterial cellulose, 25% glutaraldehyde (2%, v / v) and make 0.06% sodium fusidine and 0.1% tinrostim.

Example 6. Get a gel film of bacterial cellulose by culturing the bacterial strain Gluconacetobacter sucrofermentans VKPM B-11267 under static conditions on a medium with molasses. To remove cells and components of the culture medium, the resulting bacterial cellulose gel film is separated from the culture medium and purified. Sterilize bacterial cellulose gel film at 120 ° C for 20 minutes. Mechanical grinding of the purified gel-film of bacterial cellulose is carried out to obtain a hydromodule with a ratio of 1: 3.

To obtain a hydrogel "bacterial cellulose-chitosan" in a 50:50 ratio, mix a 2% solution of chitosan in 1% acetic acid, a hydrogel of bacterial cellulose, 25% glutaraldehyde (2%, v / v) and make 0.06% sodium fusidin, 0.1% resveratrol, 0.14% trypsin.

Example 7. Get a gel film of bacterial cellulose by culturing the bacterial strain Gluconacetobacter sucrofermentans VKPM B-11267 under static conditions on a medium with molasses. To remove cells and components of the culture medium, the resulting bacterial cellulose gel film is separated from the culture medium and purified. Sterilize bacterial cellulose gel film at 120 ° C for 20 minutes. Mechanical grinding of the purified gel-film of bacterial cellulose is carried out to obtain a hydromodule with a ratio of 1: 3.

To obtain a hydrogel "bacterial cellulose-chitosan" in a 50:50 ratio, mix a 2% solution of chitosan in 1% acetic acid, a hydrogel of bacterial cellulose, 25% glutaraldehyde (2%, v / v) and add 0.13% lysozyme and 0.1% resveratrol.

Example 8. Get a gel film of bacterial cellulose by culturing the bacterial strain Gluconacetobacter sucrofermentans VKPM B-11267 under static conditions on a medium with molasses. To remove cells and components of the culture medium, the resulting bacterial cellulose gel film is separated from the culture medium and purified. Sterilize bacterial cellulose gel film at 120 ° C for 20 minutes. Mechanical grinding of the purified gel-film of bacterial cellulose is carried out to obtain a hydromodule with a ratio of 1: 3.

To obtain a hydrogel "bacterial cellulose-chitosan" in a 50:50 ratio, mix a 2% solution of chitosan in 1% acetic acid, a hydrogel of bacterial cellulose, 25% glutaraldehyde (2%, v / v) and add 0.13% lysozyme and 2% dihydroquartsetin.

Example 9. Get a gel film of bacterial cellulose by culturing the bacterial strain Gluconacetobacter sucrofermentans VKPM B-11267 under static conditions on a medium with molasses. To remove cells and components of the culture medium, the resulting bacterial cellulose gel film is separated from the culture medium and purified. Sterilize bacterial cellulose gel film at 120 ° C for 20 minutes. Mechanical grinding of the purified gel-film of bacterial cellulose is carried out to obtain a hydromodule with a ratio of 1: 3.

To obtain a hydrogel "bacterial cellulose-chitosan" in a 50:50 ratio, mix a 2% solution of chitosan in 1% acetic acid, a hydrogel of bacterial cellulose, 25% glutaraldehyde (2%, v / v) and make 0.06% sodium fusidin, 0.1% resveratrol, 0.03% lysozyme.

Example 10. Get a gel film of bacterial cellulose by culturing the bacterial strain Gluconacetobacter sucrofermentans VKPM B-11267 under static conditions on a medium with molasses. To remove cells and components of the culture medium, the resulting bacterial cellulose gel film is separated from the culture medium and purified. Sterilize bacterial cellulose gel film at 120 ° C for 20 minutes. Mechanical grinding of the purified gel-film of bacterial cellulose is carried out to obtain a hydromodule with a ratio of 1: 3.

To obtain a hydrogel "bacterial cellulose-chitosan" in a 50:50 ratio, mix a 2% solution of chitosan in 1% acetic acid, a hydrogel of bacterial cellulose, 25% glutaraldehyde (2%, v / v) and make 0.06% sodium fusidin, 2% dihydroquarcetin, 0.15% chemotrypsin.

Example 11. Get a gel film of bacterial cellulose by culturing the bacterial strain Gluconacetobacter sucrofermentans VKPM B-11267 under static conditions on a medium with molasses. To remove cells and components of the culture medium, the resulting bacterial cellulose gel film is separated from the culture medium and purified. Sterilize bacterial cellulose gel film at 120 ° C for 20 minutes. Mechanical grinding of the purified gel-film of bacterial cellulose is carried out to obtain a hydromodule with a ratio of 1: 3.

To obtain a hydrogel "bacterial cellulose-chitosan" in a 50:50 ratio, mix a 2% solution of chitosan in 1% acetic acid, a hydrogel of bacterial cellulose, 25% glutaraldehyde (2%, v / v) and make 0.06% sodium fusidin, 10% transglutaminase solution, 2% dehydroquartsetin, 0.1% tinrostim.

Example 12. Get a gel film of bacterial cellulose by culturing the bacterial strain Gluconacetobacter sucrofermentans VKPM B-11267 under static conditions on a medium with molasses. To remove cells and components of the culture medium, the resulting bacterial cellulose gel film is separated from the culture medium and purified. Sterilize bacterial cellulose gel film at 120 ° C for 20 minutes. Mechanical grinding of the purified gel-film of bacterial cellulose is carried out to obtain a hydromodule with a ratio of 1: 3.

To obtain a hydrogel in a ratio of 5: 5: 15: 5, mix 2% chitosan solution in 1% acetic acid, 10% gelatin solution, 20% transglutaminase solution and bacterial cellulose hydrogel and add 0.1% resveratrol, 0.06% sodium fusidin. The obtained hydrogel-based biocomposite is washed with distilled water.

Example 13. Get a gel film of bacterial cellulose by cultivating the bacterial strain Gluconacetobacter sucrofermentans VKPM B-11267 under static conditions on a medium with molasses. To remove cells and components of the culture medium, the resulting bacterial cellulose gel film is separated from the culture medium and purified. Sterilize bacterial cellulose gel film at 120 ° C for 20 minutes. Mechanical grinding of the purified gel-film of bacterial cellulose is carried out to obtain a hydromodule with a ratio of 1: 3.

To obtain a hydrogel in a ratio of 5: 5: 15: 5, mix 2% chitosan solution in 1% acetic acid, 10% gelatin solution, 20% transglutaminase solution and bacterial cellulose hydrogel and add 0.1% resveratrol, 0.06% fusidin sodium, 0.14% trypsin.

The antibacterial properties of the claimed biocomposite are determined by a method based on the ability of antibiotic substances to diffuse in agar media and form zones in which microorganisms sensitive to these antibiotics do not develop. The bacteria Staphylococcus aureus 209 P are used as a test microorganism. A biocomposite disc with an antibiotic is placed in the center of a Petri dish inoculated with a test culture. The antibacterial properties of the obtained biocomposites are judged by the diameter of the growth inhibition zones of the test culture. Zone of absence of test culture growth when using biocomposites based on hydrogels of bacterial cellulose, chitosan and sodium fusidin, as well as physiologically active compounds of polyphenolic nature (dehydroquartzin, resveratrol), fements (trypsin, chymotrypsin, lysozyme), low molecular weight peptides, blood coagulation factors (thrombin, Ca ^{2+ ions} , transglutaminase) is 43.3 ± 4.1 mm.

Thus, a biocomposite based on a hydrogel of bacterial cellulose has been developed, which has a therapeutic effect, containing chitosan as an antimicrobial agent, fusidic acid as an enhancer of antimicrobial action, physiologically active polyphenolic compounds (dehydroquartsetin, resveratrol), enzymes of the hydrolase class that selectively break down tissues necrosis (trypsin, chymotrypsin), and destroying the cell walls of bacteria (lysozyme), low molecular weight peptides, in particular tinrostim. The resulting biocomposite has high antibiotic activity against Staphylococcus aureus bacteria, and regenerative properties and can be used in medicine as wound dressings.

Claims (1)

A method of obtaining a biocomposite with regenerative properties based on a hydrogel of bacterial cellulose, which consists in obtaining a gel film of bacterial cellulose by cultivating a bacterial strain Gluconacetobacter sucrofermentans VKPM B-11267 under static conditions on a medium with molasses, separating the obtained gel film of bacterial cellulose from the culture medium and its purification, mechanical grinding of the purified gel-film of bacterial cellulose for 10 min to obtain a bacterial cellulose hydrogel with a hydromodule with a ratio of 1: 3, to obtain a hydrogel bacterial cellulose: chitosan in a 50:50 ratio by mixing the bacterial cellulose hydrogel with a 2% chitosan solution in 1% acetic acid and with a 25% solution of glutaraldehyde, introduced in an amount of 2%, or obtaining a hydrogel bacterial cellulose: chitosan: gelatin: transglutaminase in a ratio of 5: 5: 15: 5, respectively, by mixing the hydrogel of bacterial cellulose with 2% chitosan solution in 1% acetic acid, 10% gelatin solution, 20% transglutaminase solution, after which 0.06% sodium fusidin is additionally added to the mass of the hydrogel and physiologically active polyphenolic compounds in the form of dehydroquartzin or resveratrol and / or enzymes in the form of trypsin, chymotrypsin, lysozyme and transglutaminase and / or low molecular weight peptides in the form of tinrostim.