Method of providing improved sealing by means of fibrin

FIELD: medicine.

SUBSTANCE: invention relates to medicine. Described are: fibrin matrix, method of its obtaining and method of processing or prevention of defect on wet tissue of object requiring processing. Described is application for hermetic sealing of defect in mucous membrane or other wet tissue.

EFFECT: fibrin matrix is efficient for sealing tissue leakages.

35 cl, 3 dwg, 1 tbl, 3 ex

 

The technical field TO WHICH the INVENTION RELATES

The present invention relates to a fibrin matrix with improved sealing properties, its access and use.

Background of the INVENTION

Fibrin sealant as a rule, is a blood product derived from industrial sources,and from some of the regional blood transfusion centres. Among the components typically used to obtain fibrin sealants are mainly fibrinogen, supplemented with different amounts of factor VIII, factor XIII, fibronectin, vitronectin and of von Willebrand factor (vWF). Fibrinogen component, typically activated last protease coagulation cascade is thrombin.

Fibrin sealant is formed by enzymatic reactions, including fibrinogen, thrombin and factor XIII. Thrombin convertsfibrinogen to fibrin by enzymatic action at a speed, determinedthe concentration of thrombin. Factor XIII, typically present in fibrinogen component of the seal and is an enzyme of the coagulation system of the blood, cross linking and stabilizing the fibrin clot. This process bypasses most of the stages of normal coagulation and simulates its last f�zu. Some manufacturers add antiproteolytic substances in the composition of fibrin sealant (see patent WO 93/05822) or specifically remove the plasminogen to stop or delay fibrinolysis (see U.S. patents Nos. 5792835 and 7125569).

In U.S. patent No. 4427650 revealed tissue adhesive that can be applied in the form of a dry powder mixture immediately and directly to the wound or surgical site. The adhesive consists of a solid powdered biologically active components and contains from 60 to 96% by weight. fibrinogen, isolated mainly from cryopreservative globulin, 0.05 to 5 wt%. inhibitor of fibrinolysis and from 0.1 to 15% by weight. thrombin and / or prothrombin.

In U.S. patent No. 5962405 disclosed lyophilized fibrinogen the drug to obtain fibrinogenous solution, used as a tissue adhesive. Fibrinogenic the product contains a substance that improves the solubility.

Some time ago it became known that fibrin sealant can be applied to injuries such as open wounds of a person, for the purpose of closing, stop the bleeding and prevent the wound with other substances, such as infectious substances.

It was reported on results of application of fibrin sealant to seal defects in the tissues, which is in constant motion, for example t�the Anh gastrointestinal tract and lungs. However, in these messages, there are differences.

One describes an approach to reduce postoperative defects such as leakage line bracket or thread (bracket/thread) weld after gastrointestinal resection, involves the strengthening bracket/thread seam. In sale there are various products to enhance bracket/thread seam. For example, doctors use neabsorbiruemye or prabaramee products to enhance bracket weld, such as Seamguard®, Peristrips Dry® and Surgisis®. However, in early experimental and clinical studies of absorbable material to strengthen the bracket weld demonstrated significant advantages compared to neabsorbiruemye or proabability material for reinforcement bracket weld (Yo et al. "Buttressing of the staple line in gastrointestinal anastomoses: overview of new technology designed to reduce perioperative complications". Dig Surg. 2006; 23:283-291). Recent reports on the use of fibrin as an absorbable amplifier bracket suture to prevent postoperative leakage at the gastrointestinal tract (Fullum et al. "Decreasing anastomotic and staple line leaks after laparoscopic Roux-en-Y gastric bypass". Surg Endosc. 2009; 23:1403-1408; Efthimiou et al. "Fibrin sealant associated with increased body temperature and leukocytosis after laparoscopic gastric bypass". Surg Real Trap Relat Dis. March 17, 2009) suggest that the existing composition of fibrin sealants neprihanita strengthening bracket/thread seams.

In U.S. patent No. 5690675 discloses the method of wound closure in the lung tissue through a two-step method, essentially consisting of a superposition of fasteners (e.g., staples, clips, pins, hooks) on coloradobuy the area where the fastening elements can form a penetrating hole. The fastening elements are covered with a preformed layer of collagen, fibrin, fibrinogen, elastin, albumin, or a combination of, and under the influence of this energy region the material is fused into the fabric, creating an airtight seal penetrating holes in the fabric.

In U.S. patent No. 5883078 revealed stable tissue adhesive containing fibrinogen and activator or preactuator of prothrombin. The adhesive may be provided in liquid or dry form. In one embodiment, the implementation of solid 2-sided tissue adhesive is prepared by blending the dry adhesive in solid form on the wound surface and the subsequent processing of the second wound surface (the second part of tissue) and intermittent squeezing the two parts together. The drug dissolves quickly with the help of present blood and (or) wound secretions and then hardens with the beginning of coagulation, while achieving a tight connection and hemostatic effect. Solid 2-sided tissue adhesive is particularly suitable for the connection of parts miah�their tissues, such as the liver or spleen.

Lillemoe et al. [(2004) J Gastrointest Surg., volume 8, No. 7, pp. 766-774 in the article entitled "Does Fibrin Glue Sealant Decrease the Rate of Pancreatic Fistula After Pancreaticoduodenectomy? Results of a Prospective Randomized Trial] showed that local application of fibrin glue sealant on the surface of pancreatic anastomosis has not contributed to the reduction in the incidence of pancreatic fistula or General complications of patients after pancreaticoduodenectomy, and came to the conclusion that the use of this substance in this case is not useful.

Another study ("The sealing effect of fibrin glue against alveolar air leakage evaluated up to 48h; comparison between different methods of application" Kawamura et al. (2005) Eur J of Cardiothorac surg. 28(1):39-42) discloses a sealing effect of fibrin glue against alveolar air leakage on the example of an animal model and evaluate the various methods of application. Used solution A, consisting of protein concentrate containing fibrinogen and a solution B containing thrombin. In the framework of the method of applying grinding and spraying solution a was applied dropwise and gently rubbed on the surface area of air leakage. Then both solutions were simultaneously sprayed in the form of a mixed aerosol. In another way received double layer by applying dropwise A solution on the surface area of air leakage, and then dropwise inflicted Rast�'or B. Within the other way in animal models (dogs) used a collagen matrix, with one side covered with dry fibrinogen and thrombin (TachoComb), and checked the results after 24 hours. According to the authors, the sealing effect of fibrin glue remains relatively volatile over the period up to 12 hours after application. In addition, the authors concluded that the method of applying grinding and spraying can increase the speed of achieving the fibrin sealant its full potential compared to other two methods.

Another study, by Yo et al. (2006) (Dig Surg 23: 283-291), discloses an attempt to reduce complications of the anastomosis, such as bleeding or leakage in lines bracket weld, when performing gastrointestinal resections. Yo notes that animal model rats with colonic anastomosis the use of fibrin glue to seal and prevent leakage does not seem appropriate. The article reveals that when gastrectomy application of fibrin glue to support and seal bracket weld seems to be more successful.

In U.S. patent No. 7196054 discloses the method of treatment of damaged tissue in a patient, comprising applying to the tissue a composition in the form of a dry powder containing fibrinogen in quantities�, sufficient for the formation of a fibrin matrix in the presence of thrombin, factor XIII, Ca2+and water solution; the introduction of the composition of thrombin, factor XIII, Ca2+and aqueous solution in amounts sufficient for the formation of a fibrin matrix in the presence of fibrinogen.

Fullum et al. (2009) reveals that the failure of the anastomosis and leakage line bracket weld (ASL) appear after laparoscopic gastric bypass with gastroenteroanastomosis Roux (LRYGB) patients are obese. Fibrin sealant was applied along the line bracket weld on the stomach. The authors concluded that the surgical technique, including suitable size of clamps, reinforcement bracket seam, stitching the incision manually fixing the seam, leakage test carried out during the operation, and fibrin sealants, has become a tool to reduce the frequency of occurrence of ASL after LRYGB. The authors were unable to isolate which components played a major role in preventing ASL.

In contrast, Efthimiou et al. (2009) reveals that the use of fibrin sealant did not affect the degree of insolvency of the anastomosis and leakage line bracket weld LRYGB.

There is an unmet need for an improved composition of fibrin sealant to seal defects in the tissues, nah�working people in constant motion, for example, tissues of the gastrointestinal tract and lungs.

BRIEF description of the INVENTION

Currently fibrin sealants are used primarily for hemostasis; this action can be enhanced by autologous blood coagulation factors such as fibrinogen present in the blood covering the wound area.

Fibrin sealant is used to seal the tissue at the leakage, for example, when the leakage of air or fluid, for example urine, a digestive fluid, serous fluid. Usually in the presence in the tissues of such leakage autologous blood coagulation factors are absent. Most of the seals, including synthetic seals are not effective to seal the leakage in the tissues. Without reference to the mechanism of the air/liquid can act as a barrier between the sealant and the tissue and thus prevents direct contact between the sealant and fabric.

Typically, surgeons most often dried wound leaking damaged tissue before applying the sealer. Typically, the drying is performed manually by means of a sponge/gauze and gas spray (Hidas et al. "Sutureless nephron-sparing surgery: use of albumin glutaraldehyde tissue adhesive (BioGlue)". Urology, 2006; 67:697-700). These drying procedures increase the risk of tissue adhesion (Kamel RM. "Prevention of postoperative peritoneal adhesions". Eur J Obstet Gynecol ReprodBiol. 2010; 150(2):111-118).

In addition, these drying procedures are also difficult to perform on inaccessible to the surgeon's sites, for example during laparoscopic operations.

The present invention provides a fibrin matrix, which is effective to seal the leakage of tissue.

In one aspect, the present invention relates to a method for applying fibrin matrix on a wet cloth, comprising: applying an effective amount of a solid fibrin sealing mixture to wet the fabric; and applying an effective amount of a liquid fibrin sealing composition over at least part of the deposited solid fibrin sealing mixture, and an effective amount of a solid fibrin sealing mixture and liquid fibrin sealing composition is sufficient for the formation of a fibrin matrix to wet the fabric with a higher sealing ability associated with the application of an effective amount of either liquid fibrin sealing compositions or solid fibrin sealing mixture.

In one embodiment of the present invention, the sealing ability is increased at least 1.2 times relative to applying liquid fibrin sealing composition.

In another embodiment of the present invention UE�unausa capacity increases approximately 1.7 times compared to applying liquid fibrin sealing composition.

In another additional embodiment of the present invention, the deposition is carried out on the tissue that is enriched, has small amount, not enough or has no blood vessels, and (or) on the fabric, slowly releasing the fluid or necropolises.

In one embodiment of the present invention, the solid fibrin sealing the mixture contains a solid component containing fibrinogen and solid component containing a proteolytic enzyme component capable of forming fibrin when interacting with fibrinogen.

In another additional embodiment of the present invention, the solid component fibrin sealing mixture comprising fibrinogen, prepared at the stage of drying, the fibrinogen-containing solution with a fibrinogen concentration of less than 25 mg/ml.

In one embodiment of the present invention, the fibrinogen concentration is approximately 20 mg/ml.

In another embodiment of the present invention, the proteolytic enzyme is a thrombin.

In another additional embodiment of the present invention, the deposition is carried out on a fabric defect.

In another embodiment of the present invention is carried out by applying �ENISA least a portion of a bracket or thread seams on the fabric.

Another aspect of the present invention relates to a method for producing a fibrin matrix on a wet surface, comprising the following stages: providing a solid component comprising fibrinogen; providing a solid component comprising a proteolytic enzyme capable of forming fibrin when interacting with fibrinogen; providing a liquid fibrin sealing composition; applying an effective amount of the solid components of at least a portion of the wet surface; and applying an effective amount of a liquid fibrin sealing composition over at least part of the deposited solid component.

In one embodiment of the present invention a wet surface does not contain fibrinogen.

In another embodiment of the present invention, the liquid fibrin sealant composition is provided in solid form and restore before applying.

In another additional embodiment of the present invention, the liquid fibrin sealant composition is provided in frozen form and thawed before use.

In another embodiment of the present invention provide a solid component in the liquid form and dried prior to application.

In one embodiment, the implementation�Oia of the present invention, the solid component provide frozen and dried before application.

In another additional embodiment of the present invention, the solid components are applied simultaneously or one after the other.

In another embodiment of the present invention, the liquid components are applied simultaneously or one after the other.

In another additional embodiment of the present invention, the solid components are presented in the form of a mixture.

In one embodiment of the present invention, the surface is a fabric.

In one embodiment of the present invention, the fabric is raw, has a small amount, not enough or does not have blood vessels and (or) slowly releases the fluid or bleed.

In another aspect, the present invention relates to a fibrin matrix, obtained in accordance with the present invention.

In another aspect the present invention provides a method of treatment or prevention of a defect on damp tissue in need of treatment of an object, comprising the following stages:

(a) providing a component comprising solid fibrinogen, providing a component comprising a solid proteolytic enzyme capable of forming fibrin when interacting with fibrinogen; and providing a liquid fibrin sealing compositions;

(b) n�carrying an effective amount of the solid components of p. a) at least a portion of the wet surface; and

c) applying an effective amount of a liquid fibrin sealing composition from step (a) over at least part of the deposited solid component.

In one embodiment of the present invention, the wet tissue is enriched, has small amount, not enough or does not have blood vessels and (or) slowly releases the fluid or bleed.

In another embodiment of the present invention, the defect is a leak in the tissue.

In another additional embodiment of the present invention, the leaking substance is enriched, has small amount, not enough or does not have a plasma or blood components.

In one embodiment of the present invention, the liquid fibrin sealant composition is provided in solid form and restore before applying.

In another additional embodiment of the present invention, the liquid fibrin sealant composition is provided in frozen form and thawed before use.

In one embodiment of the present invention provide a solid component in the liquid form and dried prior to application.

In another variant� implementation of the present invention, the solid component provide frozen and dried before application.

In another additional embodiment of the present invention, the solid components are applied simultaneously or one after the other.

In one embodiment of the present invention, the liquid components are applied simultaneously or one after the other.

In another embodiment of the present invention, the solid components of p. (a) presented in the form of a mixture.

In another additional embodiment of the present invention, the defect represents a bracket or thread.

In one aspect, the present invention provides a kit containing: (i) the container(s) contain(s) of solid components containing (a) fibrinogen component and (b) proteolytic enzyme component capable of forming fibrin when interacting with fibrinogen, wherein components (a) and (b) are in separate containers or in the same container in the form of a mixture; and (ii) at least two separate container, and at least one separate container contains a liquid, frozen or solid fibrinogen component, and at least a second separate container contains a liquid, frozen or solid proteolytic enzyme component capable of forming fibrin when interacting with fibrinogen, wherein if at least two separate container (ii) contain a solid component�you the components (a) and (b) are in the same container in the form of a mixture.

In one embodiment of the present invention, the proteolytic enzyme is a thrombin.

In another embodiment of the present invention is a Toolbox that applied to wet surfaces.

In another additional embodiment of the present invention, the kit is used to seal the leakage in tissues that are raw, have a small amount of, insufficient or do not have blood vessels, and (or) tissues, slowly releasing the fluid or necrobutcher.

Fibrin matrix, or set in accordance with the present invention can be used for treatment or prevention of a defect on a damp cloth.

BRIEF description of the DRAWINGS

Figure 1 shows the force required to separate two layers of tissue of the ileum (the force of exfoliation), connected together by means of two different fibrin sealing compositions: mixture of lyophilized fibrinogen and thrombin in comparison with two-component liquid fibrin sealing composition.

Figure 2 shows the sealing ability (measured by destructive testing) of a mixture of lyophilized fibrinogen and thrombin, two-component liquid� fibrin sealing compositions and fibrin matrix, obtained by consecutive application of lyophilized fibrinogen and thrombin and two component liquid fibrin sealant.

Figure 3 shows the sealing ability (measured by destructive testing) of fibrin obtained by applying lyophilized thrombin with subsequent atomization of the liquid fibrin sealant; fibrin, obtained by applying a lyophilized fibrinogen with subsequent atomization of the liquid fibrin sealant; fibrin, obtained by coating a mixture of lyophilized fibrinogen and thrombin with subsequent atomization of the liquid fibrin sealant; and a fibrin composition obtained by coating of dry albumin, followed by spraying of the liquid fibrin sealant.

DETAILED DESCRIPTION of embodiments of the INVENTION

The present invention relates to a method for applying fibrin matrix on a mucosa or other moist tissue, the method includes the sequential application of solid fibrin sealing mixture on the fabric surface followed by application of a liquid fibrin sealing composition over at least part of the surface. The present invention also relates to an effective sealing of the defect in the mucosa or in other�important tissue by sequentially applying a first solid fibrin sealing mixture, and then liquid fibrin sealing composition.

The basis of the present invention is based on the following research data.

The adhesion force (or the force of exfoliation) different test fibrin compositions studied by conducting research to flaking. The test allows you to measure the bond strength of fibrin glue between the layers of the submucosal membrane of the small intestine of the pig. In one embodiment, the implementation of the tested adhesive composition was coated on serous field ileum of the pig. After application of the test fibrin sealing composition fabric ileum turned (serosa to the serosa layers) and left fibrin sealing composition for polymerization. The force required to separate two glued layers of tissue of the ileum from each other (the force of exfoliation), was measured using discontinuous universal testing machines LFPLUS production Lloyd Instruments. Generally, the higher the measured force of exfoliation, the greater the force of adhesion of the test composition. In accordance with the present invention it was found that the adhesive force is significantly increased due to the use of lyophilized fibrin sealing composition is a liquid fibrin sealing composition. It was found that the strength of adhesion of the dried f�benovoy sealing composition was approximately 2.7 times higher compared to the adhesive force of the liquid fibrin sealing composition.

The sealing properties of different fibrin compositions was measured by conducting destructive tests. Destructive test is used to determine and to evaluate the ability of a sealant to prevent leakage. In short, aluminum tube connected to a water source and having holes injected into the tubular segment of the ileum of the pig, which is sealed and clamped at both ends of the tube. Then in an aluminium tube supplied with water, the water flows through the holes into the space between the ileum and aluminum tube, with a return flow of water into the aluminum tube is eliminated. Before serving water to produce a 10-mm slit perpendicular to the length of the colon, incision first stitch the thread on the middle line, and then on the incision area is applied to the test fibrin sealing composition through the stencil from parafilm area of 6 cm2placed around the incision area. After application of the test composition fibrin leave to cure (or polymerization) for 10 minutes and fill the bowel with water to study its ability to withstand pressure. As the water enters the space between the ileum and the pipe the water pressure increases until, until the sealing of the incision will not open and there is a sharp drop in pressure. The level� pressure measured using a manometer (D-logmate MRC 590 (Israel)), connected to the liquid outlet. The pressure measured before the drop-in, register and take the pressure destruction. Generally registered higher burst pressure indicates a higher sealing properties of the test composition.

In accordance with the present invention it was also found that freeze-dried fibrin sealant showed a higher adhesion force than that of liquid fibrin sealant has failed to seal. In contrast, the liquid fibrin sealing composition has a better sealing effect than freeze-dried fibrin sealant.

In addition, unexpectedly found that sequential application of lyophilized fibrin sealant and liquid fibrin sealing composition provides a synergistic improvement of the sealing properties. The observed increase sealing capacity of approximately 1.7 times compared with the sealing ability of a liquid fibrin sealing composition. However, the synergistic improvement of the sealing properties were identified during the sequential application of lyophilized fibrinogen or lyophilized thrombin with subsequent introduction of liquid fibrin sealing composition.

It was also set�Leno, the drying of the mucous membrane before applying the liquid fibrin sealing composition (e.g., by applying a protein powder, such as dry albumin, slimy layer) does not improve the sealing characteristics of the liquid fibrin sealing composition.

These data contributed to the development of an improved method of creating a fibrin matrix with a higher sealing ability to the mucosa or other moist tissue. Fibrin matrix, obtained in accordance with the principles of the present invention, can be used to seal defects in the mucosa or other moist tissue, such as tissues, are in constant motion, such as tissues of the gastrointestinal tract and lungs.

The term "fabric" refers to the Association of cells and (or) cellular components which are combined to perform specific functions. The cells in the tissue may belong to one type or more than one type. The fabric can be artificial, where cells are grown to ensure the functioning like a tissue in a living organism. The tissue may be tissue of the human or animal body.

The term "fibrin matrix" refers to fibrin, obtained by sequential application of solid fibrin sealing mixture and liquid fibrin uplo�enforcement of the composition.

The term "effective amount of solid fibrin sealing mixture" refers to the amount of the solid component containing fibrinogen, and the amount of solid component comprising a proteolytic enzyme capable of forming fibrin when interacting with fibrinogen, which after hydration for moist tissues form the fibrin clot.

The term "effective amount of a liquid fibrin sealing composition" refers to the amount of the liquid component containing fibrinogen and the quantity of the liquid component contains a proteolytic enzyme (capable of forming fibrin when interacting with fibrinogen), which ensure the formation of a fibrin clot after mixing of liquid components.

Without being bound to mechanism, it turns out that after applying the solid sealing mixture to wet the fabric is a hydration. Hydrated solid sealing compound fills the fabric, otorita liquid that is present in a humid fabric and forming a fibrin layer in close proximity to the tissue. Then put the liquid fibrin sealant composition of this tissue filled, resulting in improved sealing ability and strength.

The advantage is that the curing of the moisture on the surface without the need of using�ü sponge or gauze can reduce the risk of formation of adhesions in the postoperative period.

The term "spike" refers to an incorrect connection between tissues and / or organs. Usually adhesions formed after surgery, for example due to uneven processing of tissues; due to drying of the surface and (or) due to the presence of reactive foreign bodies (for example, suture materials, talc powder or fibre unravelling, to the operative field.

The term "mucosa" or "mucous tissue" refers to moist tissue that covers some organs and body cavities. Usually mucous tissue secretes a mucous material. Examples of mucosal tissue include, among others, the mucous membrane of the mouth, such as buccal and sublingual; nasal mucosa; mucous membrane of the eye; the mucous membrane of the genital organs; rectal mucosa; mucous membrane of the ear; the mucous membrane of the lungs; the mucous membrane of bronchus; mucous membrane of stomach; mucosa; mucous membrane of the olfactory organs; the mucous membrane of the uterus; and the lining of the esophagus. The term "mucous" is used to define raw (wet) and viscous material, such as a substance rich in mucin, albumin glycoprotein and (or) another component that contributes to the viscosity.

The term "another damp cloth" refers to the AGR�by tissue. The fabric can moistured physiological body fluids such as serum, infiltration of serum, blood and inflammatory fluid, and (or) other liquids (fluids), such as a phosphate buffer solution (FBI). In one embodiment of the present invention, the fluid does not contain coagulation factors (i.e. does not contain fibrinogen).

Within this document, the term "defect" refers to the strain, divergence, channel, fissure, puncture, wound, fistula, opening, cut, slit, perforation, fracture, puncture or rupture, leakage, etc, for example in the tissue. For example, a defect may occur as a result of anastomosis. The defect may be congenital, such as hernia; is caused by an abnormality in the body, such as seroma, hernia, infection, inflammation; to occur after the surgery, blending filament and (or) bracket weld; or due to external factors such as accidents, injuries.

The term "leakage" refers to the seepage or leakage of substances such as fluid, viscous material and / or air, for example using the strain, divergence, channel, fissure, puncture, wound, a fistula, a hole, a cut, a slit, a perforation, a crack, puncture or tear in the fabric.

The term "anastomosis" is usually used to refer to a surgical procedure�ry, used to re-connect two or more sites of an organ or tissue. The procedure can be performed after dissection of the urinary tract (urethra), throat (esophagus) or during surgical operations on the intestines. The procedure can also be performed after removal of diseased tissue (such as inflamed, cancerous tissue or tissue with another diseases, such as ulcers).

The present invention also relates to a kit containing: (i) the container(s) contain(s) of solid components containing (a) fibrinogen component and (b) proteolytic enzyme component capable of forming fibrin when interacting with fibrinogen; and (ii) at least two separate container, and at least one separate container contains a liquid, frozen or solid fibrinogen component, and at least a second separate container contains a liquid, frozen or solid proteolytic enzyme component capable of forming fibrin when interacting with fibrinogen.

Components (a) and (b) of item (i) can be in separate containers or in the same container in the form of a mixture. In one embodiment, the implementation, if at least two separate container (ii) contain solid components, components (a) and (b) are in the same container in the wee�e mixture. In another embodiment of the implementation, if at least two separate container (ii) contain solid components, at least one container contains an aqueous solution for hydration.

Unlike commercially available fibrin kits that are designed specifically for hemostasis, the set constituting the subject of the present invention, it is also possible to seal the leakage in tissues that are not enriched, have a small amount of, insufficient or do not have blood vessels.

Non-limiting examples of tissues that are not enriched, has small amount, not enough, or not having blood vessels, are: Dura mater, bladder, eyes, lungs and gall bladder.

In one embodiment of the present invention, the set is designed to seal the leakage in tissues that are not enriched, has small amount, insufficient or lacking blood vessels.

The term "hemostasis" refers to the ability of the agent to stop the bleeding from a damaged blood vessel and / or to promote retention of blood within the blood vessel.

In addition, unlike commercially available fibrin kits that are designed specifically to stop cromatiche�Oia from a damaged blood vessel and (or) to hold the blood within the blood vessel, the set constituting the subject of the present invention, it is also possible to seal the leakage of substances, not containing blood and plasma, such as cerebrospinal fluid (CSF), the air content of the intestine, bile, lymph fluid and the liquid portion of the vitreous body.

In one embodiment of the fibrin matrix or a set constituting the subject of the present invention, is intended to seal the leakage in tissues, slowly releasing the fluid or necrobutcher. The term "slow disbursement of fluid medium" refers to, for example, minor bleeding. The term "slow disbursement fluid" covers cases of bleeding, where there is relatively low blood loss at a relatively low speed.

The kit can also contain instructions for use. The kit may also contain means for cutting and / or stapling of the tissue or organ with staples or thread, such as mechanical or manual cutting and suturing device. As the container can be a vial, prefilled syringe, bottle, tube, or any other valid container containing solid or liquid components. Containers can be of different sizes and contain different amount (weight) of the compositions, for example, each liquid component may not have the volume b�than about 10 ml, and each solid component may have a mass of dry matter not more than about 3 g In a set may also be optionally included applicator for introducing solid and liquid components.

Fibrin matrix, or set in accordance with the present invention can be used for any therapeutic purposes. The term "any therapeutic purpose" refers to any type of treatment and prevention. Examples of therapeutic purposes include, among other things, seal the drilled holes made in the tissue or organ, for example in bone; anastomosis on blood vessels; the connection of tissue parts, such as parts of the soft tissue; treatment or prevention of defects of the Dura mater, such as gaps and leakage after infection of the Dura mater, fissures or cracks; stop or prevent bleeding; the elimination or prevention of air leaks, for example after resection of a lung; treatment or prevention of defects after perforation of the bowel; treatment or prevention of defects after the procedure anastomosis performed on any tissue, such as the uterus,esophagus, stomach, pancreas, pancreatic duct, gallbladder, bile duct, intestines (including the small intestine and colon) and rectum; the elimination or p�editremove postoperative leakage at any fabric, such as the uterus, esophagus, stomach, pancreas, pancreatic duct, gallbladder, bile duct, intestines (including the small intestine and colon) and rectum; preventing or reducing the incidence of postoperative leakage line bracket thread or seam, for example by applying a fibrin matrix in accordance with the present invention at least part of the defect, such as a bracket/thread seam; for solid fixation of the prosthesis, for example during an operation for hernia; to strengthen the bracket/thread seam; preventing or reducing leakage of alveolar air; treatment or prevention of renal defects; for treatment or prevention of fistula; treatment or prevention of cardiac defects, such as penetrating wounds to the heart; the strengthening of the vascular graft; and the elimination or prevention of leakage of cerebrospinal fluid.

In one embodiment of the present invention therapeutic use involves the seal for leakage in tissues that are not enriched with blood vessels or having an insufficient amount of blood vessels.

In another embodiment of the present invention therapeutic use involves strengthening bracket/thread seam.

The term "staple or filament" includes any fastening element used for closing wounds, such as, inter alia, clip, clip, pin, hook, thread, etc.

The term "at least part of the defect" in this document refers to the area that is less than, equal to or greater than the area of the defect. For example, the area may be 1, 10, 20, 30, 40, 50, 60, 70, 80, 90 or 100%, including any range between these percentages, or more from the defect area. In one embodiment of the present invention, the fibrin matrix, or a set constituting the subject of the present invention, is used to enhance a bracket/thread seam. For example, a 10 mm incision is made at the facility during a surgical operation, first sew on a thread or staples. For reinforcement in bracket/string suture is applied 100 mg of solid fibrin sealing mixture (1 000 IU of thrombin on 70 mg of fibrinogen), after which spray liquid fibrin sealant composition. Liquid fibrin is applied by spraying the composition with a total volume of 2 ml (the ratio of the two components 1:1) containing 1000 IU/ml of thrombin and 70 mg/ml of fibrinogen.

The term "object" in this document includes animals belonging to the class of mammals, including humans. In one embodiment, the implementation of the object is the patient.

In one aspect, the present invention provides a method of applying a fibrin matrix on a damp cloth. The method includes the following stages: application of an effective amount of a solid fibrin sealing mixture to wet the fabric; and applying an effective amount of a liquid fibrin sealing composition over at least part of the solid fibrin sealing mixture, and an effective amount of a solid fibrin sealing mixture and liquid fibrin sealing composition is sufficient for the formation of a fibrin matrix to wet the fabric having improved sealing ability associated with the application of an effective amount of either liquid fibrin sealing compositions or solid fibrin sealing mixture. Priority is the application of solid fibrin sealant before liquid fibrin sealant, which increases its sealing ability of a liquid fibrin sealant.

The term "wet tissue" refers to the wetted fabric and includes, for example, mucosa, mucous tissue and another damp cloth. In another embodiment of the present invention, the fabric is wetted with a fluid medium that does not contain fibrinogen. The term "not content�schy fibrinogen" refers, for example, the concentration of fibrinogen less than 1.5 g/L.

The term "over at least part of the solid fibrin sealing mixture" in the present document relates to the field ranging from the field with less surface area to a region with a greater surface area compared with the area to which applied(s) solid(s) fibrin(s) o(s) component(s). For example, a liquid fibrin sealing composition may be applied to 1, 10, 20, 30, 40, 50, 60, 70, 80, 90 or 100%, including any range between these percentages, surface area, that is covered by(s) solid(s) component(s). In an alternative embodiment of the liquid fibrin sealant composition can be applied to a larger surface area than the area to which applied(s) solid(s) component(s), for example, on the entire surface of the wet tissue or organ.

The term "liquid fibrin sealing composition" refers in the context of the present application at least two separate liquid components necessary for the formation of a fibrin sealant. At least one separate component contains fibrinogen and at least a second separate component contains a proteolytic enzyme capable of forming fibrin when interacting with fibrinogen (or fibrin�gene-containing composition). With the introduction of at least two separate components, for example by injection into the target area, the liquid components interact, simulating the final stage of the coagulation cascade of the blood and forming known to those skilled clot, fibrin.

The term "soft tissue" generally refers to structures of the body connecting, supporting, covering and (or) other surrounding structures and organs. Soft tissue can be connective tissue or nesoedinimogo cloth. Examples of soft tissue include, among others, tendons, ligaments, fibrous connective tissue, synovial membrane, connective tissue sheath, muscle, muscular wall, nerves, intestines, adipose tissue, liver, skin, spleen and blood vessels.

The term "solid fibrin sealing mixture" in the context of the present application relates to solid compositions containing a basic substance for the formation of a fibrin sealant. The composition contains a solid proteolytic enzyme capable of forming fibrin when interacting with fibrin (or fibrinogen-containing composition), fibrinogen and solid. The solid mixture may also contain other components, such as albumin. As a proteolytic enzyme can be used thrombin and / or a substance derived from snake venom. In about�nom embodiment of the present invention, the solid composition comprises fibrinogen and thrombin components are in lyophilized form. In another embodiment, the implementation of solutions containing thrombin and fibrinogen, are dried separately, for example by lyophilization, to obtain the respective solid components in the composition of the total solid fibrin sealing mixture. In another embodiment of the present invention, the resulting solid materials are ground into a powder using a mill, ultrafine grinder or mill with a cooled blade or manually by wiping with a solid material through a sieve, for example with a spatula. In one embodiment of the present invention, the material was ground by rubbing through a sieve with a mesh of 200 microns with a spatula. In another embodiment of the present invention, the solid materials were crushed into powder using the method described in international patent application WO 2008/053475, incorporated herein by reference.

The particle size of solid material after grinding can be less than 1 000 microns. In one embodiment of the present invention, the particle size of solid material after grinding is not more than 200 μm. The particle size may be in the range of 10 to 100 μm, or from 10 to 60 μm. Solid proteolytic enzyme and a solid fibrinogen can be provided together. It is possible for a firm provide the initial substance p� separately and applied to the target surface simultaneously or one after the other. In one embodiment of the present invention solid proteolytic enzyme and a solid fibrinogen provide in the form of a mixture.

The present invention also provides a method of producing a fibrin matrix wetted surface. The method includes the following stages: providing a solid component comprising fibrinogen; providing a solid component comprising a proteolytic enzyme capable of forming fibrin when interacting with fibrinogen; providing a liquid fibrin sealing composition; applying an effective amount of the solid components of at least a portion of the wetted surface; applying an effective amount of a liquid fibrin sealing composition over at least part of the solid components.

The term "wetted surface" refers to a wet surface. Surface mucosa, the mucous tissue and (or) another damp cloth. The surface can moistured biological fluids, such as infiltration of serum, blood and inflammatory fluid, and / or non-biological fluids such as the FBI.

The wetted surface may be a surface part of a patient's body, for example, any tissue that contains fluid or air. The term "surface" includes, p�addition, the area of the genital organs, including the uterus, vagina, and ovaries; the lungs; the anus; the spleen; the liver; the Dura; kidney; esophagus; stomach; pancreas; pancreatic duct; gallbladder; bile duct; intestine (including small intestine and colon); and cardiac muscle. The surface can bleed or not to bleed. In one embodiment of the present invention, the surface is not bleeding. In another embodiment of the present invention, the surface wetted by the fluid, not containing coagulation factor (i.e. does not contain fibrinogen). The surface can be any surface, such as a working surface, the surface of the prosthesis.

The term "at least part of the wetted surface" in this document refers to the area that is less than, equal to or greater wetted surface. For example, the area may be 1, 10, 20, 30, 40, 50, 60, 70, 80, 90 or 100%, including any range between these percentages, or more surface area.

Solid components in the solid fibrin mixture interact after hydration, for example after contact with damp cloth, forming a fibrin sealant. Usually fibrinogen drugs have a relatively low solubility, and mucous tissue demonstrating�should have a relatively low liquid content. The advantage is that fibrinogen component used solid fibrin sealing mixture capable of rapid dissolution and / or hydration after application to the fabric.

The term "over at least part of the solid(s) component(s)" refers to a region ranging from the field with less surface area to a region with a greater surface area compared with the area to which applied(s) solid(s) fibrin(s) o(s) component(s). For example, a liquid fibrin sealing composition may be applied to 1, 10, 20, 30, 40, 50, 60, 70, 80, 90 or 100%, including any range between these percentages, surface area, that is covered by(s) solid(s) component(s). In an alternative embodiment of the liquid fibrin sealant composition can be applied to a larger surface area than the area to which applied(s) solid(s) component(s), for example, on the entire surface.

In accordance with the present invention the solid fibrinogen obtained by lyophilization fibrinogen-containing solution with a fibrinogen concentration of 20 mg/ml, restored in aqueous solution at room temperature for a period of time less than 10 minutes. In one embodiment of the present invention fibrinogenic solid component fibrenew�th o mixture derived from a fibrinogen-containing solution with a fibrinogen concentration of not more than 150, for example, not more than 85, 25 mg/ml or less, 20 mg/ml.

Solid fibrin sealing mixture can be obtained by drying liquid fibrin sealing composition. Allowed the use of any known in this field procedures of drying, including, without limitation, lyophilization (freeze-drying) or spray drying. Drying can be performed at various temperatures using a variety of equipment for drying, such as a camera with adjustable humidity, drying oven, drying channel, vacuum drier, or any other legitimate means that does not affect clot formation.

The term "solid" in the framework of the present description relates to compositions containing liquid in an amount of less than 5%, for example less than 4, 3, 2, 1% by weight or less of the total dry weight of the composition. The term "solid" and the terms "dry" or "powder" are used interchangeably.

Solid fibrin sealant can be applied to the fabric by means of a dispenser. Examples of dispensers are presented in U.S. patent No. 1776489 and in U.S. patent No. 7455248, incorporated herein by reference.

The term "improved sealing capability" refers to, for example, sealing ability, which is at least approximately 1.2 times higher compared with the sealing ability �of the strange liquid sealer. In one embodiment of the present invention, the sealing ability is increased approximately 1.7 times compared to applying liquid fibrin sealing composition. The sealing ability can be measured, for example, by means of destructive tests described above.

The phrase "sealing ability, increased 1.2 times" refers to the sealing ability, which is 20% higher than the sealing ability is achieved through applying liquid fibrin sealing composition. The phrase "sealing ability, increased 1.7 times" refers to the sealing ability, which is 70% higher than the sealing ability is achieved through applying liquid fibrin sealing composition.

As a proteolytic enzyme can be used thrombin and / or a substance derived from snake venom. Liquid components can be frozen until use (for example, at -18°C or below) or dried, for example by lyophilization for long-term storage. Dried beans can be restored before use by adding different amount of pharmaceutically acceptable carrier. The term "pharmaceutically acceptable carrier" refers to any diluent or environment that are valid for use in relation to people�age or other animals. The carrier can be selected from among known in the field of media, such as, inter alia, phosphate buffer solution (PBS), saline, a solution of sodium chloride, calcium chloride, lactate ringer's (LR), 5% dextrose in normal saline solution and water for injections. For the recovery of lyophilized fibrinogen before use, you can use sterilized water. For the recovery of lyophilized thrombin prior to use, you can apply a sterile solution of calcium chloride or water, for example distilled water. Restored lyophilized fibrinogen and thrombin can be used in the composition of the liquid fibrin sealing composition with the subsequent formation of fibrin. Two components can be applied to the desired area at the same time or one after the other.

In one embodiment of the present invention, the fibrinogen-containing component comprises a biologically active component (BAC), which is a solution of proteins derived from blood plasma. This component may further contain tranexamic acid and arginine or lysine or mixtures of arginine and lysine, or their pharmaceutically acceptable salts.

The TANK can be obtained from cryoprecipitate, such as a concentrated cryoprecipitate. The term "crop�alpicat" refers to a component of blood, obtained from frozen plasma derived from whole blood. Cryoprecipitate can be obtained by thawing frozen plasma in the cold, typically at a temperature of 0-4°C, resulting in a precipitated supernatant containing fibrinogen and factor XIII. The precipitate can be collected, for example, by centrifugation. Usually the TANK contains factor VIII, fibronectin, VWF (vWF), vitronectin, etc., for example, as described in U.S. patent No. 6121232 and corresponding published PCT application WO 98/33533, incorporated herein by reference.

The TANK may contain stabilizers such as arginine hydrochloride. Generally, the amount of fibrinogen in the TANK is in the range from about 40 to about 60 mg/ml. the Number of tranexamic acid in the solution TANK can be in the range of from about 80 to about 110 mg/ml. Amounts of arginine hydrochloride may be in the range of from about 15 to about 25 mg/ml.

Solution optional buffer to a physiologically compatible pH. Buffer solution may consist of glycine, sodium citrate, sodium chloride, calcium chloride and water for injection, used as the medium. Glycine may be present in the composition in an amount of from about 6 to about 10 mg/ml, citrate �Atria may be present in the range from approximately 1 to approximately 5 mg/ml, sodium chloride may be present in the range from approximately 5 to approximately 9 mg/ml, and the concentration of calcium chloride may be about 0.1-0.2 mg/ml.

In another embodiment, the implementation of the concentration of plasminogen and plasmin in the composition of the TANK is reduced to a value of not more than 15 μg/ml, for example up to 5 μg/ml or less plasminogen, for example, the method described in U.S. patent No. 7125569 and corresponding published PCT application WO 02/095019, incorporated herein by reference.

Also allowed to fibrin sealing composition contained the components that contribute to the formation of a clot, such as Ca2+, factor VIII, fibronectin, vitronectin, of von Willebrand factor (vWF), which can be prepared in the form of separate components or mixed with the liquid components.

Liquid fibrin sealant composition can be applied to the fabric by means of a dispenser, which allows you to apply the sealer directly onto the fabric or other media, or work surface using a jet of air or without, for example, by adding dropwise. In one embodiment of the present invention, the sealant is sprayed under an air pressure of 103.4 kPa (15 psi). Examples of dispensers for tissue sealants, see U.S. patents№№ 4631055, 4846405, 5116315, 5582596, 565067, 5989215, 6461361 and 6585696, 6620125 and 6802822, and in PCT publications Nos. WO 96/39212, WO 2007/059801 and WO 2010/095128, incorporated herein by reference.

Before use in surgery liquid fibrin sealing composition comprises at least two components. In one embodiment of the liquid fibrin sealing composition comprises two liquid component, wherein one component contains fibrinogen and in the interaction with the second component containing a proteolytic enzyme, such as human thrombin forms fibrin clot. During surgical operations separate liquid components, for example two liquid component is applied simultaneously or one after the other, for example, using two syringes, resulting in the two components are mixed with the formation of fibrin.

In one embodiment of the present invention, the proteolytic enzyme used is human thrombin with activity in the range from approximately 2 to approximately 4 000 IU/ml. Coagulation activity of thrombin can be measured directly, for example, in accordance with the test procedure according to the European Pharmacopoeia (0903/1997), and (or) indirectly, for example by measuring the length of the migration path on an inclined surface ("drip test" as described in published�Oh patent application U.S. 2010/0203033) or in any other way, known in this field. The specialist will be clear that the composition of the fibrin glue can be specified coagulating protein is the clotting of fibrinogen.

In another embodiment of the present invention fibrinogenic and proteolytic enzyme components is applied in such a way that equal amounts of the two components are mixed and applied to the corresponding wound surface of a patient. Of course, it should be understood that the fibrin matrix, which is the subject of the present invention, can be used during surgical operations and in other situations when you need to stop the bleeding.

Solid fibrin sealing mixture can be obtained by mixing the solid fibrinogen and solid thrombin in any desired range of ratios. For example, if a firm fibrinogen component received from a fibrinogen-containing solution with a concentration of fibrinogen 40-85 mg/ml, and a solid thrombin component received from the thrombin-containing solution with a concentration of thrombin approximately 800-1 200 IU/ml, two solid components can be mixed at a ratio of 1:1, 3,2:1, 6,4:1 accordingly, etc. the Mixing can be carried out under controlled humidity (relative humidity of 25% at a temperature of 37°C). The powder mixture can be stored up to ispolzovaniem closed containers.

Before drying, the fibrinogen-containing solution can be diluted, for example, to obtain a fibrinogen concentration of 25 mg/ml or 20 mg/ml. mixing of the two components can be performed before applying the solid fibrin sealing of the mixture on a cloth or you can apply these components directly on the fabric in the desired proportions.

During application of the liquid fibrin sealing composition on the fabric can cause the fibrinogen-containing component and a thrombin-containing component in any desired range of ratios. For example, if the concentration fibrinogenous component is 40-85 mg/ml and the concentration of thrombin is approximately 800-1 200 IU/ml, the two components may be mixed in the ratio 1:1, 1:2, 1:3, 1:4, 1:5, 1:6 accordingly, etc. In one embodiment of the present invention the components of the liquid fibrin sealant is used in a 1:1 ratio.

Fibrin matrix, which is the subject of the present invention, can be used in therapeutically effective amounts. The term "therapeutically effective amount" refers to the dose required for prevention or treatment of disease, disorder or pathological condition. Effective dose can be changed depending on the age and weight of the object, the disease and its severity (for example, the amount of time�ESA) and other factors, which can be detected by specialists in this field. For example, solid fibrin sealing mixture can be applied in the amount of from 0.1 to 100 mg / cm2for example , in volume 0,5, 1, 4, 8, 16, 30, 60 or 100 mg / cm2. Liquid fibrin sealant composition can be applied in the amount of from approximately 0.01 to approximately 1 ml per cm2for example , in the amount of 0.01, 0.05, and 0.5 or 1 ml per cm2.

In one embodiment, the implementation in the context of 10 mm on an area of 6 cm2consistently applied by scattering 100 mg of solid fibrin sealing mixture (1 000 IU of thrombin on 70 mg of fibrinogen), after which spray liquid fibrin sealant composition. The liquid composition is applied by spraying a total volume of 2 ml (the ratio of the two components 1:1), where thrombin is 1 000 IU/ml and fibrinogen is 70 mg/ml.

In accordance with the methods of the present invention a liquid fibrin sealing composition may be provided in solid form and restored before application. Recovery can be accomplished by adding a pharmaceutically acceptable carrier in various amounts. The term "recover" refers to a method for the conversion of a solid in a liquid form. The reconstituted product may be a liquid product obtained by adding fluids are�and water based for dry substances, from which previously removed liquid.

In an alternative embodiment of the liquid fibrin sealing composition may be provided in frozen form, for example at temperatures
-18°C or below and thawed before use. Defrosting can be done by incubation containers containing liquid components at room temperature (from 20°C to 25°C), at a temperature between 2 to 8°C or at 37°C. In another embodiment of the liquid components are applied simultaneously. In another additional embodiment of the liquid components are applied one after the other.

The solid component may be provided in liquid form and dried prior to application, for example by lyophilization (freeze-drying) or spray drying. In an alternative embodiment of the solid component can be provided in frozen form and dried prior to application. Frozen components can be thawed and then dried, or can be converted directly from frozen in a solid form, for example by sublimation.

The solid component may be provided in suspension, for example in any acceptable solvent, including non-aqueous liquid, such as hydrofluorocarbons (HFCs), or any other liquid carrier in the cat�Roy does not dissolve the solid component, such as alcohols, ethers or other organic liquids. Solid components and liquid components can be applied simultaneously or one after another in accordance with the methods constituting the subject of the present invention. In one embodiment of the solid components provide in the form of a mixture and applied simultaneously. In another embodiment of the solid components are provided in separate containers and applied simultaneously. In another additional embodiment of the solid components are provided in separate containers and put one after the other.

Fibrin matrix, which is the subject of the present invention, can be applied by any method used by surgeons to correct or prevent defects, including, among other things, open surgery and minimally invasive surgery (m & e), such as laparoscopic surgery. In one embodiment of the present invention in the operative field perform incision and the defect is applied to a fibrin matrix. In another embodiment, the implementation at operational field perform incision, the incision is sutured with staples or thread and bracket thread or suture is applied in a fibrin matrix. The patient receives local, regional or General anesthesia. The term "open surgery" refers to surgery.�quarter intervention when the surgeon has direct access to the operating area through a relatively large incision.

Within this document, the term "minimally invasive surgery" refers to surgery wherein the surgeon gains access to the operating area through small incisions or through the cavity of the body, or through the anatomical aperture, such as by laparoscopy. For visualization of the surgical field it is possible to use special equipment, such as miniature cameras with microscopes, tiny fiber-optic flashlights and high definition monitors. In relation to the operating area, you can use those tools in "end terminals" such as forceps, cutters, needle holders, prijateli, etc.

Fibrinogen - and thrombin-containing components, supplying manufacturers such as OMRIX (for example, EVICEL®, QUIXIL®, ADHEXIL™; EVITHROM®); Baxter (e.g. TISEEL®); CSL (e.g., Beriplast®), etc. In one embodiment of the liquid fibrin sealing components prepared from total human plasma and supplied as a set for single use, comprising two vials: one vial contains biologically active component 2 (BAC) and the other ampoule contains thrombin. The kit may further include a sterile applicator, and instructions for use.

In od�ohms embodiment of the component BACK is a sterile solution with a pH of 6.7 to 7.2, essentially consisting of a concentrate of human fibrinogen. Fibrinogen - a protein in human blood that forms a clot in the interaction with thrombin. Liquid fibrin sealing composition, such as a solution BACK, may contain: concentrate of human fibrinogen (55-85 mg/ml), arginine hydrochloride, glycine, sodium chloride, sodium citrate, calcium chloride and water for injection (WFI).

In one embodiment of the thrombin component is presented in the form of a sterile solution, pH 6,8-7,2, containing human thrombin high cleaning, activating coagulation of the final combined product. Thrombin is a highly specific protease that transforms the fibrinogen contained in BACK in fibrin. A solution of thrombin may include: human thrombin (800-1 200 IU/ml), calcium chloride, human albumin, mannitol, sodium acetate and water for injection.

Cryoprecipitate, which may be the source material for BACK, and cryosupernatant plasma, which can be the source material for the production of thrombin, can be derived from total human plasma. Specialists in this field are well-known stage of cryoprecipitate and cryosupernatant plasma. In one embodiment, the implementation of thrombin is produced by chromatographic purification of prothrombin from cryosupernatant plasma pic�e its activation with calcium chloride, for example, as described in U.S. patent No. 5143838, incorporated herein by reference. In another embodiment of fibrinogen component is prepared from cryoprecipitate, in particular from concentrated cryoprecipitate.

Fibrinogen and thrombin components mentioned above as part of the liquid fibrin sealing component or lyophilized component, can be obtained from human plasma or animal. However, it is also allowed to obtain components recombinant methods. Fibrinogen components can be obtained in accordance with methods described in U.S. patent No. 6121232 and PCT published application WO 98/33533 where plasmin(just started playing) is removed, as described in published application EP 1390485, and add tranexamic acid.

Fibrin sealing components in the two component liquid fibrin sealant or solid fibrin sealing mixture derived from blood or fractions of blood, is usually subjected to at least two discrete stages of inactivation/removal of viruses. The processes of inactivation and removal of viruses can be performed, for example, in the following ways: nanofiltration; dissolution/purification; treatment at low pH, UV irradiation; by treatment with sodium thiocyanate and (or) by any other method known in this field.

Therm�n "stage of virus inactivation" refers to a situation in which the virus remains in solution, but are viable, for example by dissolving their lipid membranes. The term "stage of the virus removal" refers to a situation in which physically removes viruses from solution, for example by exclusion by size.

The term "at least two discrete stages of the inactivation/removal of viruses" includes at least two different and independent types of processing to inactivate or remove viruses. Allowed the combination of two or more of the following non-limiting examples of processing: pasteurization, dissolution/purification (R/O), nanofiltration, processing at low pH, UV irradiation and treatment with sodium thiocyanate.

The use of the TANK, obtained from concentrated cryoprecipitate, is advantageous, since this fraction, in addition to fibrinogen, also contains valuable blood components that play an important role in the process of coagulation of blood in the interaction of proteolytic enzyme, such as human thrombin, with the solution TANK. Among the valuable components include, for example, factor VIII, factor XIII, fibronectin, vitronectin, of von Willebrand factor (vWF), etc.

The application of solid fibrin sealing mixture, followed by coating a liquid fibrin sealing composition promotes the formation �Obrenovac matrix on the fabric with a higher sealing ability to achieve that is through the application of each of these sealing compositions separately is not possible. A fibrin matrix is formed in accordance with the present invention, particularly suitable for sealing defects in mucosal tissues and other intestinal or wet tissue, which is in constant motion, for example in the tissue of the gastrointestinal tract. For example, fibrin in accordance with the present invention can be used to prevent complications in the form of leakage after resection overlay bracket welds or anastomosis in surgery of the gastrointestinal tract. In addition, the use of fibrin in accordance with the present invention can be preferably used to secure the prosthesis during the operation for hernia.

The content of the applications, patents and publications mentioned above and hereinafter are hereby incorporated herein by reference.

The present invention is further described with reference to the following non-limiting examples.

EXAMPLES

Materials and methods

Lyophilization

Lyophilization was carried out in accordance with the following cycle:
StagePhase Time (h:m)Temp. (°C)Vacuum
(PA (mbar))
1Initial value--:--4NO
2Freezing1:00 PM-30NO
3Freezing1:00 PM-50NO
4Freezing5:40-50NO
5Receiving0:20-45NO
6Sublimation0:15-4220(0,2)
7Sublimation0:15-2520(0,2)
8Sublimation 25:00-2520(0,2)
9Sublimation1:00 PM-1520(0,2)
10Sublimation12:00-1520(0,2)
11Sublimation2:002020(0,2)
12Sublimation5:00 PM2020(0,2)
13Secondary drying0:302512(0,12)
14Secondary drying18:002512(0,12)

Liquid fibrinogen and thrombin

In the experiments described below as a two component liquid fibrin sealant used two liquid component fibrin sealant EVICEL® (BACK and thrombi�; Omrix Biopharmaceuticals Ltd.). Fibrinogen component (containing 70 mg of fibrinogen/ml) used in the existing condition and, unless otherwise indicated, the thrombin component used in the existing status (1 000 IU/ml) or diluted in 10-fold diluted buffer solution [of 0.04 M CaCl2in distilled, deionized water (DDW)]. In all experiments, the liquid fibrin sealant sprayed under an air pressure of 103.4 kPa (15 psi).

Obtaining a lyophilized mixture of fibrinogen and thrombin

Compositions of solutions used to obtain a lyophilized powder, was a two liquid component ofEVICEL®. Each of the components liofilizirovanny separately according to the lyophilization cycle described above. Before the lyophilization procedure fibrinogen component was diluted with DDW to obtain a fibrinogen concentration of 20 mg/ml.

After drying, the powders were rubbed with a spatula through a sieve with a mesh of 200 μm. Pureed powders of fibrinogen and thrombin were mixed in the ratio by weight of 3.2:1 (equal to the ratio of 1 000 IU of thrombin on 70 mg of fibrinogen, as part of EVICEL) 6.4:1 (equal to the ratio of 500 IU of thrombin on 70 mg of fibrinogen), respectively. Mixing was performed under controlled humidity (relative humidity of 25% at a temperature of 37°C). The powder mixture was stored until use�tion in a closed container. For each experiment was prepared fresh powder mix.

Example 1. Adhesive force lyophilized fibrin sealing compositions and liquid fibrin sealing composition.

In the next experiment measured the adhesive force lyophilized fibrin sealing compositions and liquid fibrin sealing composition.

The evaluation was performed by examining the efforts of exfoliation, essentially as described in the publication Nicoson ZR, G. Buckley, California. "Bond strength of fibrin glue between layers of porcine small intestinal submucosa (SIS)". Biomed Sci Instrum. 2002; 38:179-184. In short, the test of the adhesive composition was coated on serous field ileum of the pig (segment 4×10 cm) using a stencil from parafilm of size 4×4 cm (16 cm2) (application method see below).

After applying various fibrin sealing compositions stencil from parafilm removed, the tissue of the ileum turned (serosa to the serosa layers) and left to polymerize for 10 minutes at room temperature (approximately 20-25°C). To obtain four lines segment was cut into strips measuring 1×10 cm and measured the force required to separate two glued layers of tissue of the ileum from each other (known as the force of exfoliation), using a discontinuous universal testing machines LFPLUS production Lloyd Instruments.

Ispy�yvajemie compositions were applied as follows:

- liquid fibrinogen and thrombin components were applied by spraying the same amount of fibrinogen and thrombin a total volume of 2 ml. Thrombin was applied in two different concentrations, 100 or 1000 IU/ml, and fibrinogen in a concentration of 70 mg/ml; and

400 mg of each lyophilized mixture (a mixture containing 500 or 1000 IU of thrombin on 70 mg of fibrinogen, prepared as described in section "Materials and methods") were applied to the stencil from parafilm 16 cm2(per replica inflicted 100 mg). Then to prevent drying of the tissue on the powder sprayed by the FBI as long as the powder were wet.

The values of adhesion strength for different test compositions are shown in Fig. 1.

It was noted that in the formation of fibrin using the lyophilized composition to separate two layers of tissue of the intestine it took more force compared to the force required in the case of a liquid fibrin sealing composition (compare the efforts of exfoliation when using lyophilized composition, and in the case of liquid compositions with the same concentration of 1000 IU of thrombin on 70 mg of fibrinogen).

Thus, it was found that the strength of adhesion between two samples is significantly higher when using lyophilized composition.

In addition, it was about�noted, solid fibrinogen obtained by lyophilization fibrinogen-containing solution with a fibrinogen concentration of 20 mg/ml, restored in aqueous solution at room temperature for a period of time less than 10 minutes. The lower concentration of fibrinogen leads to the formation of more porous lyophilized residue and more porous particles after grinding of the residue. It is believed that the high porosity allows fluid easier to penetrate into the particles, resulting in reduced rehydration time.

Example 2. The sealing property is different fibrin sealing compositions.

The previous example demonstrated that the application of the lyophilized composition is increased adhesive force. During this experiment investigated the sealing properties of three different sealing compositions: mixture of lyophilized fibrinogen and thrombin, liquid fibrin sealing compositions and fibrin sealant obtained by consecutive application of a mixture of lyophilized fibrinogen and thrombin and the subsequent deposition of a liquid fibrin sealing composition. Destructive testing [essentially as described by Vilela et al. "What Is Important For Continent Catheterizable Stomas: Angulations or Extension?" Int Braz J Urol. 2007; vol. 33(2): 254-263] was performed to determine and to evaluate the ability of the�of latices to ensure a tight seal and to withstand the pressure.

Pressure destruction in this model indicates the ability of the test composition adheres to the cloth and to maintain its mechanical integrity to reach the value of pressure at which rupture of the seal, resulting in pressure drops dramatically and there is an obvious leakage of fluid.

As mentioned previously, a specially designed aluminum tube (length 27 cm) with holes injected into the tubular segment of the ileum of the pig to a depth of 25-30 cm Tubular segment has consolidated, pinning him to both ends of the tube using plastic discs, and tightened by means of metal screws. Aluminum tube connected to the water source. After the aluminum tube filed water, the water received in the space between the ileum and the aluminum tube through holes provided around the circumference of the tube, with a return flow of water into the aluminum tube was removed.

Before water using a sharp blade he performed a 10-mm slit perpendicular to the length of the colon. To more closely simulate clinical conditions first sewed up the cut threads on the middle line (using one strand 3×0; ETHICON; catalog number ss684), and then the incision through the stencil from parafilm (20×30 mm; 6 cm2placed around the incision area, struck ispytyvau�th fibrin sealing composition. After application of the test compositions (see drawing below) fibrin left for 10 minutes for curing, and the intestines were filled with water to study its ability to withstand pressure. As the water enters the space between the ileum and the pipe the water pressure increases until, until the seal is cut and there is a sharp drop in pressure. Set the pressure level was measured using a manometer D-logmate MRC 590 (Israel), connected to the supply line of the fluid, to detect a sharp decrease in the pressure curve. The pressure measured to a sharp decrease was registered and accepted for the pressure of destruction.

The test compositions were applied as follows:

Liquid fibrin was applied by spraying a total volume of 2 ml (the ratio between the two components is 1:1). During this experiment examined only one concentration of thrombin: 1 000 IU 70 mg of fibrinogen.

100 mg lyophilized mixture dispersed through the stencil size 6 cm2. Experiencing only one mixture is a mixture containing 1000 IU of thrombin 70 mg (lyophilized fibrinogen and thrombin components were mixed in the ratio of 3.2:1, as described in "Materials and methods"). To prevent drying of the fabric after the dispersion of the lyophilized composition was sprayed to the FBI.

p> - Sequential deposition was performed by scattering 100 mg of lyophilized fibrinogen and thrombin (1000 IU thrombin 70 mg) followed by spraying of liquid fibrinogenic and thrombin components (total volume 2 ml). If the mucous membrane that surrounds the intestines, not fully wet the lyophilized powder, after the dispersion of the lyophilized composition was sprayed to the FBI.

It was found that freeze-dried fibrin sealant having a higher adhesive force than the liquid fibrin sealant (see Fig. 1) has failed to maintain a seal (showed low pressure value destruction; Fig. 2).

Liquid fibrin sealant demonstrated a higher sealing effect than freeze-dried fibrin sealant. Unexpected ways consistent application of lyophilized fibrin sealant before spraying a liquid fibrin sealant gave a higher sealing effect than when combining components. Thus, it was found that the sealing property of the sequential application of powder and liquid seals higher than the sealing properties of each composition used separately.

Without being bound to mechanism, it is obvious that the lyophilized fibrinogen and thrombin powders �astonauts in the mucosa, covering the bowel tissue, thus forming a layer of fibrin in close proximity to the serous tissue. When spraying liquid fibrin sealant after application of the lyophilized powders marked synergistic seal tissue as described above (Fig. 2).

Example 3. The highest sealing effect of fibrin composition obtained by coating a solid fibrin sealing mixture, followed by coating a liquid fibrin sealing composition

In the previous examples it was shown that fibrin sealant obtained by consecutive application of a lyophilized mixture of fibrinogen and thrombin with subsequent atomization of the liquid fibrin sealant that has the most high sealing capability.

In the following example, using destructive tests exploring the sealing characteristics when applied to only one of the lyophilized components separately (either fibrinogen or thrombin) followed by application of a liquid fibrin sealant. The following example also examine the question of whether the application of the section of albumin, drying the mucous membrane covering the intestines, to promote better adhesion of fibrin sealant to the tissue and thereby improving the sealing characteristics of the gasket.

D�these goals, I conducted as described above (example 2) destructive test and evaluate the sealing property following fibrin sealants: fibrin, obtained by application of 25 mg of lyophilized thrombin (derived from a solution containing 1000 IU/ml of thrombin, where thrombin is the liquid component ofEVICEL; lyophilized 625 µl solution), followed by spraying 2 ml of liquid fibrin sealant; fibrin obtained by applying 75 mg of lyophilized fibrinogen (obtained from a solution containing 20 mg/ml of fibrinogen, as described above) followed by spraying 2 ml of liquid fibrin sealant; fibrin obtained by applying 100 mg of a mixture of lyophilized fibrinogen and thrombin (in the ratio of 3.2:1 by weight as above) followed by spraying 2 ml of liquid fibrin sealant; and a fibrin composition obtained by applying 100 mg of dry albumin (Sigma; catalog number 078k1503) followed by spraying 2 ml of liquid fibrin sealant. All powders (including dry albumin) before using rubbing with a spatula through a sieve with a mesh size of 200 microns.

The results are presented in Fig. 3. Statistical data were obtained by one-factor analysis of variance with the definition of the criterion of Tukey-Kramer. The same letters indicate the average values are not significantly different from each other (p>0.05) and different letters are used to indicate significantly different average value�n (p≤0.05). It is noted that sequential application of solid fibrin sealant and liquid fibrin sealing of the composition promotes formation of a fibrin matrix with superior sealing ability compared with fibrin matrix obtained by applying only one of the lyophilized components separately (either fibrinogen or thrombin), followed by applying a liquid composition. It was also found that the application of dry protein (e.g., albumin), drying the mucous membrane, does not increase the sealing ability of a liquid fibrin sealing composition.

1. The method of application of fibrin matrix on a damp cloth containing:
(a) applying an effective amount of a solid fibrin sealing mixture on a damp cloth, where the solid fibrin sealing the mixture contains a solid component containing fibrinogen and solid component containing thrombin; and
(b) applying an effective amount of a liquid fibrin sealing composition over at least part of the deposited solid fibrin sealing mixture, where the liquid fibrin sealing composition comprises at least two separate liquid component, wherein at least one separate component contains fibrinogen and at least a second of�practical component contains thrombin;
and an effective amount of a solid fibrin sealing mixture and liquid fibrin sealing composition sufficient to obtain a fibrin matrix to wet the fabric having improved sealing ability in relation to the application of an effective amount of either liquid fibrin sealing compositions or solid fibrin sealing mixture.

2. A method according to claim 1, wherein the sealing ability is increased at least 1.2 times relative to applying liquid fibrin sealing composition.

3. A method according to claim 1, wherein the sealing ability
increases approximately 1.7 times compared to applying liquid fibrin sealing composition.

4. A method according to claim 1, wherein the applying is carried out on the tissue that is enriched, has small amount, not enough or has no blood vessels, and (or) on the fabric, slowly releasing the fluid or necropolises.

5. A method according to claim 1, wherein the fibrinogen-containing solid component fibrin sealing mixture is prepared at the stage of drying, the fibrinogen-containing solution with a fibrinogen concentration of less than 25 mg/ml.

6. A method according to claim 5, in which the concentration of fibrinogen is approximately 20 mg/ml.

7. A method according to claim 1, wherein the applying of�Westside on a fabric defect.

8. A method according to claim 1, wherein the applying is performed by at least part of the bracket or thread seam on the fabric.

9. A method of producing a fibrin matrix into wet tissue, comprising the following stages: providing a solid component comprising fibrinogen; providing a solid component comprising thrombin; the provision of a liquid fibrin sealing composition, where the liquid fibrin sealing composition comprises at least two separate liquid component, wherein at least one separate component contains fibrinogen and at least a second separate component contains thrombin; applying an effective amount of the solid components of at least a portion of the wet surface; and applying an effective amount of a liquid fibrin sealing composition over at least part of the deposited solid component.

10. A method according to claim 9, in which a wet cloth does not contain fibrinogen.

11. A method according to claim 9, in which the liquid fibrin sealant composition is provided in solid form and regenerate before its application.

12. A method according to claim 9, in which the liquid fibrin sealant composition is provided in frozen form and thawed before its application.

13. A method according to claim 9, in which a solid component is a C�control in liquid form and allowed to dry before applying.

14. A method according to claim 9, in which the solid component provide frozen and dried before applying.

15. A method according to claim 9, in which the solid components are applied simultaneously or one after the other.

16. A method according to claim 9, in which the liquid components are applied simultaneously or one after the other.

17. A method according to claim 9, in which the solid components are presented in the form of a mixture.

18. A method according to claim 9, in which the fabric is enriched, has small amount, not enough or does not have blood vessels and (or) slowly releases the fluid or bleed.

19. Fibrin matrix prepared by the method according to claim 9.

20. Method of treatment or prevention of a defect on damp tissue in need of treatment of an object, comprising the following stages:
(a) providing a component comprising solid fibrinogen, providing a component comprising a solid thrombin; and providing a liquid fibrin sealing composition, where the liquid fibrin sealing composition comprises at least two separate liquid component, wherein at least one separate component contains fibrinogen and at least a second separate component contains thrombin;
(b) applying an effective amount of the solid components of p. a) at least a portion of the wet tissue; and
(c) on�Esenia effective quantity of a liquid fibrin sealing compositions of p. a) over at least part of the deposited solid component.

21. A method according to claim 20, in which a wet cloth is enriched, has small amount, not enough or does not have blood vessels and (or) slowly releases the fluid or bleed.

22. A method according to claim 20, wherein the defect is a leak in the tissue.

23. A method according to claim 22, in which the infiltrating substance is enriched, has small amount, not enough or does not have a plasma or blood components.

24. A method according to claim 20, in which the liquid fibrin sealant composition is provided in solid form and regenerate before its application.

25. A method according to claim 20, in which the liquid fibrin sealant composition is provided in frozen form and thawed before its application.

26. A method according to claim 20, in which a solid component is provided in liquid form and allowed to dry before applying.

27. A method according to claim 20, in which a solid component is provide frozen and dried before applying.

28. A method according to claim 20, in which the solid components are applied simultaneously or one after the other.

29. A method according to claim 20, in which the liquid components are applied simultaneously or one after the other.

30. A method according to claim 20, in which the solid components of p. (a) performance�cies in the form of a mixture.

31. A method according to claim 20, wherein the defect is a bracket or thread.

32. Set to be applied on a damp cloth the needy in the treatment of an object, comprising: (i) the container(s) contain(s) solid components comprising (a) fibrinogen component and (b) thrombin, wherein components (a) and (b) are in separate containers or in the same container in the form of a mixture; and (ii) at least two separate container, and at least one separate container contains a liquid, frozen or solid fibrinogen component and at least a second separate container contains a liquid, frozen or solid thrombin, and if at least two separate container of clause (ii) contain solid components, components (a) and (b) are in the same container in the form of a mixture.

33. The kit according to claim 32 for application to a wet surface.

34. The kit according to claim 32 to seal the leakage in tissues that are raw, have a small amount of, insufficient or do not have blood vessels, and (or) tissues, slowly releasing the fluid or necrobutcher.

35. Method of treatment or prevention of a defect on damp tissue in need of treatment with the introduction of a therapeutically effective amount of a fibrin matrix according to claim 19.



 

Same patents:

Biogel // 2503464

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to pharmaceutics. There are developed agent for forming a biogel, biogels for hemostasis, wound closure, tissue engineering and targeted drug delivery. The agent contains a soluble carrier whereon a number of fibrinogen-binding groups is immobilised. The biogel that contains fibrinogen molecules and a number of soluble carriers applicable for intravenous and/or local administration; each carrier comprises a number of fibrinogen-binding groups immobilised on the carrier, and each fibrinogen molecule is bound to at least two fibrinogen-binding groups so that the fibrinogen molecules occurs to be bound to each other through the carriers by non-covalent bonds between the fibrinogen-binding groups and the fibrinogen molecules. The biogel containing fibrin monomers and a number of soluble carriers applicable for intravenous and/or local administration, wherein each carrier comprises a number of fibrinogen-binding groups immobilised on the carrier, while the fibrin monomers are bound to each other through the carriers by non-covalent bonds between the fibrinogen-binding groups and the fibrinogen monomers. The biogel containing fibrin and a number of soluble carriers applicable for intravenous and/or local administration, wherein each carrier comprises a number of fibrinogen-binding groups immobilised on the carrier with the fibrin monomers in fibrin are covalently bound to each other by peptide bonds, and the fibrin monomers in fibrin are bound to each other through the carriers by non-covalent bonds between the fibrinogen-binding groups and the fibrinogen monomers. A method for forming the biogel involving a contact of the fibrinogen molecules with a number of soluble carriers. A method for hemostasis by topical administration of the biogel at a haemorrhage or a wound. Using a number of soluble carriers applicable for intravenous and/or local administration. A pharmaceutical formulation for topical administration containing the biogel, agent or a number of soluble carriers.

EFFECT: using the declared invention enables preparing the agents requiring no toxic reagents to be used, have a minimal risk of allergic reactions, and are easy to prepare and use.

2 tbl, 4 dwg, 5 ex

FIELD: medicine.

SUBSTANCE: invention relates to medicine. Described is method of obtaining polymeric cement of medical purpose, which includes mixing of prescription quantity of powdery polymer, liquid monomer, antibacterial component and mixture mixing, as antibacterial componen applied is solution, which contains bacteriophage or mixture of several different bacteriophages, which have lytic activity, in concentration not less than 106 PFU/ml, in amount 5.0-6.5 wt %, which is introduced into cement after mixture cooling to temperature 4-10°C. Mainly applied is/are bacteriophage(s) against bacteria of genus Pseudomonas, Staphylococcus, Bacillus. In particular case, applied is/are bacteriophage(s), individually selected for particular pathogen, for this purpose typing of causative agent of infection process of bacterial etiology agent in particular patient is preliminarily carried out.

EFFECT: method insures incretion quality of target product by incretion of antibacterial properties of polymeric cement.

4 cl, 1 dwg, 6 ex

The invention relates to medicine

FIELD: chemistry.

SUBSTANCE: invention relates to bioadhesives and provides an adhesive complex coacervate or a polyelectrolyte complex containing at least one polycation which contains a positively charged protein produced by P. Californica, or a polymer containing at least one fragment of formula I, at least one polyanion which contains a polyphosphate compound, polyphosphoserine or a polymer containing at least one fragment of formula II, and at least one polyvalent cation, wherein at least one polycation or polyanion is a synthetic compound and the polycation contains at least one cross-linking group and/or the polyanion contains at least one cross-linking group, where said cross-linking groups are capable of cross-linking with each other.

EFFECT: invention enables to obtain adhesive complex coacervates having low surface tension in water and high cohesion strength.

41 cl, 19 dwg, 1 tbl

FIELD: medicine.

SUBSTANCE: smaller pectoral muscle is transected at an attachment point to the 3-5th ribs. That is followed by separating, drying, and applying a latex fibrous adhesive in a layer of 0.1 mm. A muscle flap is laid on the subclavicular-axillary vessels. The flap is retained for 5-7 minutes. The flap is fixed with three interrupted absorbable sutures 3.0. An edge of the separated broadest muscle of the back is fixed to the serratus anterior and smaller pectoral muscle with single sutures.

EFFECT: reduced volume and length of lymphorrhoea, prevented lymphocele and oedema of an upper extremity following breast eradication or separate subclavian axillosubcapular lymphadenectomy.

3 ex

Medical glue // 2526188

FIELD: medicine.

SUBSTANCE: medical glue "Neosulphacrylate" contains, wt.p.: ethyl-α-cyanacrylate 76.5-85.0, 3-methacrylcarboxysulpholane 9.0-12.5, decylmetacrylate 6.0-11.0.

EFFECT: glue extends an assortment of compositions of the stated purpose, possesses improved organoleptic properties, increased adhesion and reduced neurotoxicity in tissue gluing, provides an increase of anti-inflammatory action with preservation of such physic-chemical characteristics as rupture strength and elasticity at the level of its prototype.

2 tbl, 6 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to a technology for medical adhesives for biological tissues. The preparation is applied locally on the skin for the operations in surgery and combustiology. The medical adhesion for the sutureless joint of biological tissues forming an adhesion layer by a polymerisation reaction in the presence of blood iron and hydrogen peroxide, contains acrylic acid and polyvinyl pyrrolidone that is a graft polymerisation matrix when forming the adhesion layer.

EFFECT: medical adhesion according to the invention is characterised by the high adhesion, antiseptic and detoxification properties.

FIELD: medicine, pharmaceutics.

SUBSTANCE: present invention aims at improved cement flour effective as a bone cement containing an organic ingredient consisting of one or more biologically acceptable and bioresorptive polymers, and an inorganic ingredient consisting of one or more calcium-phosphate compounds wherein the above calcium-phosphate compounds contains at least approximately 70% α-TCP. The biologically acceptable and bioresorptive polymers are specified in a group consisting of polysaccharides and salts thereof in the form of microparticles. The invention is related to the apatite calcium-phosphate cement (CPC) prepared by mixing the above cement flour with a liquid phase and curing. The invention also refers to using the injectable CPC for preparing the medical agent for treating bone defects as a tissue engineering frame, for making dental or bone implants consisting of a CPC cast.

EFFECT: preparing the improved cement flour.

19 cl, 6 tbl, 11 ex, 6 dwg

FIELD: medicine.

SUBSTANCE: invention refers to medicine. What is disclosed is a porous calcium phosphate hydraulic cement for bone tissue restoration, containing p-tricalcium phosphate powder, monocalcium phosphate monohydrate, and a tempering liquid representing 7-9% aqueous solution of citric acid; also, a cement powder comprises calcium carbonate granules in the following proportions (wt %): β-tricalcium phosphate - 67-75, monocalcium phosphate monohydrate -20-22, calcium carbonate granules - 3-13. The tempering liquid and cement powder are related as 0.75.

EFFECT: calcium phosphate cements are characterised by a combination of an ability to reaction hardening, an ability to be shaped, biocompatibility, the absence of toxic side effects, as well as a potential to be replaced by the newly formed bone tissue.

4 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to medicine, particularly surgical instruments, and may be used for adhesion of body soft tissues. A biological adhesive contains 3-methacryloxysulpholane, N-butyl-2-cyanacrylate and at least one stabiliser.

EFFECT: adhesive may be used for adhesion of body soft tissues.

7 cl, 1 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to chemical-pharmaceutical industry and represents method of preparing injected calcium-phocphatic bone cement in form of apatite, releasing gembisphosphonic compound, and claimed method includes addition of bisphosphonic acid or its salt or solid calcium precursor, modified by bisphosphonic acid or its salt, in solid phase of cement or in liquid phase of cement, and claimed solid calcium precursor is selected from calcium orthophosphates and non-phosphate calcium salts, such as CaCO3 or CaSO4.

EFFECT: invention ensures obtaining calcium-phosphatic bone cement, characterised by time of hardening, suitable for surgical application; which has strength for compression, close to strength for bone compression; which is resolvable for its replacement by new bone material; which ensures release of inhibitor of bone tissue resorption.

21 cl, 10 ex, 5 dwg

FIELD: medicine.

SUBSTANCE: compositions, methods and sets applied to sealing of injured tissues are described herein; the compositions are produced by combining of the first transversally cross-linked component with the second transversally cross-linked component to the effect of formation of the porous matrix with spans, and combining of the porous matrix with the hydrocarbon gel forming component applied to filling-in at least some of the spans.

EFFECT: compositions have minimal turgescing properties.

15 cl, 15 tbl, 14 dwg, 26 ex

FIELD: medicine.

SUBSTANCE: invention refers to medicine, and can be used in oncology (oncogynaecology, oncoproctology), abdominal surgeries, urology, otolaryngology, proctology (hemorrhoids, proctitis, anal fissures), gynaecology (erosion, colpitis, vulvovaginitis), and also active drainage in surgery. A bioactive-coated drainage system for treating abdominal diseases represents a hollow shape element in the form of a cylinder made of silicon elastomers. A cylinder exterior comprises a bioactive coating tissue made of a hydrophilic textile material coated with at least one layer of a complex bioactive substance and biocompatible biodegradable polymer. The bioactive coating is attached to the shape element with a medical-grade polymer adhesive, and at least one end of the bioactive coating is sutured.

EFFECT: invention is easy-to-manufacture, practically feasible, exhibits prolonged medical action and less traumatic.

11 cl, 6 dwg

FIELD: medicine.

SUBSTANCE: trombolytic is administered intranodularly into a thrombosed haemorrhoid. After the haemorrhoidal contents gets soft, it is punctured with removing lysed blood, and an anti-inflammatory glucocorticosteroid is administered into the haemorrhoid cavity, which is followed by a tight external perianal tamponade.

EFFECT: method enables providing the clinical effectiveness of acute haemorrhoid.

3 cl, 1 ex

FIELD: veterinary medicine.

SUBSTANCE: solution is administered intravenously to chinchilla male rabbits one hour prior to surgical interference. The solution is prepared as follows: sterile distilled water for injection is added to the lyophilised fibrin-monomer with urea, so that the resulting solution contains fibrin monomer at a concentration of 11 mg/ml and urea at a concentration of 150 mg/ml, and stirred until complete dissolution of the substance. At that the dose of fibrin-monomer is 0.25 mg/kg.

EFFECT: method is highly effective the prevention of bleeding caused by the use of streptokinase, prior to surgical interferences.

1 ex, 2 dwg

FIELD: veterinary medicine.

SUBSTANCE: solution is administered intravenously to chinchilla male rabbits one hour prior to surgical interference. The solution is prepared as follows: sterile distilled water for injections is added to lyophilised fibrin-monomer with urea, so that the resulting solution contains fibrin-monomer at a concentration of 11 mg/ml and the urea at a concentration of 150 mg/ml, and stirred until complete dissolution of the substance. The dose of fibrin-monomer is 0.25 mg/kg.

EFFECT: method is highly effective for prevention of bleeding caused by the use of dabigatran etexilate in the experiment.

2 dwg, 1 ex

FIELD: medicine.

SUBSTANCE: solution containing fibrin monomer in the concentration of 11 mg/ml and urea in the concentration of 150 mg/ml is introduced intravenously into male rabbits experimentally one hour before the surgical intervention. A dose of fibrin monomer is 0.25 mg/kg.

EFFECT: method possess the high effectiveness as both reduces the bleeding volume, but causes the severe haemodynamic disturbances and allergic reactions.

2 dwg

FIELD: chemistry.

SUBSTANCE: invention relates to a method of lyophilisation of a composition, which contains purified antithrombin III (AT III) and a crystallised substance, selected from alanine, mannitol, glycine or NaCl. The claimed method includes freezing the composition at a temperature from -52°C to -60°C for 6-15 hours, annealing the composition at -30°C for 1 hour, re-freezing the composition at a temperature from -52°C to -60°C for 2-15 hours at keeping the product temperature between -48°C and -52.7°C for 4-10 before lyophilisation and drying the composition with obtaining a lyophilised cake. The invention also relates to a pharmaceutical set, which contains the said lyophilised cake and a liquid reagent.

EFFECT: invention provides obtaining the lyophilised composition, containing AT III, which preserves its activity and stability.

14 cl, 24 dwg, 5 tbl, 2 ex

FIELD: medicine.

SUBSTANCE: invention refers to biotechnology, more specifically to modified von Willebrand factor (VWF), and can be used in medicine. A recombinant method is used to preparing modified VWF fused in C-terminal of its primary translation product with N-terminal of albumin by the linker SSGGSGGSGGSGGSGGSGGSGGSGGSGGSGS. The prepared modified VWF is used as a part of the pharmaceutical composition for treating or preventing coagulation failure.

EFFECT: invention enables preparing the modified VWF which maintains its ability to N-terminal dimerisation and C-terminal multimerisation with a prolonged half-period of functional blood plasma occurrence as compared to the half-period of functional VWF occurrence.

17 cl, 5 dwg, 4 tbl, 11 ex

FIELD: medicine.

SUBSTANCE: to correct haemostatic disorders in case of chronic calculous cholecystitis at the background of chronic hepatitis or cirrhosis of liver, preoperative correction of haemostasis with the preparation prothromplex 600, introduced intravenously at a rate not higher than 2 ml/min in a dose of 20 IU/kg of the patient's body weight, is carried out 3 days before endosurgical treatment. After that, on the first day after operation 20 IU/kg of prothromplex 600 and additionally 10 mg/kg of etamsylate and 25 mg/kg of aminomethyl are introduced intravenously. The preparations are introduced 1 time per day for 3 days with obligatory control of haemostasis indices.

EFFECT: method makes it possible to enhance the clinical effect due to an improvement of haemostasis system indices, connected with an increase of quantitative indices of a coagulation link, at the background of normalisation of the aggregation activity of platelets, as well as an arrest of hypocoagulation due to normalisation of the prothrombin complex, which makes it possible to twice reduce the treatment duration.

2 tbl, 1 ex

FIELD: chemistry.

SUBSTANCE: group of inventions relates to the field of biotechnology. Claimed is a method of purification of a factor, contributing to wound healing, which represents a hepatocyte growth factor (HGF). All stages of purification are carried out in the presence of antithrombin III (AT-III). In accordance with the claimed method carried out are: defrosting of the frozen HGF-containing source and removal of sediment from the defrosted source. After that, the obtained solution, which contains a supernatant and AT-III, is brought in contact with a carrier for affinity chromatography on an immobilised heparin. Then, the solution is separated from the carrier for affinity chromatography. The carrier is brought in contact with a desorption buffer with ionic strength sufficient for HGF desorption. The desorption buffer, containing HGF, AT-III and histidine-rich glycoprotein (HRGP) is collected. Also claimed are wound-healing compositions, which contain HGF, AT-III and/or HRGP, purified by the claimed method.

EFFECT: inventions make it possible to increase step-by-step output of the hepatocyte growth factor, with the hepatocyte growth factor being concentrated in eluate in the presence of AT-III.

26 cl, 2 tbl, 6 ex

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