Biological adhesive glue

 

(57) Abstract:

The invention relates to the field of medicine and relates to adhesive compositions to stop bleeding on the basis of factor VIII. The essence of the invention lies in the fact that the proposed composition contains cryoprecipitate obtained by refrigerated centrifugation at a speed of 2500-3000 rpm for 1-1 .5 h, the dose of which contains factor VIII activity from 150 to 220 units , thrombin activity from 160 to 700 units in 7-15 ml of isotonic solution of sodium chloride and 3-10 ml of 10% calcium chloride or calcium gluconate. The technical result is the creation of adhesive compositions, the application of which allows you to stop bleeding not only from the capillaries, but vessels of small and medium caliber, and extensive tissue surfaces.

The invention relates to the field of medicine and relates to adhesive compositions based on fractional plasma protein (factor VIII) to stop the bleeding and restore the biological integrity of tissues (reconstructive and trauma surgery, traumatology, emergency medicine, obstetrics, gynecology, ophthalmology, otolaryngology (ENT).

Adhesive compositions based on biological substances, ear surgical field;

- slow tissue regeneration;

- toxic effect on the surrounding tissue.

Fibrin glue is not only stops the bleeding but also promotes tissue regeneration due to the unique stimulating properties of fibrinogen, a multifunctional protein, which serves as a matrix for the regulation of fibrinolysis, attachment of the cells in the repair process, activates the production of leukocyte germ angiogenic factors.

Known fibrin-containing adhesives, manufactured by companies like drugs Beriplast (Germany), Tissucol (Austria) [1], Biocol (France) [2].

They are similar in composition. For example, glue Beriplast consists of: 1) lyophilized fibrinogen human blood 115-232 mg; 2) thrombin (bovine) 400-500 units; 3) placental protein fraction containing 40-80 units of factor XIII; 4) solution of Aprotinin - up to 1000 KE; 5) solution of calcium chloride, 40 mmol/l 25 ml.

The development of such domestic product is complex due to the lack of technology for factor XIII, no drug Aprotinin and the high cost of the final product.

The closest in technical essence and the achieved result is fibrin glue, developed in the PCR CCH 7 Moscow [3]. His Doc is t in the usual way, to it was added a solution of PEG - 6000 and 1 ml of aminocaproic acid (sterile solutions seamed in sterile pharmacy ware, pull through a puncture of the tube by vacuum aspiration).

2). The vial is shaken, defend for 30 minutes and centrifuged for 15 minutes at 4oC.

3). The supernatant is sucked off, and the residue is dissolved in sodium citrate and collect in a sterile rolled penicillin vial (about 4 ml of the fibrinogen content of the last 40-60 g/l) and additionally paraffinicity. For polymerization on the bleeding surface to it add 500 units of thrombin in 4 ml of 10% solution l2.

The disadvantages of the adhesive obtained in this way are:

- the possibility of contamination of the plasma by viruses of different origin, including AIDS and hepatitis C;

- the complexity of the composition of the first component adhesive consisting of cryoplasty, PEG - 6000 and - aminocaproic acid;

- the use of expensive sodium citrate for pH regulation;

- narrow the scope of its application: to stop bleeding only from the capillaries;

- receiving adhesive directly on the wound surface, alternately introducing the components of the adhesive composition is LASS="ptx2">

The present invention is the creation of a fibrin-containing safe glue based keratinizing cryoprecipitate, which eliminates viral contamination. Glue produced from domestic raw materials and it is designed to stop bleeding from parenchymatous, hollow organs, extensive bleeding tissue surfaces and vessels of small and medium caliber, to commit their own biological tissues (autotransplants) and to improve reparative processes in the application of the adhesive mass.

The solution of the problem is the creation of a biological adhesive glue (LHC), including cryoprecipitate, in the amount of 1 dose containing factor VIII activity from 150 to 220 units, thrombin activity from 160 to 700 units in isotonic sodium chloride (7-15 ml) and calcium chloride or calcium gluconate (3-10 ml).

Preferably the TANK contains: thrombin activity from 240 to 560 units

Preferably the TANK contains factor VIII activity from 160 to 200 units

The advantages of the present invention:

1. Use keratinizing cryoprecipitate, which eliminates viral contamination.

2. Ease of access (the user.

4. Economic benefit.

5. Use to stop bleeding not only from the capillaries, but also vessels of small and medium caliber, and extensive tissue bleeding surfaces.

6. Reliable acceleration and stimulation of tissue regeneration.

7. The possibility of applying for the manufacture of biological prostheses, since the adhesive composition "in vitro" is able to continuously retain a given shape and fully biologically compatible with body tissues.

8. The ability to maintain in vivo in humans adhesive properties within 5-7 days, forming on the damaged surface of the fibrin film, followed by granulation and epithelialization.

The invention is illustrated in the following examples.

EXAMPLE 1.

To obtain cryoprecipitate karantinirovaniya (within 6 months) fresh frozen plasma (donor or AUTOLOGOUS) is thawed at room temperature to form a "wet sugar", in the amount of 300 ml, then plasma substance centrifuged in a refrigerated centrifuge for 1-1 .5 hours at a speed of 2500-3000 rpm the Supernatant part of the plasma (supernatant) and the average fraction of plasma (concentrate NAT is ATiM sediment (cryoprecipitate) leave it in the main bag and freeze again. Received cryoprecipitate contains factor VIII activity from 150 to 220 units Store cryoprecipitate at a temperature of from 28 to 73oC.

EXAMPLE 2.

To obtain biological adhesive glue:

1. One dose of cryoprecipitate containing 160 units of antihemophilic globulin (factor VIII), thawed at room temperature under the conditions of shaking until a homogeneous mass (to avoid the formation of lumps).

2. Thrombin, taken from activity in the amount of 240 units, dissolved in equal amounts in the two test tubes containing 10 ml of isotonic NaCl solution and 5 ml of 10% l2respectively, then mixed.

3. Cryoprecipitate and thrombin solution are mixed immediately prior to use.

EXAMPLE 3. To obtain biological adhesive glue:

1. One dose of cryoprecipitate containing 160 units of antihemophilic globulin (factor VIII), thawed at room temperature under the conditions of shaking until a homogeneous mass (to avoid the formation of lumps).

2. Thrombin, taken from activity a quantity of 700 units, dissolved in equal amounts in the two test tubes containing 10 ml of isotonic NaCl solution and 5 ml of 10% gluco is directly before use.

EXAMPLE 4.

Differs from example 2 in that cryoprecipitate is taken from activity antihemophilic globulin (factor VIII) 180 units

EXAMPLE 5.

Patient R. , 75 years enrolled in the MSC 1 ZIL with clinical signs of gastrointestinal bleeding and low hemodynamic indices in three days from the onset of the disease. When emergency gastroduodenoscopy revealed extensive (up to 3.5-4 cm) ulcer of the gastric body with blood clot under which actively received fresh blood. The situation is regarded as an ongoing gastrointestinal bleeding (Forrest 16). Due to the fact that the ulcer was located on the lesser curvature of the stomach, it has not been technically possible to perform coagulation and injection methods of stopping bleeding, and the risk of emergency surgery was very high in patients of advanced age with severe concomitant somatic pathology. It was decided to treat the surface of ulcers Tank. Two component adhesive were mixed ex tempora and applicaany through the catheter on the ulcer surface. Applied composition, filling ulcerative defect, formed matte gel fibrin film, intimately adjacent to the mucosa. Already during manimala, where she conducted infusion-transfusion correction of blood loss circulatory disorders in combination with antiulcer drug treatment. When the control gastroduodenoscopy 24 hours bleeding is not identified, ulcerative defect was still filled with fibrin film. After stabilization of the patient was transferred for further treatment in the surgery Department, where she continued combination therapy. After three weeks at endoscopy ulcer defect is represented by a red scar. In a satisfactory condition the patient was discharged home.

EXAMPLE 6.

Patient K., aged 65, was admitted to the vascular Department of MSU 1 ZIL with a diagnosis of atherosclerosis of the lower extremities, Leriche syndrome, chronic arterial insufficiency stage 2B. During angiographic examination, it was found 50% stenotic disease aortic aneurysm and occlusion of the ileo-femoral segment on both sides.

In order to restore blood flow in the lower extremities, the patient was shown reconstructive surgery of the aortic-bifemoral bypass. In the process of operation was used vysokoporodny Dacron prosthesis "North". To minimize blood loss n in the TANK before implantation for 5 minutes. According to our data, the estimated blood loss without the use of the TANK enabling blood flow ranged from 600 to 900 ml, which subsequently caused various complications. When using TANK hemorrhage along the line of anastomosis and through the pores of the prosthesis was not observed. The patient was discharged 10 days after surgery in satisfactory condition.

Sources of information

1. Gastpar h Local application of human fibrin seal (Tissucol/Tissel) in tonsillectomies and adenectorrues in patients with bleeding disorders. Ten years clinical experience // Fibrin sealant in operative medicine / Shlag G. Readi H (Eds). - Springer Verlag: Berlin - Heidelberg, 1986. - P. 128-131.

2. Kraum H. B., Nathan R. C, MacKable J. R. Clinical use of noneautologous fibrin glue // Amer. Surg. - 1988. - Vol. 54, 9. - P. 570-573.

3. So Century. Horobryh, A. N. Antonov, G. K. Orlov, I. M. Soloviev. Receiving fibrin glue in clinical conditions //M - 1994. Scientific achievements into practice, vol. 7, pp. 133-136.

Adhesive composition to stop the bleeding and restore the biological integrity of tissues, characterized in that it contains cryoprecipitate obtained by refrigerated centrifugation at a speed of 2500-3000 rpm for 1-1 .5 h, the dose of which contains factor VIII activity from 150 to 220 units, thrombin activity from 160 to 700 units in 7-15 ml of isotonic solution of sodium chloride and 3-10 ml of 10%

 

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2 tbl, 3 ex

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