WO2017117995A1 - Chitosan-containing fibrin glue and use method thereof - Google Patents

Abstract

A chitosan-containing fibrin glue and use method thereof. The chitosan-containing fibrin glue comprises first lyophilized powder and second lyophilized powder, wherein components of the first lyophilized powder comprise fibrinogen, factor XIII, and chitosan or its derivatives; components of the second lyophilized powder comprise thrombin and chitosan or its derivatives. In use, the two lyophilized powders are dissolved in solvents, and the solution mixture is sprayed on a wound surface to accelerate wound healing. By adding chitosan into the fibrin glue, the hemostatic effects of both fibrin and chitosan can be achieved, thereby making the stoppage of bleeding faster and achieving a better wound healing effect after use.

Description

Protein gum containing chitosan and use method thereof Technical field

The invention relates to the technical fields of biopharmaceuticals, biological products, biological materials and medical instruments, in particular to a protein gel containing chitosan, which can be used for hemostasis, adhesion and sealing in surgery.

Background technique

Rapid and complete hemostasis during surgery, reducing the amount of bleeding, and keeping the field clear, is one of the core operations of the basic operation of surgery. In various kinds of first-aid, such as frequent traffic accidents and accidents, it is also generally faced with problems such as rapid and effective hemostasis, alleviating patient suffering and saving people's lives.

Local hemostatic materials are widely used for surgical hemostasis, and their hemostatic effects have been fully affirmed in animal experiments and clinical practice. The use of topical hemostatic agents not only reduces the amount of bleeding, simplifies the operation, shortens the operation time, and sometimes promotes wound healing.

At present, commonly used local hemostatic drugs include fibrin glue, gelatin sponge, chitosan, oxidized cellulose, microfiber collagen, zeolite powder, thrombin and alginic acid fiber. Different types of local hemostatic drugs have different physical and chemical properties due to their different structures and compositions. Therefore, their mechanism of action and methods of use are not the same, and the effect of hemostasis is also very different.

The history of fibrin (original) as a medical adhesive dates back to the beginning of the last century. In 1909, Bergel reported that fibrin powder has a hemostatic function, and Grey immediately used it to stop bleeding in the liver and brain. In 1940, Young attempted to use the coagulation reaction to bind the severed peripheral nerves, and Tidrick and Cronkile used it to fix the transplanted skin. However, fibrin was not established until 1972 by Matras. For the first time, Matras extracted high-concentration human fibrinogen by cryoprecipitation, and high-concentration bovine thrombin and XIII coagulation factor were used as binders to achieve peripheral nerve anastomosis. Fibrinogen is hydrolyzed to fibrin monomer under the action of thrombin, and then cross-linked to form a solidizable fibrin solid, thereby adhering to the surface of the bleeding wound and functioning to stop bleeding.

Summary of the invention

The technical problem to be solved by the present invention is to provide a protein gum containing chitosan and a method for using the same, Fast wound healing.

In order to solve the above technical problem, the technical solution of the present invention is: a chitosan-containing protein gel comprising a first lyophilized powder and a second lyophilized powder, the components of the first lyophilized powder comprising: fibrin Original, XIII coagulation factor and chitosan or a derivative thereof; components of the second lyophilized powder include: thrombin and chitosan or a derivative thereof.

Chitosan has a good hemostatic effect. The hemostasis mechanism is to form red blood cell agglomeration through the combination of polycations carried by chitosan and anion on the surface of erythrocyte membrane, and activate platelet aggregation and activate thrombin to achieve rapid hemostasis. In addition, when the chitosan is in contact with the wound, it can also absorb a large amount of exudate from the wound surface and accelerate hemostasis. Chitosan has the effect of promoting wound healing, often accompanied by low levels of scar formation and good healing appearance. Cellular and molecular studies have shown that this phenomenon is likely to be related to the regulation of collagen synthesis and secretion in the wound by carboxymethyl chitosan. Chitosan can promote fibroblast proliferation, activate macrophages, increase the content of TGF-β1 and IL-6, and promote wound healing.

As a modification, the fibrinogen, XIII coagulation factor and thrombin are derived from a mammal.

In order to solve the above technical problem, the method of using the technical solution of the present invention: a method for using a chitosan-containing protein gel, comprising the following steps:

(1) dissolving the first lyophilized powder in the first solution, and dissolving the second lyophilized powder in the second solution;

(2) mixing and spraying the first solution and the second solution onto the wound;

(3) The mixture is used for hemostasis, adhesion and closure.

As an improvement, the content of chitosan or its derivative in the first solution is 0.1-20 mg/ml, and the content of chitosan or its derivative in the second solution is 0.1-20 mg/ml. .

As an improvement, the content of chitosan or its derivative in the first solution is 0.2 to 10 mg/ml, and the content of chitosan or its derivative in the second solution is 0.2 to 10 mg/ml. .

As an improvement, the content of chitosan or its derivative in the first solution is 0.5 to 5 mg/ml, and the content of chitosan or its derivative in the second solution is 0.5 to 5 mg/ml. .

The beneficial effects of the present invention compared to the prior art are:

After adding chitosan to the protein gel, the blood coagulation effect of both fibrin and chitosan can be exerted, so that the hemostasis speed is faster, and the wound healing effect after use is better.

detailed description

Example 1

The human fibrinogen intermediate was dissolved in an appropriate amount of sodium chloride (3 g/L, sodium citrate 1.5 g/L, sodium hydrogencarbonate 0.8 g/L, L-arginine hydrochloride 3.3 g/L). The protein concentration was 4%, and then chitosan was added to 5 g/L to adjust the pH to 7.2. After centrifugation and filtration, the mixture was lyophilized to obtain the first lyophilized powder. The human thrombin intermediate was formulated into 550 IU/ml, glycine 5 g/L, human albumin 2 g/L, chitosan 20 g/L, centrifuged, and lyophilized to obtain a second lyophilized powder.

After dissolving the two lyophilized powders with a solution, they were sprayed using a dispenser, and the adhesion strength was measured to be greater than 460 gf/cm ² according to the spray area of 10 cm ² /ml. On the liver surface of 20 rats, the wounds were 0.2 cm deep, 1.0 cm long and 0.5 cm wide. Ten of them were treated with protein gel without chitosan, and 10 gelatin with chitosan was added to stop bleeding. The proportion of rats that stopped bleeding in 1 minute was 50% for the former and 90% for the latter. Chitosan-containing protein gel has a better hemostatic effect.

Example 2

The porcine fibrinogen intermediate was dissolved in the protein with an appropriate amount of sodium chloride (3 g/L, sodium citrate 2 g/L, polysorbate 800.2 g/L, L-arginine hydrochloride 3.3 g/L). The concentration was 3%, and then chitosan was added to 8.5 g/L, and the pH was adjusted to 7.5. After centrifugation and filtration, the mixture was lyophilized to obtain the first lyophilized powder. The pig-derived thrombin intermediate was formulated into 550 IU/ml, glycine 5 g/L, human albumin 2 g/L, chitosan 10 g/L, centrifuged, and lyophilized to obtain a second lyophilized powder.

After dissolving the two lyophilized powders with a solution, they were sprayed using a dispenser, and the adhesion strength was measured to be greater than 250 gf/cm ² according to the spray area of 10 cm ² /ml.

Example 3

The porcine fibrinogen intermediate was dissolved in the protein with an appropriate amount of sodium chloride (2 g/L, sodium citrate 2 g/L, polysorbate 800.2 g/L, L-arginine hydrochloride 3.3 g/L). The concentration was 3.5%, and then hydroxybutyl chitosan was added to 2.5 g/L to adjust the pH to 7.5. After centrifugation and filtration, the mixture was lyophilized to obtain the first lyophilized powder. The pig-derived thrombin intermediate was formulated into 550 IU/ml, glycine 15 g/L, human albumin 2 g/L, chitosan 1 g/L, centrifuged, and lyophilized to obtain a second lyophilized powder.

After dissolving the two lyophilized powders with a dissolving solution, they were sprayed into a dispenser to measure the adhesive strength of more than 250 gf/cm ² and an elastic height of 6.2 cm according to a spray area of 10 cm ² /ml.

Claims (6)

A protein gum containing chitosan, comprising: a first lyophilized powder and a second lyophilized powder, the components of the first lyophilized powder comprising: fibrinogen, XIII coagulation factor and chitosan Or a derivative thereof; The components of the second lyophilized powder include: thrombin and chitosan or a derivative thereof. A chitosan-containing protein gel according to claim 1, wherein the fibrinogen, XIII coagulation factor and thrombin are derived from a mammal. The method for using a chitosan-containing protein gel according to claim 1, comprising the steps of: (1) dissolving the first lyophilized powder in the first solution, and dissolving the second lyophilized powder in the second solution; (2) mixing and spraying the first solution and the second solution onto the wound; (3) The mixture is used for hemostasis, adhesion and closure. The method for using a chitosan-containing protein gel according to claim 3, wherein the content of chitosan or a derivative thereof in the first solution is 0.1 to 20 mg/ml, The content of chitosan or its derivative in the second solution is 0.1 to 20 mg/ml. The method for using a chitosan-containing protein gel according to claim 3, wherein the content of chitosan or a derivative thereof in the first solution is 0.2 to 10 mg/ml, In the second solution, the content of chitosan or a derivative thereof is 0.2 to 10 mg/ml. The method for using a chitosan-containing protein gel according to claim 3, wherein the content of chitosan or a derivative thereof in the first solution is 0.5 to 5 mg/ml, In the second solution, the content of chitosan or a derivative thereof is 0.5 to 5 mg/ml.