Shapeable hemostatic putty as well as preparation method and application thereof

Abstract

The invention provides moldable hemostatic putty as well as a preparation method and application thereof. The hemostatic putty comprises hemostatic powder and a solvent, wherein the hemostatic powder comprises gelatin particles and a polysaccharide compound containing an aldehyde group or a succinimide ester group, and the gelatin particles are cross-linked gelatin particles coated with an active substance. The hemostatic putty disclosed by the invention has good flexibility, moldability, adhesion plugging property and degradability, can be kneaded and molded into any shape to be filled at a bleeding part of a bone wound surface, and forms adhesion plugging on the bleeding part, so that a good plugging hemostatic effect is realized. Active substances are wrapped in a network structure of crosslinked gelatin to form a stable core-shell structure, so that the active substances can be prevented from influencing the adhesion plugging property and hemostatic effect of a final product, and the active substances can be slowly released along with degradation of the gelatin, so that generation of new callus is induced or accelerated, and bone healing is accelerated.

Description

Technical Field

[0001] The invention relates to a plastic hemostatic putty and a preparation method and application thereof, belonging to the field of medical materials. Background Art

[0002] Due to its loose structure and rich blood supply, cancellous bone has poor vascular contractility in the tissue of the bone wound, platelets are difficult to aggregate, blood clots are difficult to adhere to the bone wound, and it is difficult to stop bleeding by blood vessel self-contraction. At the same time, it is difficult to effectively stop bleeding by conventional methods such as electrocoagulation, ligation, clamping, and gauze compression during surgery. The ideal cancellous bone hemostatic material should have the following characteristics: ① It can be degraded and absorbed in the human body and does not hinder bone healing; ② It has good biocompatibility, no toxic side effects, and small tissue reaction; ③ It has good adhesion, can firmly adhere to the bone wound, and stop bleeding quickly and effectively; ④ It has good plasticity and can be used for bone wounds of various shapes.

[0003] Bone wax is commonly used in clinical practice to stop bleeding from cancellous bone wounds. Its main ingredients are beeswax, vaseline, etc. It becomes plastic after being rubbed soft by hand, and stops bleeding by physically blocking capillary bleeding from bone wounds. However, bone wax has poor biocompatibility and is difficult to be degraded and absorbed by the human body after use. It will remain in the body as a foreign body for a long time, which will hinder bone healing and increase the risk of postoperative infection.

[0004] In the current development of existing bone hemostatic materials, domestic and foreign scholars have tried some solutions to replace bone wax hemostasis, such as using oxidized cellulose, porous sponges, polysaccharides and other alternative bone wax materials. Although these materials have good biocompatibility, most of them are powdery or gelatinous, have poor adhesion to the wound surface, are difficult to operate during surgery, and are easily washed away by the blood flow.

[0005] Currently, active groups that can react with groups on the surface of blood and tissues are also used to improve the tissue adhesion of the product. Commonly used active groups such as succinimidyl ester groups, iminoester groups, aldehyde groups, etc. are usually grafted onto multi-arm polyethylene glycol (PEG) to react with other components to form a gel, which is then adhered and blocked on the tissue to achieve a good hemostatic effect. Although this type of product can improve the adhesion to tissues, the active groups are located at the end of the multi-arm structure of the PEG-based material. The multi-arm PEG material has a smaller steric hindrance effect due to its linear stretch structure, which leads to a higher cross-linking reaction ratio of the active groups, making the final colloid brittle and poor in plasticity, resulting in the inability to perform secondary processing during use to meet the use requirements caused by differences in the shape of the bone wound surface. Summary of the invention

[0006] Problem that the invention aims to solve

[0007] In view of the technical problems existing in the prior art, the purpose of the present invention is to provide a hemostatic putty, which has good flexibility, plasticity, adhesion and blocking properties, and degradability, and can be kneaded and molded into any shape to fill the bleeding site of the bone wound, and form adhesion and blocking on the bleeding site to achieve a good blocking and hemostatic effect. In addition, by encapsulating the active substance in the network structure of cross-linked gelatin to form a stable core-shell structure, the active substance can be prevented from affecting the adhesion and blocking properties and hemostatic effect of the final product, and the active substance can be slowly released with the degradation of gelatin, inducing or accelerating the formation of new callus, and accelerating bone healing.

[0008] The hemostatic putty of the present invention comprises hemostatic powder and solvent. The hemostatic powder is in the form of dry powder, which is convenient for storage before use and has a stable product state. Before use, the hemostatic powder is mixed with the solvent to form a hemostatic putty with plasticity, which can be filled in bone wounds of any shape, has no special requirements on the position and shape of the wound, and is easy to use.

[0009] The invention also provides a method for preparing the hemostatic putty. The preparation method is simple and easy to implement, the raw materials are easy to obtain, and the method is suitable for mass production.

[0010] Solutions for solving problems

[0011] The present invention provides a plastic hemostatic putty, which comprises hemostatic powder and a solvent; wherein the hemostatic powder comprises gelatin particles and a polysaccharide compound containing an aldehyde group or a succinimide ester group, and the gelatin particles are cross-linked gelatin particles encapsulating an active substance;

[0012] Preferably, the particle size of the gelatin particles is below 500 μm.

[0013] Furthermore, in the polysaccharide compound, the degree of substitution of aldehyde groups and / or succinimide ester groups is greater than or equal to 10%;

[0014] Furthermore, the mass ratio of the gelatin particles to the polysaccharide compound containing an aldehyde group or a succinimide ester group is (1:2) to (2:1).

[0015] Furthermore, the particle size of the active substance is less than 150 μm;

[0016] Preferably, the active substance comprises one or a combination of two or more of bioactive glass material, degradable metal, degradable metal oxide, degradable metal salt, hydroxyapatite, tricalcium phosphate, degradable metal alloy, and recombinant human bone morphogenetic protein.

[0017] Furthermore, the mass ratio of the hemostatic powder to the solvent is 1:1 to 1:5;

[0018] Preferably, the solvent includes one or a combination of two or more of water, physiological saline, buffer solution, and water for injection; more preferably, a drug is added to the solvent, and the drug preferably includes one or a combination of two or more of thrombin, fibrin, and growth factor.

[0019] Furthermore, the adhesion and sealing strength of the hemostatic putty is greater than 10 kPa; and / or the hemostatic time of the hemostatic putty is less than 8 s.

[0020] Furthermore, the preparation process of the gelatin particles includes:

[0021] mixing gelatin with the active substance and then chemically cross-linking the gelatin;

[0022] The cross-linked gelatin mixed with the active substance is freeze-dried to obtain a porous sponge;

[0023] The porous sponge is subjected to physical cross-linking and pulverization treatments.

[0024] Further, the active substance is added to the gelatin solution to obtain a mixture, and a cross-linking agent is added to chemically cross-link the gelatin to obtain cross-linked gelatin mixed with the active substance;

[0025] Preferably, based on the total mass of the mixture as 100%, the content of gelatin is 5-20%, and the content of the active substance is 0.5-10%;

[0026] Preferably, the cross-linking agent comprises carbodiimide and / or transglutaminase, and the mass ratio of the cross-linking agent to the gelatin is 1:25-1000.

[0027] Furthermore, during the preparation process, the porous sponge is first crushed and then physically cross-linked, or the porous sponge is first physically cross-linked and then crushed, or the porous sponge is sequentially physically cross-linked, crushed, and then physically cross-linked;

[0028] Preferably, the temperature of the physical crosslinking is 140-170° C., and the total time of the physical crosslinking is 4-10 h.

[0029] The present invention also provides a method for preparing the hemostatic putty according to the present invention, the preparation method comprising the step of mixing the hemostatic powder and a solvent to form a paste.

[0030] The present invention also provides a kit for preparing the hemostatic putty of the present invention, the kit comprising hemostatic powder and a solvent, and the hemostatic powder and the solvent are stored separately;

[0031] Preferably, in the hemostatic powder, the gelatin particles and the polysaccharide compound containing an aldehyde group or a succinimide ester group are stored separately.

[0032] The present invention also provides an application of the above-mentioned hemostatic putty in preparing a bone wound hemostatic product; the bone wound hemostasis is preferably cancellous bone wound hemostasis.

[0033] Effects of the Invention

[0034] The hemostatic putty of the present invention has good flexibility, plasticity, adhesion and blocking properties, and degradability, and can be kneaded and molded into any shape to fill the bleeding site of the bone wound, and form adhesion and blocking on the bleeding site to achieve a good blocking and hemostatic effect. In addition, by encapsulating the active substance in the network structure of the cross-linked gelatin to form a stable core-shell structure, the active substance can be prevented from affecting the adhesion and blocking properties and hemostatic effect of the final product, and the active substance can be slowly released as the gelatin degrades, inducing or accelerating the formation of new bone callus, and accelerating bone healing.

[0035] In addition, the hemostatic putty of the present invention has a hemostatic powder in the form of a dry powder, which is easy to store before use and has a stable product state. Before use, the hemostatic powder is mixed with a solvent to form a hemostatic putty with plasticity, which can be filled in bone wounds of any shape, has no special requirements on the position and shape of the wound, and is easy to use.

[0036] Furthermore, the preparation method of the hemostatic putty of the present invention is simple and easy, the raw materials are easy to obtain, and it is suitable for mass production. BRIEF DESCRIPTION OF THE DRAWINGS

[0037] Figure 1 The product photo of the shapeable hemostatic putty of Example 1 and the shapeable effect diagram are shown;

[0038] Figure 2 A schematic diagram of a test device for plugging adhesion strength is shown;

[0039] Figure 3 Schematic diagram of the testing device showing the hemostasis time. DETAILED DESCRIPTION

[0040] Various exemplary embodiments, features and aspects of the present invention will be described in detail below. The word "exemplary" used here means "used as an example, embodiment or illustrative". Any embodiment described here as "exemplary" is not necessarily interpreted as being superior or better than other embodiments.

[0041] In addition, in order to better illustrate the present invention, numerous specific details are provided in the following specific embodiments. It should be understood by those skilled in the art that the present invention can be implemented without certain specific details. In other examples, methods, means, equipment and steps well known to those skilled in the art are not described in detail in order to highlight the subject matter of the present invention.

[0042] Unless otherwise stated, the units used in this specification are all international standard units, and the numerical values ​​and numerical ranges appearing in the present invention should be understood to include the inevitable systematic errors in industrial production.

[0043] In this specification, the word "may" means both performing a certain process and not performing a certain process.

[0044] In this specification, the references to "some specific / preferred embodiments", "other specific / preferred embodiments", "embodiments", etc., mean that the specific elements (e.g., features, structures, properties and / or characteristics) described in connection with the embodiments are included in at least one embodiment described herein, and may or may not exist in other embodiments. In addition, it should be understood that the elements may be combined in various embodiments in any suitable manner.

[0045] In this specification, the numerical range expressed using "a numerical value A to a numerical value B" means a range including the endpoints numerical values ​​A and B.

[0046] <First aspect>

[0047] The first aspect of the present invention provides a shapeable hemostatic putty, wherein the hemostatic putty comprises hemostatic powder and a solvent; wherein,

[0048] The hemostatic powder comprises gelatin particles and a polysaccharide compound containing an aldehyde group or a succinimide ester group, and the gelatin particles are cross-linked gelatin particles encapsulating active substances.

[0049] The succinimide ester group of the present invention may be optionally substituted, for example, substituted by a hydroxyl group, such as, for example, N-hydroxysuccinimide ester group.

[0050] The hemostatic putty of the present invention has good flexibility, plasticity, adhesion and blocking properties and degradability, and can be kneaded and molded into any shape to fill the bleeding part of the bone wound surface; after filling the wound surface, it can quickly absorb tissue fluid or blood, concentrate blood cells, coagulation factors, etc., and promote blood coagulation. At the same time, the aldehyde group or succinimide ester group of the polysaccharide compound in the hemostatic putty reacts with gelatin and amino groups on the tissue to form a gel, and can form adhesion and blocking on the bleeding part without pressing, slowing down bleeding, preventing blood from continuously flowing out of the bleeding hole, and achieving a good blocking and hemostasis effect.

[0051] In addition, the hemostatic putty of the present invention comprises hemostatic powder and a solvent. The hemostatic powder is in the form of a dry powder, which is convenient for storage before use and has a stable product state. The hemostatic putty is mixed with the solvent before use to form a plastic hemostatic putty, which can be filled in bone wounds of any shape, has no special requirements on the position and shape of the wound, and is easy to use.

[0052] Specifically, in the present invention, the adhesion and sealing strength of the hemostatic putty is greater than 10 kPa; and / or the hemostatic time of the hemostatic putty is less than 8 s.

[0053] Gelatin Granules

[0054] The hemostatic powder of the present invention comprises coated gelatin particles and a polysaccharide compound containing an aldehyde group or a succinimide ester group, wherein the gelatin particles are cross-linked gelatin particles coated with an active substance; specifically, the particle size of the gelatin particles is less than 500 μm. The particle size of the gelatin particles is mainly to match the pore size of cancellous bone.

[0055] In the present invention, the particle size of the active substance is less than 150 μm. Specifically, the active substance includes one or a combination of two or more of bioactive glass materials, degradable metals, degradable metal oxides, degradable metal salts, hydroxyapatite, tricalcium phosphate, degradable metal alloys, and recombinant human bone morphogenetic protein. Among them, the degradable metal can be magnesium or zinc; the degradable metal salt can be a degradable metal chloride, such as zinc chloride, magnesium oxide, etc. In the gelatin particles, the mass ratio of the active substance to the gelatin is 1: (0.5-5).

[0056] The preparation process of the gelatin particles comprises:

[0057] mixing gelatin with the active substance and then chemically cross-linking the gelatin;

[0058] The cross-linked gelatin mixed with the active substance is freeze-dried to obtain a porous sponge;

[0059] The porous sponge is subjected to physical cross-linking and pulverization treatments.

[0060] In some specific embodiments, after the active substance is added to the gelatin solution to obtain a mixture, a cross-linking agent is added to chemically cross-link the gelatin to obtain cross-linked gelatin mixed with the active substance.

[0061] Specifically, gelatin is dissolved in a solvent, and then the active substance is added to the gelatin solution, and stirred at high speed to disperse the active substance in the gelatin solution system, thereby obtaining a mixture. Preferably, based on the total mass of the mixture as 100%, the content of the gelatin is 5-20%, and the content of the active substance is 0.5-10%.

[0062] In some specific embodiments, the crosslinking agent is added during mechanical stirring of the mixture. Specifically, the stirring speed can be 100 to 2000 r / min, preferably 500 to 1000 r / min. Preferably, the crosslinking agent is added in the form of a crosslinking agent solution, specifically, the addition speed of the crosslinking agent is 0.1 to 1 mL / s, preferably 0.2 to 0.5 mL / s. In the crosslinking agent solution, the mass concentration of the crosslinking agent is 0.05 to 2%.

[0063] In the present invention, the cross-linking agent includes carbodiimide (EDC) and / or transglutaminase (TGase), and the mass ratio of the cross-linking agent to the gelatin is 1:25 to 1000. By chemically cross-linking gelatin, its cross-linking degree can be increased, the degradation time can be prolonged, and the stability of the composite structure of gelatin particles can be improved.

[0064] Furthermore, the cross-linked gelatin mixed with the active substance is freeze-dried to remove water, thereby obtaining a porous sponge. Next, the porous sponge is physically cross-linked and pulverized to obtain gelatin particles encapsulating the active substance.

[0065] In some specific embodiments, the porous sponge can be first crushed and then physically cross-linked, or first physically cross-linked and then crushed, or the porous sponge can be physically cross-linked, crushed, and then physically cross-linked in sequence.

[0066] In the present invention, the pulverization may be performed by mechanical pulverization.

[0067] Preferably, the physical crosslinking includes high temperature crosslinking. Specifically, the temperature of the physical crosslinking is 140-170° C., and the total time of the physical crosslinking is 4-10 hours.

[0068] Furthermore, before freeze-drying, the step of removing the cross-linking agent is also included. Specifically, the cross-linked gelatin mixed with active substances is soaked in purified water to fully swell, divided into small pieces and crushed into particles, and then placed in a dialysis bag for extraction and dialysis for 48 to 72 hours to remove the cross-linking agent. Wherein, the pulverization is performed by a water-phase material pulverizer. The present invention uses a cross-linking agent EDC or TG enzyme that does not participate in the reaction, and the cross-linked gelatin is fully swollen with water after the cross-linking treatment, and then crushed into micron-sized particles, and then the cross-linking agent can be well removed by extraction and dialysis treatment, thereby achieving a good purification effect and effectively avoiding the residue and toxicity of irritating substances. Because the content of the cross-linking agent used in the present invention is relatively low, the step of removing the cross-linking agent may not be included.

[0069] In the present invention, the active substance is encapsulated in the cross-linked network structure of gelatin by combining chemical cross-linking and physical cross-linking to form a stable core-shell structure, which can prevent the active substance from affecting the cross-linking reaction between gelatin and polysaccharide compounds and the reaction between gelatin, polysaccharide compounds and tissues, thereby affecting the adhesion, blocking and hemostatic effects of the final product. In addition, the active substance can be slowly released as the gelatin degrades, which can induce or accelerate the formation of new callus and accelerate bone healing.

[0070] Polysaccharide compounds

[0071] In the present invention, the polysaccharide compound contains an aldehyde group or a succinimide ester group. After the hemostatic putty of the present invention is filled on the wound surface, it can quickly absorb tissue fluid or blood. At the same time, the aldehyde group or succinimide ester group of the polysaccharide compound can react with the amino group of gelatin and tissue to form a gel, which can form an adhesion blockage on the bleeding site without pressing, slowing down bleeding, preventing blood from continuously flowing out of the bleeding site, and achieving a good blocking and hemostasis effect.

[0072] In the present invention, the inventors of the present invention have found that, based on the molecular chain structure and the repeating unit structure, the reaction efficiency of the active functional groups in the aldehyde- or succinimidyl-esterified polysaccharide compounds is lower than that in the aldehyde- or succinimidyl-esterified polyethylene glycol substances, so that the reaction ratio of the active groups in the polysaccharide compounds is lower than that of the polyethylene glycol substances. Therefore, the gel formed by the final cross-linking of the aldehyde- or succinimidyl-esterified polysaccharide compounds and gelatin has good flexibility and plasticity.

[0073] Preferably, in the polysaccharide, the degree of substitution of aldehyde and / or succinimide ester is greater than or equal to 10%; in the present invention, the higher the degree of substitution of aldehyde and / or succinimide ester, the better, which can improve the adhesion of the hemostatic putty; when the degree of substitution of aldehyde and / or succinimide ester is low, it is more difficult to form a plastic putty. The polysaccharide compound includes one or a combination of two or more of chitosan, chitin, sodium alginate, and carboxycellulose; preferably, the polysaccharide compound is hyaluronic acid.

[0074] In some specific embodiments, the mass ratio of the gelatin particles to the polysaccharide compound containing an aldehyde group or a succinimide ester group is (1:2) to (2:1). When the mass ratio of the gelatin particles to the polysaccharide compound containing an aldehyde group or a succinimide ester group is (1:2) to (2:1), a hemostatic putty with good flexibility, plasticity, and adhesion and sealing properties can be formed.

[0075] Solvents

[0076] In the present invention, the mass ratio of the hemostatic powder to the solvent is 1:1 to 1:5. Preferably, the solvent includes one or a combination of two or more of water, physiological saline, buffer solution, and water for injection; more preferably, a drug is added to the solvent, and the drug preferably includes one or a combination of two or more of thrombin, fibrin, and growth factor. When the mass ratio of the hemostatic powder to the solvent is 1:1 to 1:5, it is easier to form a plastic hemostatic putty.

[0077] Furthermore, drugs may be added to the solvent, and the drugs preferably include one or a combination of two or more of thrombin, fibrin, and growth factors.

[0078] <Second Aspect>

[0079] A second aspect of the present invention provides a method for preparing the hemostatic putty according to the first aspect of the present invention, the preparation method comprising the step of mixing hemostatic powder and a solvent to form a paste.

[0080] Specifically, before use, the hemostatic powder and the solvent are mixed and quickly stirred to form a paste, namely the hemostatic putty.

[0081] The hemostatic putty of the present invention comprises hemostatic powder and solvent. The hemostatic powder is in the form of dry powder, which is convenient for storage before use and has a stable product state. The hemostatic putty is mixed with the solvent before use to form a plastic hemostatic putty. The putty has no special requirements on the position and shape of the wound surface and is easy to use. The putty can be kneaded and molded into any shape and filled in the bleeding part of the bone wound surface, and the bleeding part can be adhered and blocked to achieve a good blocking and hemostatic effect. The hemostatic putty is degradable and has good biocompatibility. Other ingredients such as drugs can also be added to the solvent, providing a new way for drug delivery, wound treatment, etc.

[0082] <Third Aspect>

[0083] The third aspect of the present invention provides a kit for preparing the hemostatic putty according to the first aspect of the present invention, the kit comprising a hemostatic powder and a solvent; wherein the hemostatic powder comprises gelatin particles and a polysaccharide compound containing an aldehyde group or a succinimide ester group; and the hemostatic powder and the solvent are stored separately;

[0084] In some specific embodiments, in the hemostatic powder, gelatin particles and polysaccharide compounds containing aldehyde groups or succinimide ester groups are stored separately. When used, the gelatin particles and the polysaccharide compounds containing aldehyde groups or succinimide ester groups are mixed. In the process of actual application, the gelatin particles and the polysaccharide compounds containing aldehyde groups or succinimide ester groups can be stored separately. The present invention does not limit the storage method of each component in the kit, and those skilled in the art can select a specific storage method as needed, all within the scope of the present invention.

[0085] <Fourth Aspect>

[0086] The fourth aspect of the present invention provides a use of the hemostatic putty according to the first aspect of the present invention in the preparation of a bone wound hemostasis product; the bone wound hemostasis is preferably cancellous bone wound hemostasis.

[0087] Example

[0088] The embodiments of the present invention will be described in detail below in conjunction with the examples, but those skilled in the art will appreciate that the following examples are only used to illustrate the present invention and should not be considered to limit the scope of the present invention. If no specific conditions are specified in the examples, they are carried out according to normal conditions or the conditions recommended by the manufacturer. If the manufacturer is not specified for the reagents or instruments used, they are all conventional products that can be obtained commercially.

[0089] Example 1

[0090] (1) Take 10 g of gelatin, add it to 85 ml of 60°C purified water, stir and dissolve to obtain a gelatin solution.

[0091] (2) Take hydroxyapatite (HA) particles and β-tricalcium phosphate (β-TCP), grind or crush them separately, and then sieve them to retain particles of 8 to 53 μm. Take 3 g of HA and 2 g of β-TCP particles and add them to the gelatin solution. Stir at high speed to disperse them in the gelatin solution system to obtain a mixture.

[0092] (3) Take 0.01 g of carbodiimide and dissolve it in 15 ml of purified water. Add the crosslinking agent solution to the mixture in step (2) while stirring. The stirring speed is 1000 r / min and the crosslinking agent solution is added at a rate of 0.2 mL / s to allow the gelatin to undergo a crosslinking reaction.

[0093] (4) The cross-linked gelatin mixed with active substances obtained in step (3) is soaked in purified water to fully swell, divided into small pieces and crushed into particles, and then placed in a dialysis bag for extraction and dialysis for 48 hours, followed by freeze-drying to remove moisture to obtain a porous sponge.

[0094] (5) The porous sponge is subjected to a primary heat cross-linking treatment in a nitrogen environment at 150°C for 6 hours, and then crushed and sieved to obtain particles with a size of 53 to 104 μm. The porous sponge is then subjected to a secondary heat cross-linking treatment in a nitrogen environment at 150°C for 3 hours. After irradiation sterilization, gelatin particles encapsulating the active substance are obtained.

[0095] Before use, take 1 g of gelatin particles encapsulating the active substance, 1 g of hyaluronic acid esterified with N-hydroxysuccinimide (degree of substitution 40%), add 6 g of sterile water for injection, and stir quickly to obtain a paste, which is the hemostatic putty.

[0096] Example 2

[0097] (1) Take 10 g of gelatin, add it to 85 ml of 40°C purified water, and stir to dissolve.

[0098] (2) Take hydroxyapatite (HA) particles and β-tricalcium phosphate (β-TCP), grind or crush them separately, and then sieve them to retain particles of 8 to 15 μm. Take 3 g of HA and 2 g of β-TCP particles and add them to the gelatin solution. Stir at high speed and disperse them in the gelatin solution system to obtain a mixture.

[0099] (3) 0.1 g of carbodiimide and 0.1 g of glutamine transaminase were dissolved in 15 ml of purified water, and a cross-linking agent was added to the mixture of step (2) while stirring, wherein the stirring speed was 1000 r / min and the cross-linking agent addition rate was 0.2 mL / s, so that gelatin was cross-linked.

[0100] (4) The cross-linked gelatin mixed with active substances obtained in step (3) is soaked in purified water to fully swell, divided into small pieces and crushed into particles, and then placed in a dialysis bag for extraction and dialysis for 60 hours, followed by freeze-drying to remove moisture to obtain a porous sponge.

[0101] (5) The porous sponge is subjected to a primary heat cross-linking treatment in a nitrogen environment at 160°C for 4 h, and then crushed and sieved to obtain particles with a size of 37 to 61 μm. The porous sponge is then subjected to a secondary heat cross-linking treatment in a nitrogen environment at 160°C for 4 h, and then irradiated and sterilized to obtain gelatin particles encapsulating the active substance.

[0102] Before use, take 2 g of gelatin particles encapsulating active substances, 1 g of hyaluronic acid esterified with N-hydroxysuccinimide (substitution degree 60%), add 9 g of sterile water for injection, and stir quickly to obtain a paste, which is the hemostatic putty.

[0103] Example 3

[0104] (1) Take 15 g of gelatin, add it to 75 ml of 60°C purified water, and stir to dissolve.

[0105] (2) Grind or crush hydroxyapatite (HA) particles and β-tricalcium phosphate (β-TCP) particles, respectively, and sieve to retain particles smaller than 150 μm. Add 8 g of HA and 2 g of β-TCP particles to a gelatin solution, stir at high speed, and disperse them in the gelatin solution system to obtain a mixture.

[0106] (3) Take 0.12 g of carbodiimide and dissolve it in 25 ml of purified water. Add a crosslinking agent to the mixture in step (2) while stirring. The stirring speed is 1000 r / min and the crosslinking agent is added at a rate of 0.2 mL / s to allow gelatin to undergo a crosslinking reaction.

[0107] (4) The cross-linked gelatin mixed with active substances obtained in step (3) is soaked in purified water to fully swell, divided into small pieces and crushed into particles, and then placed in a dialysis bag for extraction and dialysis for 56 hours, followed by freeze-drying to remove moisture to obtain a porous sponge.

[0108] (5) The porous sponge is subjected to a thermal cross-linking treatment in a nitrogen environment at 150° C. for 10 h, and then crushed and sieved to obtain particles with a particle size of less than 500 μm. After irradiation sterilization, gelatin particles encapsulating the active substance are obtained.

[0109] Before use, take 1 g of gelatin particles encapsulating active substances, 1.5 g of aldehyded hyaluronic acid (substitution degree 30%), add 6 g of sterile water for injection, and stir quickly to obtain a paste, which is the hemostatic putty.

[0110] Example 4

[0111] (1) Take 5 g of gelatin, add it to 85 ml of 40°C purified water, and stir to dissolve;

[0112] (2) Grinding or crushing hydroxyapatite (HA) particles and β-tricalcium phosphate (β-TCP) particles, respectively, and sieving to retain particles of 2.5 to 8 μm. Adding 4 g of HA and 6 g of β-TCP particles to a gelatin solution, stirring at high speed, and dispersing them in the gelatin solution system to obtain a mixture;

[0113] (3) taking 0.2 g of glutamine aminotransferase and dissolving it in 15 ml of purified water, adding a crosslinking agent to the mixture in step (2) while stirring, wherein the stirring speed is 1000 r / min, and the crosslinking agent is added at a speed of 0.2 mL / s, so that the gelatin undergoes a crosslinking reaction;

[0114] (4) freeze-drying the cross-linked gelatin mixed with the active substance obtained in step (3) to remove moisture and obtain a porous sponge;

[0115] (5) The porous sponge is subjected to a thermal cross-linking treatment in a nitrogen environment at 150° C. for 8 h, and then crushed and sieved to obtain particles with a particle size of 8-37 μm. After irradiation sterilization, gelatin particles encapsulating the active substance are obtained.

[0116] Before use, take 1.5 g of gelatin particles encapsulating active substances, 1 g of aldehyded hyaluronic acid (substitution degree 20%), add 8 g of sterile water for injection, and stir quickly to obtain a paste, which is the hemostatic putty.

[0117] Example 5

[0118] (1) Take 10 g of gelatin, add it to 80 ml of 40°C purified water, and stir to dissolve;

[0119] (2) Grinding or crushing hydroxyapatite (HA) particles and β-tricalcium phosphate (β-TCP) particles, respectively, and sieving to retain particles of 104 to 150 μm. Adding 6 g of HA and 4 g of β-TCP particles to a gelatin solution, stirring at high speed, and dispersing them in the gelatin solution system to obtain a mixture;

[0120] (3) dissolving 0.4 g of glutamine aminotransferase in 20 ml of purified water, adding a crosslinking agent to the mixture of step (2) while stirring, wherein the stirring speed is 1000 r / min, and the crosslinking agent is added at a speed of 0.2 mL / s, so that the gelatin undergoes a crosslinking reaction;

[0121] (4) freeze-drying the cross-linked gelatin mixed with the active substance obtained in step (3) to remove moisture and obtain a porous sponge;

[0122] (4) The porous sponge is subjected to a primary heat cross-linking treatment in a nitrogen environment at 150°C for 6 hours, and then crushed and sieved to obtain particles with a particle size of 104 to 500 μm. The porous sponge is then subjected to a secondary heat cross-linking treatment in a nitrogen environment at 150°C for 4 hours. After irradiation sterilization, gelatin particles encapsulating the active substance are obtained.

[0123] Before use, take 1 g of gelatin particles encapsulating active substances, 2 g of aldehyded hyaluronic acid (substitution degree 10%), add 12 g of sterile water for injection, and stir quickly to obtain a paste, which is the hemostatic putty.

[0124] Comparative Example 1

[0125] Compared with Example 1, the preparation method of the gelatin particles encapsulating the active substance without N-hydroxysuccinimide esterified hyaluronic acid is the same as that of Example 1. When used, 2 g of the gelatin particles encapsulating the active substance are taken, 6 g of sterile injection water is added, and the mixture is quickly stirred evenly. The gelatin particles absorb water and swell, and the obtained gelatin particles are discontinuous gel particles, which cannot form a plastic hemostatic putty.

[0126] Comparative Example 2

[0127] Compared with Example 3, the gelatin particles encapsulating the active substance are not contained. When used, 2.5 g of aldehyde-modified hyaluronic acid (substitution degree 30%) is taken, 6 g of sterile injection water is added, and the mixture is quickly stirred evenly. The aldehyde-modified hyaluronic acid absorbs water and swells to obtain viscous particles, which cannot form a plastic hemostatic putty.

[0128] Performance Testing

[0129] 1. Plasticity

[0130] like Figure 1 As shown, the hemostatic putty of Example 1 can be molded into three shapes, and can also be molded into any other shape, having good plasticity. The hemostatic putties of Examples 2-5 of the present invention can also be molded into any shape.

[0131] 2. Sealing adhesion strength test

[0132] Take 91 amino glass capillaries with an inner diameter of 0.5mm, an outer diameter of 1.0mm, and a length of 30mm. Fix the middle part and tighten it with hexagonal close packing. Use polydimethylsiloxane (PDMS) to seal and fix the middle 20mm position. Connect the cured part to an elastic connecting hose with an inner diameter of 1cm. The above part is used as a test tool. The other end of the elastic connecting hose is connected to a hard tube. The test tool can be disassembled and replaced after each test.

[0133] Connect the non-deformable hard pipe to the three-way structure, and connect the three ends to the liquid propulsion pump, pressure gauge and test fixture respectively. The specific device is as follows: Figure 2 shown.

[0134] Testing process:

[0135] Fill the liquid propulsion pump with water for injection and propel the liquid at a rate of 2 ml / min. Start the test after there is no air remaining in the pipeline and liquid continuously flows out from the outer end of the test fixture.

[0136] Take 0.05g of the sample to be tested, use a sharp flat tool to quickly and evenly spread the sample and fill it on the outer end of the test tool, ensuring that the filling thickness is within the range of 2±0.2mm;

[0137] At the same time, observe the changes in the pressure gauge. When liquid seeps out from the outer end of the test fixture, record the maximum pressure value of the pressure gauge.

[0138] The test fixture was replaced and parallel tests were performed. The test results are shown in Table 1 below.

[0139] 3. Hemostasis time test

[0140] Take 91 amino glass capillaries with an inner diameter of 0.5mm, an outer diameter of 1.0mm, and a length of 30mm. Fix the middle part and tighten it with hexagonal close packing. Use polydimethylsiloxane (PDMS) to seal and fix the middle 20mm position. Connect the cured part to an elastic connecting hose with an inner diameter of 1cm. The above part is used as a test tool. The other end of the elastic connecting hose is connected to a hard tube. The test tool can be disassembled and replaced after each test.

[0141] The liquid propulsion pump and the test fixture are connected by a non-deformable hard pipe. The specific device is as follows: Figure 3 shown.

[0142] Testing process:

[0143] Fill the liquid propulsion pump with fresh heparinized pig blood and propel the liquid at a rate of 1 ml / min. Start the test after there is no air remaining in the pipeline and blood continuously flows out from the outer end of the test tooling.

[0144] Take 0.05g of the sample to be tested, use a sharp flat tool to quickly and evenly spread the sample and fill it on the outer end of the test tool, ensuring that the filling thickness is within the range of 2±0.2mm;

[0145] After the packing is completed, start timing and observe whether there is blood seepage at the outer end of the test tool. If there is no blood seepage, hemostasis is completed;

[0146] The test fixture was replaced and parallel tests were performed. The test results are shown in Table 1 below.

[0147] 4. In vitro simulated degradation test

[0148] Take 91 amino glass capillaries with an inner diameter of 0.5 mm, an outer diameter of 1.0 mm, and a length of 30 mm, fix the middle part and tighten it with hexagonal close packing, use polydimethylsiloxane (PDMS) to seal and fix the middle 20 mm position, and connect the cured part to an elastic connecting hose with an inner diameter of 1 cm. The above parts are used as test fixtures, and the test fixtures can be replaced and updated after each test;

[0149] The test tool is sealed and packaged and then sterilized by moist heat, epoxy or irradiation;

[0150] Testing process:

[0151] Take 0.05g of the sample to be tested, use a sharp flat tool to evenly spread the sample and fill it on the outer end of the test fixture, ensuring that the filling thickness is within the range of 2±0.2mm;

[0152] Place the completed plugging test tool in a beaker containing 10 ml of PBS buffer solution, ensuring that the PBS buffer solution completely immerses the sample and the whole thing remains stationary in a sterile environment at 37°C;

[0153] Observe the sample every 24 hours until the sample in the capillary tube is completely invisible, then the degradation is considered complete and the degradation time is recorded;

[0154] The test fixture was replaced and parallel tests were performed. The results are shown in Table 1 below.

[0155] Table 1

[0156] Test samples Sealing adhesion strength / Kpa Hemostasis time / s In vitro degradation time / week Example 1 37 2 6 Example 2 21 2 5 Example 3 25 2 5 Example 4 14 4 5 Example 5 33 5 4 Comparative Example 1 3 6 3 Comparative Example 2 6 7 1

[0157] As can be seen from Table 1, the plugging adhesion strength of the hemostatic putties of Examples 1 to 5 of the present invention is higher than that of the hemostatic products of Comparative Examples 1 to 2, and the adhesion plugging strength is above 10 kPa, and the hemostatic time is shorter, and the in vitro degradation time can reach more than 4 weeks. Comparative Examples 1 to 2 cannot form a plastic hemostatic putty, so the plugging adhesion strength is lower, the hemostatic time is longer, and the in vitro degradation time is shorter.

[0158] It should be noted that, although the technical solution of the present invention is introduced with specific examples, those skilled in the art will appreciate that the present invention should not be limited thereto.

[0159] The embodiments of the present invention have been described above, and the above description is exemplary, not exhaustive, and is not limited to the disclosed embodiments. Many modifications and variations will be apparent to those of ordinary skill in the art without departing from the scope and spirit of the described embodiments. The selection of terms used herein is intended to best explain the principles of the embodiments, practical applications, or technical improvements in the market, or to enable other persons of ordinary skill in the art to understand the embodiments disclosed herein.

Claims

1. A plastic hemostatic putty, characterized in that: The hemostatic putty comprises hemostatic powder and solvent; wherein, The hemostatic powder comprises gelatin particles and a polysaccharide compound containing an aldehyde group or a succinimide ester group, wherein the gelatin particles are cross-linked gelatin particles encapsulating an active substance; Preferably, the particle size of the gelatin particles is below 500 μm.

2. The hemostatic putty according to claim 1, characterized in that: In the polysaccharide compound, the degree of substitution of aldehyde groups and / or succinimide ester groups is greater than or equal to 10%; Preferably, the mass ratio of the gelatin particles to the polysaccharide compound containing an aldehyde group or a succinimide ester group is (1:2) to (2:1).

3. The hemostatic putty according to claim 1 or 2, characterized in that: The particle size of the active substance is less than 150 μm; Preferably, the active substance comprises one or a combination of two or more of bioactive glass material, degradable metal, degradable metal oxide, degradable metal salt, hydroxyapatite, tricalcium phosphate, degradable metal alloy, and recombinant human bone morphogenetic protein.

4. The hemostatic putty according to any one of claims 1 to 3, characterized in that: The mass ratio of the hemostatic powder to the solvent is 1:1 to 1:5; Preferably, the solvent includes one or a combination of two or more of water, physiological saline, buffer solution, and water for injection; more preferably, a drug is added to the solvent, and the drug preferably includes one or a combination of two or more of thrombin, fibrin, and growth factor.

5. The hemostatic putty according to any one of claims 1 to 4, characterized in that: The adhesion and sealing strength of the hemostatic putty is greater than 10 kPa; and / or, The hemostatic putty has a hemostatic time of less than 8 seconds.

6. The hemostatic putty according to any one of claims 1 to 5, characterized in that: The preparation process of the gelatin particles comprises: mixing gelatin with the active substance and then chemically cross-linking the gelatin; The cross-linked gelatin mixed with the active substance is freeze-dried to obtain a porous sponge; The porous sponge is subjected to physical cross-linking and pulverization treatments.

7. The hemostatic putty according to any one of claim 6, characterized in that: The active substance is added to the gelatin solution to obtain a mixture, and a cross-linking agent is added to chemically cross-link the gelatin to obtain cross-linked gelatin mixed with the active substance; Preferably, based on the total mass of the mixture as 100%, the content of gelatin is 5-20%, and the content of the active substance is 0.5-10%; Preferably, the cross-linking agent comprises carbodiimide and / or transglutaminase, and the mass ratio of the cross-linking agent to the gelatin is 1:25-1000.

8. The hemostatic putty according to claim 6 or 7, characterized in that: The porous sponge is first crushed and then physically cross-linked, or the porous sponge is first physically cross-linked and then crushed, or the porous sponge is sequentially physically cross-linked, crushed, and then physically cross-linked; Preferably, the temperature of the physical crosslinking is 140-170° C., and the total time of the physical crosslinking is 4-10 h.

9. A method for preparing the hemostatic putty according to any one of claims 1 to 8, characterized in that: The preparation method comprises the step of mixing hemostatic powder and a solvent to form a paste.

10. Use of the hemostatic putty according to any one of claims 1 to 8 in preparing a bone wound hemostasis product; the bone wound hemostasis is preferably cancellous bone wound hemostasis.

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