Self-aggregation composite hemostatic material and preparation method thereof

Abstract

The invention relates to a preparation method of a self-aggregation composite hemostatic material, which comprises the following steps: dissolving sodium carboxymethyl starch in pure water to prepare an anionic polysaccharide solution, mixing chitosan and calcium salt, dissolving in an acid solution to prepare a cationic polysaccharide solution, quantitatively mixing the anionic polysaccharide solution and the cationic polysaccharide solution, and preparing the self-aggregation composite hemostatic material. A compact, stable and high-mechanical-strength flocculent precipitate with a three-dimensional network structure can be formed through electrostatic complexing and ionic cross-linking reaction, finally the flocculent precipitate is washed, dried, crushed and sieved, the self-aggregation composite hemostatic material is prepared, the whole preparation process is mild in reaction condition, a toxic cross-linking agent does not need to be used, the process is simple and environmentally friendly, and the cost is low. The preparation method is simple and easy for large-scale production, meanwhile, the prepared self-aggregation composite hemostatic material has unique hydrophilicity, can swell but not dissolve, can quickly absorb liquid, permeate and diffuse, can quickly form a hemostatic physical barrier after absorbing liquid, and has a synergistic hemostatic effect.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of biomedical materials, and particularly relates to a preparation method of a self-aggregation composite hemostatic material. BACKGROUND

[0002] In surgical operation and trauma treatment, effective hemostasis is crucial, which is mainly achieved through mechanical hemostasis, drug hemostasis and biological hemostasis. Effective hemostasis can quickly control bleeding, maintain blood volume stability, prevent the occurrence of hemorrhagic shock, and reduce the occurrence of complications such as local tissue ischemia, necrosis and anemia. By reducing blood loss, the patient's physical recovery is promoted, the recovery time is shortened, and the surgical field is kept clear, which is beneficial to precise operation of the doctor and improves the success rate of operation.

[0003] In the biological hemostasis mode, biological materials are usually used for hemostasis. An ideal hemostatic material should have the characteristics of rapid onset, good biocompatibility, convenient use, high safety, etc. The commonly used hemostatic materials in the clinic at present include gelatin sponge, oxidized cellulose and fibrin glue, but they each have some limitations, for example, the gelatin sponge is easy to collapse after absorbing blood, has the problem of insufficient mechanical strength, the oxidized cellulose has strong acidity and is easy to stimulate the surrounding tissue and cause inflammatory reaction, and the fibrin glue has the problems of virus transmission risk and high cost. These limitations make it difficult for the existing commonly used hemostatic materials to meet the hemostatic needs in the clinic. SUMMARY

[0004] Therefore, it is necessary to provide a preparation method of a self-aggregation composite hemostatic material. The technical scheme for solving the above technical problems is as follows:

[0005] A preparation method of a self-aggregation composite hemostatic material, comprising the following steps:

[0006] Dissolving sodium carboxymethyl starch in pure water to obtain an anionic polysaccharide solution;

[0007] Mixing and dissolving chitosan and calcium salt in an acidic solution to obtain a cationic polysaccharide solution;

[0008] Using a quantitative mixing device to quantitatively mix the anionic polysaccharide solution and the cationic polysaccharide solution, and filtering the flocculent precipitate obtained after electrostatic complexation and ion crosslinking reaction;

[0009] Washing, drying and crushing and sieving the flocculent precipitate to obtain a self-aggregation composite hemostatic material.

[0010] In one embodiment, when the sodium carboxymethyl starch is dissolved in pure water, the preparation concentration is controlled to be 0.5%-2.0% (w / v), and the pH value is adjusted to 4.0-7.0.

[0011] In one embodiment, the sodium carboxymethyl starch has a degree of substitution of 0.6-1.0.

[0012] In one embodiment, when the chitosan and the calcium salt are dissolved in the acidic solution, the concentration is controlled to be 0.5%-2.0% (w / v), and the pH value is controlled to be 3.0-5.5.

[0013] In one embodiment, the mass ratio of the chitosan to the calcium salt is 10-50:1.

[0014] In one embodiment, the degree of deacetylation of the chitosan is greater than or equal to 95%.

[0015] In one embodiment, the calcium salt is calcium chloride, calcium gluconate or calcium lactate, and the acidic solution is an acetic acid solution, a hydrochloric acid solution or a citric acid solution.

[0016] In one embodiment, when the anionic polysaccharide solution and the cationic polysaccharide solution are quantitatively mixed using the quantitative mixing device, the volume ratio of the anionic polysaccharide solution to the cationic polysaccharide solution is (0.8-1.2):1.

[0017] In one embodiment, when the flocculent precipitate is washed, dried and crushed and sieved, the flocculent precipitate is beaten and washed using an ethanol solution or an acetone solution, dried by vacuum drying or freeze drying after washing, crushed and sieved to obtain the self-aggregation composite hemostatic material with a mesh number of 80-200.

[0018] The present application also provides a self-aggregation composite hemostatic material prepared by the preparation method of the self-aggregation composite hemostatic material according to any one of the above embodiments.

[0019] The application has the beneficial effects that: the self-aggregation composite hemostatic material preparation method provided by the application can form a flocculent precipitate with a dense, stable and high mechanical strength three-dimensional network structure through electrostatic complexation and ion crosslinking reaction by dissolving carboxymethyl starch sodium in pure water to prepare an anionic polysaccharide solution, dissolving chitosan and calcium salt in an acidic solution to prepare a cationic polysaccharide solution, and then quantitatively mixing the anionic polysaccharide solution and the cationic polysaccharide solution, the whole preparation process has mild reaction conditions, does not need to use toxic crosslinking agents, has a simple and environmentally friendly process, is easy to scale up, and the self-aggregation composite hemostatic material prepared has unique hydrophilicity, can swell without dissolving, can quickly absorb liquid, permeate and diffuse, and will not appear as a gel package, after the self-aggregation composite hemostatic material absorbs liquid, the particles can quickly aggregate into a mass through electrostatic action and settle to the bottom of the blood, can directly block the bleeding wound surface, form a physical barrier for hemostasis, has good biocompatibility between components, and has a synergistic hemostatic effect. BRIEF DESCRIPTION OF DRAWINGS

[0020] In order to more clearly illustrate the technical solutions of the embodiments of the present application, the drawings needed in the embodiments will be briefly introduced below, and it should be understood that the following drawings only show some embodiments of the present application, and should not be regarded as a limitation on the scope, and for those skilled in the art, other related drawings can also be obtained without creative labor on the basis of these drawings.

[0021] Figure 1 The flowchart of the self-aggregation composite hemostatic material preparation method of the present application is shown in the figure.

[0022] Figure 2 The electron microscope image of the self-aggregation composite hemostatic material of the present application is shown in the figure.

[0023] Figure 3 The rabbit femoral artery hemostasis effect diagram of the self-aggregation composite hemostatic material of the present application is shown in the figure.

[0024] Figure 4 The rabbit liver hemostasis effect diagram of the self-aggregation composite hemostatic material of the present application is shown in the figure.

[0025] Figure 5 The rabbit femoral artery hemostasis effect diagram of the self-aggregation composite hemostatic material of the present application is shown in the figure. DETAILED DESCRIPTION

[0026] For the purposes of promoting an understanding of the principles of the application, reference will now be made to the embodiments illustrated in the drawings. There is shown by way of illustration a preferred embodiment of the application. It is to be understood that the application can be practiced with modification and alteration, and can take on various forms. Accordingly, the embodiments are set forth only for the purpose of illustration and are not intended as a limitation on the application set forth in the claims. As used herein, the term "and / or" includes any and all combinations of one or more of the associated listed items.

[0027] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. The terminology used in the description of the application herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the application. As used herein, the term "and / or" includes any and all combinations of one or more of the associated listed items.

[0028] In one embodiment, as shown in FIG. 1, a method for preparing a self-aggregated composite hemostatic material includes the following steps: Figure 1

[0029] Step 110, dissolving sodium carboxymethyl starch in pure water to obtain an anionic polysaccharide solution.

[0030] In this embodiment, when preparing the anionic polysaccharide solution, sodium carboxymethyl starch is dissolved in pure water, the preparation concentration is controlled to be 0.5%-2.0% (w / v), i.e. the mass of the solute in 100 mL of the solution is 0.5 g-2 g, and acetic acid, hydrochloric acid or citric acid is used to adjust the pH value to 4.0-7.0.

[0031] In one embodiment, the degree of substitution of the sodium carboxymethyl starch is 0.6-1.0. Specifically, sodium carboxymethyl starch is an anionic polysaccharide with extremely strong water absorption performance. Sodium carboxymethyl starch with a high degree of substitution is easily soluble in water, can quickly form a physical barrier on the surface of a wound, slow down blood flow, and play a role of a physical barrier. At the same time, the sodium carboxymethyl starch particles can quickly swell after absorbing water, can quickly absorb the water in the blood, concentrate the blood components, promote the adhesion and aggregation of platelets and coagulation factors, and achieve the promotion of coagulation and the formation of a stable blood clot.

[0032] Step 120, dissolving chitosan and calcium salt in an acid solution to obtain a cationic polysaccharide solution.

[0033] In this embodiment, when preparing the cationic polysaccharide solution, chitosan and calcium salt are dissolved in an acetic acid solution, a hydrochloric acid solution or a citric acid solution, the mass ratio of chitosan to calcium salt is 10-50:1, the preparation concentration is controlled to be 0.5%-2.0% (w / v), i.e. the mass of the solute in 100 mL of the solution is 0.5 g-2 g, and the pH value of the cationic polysaccharide solution is controlled to be 3.0-5.5.

[0034] ​In one embodiment, the degree of deacetylation of the chitosan is greater than or equal to 95%. Specifically, chitosan is a natural cationic polysaccharide with good biocompatibility, antibacterial properties and procoagulant activity, and can adsorb negatively charged red blood cells and platelets through its positive charge to promote the formation of cell clots or hemostatic plugs. The higher the degree of deacetylation of chitosan, the higher the proportion of amino groups on the chitosan molecular chain, and the higher the positive charge density, which is conducive to the rapid adhesion of positively charged chitosan to negatively charged substances on the surface of red blood cells such as sialic acid through electrostatic interaction to form cell clots or hemostatic plugs, and to the activation of negatively charged substances on the surface of platelets such as phosphatidylserine to promote platelet aggregation through electrostatic attraction. At the same time, the amino and hydroxyl groups on the molecular chain of chitosan can activate the complement system, further bind to coagulation factors, accelerate the coagulation cascade reaction, and form a stable blood clot. At the same time, the antibacterial performance of chitosan with high degree of deacetylation is optimized, which can better and more effectively reduce the risk of wound infection and create a sterile environment for hemostasis.

[0035] In one embodiment, the calcium salt is calcium chloride, calcium gluconate or calcium lactate. Specifically, the use of calcium chloride, calcium gluconate and calcium lactate as calcium salts can participate in the coagulation process by providing calcium ions to assist hemostasis. Calcium ions bind to coagulation factors VII, IX and X to promote their activation to active forms, accelerate the conversion of prothrombin to thrombin, enhance the adhesion and aggregation capacity of platelets, promote platelet thrombus formation, and inhibit the release of histamine from mast cells to reduce the vascular dilation and plasma exudation caused by allergic reactions, further assisting hemostasis, and promoting fibroblast proliferation and collagen synthesis to accelerate wound healing and reduce scar formation.

[0036] Step 130, using a quantitative mixing device to quantitatively mix the anionic polysaccharide solution and the cationic polysaccharide solution, and filtering the flocculent precipitate after electrostatic complexation and ionic crosslinking reaction.

[0037] In this embodiment, the prepared anionic polysaccharide solution and cationic polysaccharide solution are quantitatively mixed at room temperature using a quantitative mixing device. The volume ratio of the anionic polysaccharide solution to the cationic polysaccharide solution is (0.8-1.2):1. In this way, positively charged chitosan and negatively charged carboxymethyl starch sodium can form intermolecular complexes through electrostatic interaction, and calcium ions can form ionic crosslinks with carboxymethyl starch sodium. The electrostatic complexation and ionic crosslinking reaction can form a flocculent precipitate with a dense, stable and high mechanical strength three-dimensional network structure, so that the flocculent precipitate not only has the characteristics of chitosan positive adsorption procoagulant and carboxymethyl starch sodium hydrophilicity, but also the crosslinked calcium ions can act as coagulation factor IV to directly participate in multiple links of the endogenous coagulation pathway, produce significant synergistic effect, play a synergistic hemostatic role, and have better hemostatic speed.

[0038] Step 140, the flocculent precipitate is washed, dried and broken and sieved to obtain the self-aggregation composite hemostatic material.

[0039] In this embodiment, when the flocculent precipitate is washed, dried and broken and sieved, the flocculent precipitate is beaten and washed with an ethanol solution or an acetone solution, after washing, vacuum drying or freeze drying is used for drying treatment, the drying temperature of vacuum drying is set to 40-60°C, after drying treatment, breaking and sieving are performed to obtain the self-aggregation composite hemostatic material with a mesh number of 80-200.

[0040] In this embodiment, as shown in Figure 2 and Figure 3 The self-aggregation composite hemostatic material prepared by the double action has a unique hydrophilicity, can swell without dissolving, can quickly absorb liquid and permeate and diffuse, and cannot form a gel package. After absorbing liquid, the self-aggregation composite hemostatic material can quickly aggregate into a mass through electrostatic action, settle to the bottom of the blood, directly block the bleeding wound surface, not easily washed away by flowing blood, more effectively form a physical barrier for hemostasis, and play a synergistic hemostatic effect. At the same time, the components of the self-aggregation composite hemostatic material are polysaccharides or inorganic salts of natural origin, have good biocompatibility, the degradation products are non-toxic and have no side effects, and the suitable pH value avoids potential irritation of a strong acidic environment to tissues.

[0041] In one embodiment, compared with chitosan and sodium carboxymethyl starch, the use of sodium carboxymethyl starch and carboxymethyl chitosan after blending does not have a self-aggregation effect. Carboxymethyl chitosan is a derivative of chitosan, and carboxymethyl and residual acetyl amino or amino groups exist on the molecular chain. The protonation of the amino group makes the carboxymethyl chitosan molecule have a double-charge characteristic, which reduces the overall aggregation ability. At the same time, in the carboxymethyl chitosan molecule, the carboxymethyl and acetyl amino or amino groups can form intramolecular hydrogen bonds, making the molecular chain have a rigid conformation, which further limits the flexible arrangement between molecules and inhibits self-aggregation.

[0042] In one embodiment, the quantitative mixing of the anionic polysaccharide solution and the cationic polysaccharide solution is a quantitative precision control based on the dual reaction requirements. When the positively charged chitosan molecular chain is quantitatively mixed with the negatively charged carboxymethyl starch sodium molecular chain, it can quickly complex through charge attraction, form a basic skeleton of a three-dimensional network, calcium ions simultaneously ionically crosslink with the carboxyl groups of the carboxymethyl starch sodium, further form a dense, high-mechanical-strength flocculent precipitate, and the quantitative control ensures the reaction balance, and finally the formed structure has the characteristics of balance between hydrophilicity and stability and strong self-aggregation ability, that is, it can swell without dissolving and the particles can quickly agglomerate after absorbing liquid, which can better cope with the clinical pain points of critical bleeding. The other multi-component random coexistence mixing system of hemostatic materials, that is, by sequentially adding each component without control, its unassociated reaction mechanism and functional requirements, although there are electrostatic effects, ion chelation and hydrogen bonding, there is mutual interference and cannot form a unified structure, the crosslinking sites are chaotic, the existence of multiple components disperses the intermolecular forces, and only a loose gel network structure can be formed, which cannot withstand the impact of flowing blood and can only achieve basic hemostasis by physically absorbing water to concentrate blood, lacks active plugging and rapid coagulation ability, and cannot achieve stable physical barrier function.

[0043] In one embodiment, the quantitative mixing device comprises a first liquid storage tank, a second liquid storage tank and a double liquid valve, the first liquid storage tank is connected with the first end of the double liquid valve through a first feeding pipe, the second liquid storage tank is connected with the second end of the double liquid valve through a second feeding pipe, a discharge mixing pipe is arranged on the double liquid valve, the discharge mixing pipe communicates with the first feeding pipe and the second feeding pipe respectively, a first pressure regulating valve is arranged on the first liquid storage tank, and a second pressure regulating valve is arranged on the second liquid storage tank, which can well quantitatively mix the anionic polysaccharide solution and the cationic polysaccharide solution, thereby achieving quantitative precision control based on the dual reaction requirements.

[0044] The self-aggregation composite hemostatic material preparation method provided by the application can form a dense, stable and high-mechanical-strength flocculent precipitate with a three-dimensional network structure through electrostatic complexation and ion crosslinking reaction by dissolving carboxymethyl starch sodium in pure water to prepare an anionic polysaccharide solution, dissolving chitosan and calcium salt in an acidic solution to prepare a cationic polysaccharide solution, and then quantitatively mixing the anionic polysaccharide solution and the cationic polysaccharide solution. Finally, the flocculent precipitate is washed, dried and crushed and sieved to prepare the self-aggregation composite hemostatic material. The entire preparation process is carried out in an aqueous phase, the reaction conditions are mild, no toxic crosslinking agent such as glutaraldehyde needs to be used, the biological toxicity risk caused by the residual chemical crosslinking agent is avoided, the process is simple and environmentally friendly, and the process is easy to scale up.

[0045] The application also provides a self-aggregation composite hemostatic material prepared by the preparation method of the self-aggregation composite hemostatic material in any of the above embodiments. Specifically, the self-aggregation composite hemostatic material prepared by the dual action can combine the advantages of both chitosan and sodium carboxymethyl starch and produce a synergistic enhancement effect through intermolecular interaction. Compared with sodium carboxymethyl starch, chitosan or simply physically mixed or chemically cross-linked sodium carboxymethyl starch and chitosan, the self-aggregation composite hemostatic material has faster hemostatic speed, better biocompatibility and better mechanical properties. The self-aggregation composite hemostatic material has unique hydrophilicity, can swell without dissolving, can quickly absorb liquid, permeate and diffuse, and will not appear as a gel package. After absorbing liquid, the self-aggregation composite hemostatic material can quickly aggregate into a mass through electrostatic action, settle to the bottom of the blood, directly block the bleeding wound, not easily washed away by flowing blood, more effectively form a physical barrier for hemostasis, play a synergistic hemostatic role, and can be well used as a biological hemostatic material for surgical or traumatic hemostasis, and meet the hemostatic needs in clinical practice.

[0046] The application will be further described below with specific embodiments.

[0047] Embodiment 1

[0048] A preparation method of a self-aggregation composite hemostatic material, comprising the following steps: dissolving sodium carboxymethyl starch in pure water, controlling the preparation concentration to be 0.5% (w / v), and using a hydrochloric acid solution to adjust the pH value to 4.0; then dissolving chitosan and calcium salt in the hydrochloric acid solution, the mass ratio of chitosan and calcium salt being 10:1, controlling the preparation concentration to be 0.5% (w / v), and controlling the pH value of the cationic polysaccharide solution to be 3.0; using a quantitative mixing device to quantitatively mix the prepared anionic polysaccharide solution and cationic polysaccharide solution at room temperature, the volume ratio of the anionic polysaccharide solution and cationic polysaccharide solution being 0.8:1; filtering the flocculent precipitate obtained after electrostatic complexation and ion cross-linking reaction; washing, drying and crushing the flocculent precipitate, washing the flocculent precipitate by beating with an ethanol solution, drying the flocculent precipitate by vacuum drying after washing, setting the drying temperature of the vacuum drying to be 50°C, crushing and sieving the self-aggregation composite hemostatic material after drying treatment to obtain a self-aggregation composite hemostatic material with a mesh number of 80.

[0049] In the formula, the degree of substitution of the sodium carboxymethyl starch is 0.6, the degree of deacetylation of the chitosan is 95%, and the calcium salt is calcium chloride.

[0050] Embodiment 2

[0051] A preparation method of a self-aggregation composite hemostatic material, comprising the following steps: dissolving sodium carboxymethyl starch in pure water, controlling the preparation concentration to be 1% (w / v), and using a citric acid solution to adjust the pH value to 6.0, then dissolving chitosan and calcium salt in the citric acid solution, the mass ratio of chitosan and calcium salt being 30:1, controlling the preparation concentration to be 1% (w / v), and controlling the pH value of the cationic polysaccharide solution to be 4.0, using a quantitative mixing device to quantitatively mix the prepared anionic polysaccharide solution and cationic polysaccharide solution at room temperature, the volume ratio of the anionic polysaccharide solution and cationic polysaccharide solution being 1:1, filtering the flocculent precipitate after electrostatic complexation and ionic crosslinking reaction, washing, drying and crushing and sieving the flocculent precipitate, using an acetone solution to beat and wash the flocculent precipitate, drying the flocculent precipitate by vacuum drying after washing, the drying temperature of the vacuum drying being set to be 50 DEG C, and crushing and sieving the flocculent precipitate after drying treatment to obtain the self-aggregation composite hemostatic material with a mesh number of 120.

[0052] In the formula, the degree of substitution of the sodium carboxymethyl starch is 0.8, the degree of deacetylation of the chitosan is 95%, and the calcium salt is calcium gluconate.

[0053] Example 3

[0054] A preparation method of a self-aggregation composite hemostatic material, comprising the following steps: dissolving sodium carboxymethyl starch in pure water, controlling the preparation concentration to be 2% (w / v), and using an acetic acid solution to adjust the pH value to 7.0, then dissolving chitosan and calcium salt in the acetic acid solution, the mass ratio of chitosan and calcium salt being 50:1, controlling the preparation concentration to be 2% (w / v), and controlling the pH value of the cationic polysaccharide solution to be 5.5, using a quantitative mixing device to quantitatively mix the prepared anionic polysaccharide solution and cationic polysaccharide solution at room temperature, the volume ratio of the anionic polysaccharide solution and cationic polysaccharide solution being 1.2:1, filtering the flocculent precipitate after electrostatic complexation and ionic crosslinking reaction, washing, drying and crushing and sieving the flocculent precipitate, using an ethanol solution to beat and wash the flocculent precipitate, drying the flocculent precipitate by vacuum drying after washing, the drying temperature of the vacuum drying being set to be 50 DEG C, and crushing and sieving the flocculent precipitate after drying treatment to obtain the self-aggregation composite hemostatic material with a mesh number of 200.

[0055] In the formula, the degree of substitution of the sodium carboxymethyl starch is 1.0, the degree of deacetylation of the chitosan is 95%, and the calcium salt is calcium lactate.

[0056] Comparative Example 1

[0057] A commercially available crosslinked starch hemostatic powder, mainly comprising starch, carboxymethyl chitosan and an ionic crosslinking agent.

[0058] The application will be further described in detail by specific tests.

[0059] Hemostatic effect test: New Zealand white rabbits were selected to make liver wound bleeding model and femoral artery bleeding model, respectively. The self-aggregation composite hemostatic material prepared in the examples and the commercially available cross-linked starch hemostatic powder of Comparative Example 1 were used for hemostasis, and the self-aggregation composite hemostatic material prepared in the examples and the commercially available cross-linked starch hemostatic powder of Comparative Example 1 were used for in vitro blood coagulation test.

[0060] Test 1: Taking Example 1 and Comparative Example 1 as examples, liver hemostasis test of rabbits was carried out.

[0061] New Zealand white rabbits were selected to make liver wound bleeding model: After anesthesia, the rabbits were fixed on the operating board in a supine position, the skin was prepared, disinfected, the abdominal cavity was opened, and the liver was fully exposed. Then a cross-shaped incision with a size of 2.0x2.0 cm was made on the same part of the left, middle and right liver of the rabbit with a surgical knife, and the cutting wound was 0.5 cm deep. After the bleeding model was established, the cutting wound was allowed to bleed freely for 30 seconds, and the blood on the wound was absorbed with weighed absorbent cotton gauze. After the free bleeding time reached, the self-aggregation composite hemostatic material prepared in Example 1 or the commercially available cross-linked starch hemostatic powder of Comparative Example 1 was immediately used for hemostasis on the bleeding wound, and after completion, timing was started. The wound was compressed with absorbent cotton gauze for 1 min, and after the absorbent cotton gauze was removed, it was observed whether there was bleeding, so as to evaluate the hemostatic effect.

[0062] Test 2: Taking Example 2 and Comparative Example 1 as examples, femoral artery hemostasis test of rabbits was carried out.

[0063] New Zealand white rabbits were selected to make femoral artery bleeding model: After anesthesia, the rabbits were fixed on the operating board in a supine position, the skin was prepared, disinfected, and the distal end of the left femoral artery was dissected and separated with a sterile surgical suture knot ready for use. Then the distal end of the femoral artery was ligated, the femoral artery was cut at the proximal end with an ophthalmic scissors, the blood at the proximal end was sprayed out, and the gauze was immediately pressed and wiped. Then the gauze was loosened, and the self-aggregation composite hemostatic material prepared in Example 2 or the commercially available cross-linked starch hemostatic powder of Comparative Example 1 was immediately used for hemostasis on the bleeding wound. After completion, timing was started. The wound was compressed with absorbent cotton gauze for 1 min, and after the absorbent cotton gauze was removed, it was observed whether there was bleeding, so as to evaluate the hemostatic effect.

[0064] Test 3: Taking Example 3 and Comparative Example 1 as examples, in vitro blood coagulation test of rabbits was carried out.

[0065] New Zealand white rabbits were selected, 7 mL of abdominal aortic blood was taken and placed into a 90.9 U / mL heparin anticoagulant tube, shaken well, and then 0.4 g of the self-aggregation composite hemostatic material prepared in Example 3 or the commercially available cross-linked starch hemostatic powder of Comparative Example 1 was placed. The coagulation time was recorded, and a blank control group was set up. 7 mL of abdominal aortic blood was taken and placed into a 90.9 U / mL heparin anticoagulant tube, shaken well, and placed for 18 minutes. The coagulation time was recorded.

[0066] Test results: The results of the liver hemostasis test of rabbits, the femoral artery hemostasis test of rabbits and the rabbit blood in vitro procoagulant test are shown in Table 1, Table 2 and Table 3.

[0067] Table 1 Results of liver hemostasis test of rabbits

[0068] Test subject Free bleeding amount Pressing time Hemostasis time Use material Rabbit 0.3-0.45g 1 minute Successful hemostasis Self-aggregating complex hemostatic material of Example 1 Rabbit 0.35-0.47g 1 minute Failed hemostasis Commercially available crosslinked starch hemostatic powder of Comparative Example 1

[0069] Table 2 Results of femoral artery hemostasis test of rabbits

[0070] Test subject Hemostasis measure Pressing time Hemostasis effect Rabbit Self-aggregating complex hemostatic material of Example 2 1 minute Successful hemostasis Rabbit Commercially available crosslinked starch hemostatic powder of Comparative Example 1 1 minute Failed hemostasis

[0071] Table 3 Results of rabbit blood in vitro procoagulant test

[0072] Coagulation measure Coagulation sample addition amount Coagulation result Self-aggregating complex hemostatic material of Example 3 0.4g 1 minute 30 seconds blood no longer flows Commercially available crosslinked starch hemostatic powder of Comparative Example 1 0.4g 2 minutes 30 seconds blood no longer flows Blank control / No coagulation

[0073] As shown in Table 1, Table 2 and Table 3, the self-aggregated composite hemostatic material prepared in Example 1 has good hemostatic effect on the liver wound of rabbits, can effectively control the bleeding of parenchymal organs within 1 minute, and the hemostatic effect is as shown in Figure 4 The self-aggregated composite hemostatic material prepared in Example 2 can quickly take effect in the femoral artery hemorrhage model of rabbits, successfully stop the femoral artery bleeding within 1 minute, and the formed blood clots have strong mechanical stability with the self-aggregated composite hemostatic material, and there is no re-bleeding after removing the compression, which has outstanding effectiveness and reliability for critical bleeding scenes, and the hemostatic effect is as shown in Figure 5 The self-aggregated composite hemostatic material prepared in Example 3 can significantly accelerate the blood coagulation process, and the coagulation speed is faster than that of the commercially available cross-linked starch hemostatic powder, and has excellent procoagulant activity.

[0074] The technical features of the above-described embodiments can be combined arbitrarily, and in order to make the description simple, all possible combinations of the technical features in the above-described embodiments are not described, however, as long as the combination of the technical features does not exist contradictory, it should be considered as the scope of the description.

[0075] The above-described embodiments only express several embodiments of the present application, and the description is more specific and detailed, but it should not be understood as a limitation on the scope of the patent. It should be noted that for ordinary skilled in the art, without departing from the concept of the present application, a number of modifications and improvements can be made, which are within the scope of the present application. Therefore, the scope of protection of the patent of the present application should be subject to the appended claims.

Claims

1. A method for preparing a self-aggregating composite hemostatic material, characterized in that, Includes the following steps: Sodium carboxymethyl starch was dissolved in pure water to obtain an anionic polysaccharide solution; Chitosan and calcium salt were mixed and dissolved in an acidic solution to obtain a cationic polysaccharide solution; The anionic polysaccharide solution and the cationic polysaccharide solution were quantitatively mixed using a quantitative mixing device. After electrostatic complexation and ionic cross-linking reactions, the mixture was filtered to obtain a flocculent precipitate. The flocculent precipitate was washed, dried, crushed, and sieved to obtain a self-aggregating composite hemostatic material.

2. The method for preparing the self-aggregating composite hemostatic material according to claim 1, characterized in that, When dissolving sodium carboxymethyl starch in pure water, the concentration is controlled at 0.5%-2.0% (w / v), and the pH value is adjusted to 4.0-7.

0.

3. The method for preparing the self-aggregating composite hemostatic material according to claim 2, characterized in that, The degree of substitution of the sodium carboxymethyl starch is 0.6-1.

0.

4. The method for preparing the self-aggregating composite hemostatic material according to claim 1, characterized in that, When chitosan and calcium salt are mixed and dissolved in an acidic solution, the concentration is controlled to be 0.5%-2.0% (w / v), and the pH value is controlled to be 3.0-5.

5.

5. The method for preparing the self-aggregating composite hemostatic material according to claim 4, characterized in that, The mass ratio of chitosan to calcium salt is 10-50:

1.

6. The method for preparing the self-aggregating composite hemostatic material according to claim 5, characterized in that, The degree of deacetylation of the chitosan is greater than or equal to 95%.

7. The method for preparing the self-aggregating composite hemostatic material according to claim 6, characterized in that, The calcium salt is calcium chloride, calcium gluconate, or calcium lactate, and the acidic solution is acetic acid solution, hydrochloric acid solution, or citric acid solution.

8. The method for preparing the self-aggregating composite hemostatic material according to claim 1, characterized in that, When the anionic polysaccharide solution and the cationic polysaccharide solution are quantitatively mixed using a quantitative mixing device, the volume ratio of the anionic polysaccharide solution to the cationic polysaccharide solution is (0.8-1.2):

1.

9. The method for preparing the self-aggregating composite hemostatic material according to claim 1, characterized in that, When washing, drying, crushing and sieving the flocculent precipitate, the flocculent precipitate is pulped and washed with ethanol solution or acetone solution. After washing, it is dried by vacuum drying or freeze drying. After drying, it is crushed and sieved to obtain a self-aggregating composite hemostatic material with a mesh size of 80-200 mesh.

10. A self-aggregating composite hemostatic material, characterized in that, It is prepared by the method for preparing the self-aggregating composite hemostatic material according to any one of claims 1-9.

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