CN104162183A - Chitosan and halloysite nanotube compound styptic powder and preparation method thereof - Google Patents

Abstract

The invention discloses chitosan and halloysite nanotube compound styptic powder and a preparation method thereof. The preparation method comprises the following steps: dissolving chitosan in an acetic acid aqueous solution to prepare a chitosan solution; adding halloysite nanotubes, and stirring for 20-30 hours at room temperature; adding a sodium hydroxide aqueous solution to regulate the pH to be 7 so as to prepare a suspension, or adding an organic precipitant to prepare the suspension, or adding a crosslinking agent to prepare the suspension; centrifugally separating the suspension, washing precipitate by deionized water, and freeze-drying the precipitate to obtain the chitosan and halloysite nanotube compound styptic powder. The preparation method is simple and easy for industrialization; the prepared compound styptic powder has a good styptic effect, and is capable of solving the problems of low gel strength and water residues after gelation of chitosan styptic materials and poor biocompatibility of traditional inorganic styptic materials.

Description

A kind of composite hemostatic powder of chitosan and halloysite nanotube and preparation method thereof

technical field

The invention belongs to the field of biomedical materials, and in particular relates to a composite hemostatic powder of chitosan and halloysite nanotubes and a preparation method thereof.

Background technique

Hemostasis is an important part of clinical treatment. Effective hemostasis is a necessary guarantee for the life safety of surgical patients. Hemostatic materials have an important impact on reducing bleeding, shortening operation time and healing of patients in various surgical operations. In recent years, medical absorbable hemostatic materials have attracted great attention from the medical and industrial circles of various countries, and new hemostatic materials have become one of the research hotspots in the field of surgical hemostasis. An ideal medical hemostatic material should meet the following characteristics: (1) good biocompatibility and no obvious toxic and side effects; (2) rapid hemostasis and obvious hemostatic effect; Blood under anticoagulant treatment also has good hemostatic function; (3) easy to use, can adapt to irregular wounds, and truly completely cover irregular wounds; (4) has good tissue adhesion and is not easy to fall off; (5) The wound does not generate local pressure, which promotes the rapid healing of the wound.

The commonly used hemostatic equipment clinically at present is tourniquet and hemostatic sponge, but the hemostatic effect on ruptured arteries of organs such as the chest cavity, abdominal cavity and pelvic cavity with irregular shapes is very unsatisfactory. Therefore, the development of fast, safe and effective new hemostatic materials for these bleeding sites has become an important topic in the field of biomaterials. At present, the commonly used absorbable hemostatic materials in clinical practice mainly include fibrin glue, gelatin sponge, starch-based microspheres, etc. These hemostatic materials have improved the hemostasis of surgical operations to a certain extent, but there are still many shortcomings, such as the hemostasis of large blood vessels. The efficiency is low, the effect on patients with coagulation disorders is not good, the combination with the wound is not sufficient, it may infect human or animal blood-borne diseases, and the cost is high. While the existing medical equipment and environment are constantly improving, the medical field at home and abroad also continuously improves the requirements for surgery, and a new multifunctional hemostatic material is bound to be needed.

In view of the shortage of surgical hemostatic materials currently used clinically, the development of fast, efficient, stable, safe and convenient new hemostatic materials is of great significance for material development and clinical application. In recent years, medical absorbable hemostatic materials have attracted great attention from the medical and industrial circles of various countries, and new hemostatic materials have become one of the research hotspots in the field of surgical hemostasis. Chitosan is the product of partial deacetylation of chitin, which widely exists in the crustaceans of shrimps, crabs, insects, fungi and plant cell walls, and is the only positively charged alkaline polysaccharide in nature. In 1983, Malette et al first published a study on the hemostatic function of chitosan, and then the hemostatic function of chitosan has attracted more and more attention. Numerous studies at home and abroad have confirmed that chitosan has excellent hemostatic function. On the one hand, through protein-mediated adhesion to platelets, the complex formed accelerates the polymerization of fibrin monomers and forms clots together; on the other hand, it induces red blood cell aggregation and stimulates The blood vessels constrict and eventually a thrombus forms, sealing the wound. Its hemostatic effect does not depend on the normal coagulation system and platelets, so it is increasingly becoming a research hotspot of biomaterials. However, the chitosan-based hemostatic powder material has low strength after forming a gel, and there are some problems such as water residue after gelation.

The mechanism of action of inorganic hemostatic materials is to use their high specific surface area to quickly absorb water in the blood, thereby locally concentrating blood coagulation factors and platelets, and finally achieving the effect of promoting blood coagulation. Typical representatives are zeolite (product QuikClot), kaolin (product Combat Gauze), montmorillonite (product WoundStat), and mesoporous silicon materials (product TraumaStat). However, these materials also have some problems, mainly because of their poor biological safety and easy to cause inflammation and a certain degree of toxicity. Halloysite is curled from more than 20 kaolinite sheets. The inner diameter of the tube is about 15-20nm, the outer diameter is about 50nm, and the length is about 100-1500nm. It is a natural multi-walled nanotube with a very high specific surface area. At present, the application of halloysite mainly focuses on ceramics, drug slow-release systems, templates, microreactors, catalysts, etc. It has also attracted the attention of scientists in the field of biomedicine, and it has good biocompatibility. There are reports of hemostatic sponges in the field of hemostasis.

Contents of the invention

In order to solve the shortcomings and deficiencies of the prior art, the primary purpose of the present invention is to provide a preparation method of a composite hemostatic powder of chitosan and halloysite nanotubes.

Another object of the present invention is to provide the composite hemostatic powder of chitosan and halloysite nanotubes obtained by the above preparation method. The composite hemostatic powder overcomes the problems of low gel strength and water residue after gelation of chitosan hemostatic materials on the one hand, and overcomes the biocompatibility problems caused by traditional inorganic hemostatic materials on the other hand.

In order to realize the above-mentioned purpose of the invention, the present invention adopts following technical scheme:

A preparation method of a composite hemostatic powder of chitosan and halloysite nanotubes, comprising the following steps:

(1) Dissolve chitosan in acetic acid aqueous solution to prepare a chitosan solution, then add halloysite nanotubes, and stir at room temperature for 20-30 hours, then add aqueous sodium hydroxide solution to adjust the pH to 7 to prepare a suspension , or adding an organic precipitant to obtain a suspension, or adding a cross-linking agent to obtain a suspension;

(2) Centrifuge the suspension obtained in step (1), wash the obtained precipitate with deionized water, repeat the operation of centrifugal separation and deionized water washing for 2 to 5 times, and then freeze-dry the precipitate to obtain the described Composite hemostatic powder of chitosan and halloysite nanotubes.

Preferably, when the suspension is prepared by adding a cross-linking agent in step (1), after the suspension is obtained, the solvent is removed by spray drying to obtain the composite hemostatic compound of chitosan and halloysite nanotubes pink.

More preferably, when spray drying, the suspension is pumped into the spray dryer by a peristaltic pump for spraying, and the solvent is volatilized when it meets the hot air flow; the feed flow rate of the peristaltic pump is 15-50mL/h, and the inlet temperature is 100 ~500°C; the nozzle diameter of the spray dryer is 0.7mm, and the hot gas flow is nitrogen.

Preferably, the chitosan in step (1) has a molecular weight of 1-300,000 and a deacetylation degree of 60-98%; the mass concentration of the chitosan solution is 0.2-3.0%.

Preferably, the mass ratio of chitosan to halloysite nanotubes in step (1) is 1.00:(0.05-50.00).

Preferably, the organic precipitant in step (1) is one of absolute ethanol, acetone, ether and tetrahydrofuran; the mass ratio of the organic precipitant to the chitosan solution is (2-5):1.

Preferably, the linking agent described in step (1) is one of glyoxal, glutaraldehyde, sodium tripolyphosphate, sodium tetrapolyphosphate and genipin; the mass ratio of linking agent and chitosan is (0～0.1):1.0.

More preferably, the mass ratio of the cross-linking agent to chitosan is (0.03-0.10):1.00.

Preferably, the speed of centrifugation in step (2) is 3000rpm, and the time is 30min.

The composite hemostatic powder of chitosan and halloysite nanotube obtained by the above preparation method.

Compared with the prior art, the present invention has the following advantages and beneficial effects:

The preparation method of the present invention is simple, combines the advantages of chitosan and inorganic hemostatic materials, the hemostatic effect of the prepared composite hemostatic powder is good, on the one hand overcomes the lower gel strength and gelation of chitosan hemostatic materials On the other hand, it overcomes the problem of poor biocompatibility caused by traditional inorganic hemostatic materials.

Detailed ways

The present invention will be further described in detail below in conjunction with examples, but the embodiments of the present invention are not limited thereto.

Example 1

A preparation method of a composite hemostatic powder of chitosan and halloysite nanotubes, comprising the following steps:

(1) Chitosan (chitosan molecular weight is 50,000, deacetylation degree is 91%) is dissolved in acetic acid aqueous solution and is mixed with the chitosan solution that mass concentration is 2%, then adds halloysite nanotube (shell The mass ratio of polysaccharides to halloysite nanotubes is 1:2), and after stirring at room temperature for 24 hours, adding 0.1M aqueous sodium hydroxide solution to adjust the pH to 7 to prepare a suspension;

(2) The suspension obtained in step (1) was centrifuged at a speed of 3000rpm for 30min, after the gained precipitate was washed with deionized water, the operation of centrifugation (3000rpm, 30min) and deionized water washing was repeated for 3 times, and then The precipitate is freeze-dried (the temperature of the freezer is -80 degrees Celsius, and the pressure of the drying chamber is 100 Pa), and the composite hemostatic powder of the chitosan and halloysite nanotubes is obtained.

The composite hemostatic powder of chitosan and halloysite nanotubes obtained in the present embodiment was subjected to a rabbit hemostatic test: select 3 New Zealand white rabbits, after routine operation anesthesia, the artery of the thigh was cut, and the composite hemostatic powder was applied, and recorded The hemostasis time was 19.8±1.4 seconds. The results show that the compound hemostatic powder prepared according to the invention has good hemostatic effect.

Example 2

A preparation method of a composite hemostatic powder of chitosan and halloysite nanotubes, comprising the following steps:

(1) Chitosan (chitosan molecular weight is 50,000, deacetylation degree is 60%) is dissolved in acetic acid aqueous solution and is mixed with the chitosan solution that mass concentration is 3%, then adds halloysite nanotube (shell The mass ratio of polysaccharide and halloysite nanotube is 1:50), and after stirring at room temperature for 24h, adding acetone makes suspension (the mass ratio of acetone and chitosan solution is 1:1);

(2) The suspension obtained in step (1) was centrifuged at a speed of 3000rpm for 30min, after the gained precipitate was washed with deionized water, the operation of centrifugation (3000rpm, 30min) and deionized water washing was repeated for 3 times, and then The precipitate is freeze-dried (the temperature of the freezer is -80 degrees Celsius, and the pressure of the drying chamber is 100 Pa), and the composite hemostatic powder of the chitosan and halloysite nanotubes is obtained.

The composite hemostatic powder of chitosan and halloysite nanotubes obtained in the present embodiment was subjected to a rabbit hemostatic test: select 3 New Zealand white rabbits, after routine operation anesthesia, the artery of the thigh was cut, and the composite hemostatic powder was applied, and recorded The hemostasis time was 23.8±2.1 seconds. The results show that the compound hemostatic powder prepared according to the invention has good hemostatic effect.

Example 3

A preparation method of a composite hemostatic powder of chitosan and halloysite nanotubes, comprising the following steps:

(1) Chitosan (chitosan molecular weight is 300,000, deacetylation degree is 98%) is dissolved in the acetic acid aqueous solution and is mixed with the chitosan solution that mass concentration is 0.2%, then adds halloysite nanotube (shell The mass ratio of polysaccharide to halloysite nanotubes is 1:20), and after stirring at room temperature for 24 hours, add sodium tripolyphosphate to obtain a suspension (the mass ratio of sodium tripolyphosphate and chitosan is 0.1:1) ;

(2) Then use a spray drying instrument to spray the suspension prepared in step (1) by a peristaltic pump, meet the hot air flow, and volatilize the solvent to obtain the composite hemostatic powder of chitosan and halloysite nanotubes ; The technical parameters of the spray drying are: the feed flow rate of the peristaltic pump is 20mL/h., the inlet temperature is 300 degrees Celsius; the nozzle aperture is 0.7mm; the hot gas flow is nitrogen.

The composite hemostatic powder of chitosan and halloysite nanotubes obtained in the present embodiment was subjected to a rabbit hemostatic test: select 3 New Zealand white rabbits, after routine operation anesthesia, the artery of the thigh was cut, and the composite hemostatic powder was applied, and recorded The hemostasis time was 18.7±1.8 seconds. The results show that the compound hemostatic powder prepared according to the invention has good hemostatic effect.

Example 4

A preparation method of a composite hemostatic powder of chitosan and halloysite nanotubes, comprising the following steps:

(1) Chitosan (chitosan molecular weight is 100,000, deacetylation degree is 85%) is dissolved in acetic acid aqueous solution and is mixed with the chitosan solution that mass concentration is 3%, then adds halloysite nanotube (shell The mass ratio of polysaccharide to halloysite nanotubes is 1:1), and after stirring at room temperature for 24 hours, sodium tripolyphosphate was added to obtain a suspension (the mass ratio of sodium tripolyphosphate and chitosan was 0.3:1) ;

(2) The suspension obtained in step (1) was centrifuged at a speed of 3000rpm for 30min, after the gained precipitate was washed with deionized water, the operation of centrifugation (3000rpm, 30min) and deionized water washing was repeated for 3 times, and then The precipitate is freeze-dried (the temperature of the freezer is -80 degrees Celsius, and the pressure of the drying chamber is 100 Pa), and the composite hemostatic powder of the chitosan and halloysite nanotubes is obtained.

The composite hemostatic powder of chitosan and halloysite nanotubes obtained in this embodiment was subjected to a rabbit hemostatic test: select 3 New Zealand white rabbits, after routine operation anesthesia, the artery of the thigh was cut off, and the composite hemostatic powder was applied, and recorded The hemostasis time was 25.7±3.8 seconds. The results show that the compound hemostatic powder prepared according to the invention has good hemostatic effect.

The above-mentioned embodiment is a preferred embodiment of the present invention, but the embodiment of the present invention is not limited by the above-mentioned embodiment, and any other changes, modifications, substitutions, combinations, Simplifications should be equivalent replacement methods, and all are included in the protection scope of the present invention.

Claims (10)

1. a preparation method of the composite hemostatic powder of chitosan and halloysite nanotube, is characterized in that, comprises the following steps: (1) Dissolve chitosan in acetic acid aqueous solution to prepare a chitosan solution, then add halloysite nanotubes, and stir at room temperature for 20-30 hours, then add aqueous sodium hydroxide solution to adjust the pH to 7 to prepare a suspension , or adding an organic precipitant to obtain a suspension, or adding a cross-linking agent to obtain a suspension; (2) Centrifuge the suspension obtained in step (1), wash the obtained precipitate with deionized water, repeat the operation of centrifugal separation and deionized water washing for 2 to 5 times, and then freeze-dry the precipitate to obtain the described Composite hemostatic powder of chitosan and halloysite nanotubes. 2. preparation method according to claim 1, is characterized in that, when preparing suspension by adding the mode of cross-linking agent in step (1), remove solvent by the mode of spray drying after obtaining suspension, obtain obtained The composite hemostatic powder of chitosan and halloysite nanotubes. 3. preparation method according to claim 2, is characterized in that, when carrying out spray-drying, by peristaltic pump, suspension is pumped into spray dryer spray, meets hot air flow and solvent is volatilized; The feed flow rate of peristaltic pump 15-50mL/h, the inlet temperature is 100-500°C; the nozzle aperture of the spray dryer is 0.7mm, and the hot gas flow is nitrogen. 4. preparation method according to claim 1 is characterized in that, the molecular weight of chitosan described in step (1) is 1～300,000, and deacetylation degree is 60～98%; The quality of described chitosan solution The concentration is 0.2-3.0%. 5. The preparation method according to claim 1, characterized in that the mass ratio of chitosan and halloysite nanotubes in step (1) is 1.00:(0.05～50.00). 6. preparation method according to claim 1, is characterized in that, the organic precipitating agent described in step (1) is the one in dehydrated alcohol, acetone, ether and tetrahydrofuran; The quality of organic precipitating agent and chitosan solution The ratio is (2～5):1. 7. preparation method according to claim 1, is characterized in that, step (1) described linking agent is one in glyoxal, glutaraldehyde, sodium tripolyphosphate, sodium tetrapolyphosphate and genipin species; the mass ratio of cross-linking agent to chitosan is (0～0.1):1.0. 8. preparation method according to claim 7, is characterized in that, the mass ratio of described linking agent and chitosan is (0.03～0.10):1.00. 9. The preparation method according to claim 1, characterized in that, the speed of centrifugation in step (2) is 3000rpm, and the time is 30min. 10. The composite hemostatic powder of chitosan and halloysite nanotube obtained by the preparation method described in any one of claims 1 to 9.