CN103230617A - Collagen/chitosan micro-nano fiber composite hemostatic membrane material and preparation method thereof - Google Patents

Abstract

The invention discloses a collagen/chitosan micro-nano fiber composite hemostatic film material and a preparation method thereof, which is characterized in that type I collagen is extracted and prepared from medical biological skin slices by using ultrasonic technology. Mix hexafluoroisopropanol and acetic acid in different volume ratios to prepare a spinning solvent, then mix the prepared type I collagen with a certain mass ratio of chitosan and add it to the spinning solvent. Stir in medium until transparent, prepare an electrospinning mother liquor with a concentration of 4% to 10%, inject the electrospinning mother liquor into an electrospinning machine for electrospinning, and obtain a collagen/chitosan micro-nano fiber composite film. Next, the collagen/chitosan micro-nano fiber composite membrane is soaked in natural biological macromolecules and Chinese herbal medicine successively, and finally freeze-dried to prepare the collagen/chitosan micro-nano fiber composite hemostatic membrane material. The material has excellent properties such as good biocompatibility, biodegradability, and adhesion. It can quickly stop bleeding, anti-inflammatory and analgesic, and promote wound healing. It can be used for wound hemostatic repair and general civil hemostasis first aid.

Description

A collagen/chitosan micro-nano fiber composite hemostatic membrane material and preparation method thereof

technical field

The invention relates to a hemostatic agent which has excellent properties such as good biocompatibility, biodegradability and adhesion, and can quickly stop bleeding, anti-inflammatory and analgesic, and promote wound compounding. the

Background technique

Bleeding poses a great danger to human life, and the search for fast and effective hemostatic agents has always been a research hotspot at home and abroad. In various surgical operations or trauma treatments, wound bleeding and isolation from the outside world to avoid infection have a great impact on the wound healing of patients. In war, according to statistics, bleeding is the main cause of death within 48 hours after injury, accounting for 80% of all traumatic accidents. Excessive traumatic blood loss will cause pain, shock, coma and even death of the wounded. Therefore, hemostatic materials need to stop bleeding quickly and effectively, and must also have anti-inflammatory and analgesic effects, and promote wound healing. the

Traditional hemostatic materials are mainly first aid kits, four-head bands, tourniquets and bandages, etc. These materials are not easy to carry and sterilized and stored, and the hemostasis is almost ineffective by mechanical action. In recent years, people have developed a class of bioabsorbable hemostatic materials, among which collagen-based hemostatic materials have attracted widespread attention. Collagen has good biocompatibility, biodegradability, low toxicity, weak antigenicity, and can activate cells. The expression of characteristic groups maintains the normal characteristic expression of cells, which is beneficial to the adhesion, growth, proliferation and differentiation of cells, and its hemostatic performance is also outstanding, which has been reported in a large number of literatures. the

However, Wei Hua and others presided over and undertook the "Tenth Five-Year" National High-tech Research and Development Program (863 Program) project, and developed a "medical bio-skin patch" with excellent comprehensive performance, which has been industrialized. This "medical bio-skin" has good biocompatibility and good hemostatic performance. Liang Peihong et al. used pig tendon collagen and chitosan to polymerize in a certain proportion, cross-linked with glutaraldehyde, and freeze-dried to make a composite sponge. Studies have shown that its hemostasis time is 2 minutes, which can promote wound healing [(Liang Peihong, Ye Chunting, Li Siming, etc. Animal experiments on wound hemostasis with composite type Ι collagen sponge [J]. Journal of Trauma Surgery, 2002, 4(5): 274-275)]. However, if the above-mentioned materials or similar collagen-based products are not cross-linked with glutaraldehyde, the mechanical properties cannot meet the requirements. After cross-linking, the biocompatibility of the materials will have problems again; Good removal will cause certain immunogenicity, which will cause discomfort to the wounded, and this type of product cannot relieve the pain of the wounded, and it is difficult to meet people's requirements for such products. In order to solve the problem of the single function of the above-mentioned materials, the materials can be compounded with some anti-inflammatory and analgesic traditional Chinese medicines. Many scholars have done this type of research. Zhu Chuhong and others organically compounded collagen, chitosan and chrysophanol, an active ingredient in rhubarb, and prepared a new type of dressing with multiple functions such as rapid hemostasis, anti-inflammatory and analgesic [(Zhu Chuhong, Jiang Bo, Zeng Wen , Li Li, Mi Jianhong. A new multifunctional hemostatic dressing. Application publication number: CN 102258802A)]. It can be seen that the previous hemostatic materials still have disadvantages such as long hemostatic time and poor hemostatic effect, and even some hemostatic materials have certain immunogenicity. the

In summary, there are following deficiencies in the prior art:

1. The existing collagen-based hemostatic agent has insufficient mechanical properties and poor adhesion, which causes unnecessary troubles to the operation;

2. The existing collagen-based hemostatic agents have general hemostatic performance and single function, which can no longer meet the requirements of the market;

3. The type I collagen in the existing collagen-based hemostatic agent monomer material is not pure enough, and there is also a certain degree of immunogenicity, which may cause immeasurable harm to patients when used in clinical practice.

Contents of the invention

The object of the present invention is to provide a collagen/chitosan micro-nano fiber composite hemostatic membrane material for the deficiencies of the prior art. Under the action of ultrasonic waves, the material uses medical biological skin sheets as raw materials to extract type I collagen. The obtained type I collagen has higher purity, higher extraction rate, and lower antigenicity. After electrospinning, compounding natural polymers and The active ingredient of Panax notoginseng is used to prepare the collagen/chitosan micro-nano fiber composite hemostatic membrane material. Using this material can achieve rapid and effective hemostasis, and also has the advantages of outstanding mechanical properties, good adhesion, anti-inflammatory and analgesic, and promotion of wound healing. the

The technical solution adopted to achieve the above-mentioned purpose of the present invention is such that a collagen/chitosan micro-nano fiber composite hemostatic film material can quickly and effectively stop bleeding, reduce inflammation and analgesia, and can promote wound healing. Yes, the preparation method steps are as follows:

(1) Type Ⅰ collagen extraction: Take a certain amount of medical biological skin, cut it into small pieces of 0.5cm×0.5cm, wash with ultrapure water for 3 to 5 times, and then soak in 0.05mol/L Tris, 1mol /L NaCl, in a Tris-NaCl buffer solution with a pH of 7.5, place it in an ultrasonic cleaner with a frequency of 20-40kHz, and act for 2-4 hours; pour off the buffer solution, and rinse the skin with distilled water for 2-5 times; add 30-50 times of 0.5-1M acetic acid solution, soaked for 2-4 hours, crushed and homogenized with a homogenizer at a constant temperature of 4°C, then transferred to the reaction kettle, and added the corresponding quality of gastric juice at 2% of the tare weight Protease; under ultrasonic conditions, 4°C constant temperature and slow stirring enzymolysis for 24-36 hours, every 3-5 hours for 1-2 hours; The concentration is 1.5mol/L ammonium sulfate powder, let it stand for 10-20h; centrifuge the collagen solution at 10000-20000rpm for 10-30min, remove the supernatant, dissolve the precipitate in 0.1-0.5M acetic acid solution, 10000-20000rpm Centrifuge for 10-30 minutes, remove the supernatant, dissolve and centrifuge repeatedly for 3-5 times, and finally wash the precipitate with ultrapure water, centrifuge, and freeze-dry to obtain high-purity type I collagen;

(2) Preparation of collagen/chitosan micro-nano fiber composite membrane by electrospinning: mix hexafluoroisopropanol and acetic acid in different volume ratios, the volume percentage of hexafluoroisopropanol is 90% to 99%, and the above I Type collagen is mixed with a certain proportion of chitosan, stirred under ultrasonic waves at room temperature until transparent, and prepared into a 4% to 10% mixed solution, which is the electrospinning mother solution, wherein the mass percentage of chitosan is 10% to 50% %; Inject the electrospinning mother liquor into the electrospinning machine and carry out electrospinning to obtain a collagen/chitosan micro-nano fiber composite film;

(3) Extraction of Panax notoginseng, the active ingredient of Panax notoginseng powder: Under ultrasonic conditions, soak Sanqi powder in 10-20 times of 95% ethanol solution for 15-20 hours, then dip in 5-10 times of 95% ethanol solution Put in ethanol solution for 5-10 hours, after standing still, combine the ethanol extract twice, and keep it for other use; dry the residue, add 10-15 times of water to extract for 15-20 hours; then soak in 2-5 times of water for 6-20 hours After 10 hours, centrifuge to obtain the filtrate, concentrate the filtrate, and dry to obtain crude notoginseng powder;

(4) Preparation of collagen/chitosan micro-nanofiber composite hemostatic membrane material: Under ultrasonic conditions, soak the above-prepared collagen/chitosan micro-nanofiber composite membrane in polyethylene glycol, alginate , hyaluronic acid, thrombin, fibrin, or one or more of chondroitin sulfate; then impregnated in the crude notoginsin prepared by the above method; finally freeze-dried, the dose is 6-30KGy/ The gamma ray produced by h ⁶⁰ Co is sterilized and sterilized, and then molded and packaged, which is the finished product.

In the above preparation method, pepsin activity of 3000u/g was used in step (1), which was purchased from Sigma; hexafluoroisopropanol was analytically pure in step (2), and was purchased from Beijing Huawei Raycus; step (2) Chitosan with different degrees of deacetylation is prepared by controlling the degree of deacetylation of chitin; the concentration and soaking time of the soaking solution in step (4) vary depending on the material, such as 2% sodium alginate, soaking The time is 10 to 20 hours, etc., and the soaking temperature is room temperature. the

As claimed in claim 1, the performance parameter index determination method that the collagen/chitosan micro-nano fiber composite hemostatic membrane material has is as follows:

The method for measuring the hemostatic performance of the collagen/chitosan micro-nano fiber composite hemostatic membrane material as described in the claims is as follows:

Take a glass clean test tube with an inner diameter of 8 mm, add 0.1 g of the above materials, and then add 0.1 ml of calcium chloride; shake well, add 0.5 ml of blood into the test tube, shake gently, mix well, immediately record the time, and place in a water bath at 37 °C Observe, gently tilt the test tube once every 30s, and observe whether there is blood coagulation. From the time when blood is added to the time when the blood clot in the inclined test tube coagulates at the bottom of the tube and does not flow, that is, the coagulation time, repeat six times.

The present invention has the following advantages:

(1) Complementary and synergistic properties between monomers of composite materials: Combining material monomers together can give full play to the advantages of material monomers themselves, and composite materials can produce material monomers that do not have or have better performance than monomers. Improved overall performance. For example, the collagen/chitosan micro-nano fiber composite film prepared by electrospinning is composited with natural polymer materials and the active ingredients of Panax notoginseng. The composite material has good biocompatibility, good biodegradability, and promotes Wound healing, etc., not only has the excellent hemostatic properties, antibacterial/bacteriostatic properties of chitosan, but also has the hemostatic, anti-inflammatory and analgesic effects of Panax notoginseng. After compounding, the mechanical properties, hemostatic performance, and wound healing ability of the material have been improved, and it has become a hemostatic agent with excellent comprehensive performance;

(2) The collagen/chitosan micro-nano fiber composite membrane prepared by electrospinning technology enhances the mechanical properties and hemostatic properties of collagen without adding a cross-linking agent;

(3) The type I collagen in the composite material is extracted from medical bio-skin slices under ultrasonic conditions by acid-enzyme combination method. The obtained collagen has higher purity and extraction rate, and lower immunogenicity;

(4) Ultrasound is used throughout the preparation process, which fully exerts the cavitation effect, mechanical effect, and thermal effect of ultrasonic waves, which has a great impact on the preparation of materials;

(5) The prepared collagen-based micro-nano fiber composite membrane material has excellent comprehensive properties, diverse functions, and is very convenient to carry, use and store.

Description of drawings

Figure 1. SEM scanning electron micrographs of collagen/chitosan micro-nanofiber composite membrane. the

Detailed ways

The present invention will be specifically described below by implementing it. It is necessary to point out that this embodiment is only used for further illustration of the present invention, and can not be interpreted as limiting the protection scope of the present invention. Those skilled in the art can use the above-mentioned invention Make non-essential improvements and adjustments to the content. the

Example 1

(1) Type I collagen extraction: Take 10 parts by weight of medical biological skin, cut it into small pieces of 0.5cm×0.5cm, wash it with ultrapure water for 5 times, and then soak it in 0.05mol/L Tris, 1mol/L L NaCl, in a Tris-NaCl buffer solution with a pH of 7.5, put it in an ultrasonic cleaner with a frequency of 20 kHz, and act for 2 hours; pour off the buffer solution, and rinse the skin twice with distilled water; add 30 times of 0.5M acetic acid solution, soaked for 2 hours, crushed and homogenized with a homogenizer at a constant temperature of 4°C, and then transferred to a reaction kettle, adding 0.2 parts by weight of pepsin; under ultrasonic conditions with a frequency of 30kHz, slowly stirred at 4°C Enzyme hydrolysis for 36 hours, and act for 1 hour every 3 hours; after the reaction is completed, stop stirring, filter the reaction solution, adjust the pH of the filtrate to 7.0, add ammonium sulfate powder with a final concentration of 1.5mol/L, and let it stand for 10 hours; put the collagen solution at 10000rpm Centrifuge for 30min, remove the supernatant, dissolve the precipitate in 0.5M acetic acid solution, centrifuge at 10000rpm for 30min, remove the supernatant, dissolve and centrifuge 5 times repeatedly, finally wash the precipitate with ultrapure water, centrifuge, and freeze-dry to obtain High purity type I collagen;

(2) Preparation of collagen/chitosan micro-nano fiber composite membrane by electrospinning: Mix 49 parts by volume of hexafluoroisopropanol with 1 part by volume of acetic acid and stir evenly; mix 4.5 parts by weight of type I collagen and 0.5 parts by weight Parts of chitosan with a deacetylation degree of 80% were dissolved in 10% collagen/chitosan mixed solution, stirred at room temperature under ultrasonic conditions with a frequency of 20kHz until transparent, and made into an electrospinning mother solution ; Inject the electrospinning mother liquor into the electrospinning machine, and perform electrospinning under the conditions of a voltage of 18kv, a spinning rate of 0.03mm/min, and a receiving distance of 10cm to obtain a collagen/chitosan micro-nano fiber composite film;

(3) Extraction of notoginseng, the active ingredient of notoginseng powder: Under ultrasonic conditions with a frequency of 30kHz, 10 parts by weight of notoginseng powder was immersed in 200 parts by volume of 95% ethanol solution for 20 hours, and then immersed in 100 parts by volume Parts of 95% ethanol solution for 10h, after standing still, combine the alcohol extract twice, and keep it for other use; dry the residue, add 150 parts by volume of ultrapure water for leaching for 20h; then use 50 parts by volume of ultrapure water Immerse for 10 hours, centrifuge to obtain the filtrate, concentrate the filtrate, and dry to obtain crude notoginseng powder;

(4) Collagen/chitosan micro-nano fiber composite hemostatic membrane material: Under the condition of ultrasonic frequency of 20kHz, soak the collagen-based micro-nano fiber composite membrane prepared above in 2% sodium alginate and 0.1M calcium chloride and then soaked in the crude notoginsin prepared by the above method for 8 hours; finally freeze-dried, sterilized and sterilized by γ-rays produced by 15KGy/h ⁶⁰ Co, and molded and packaged, which is the finished product.

The main parameter properties of the collagen/chitosan micro-nano fiber composite hemostatic membrane material prepared by the above method are as follows:

Example 2

(1) Type I collagen extraction: Take 10 parts by weight of medical biological skin, cut it into small pieces of 0.5cm×0.5cm, wash it with ultrapure water for 5 times, and then soak it in 0.05mol/L Tris, 1mol/L L NaCl, in a Tris-NaCl buffer solution with a pH of 7.5, put it in an ultrasonic cleaner with a frequency of 30 kHz, and act for 2 hours; pour off the buffer solution, rinse the skin twice with distilled water; add 40 times of 0.5M acetic acid Solution, soaked for 2 hours, crushed and homogenized with a homogenizer at a constant temperature of 4°C, then transferred to a reaction kettle, added 0.2 parts by weight of pepsin, and slowly stirred the enzyme at 4°C under ultrasonic conditions with a frequency of 30kHz Solution for 24 hours, every 3 hours for 1 hour. After the reaction is completed, stop stirring, filter the reaction solution with suction, adjust the pH of the filtrate to 7.5, add ammonium sulfate powder with a final concentration of 1.5mol/L, and let stand for 15h; centrifuge the collagen solution at 20000rpm for 10min, remove the supernatant, and The precipitate was dissolved in 0.5M acetic acid solution, centrifuged at 20,000 rpm for 10 minutes, the supernatant was removed, dissolved and centrifuged repeatedly for 5 times, and finally the precipitate was washed with ultrapure water, centrifuged, and freeze-dried to obtain high-purity type I collagen;

(2) Preparation of collagen/chitosan micro-nano fiber composite membrane by electrospinning: mix 45 parts by volume of hexafluoroisopropanol and 5 parts by volume of acetic acid, and stir evenly; mix 2 parts by weight of type I collagen and 1 part by weight Parts of chitosan with a deacetylation degree of 80% were dissolved in cellulose, prepared into a mixed solution of 6% collagen/chitosan, stirred at room temperature under ultrasonic conditions with a frequency of 20kHz until transparent, and made into an electrospinning mother solution ; Inject the electrospinning mother liquor into the electrospinning machine, and perform electrospinning under the conditions of voltage 20kv, spinning rate 0.01mm/min, and receiving distance 5cm, to obtain collagen/chitosan micro-nano fiber composite membrane;

(3) Extraction of notoginseng, the active ingredient of notoginseng powder: Under ultrasonic conditions with a frequency of 30 kHz, 10 parts by weight of notoginseng powder was immersed in 150 parts by volume of 95% ethanol solution for 15 hours, and then immersed in 100 volume parts part of 95% ethanol solution for 6 hours, after standing still, combine the two ethanol extracts, and keep it for other use; dry the residue, add 200 parts by volume of ultrapure water for leaching for 20 hours; then use 150 parts by volume of ultrapure water Immerse for 15 hours, centrifuge to obtain the filtrate, concentrate the filtrate, and dry to obtain crude notoginseng powder;

(4) Preparation of collagen/chitosan micro-nanofiber composite hemostatic membrane material: Under ultrasonic conditions with a frequency of 25 kHz, soak the collagen-based micro-nanofiber composite membrane prepared above in 2% sodium alginate, 4% polystyrene In the mixture of ethylene glycol and 0.1M calcium chloride for 8 hours; then immersed in the crude notoginsin prepared by the above method for 6 hours; finally freeze-dried, the dose is 20KGy/h γ-ray sterilization produced by ⁶⁰ Co , Formed and packaged, that is the finished product.

The main parameter properties of the collagen/chitosan micro-nano fiber composite hemostatic membrane material prepared by the above method are as follows:

Example 3

(1) Type I collagen extraction: Take 10 parts by weight of medical biological skin, cut it into small pieces of 0.5cm×0.5cm, wash it with ultrapure water for 5 times, and then soak it in 0.05mol/L Tris, 1mol/L L NaCl, in a Tris-NaCl buffer solution with a pH of 7.5, place it in an ultrasonic cleaner with a frequency of 30kHz, and act for 2h; pour off the buffer solution, rinse the skin twice with distilled water; add 50 times of 0.5M acetic acid solution, soaked for 2 hours, crushed and homogenized with a homogenizer at a constant temperature of 4°C, and then transferred to a reaction kettle, adding 0.2 parts by weight of pepsin; under the condition of ultrasonic waves with a frequency of 20kHz, the enzyme was slowly stirred at 4°C Decompose for 36 hours, and act for 1 hour every 3 hours; after the reaction is completed, stop stirring, filter the reaction solution with suction, adjust the pH of the filtrate to 7.3, add ammonium sulfate powder with a final concentration of 1.5mol/L, and let it stand for 20 hours. Centrifuge the collagen solution at 15000rpm for 20min, remove the supernatant, dissolve the precipitate in 0.5M acetic acid solution, centrifuge at 15000rpm for 20min, remove the supernatant, dissolve and centrifuge 5 times repeatedly, and finally wash the precipitate with ultrapure water, centrifuge, Freeze-drying to obtain high-purity type I collagen;

(2) Preparation of collagen/chitosan micro-nano fiber composite membrane by electrospinning: Mix 48 parts by volume of hexafluoroisopropanol with 2 parts by volume of acetic acid and stir evenly; mix 2.5 parts by weight of type I collagen and 0.5 parts by weight Parts of chitosan with a deacetylation degree of 70% were dissolved in cellulose, prepared into a mixed solution of 6% collagen/chitosan, stirred under ultrasonic conditions at room temperature until transparent, and made into an electrospinning mother liquor; The silk mother liquor was injected into an electrospinning machine, and electrospinning was performed under the conditions of a voltage of 19kv, a spinning rate of 0.05mm/min, and a receiving distance of 10cm to obtain a collagen/chitosan micro-nano fiber composite film;

(3) Extraction of Panax notoginseng, the active ingredient of Panax notoginseng powder: Under ultrasonic conditions with a frequency of 25kHz, 10 parts by weight of Notoginseng powder was immersed in 200 parts by volume of 95% ethanol solution for 15 hours, and then immersed in 100 parts by volume part of 95% ethanol solution for 10 hours, after standing still, combine the alcohol extracts twice, and keep them for other use; dry the residue, add 150 parts by volume of ultrapure water for extraction for 15 hours; then use 100 parts by volume of ultrapure water Immerse for 10 hours, centrifuge to obtain the filtrate, concentrate the filtrate, and dry to obtain crude notoginseng powder;

(4) Preparation of collagen/chitosan micro-nano fiber composite hemostatic membrane material: Under ultrasonic conditions with a frequency of 25 kHz, soak the collagen-based micro-nano fiber composite membrane prepared above in 3% hyaluronic acid and 1% sulfuric acid Chondroitin in a 40% ethanol mixed solution for 8 hours; then soaked in the crude notoginseng prepared by the above method for 8 hours; finally freeze-dried, sterilized by gamma rays produced by 30KGy/h ⁶⁰ Co, and molded and packaged , which is the finished product.

The main parameter properties of the collagen/chitosan micro-nano fiber composite hemostatic membrane material prepared by the above method are as follows:

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Claims (8)

1. A collagen/chitosan micro-nano fiber composite hemostatic membrane material, characterized in that the composite hemostatic membrane material possesses the following physical and chemical properties: (1) Sodium chloride content: ≤3% (m/m); (2) Heavy metal content: ≤10μg/g(m/m); (3) Hydroxyproline content: not less than 8% of the total protein content (m/m); (4) Cytotoxicity: The cytotoxic reaction is not greater than Grade 1. 2. the preparation method of collagen/chitosan micro-nano fiber composite hemostatic membrane material as claimed in claim 1, is characterized in that, comprises the steps: (1) Type Ⅰ collagen extraction: Take a certain amount of medical biological skin, cut it into small pieces of 0.5cm×0.5cm, wash with ultrapure water for 3 to 5 times, and then soak in 0.05mol/L Tris, 1mol /L NaCl, in a Tris-NaCl buffer solution with a pH of 7.5, place it in an ultrasonic cleaner with a frequency of 20-40kHz, and act for 2-4 hours; pour off the buffer solution, and rinse the skin with distilled water for 2-5 times; add 30-50 times of 0.5-1M acetic acid solution, soaked for 2-4 hours, crushed and homogenized with a homogenizer at a constant temperature of 4°C, then transferred to the reaction kettle, and added the corresponding quality of gastric juice at 2% of the tare weight Protease; under ultrasonic conditions, 4°C constant temperature and slow stirring enzymolysis for 24-36 hours, every 3-5 hours for 1-2 hours; The concentration is 1.5mol/L ammonium sulfate powder, let it stand for 10-20h; centrifuge the collagen solution at 10000-20000rpm for 10-30min, remove the supernatant, dissolve the precipitate in 0.1-0.5M acetic acid solution, 10000-20000rpm Centrifuge for 10-30 minutes, remove the supernatant, dissolve and centrifuge repeatedly for 3-5 times, and finally wash the precipitate with ultrapure water, centrifuge, and freeze-dry to obtain high-purity type I collagen; (2) Preparation of collagen/chitosan micro-nano fiber composite membrane by electrospinning: mix hexafluoroisopropanol and acetic acid in different volume ratios, the volume percentage of hexafluoroisopropanol is 90% to 99%, and the above I Type collagen is mixed with a certain proportion of chitosan, stirred under ultrasonic waves at room temperature until transparent, and prepared into a 4% to 10% mixed solution, which is the electrospinning mother solution, wherein the mass percentage of chitosan is 10% to 50% %; Inject the electrospinning mother liquor into the electrospinning machine and carry out electrospinning to obtain a collagen/chitosan micro-nano fiber composite film; (3) Extraction of Panax notoginseng, the active ingredient of Panax notoginseng powder: Under ultrasonic conditions, soak Sanqi powder in 10-20 times of 95% ethanol solution for 15-20 hours, then dip in 5-10 times of 95% ethanol solution Put in ethanol solution for 5-10 hours, after standing still, combine the ethanol extract twice, and keep it for other use; dry the residue, add 10-15 times of water to extract for 15-20 hours; then soak in 2-5 times of water for 6-20 hours After 10 hours, centrifuge to obtain the filtrate, concentrate the filtrate, and dry to obtain crude notoginseng powder; (4) Preparation of collagen/chitosan micro-nanofiber composite hemostatic membrane material: Under ultrasonic conditions, soak the above-prepared collagen/chitosan micro-nanofiber composite membrane in polyethylene glycol, alginate , hyaluronic acid, thrombin, fibrin, or one or more of chondroitin sulfate; then impregnated in the crude notoginsin prepared by the above method; finally freeze-dried, the dose is 6-30KGy/ The gamma ray produced by h ⁶⁰ Co is sterilized and sterilized, and then molded and packaged, which is the finished product. 3. The ultrasonic frequency of implementing described in claim 2 is 20～40 kHz, and power is 50～120w. 4. The preparation method of the collagen/chitosan micro-nano fiber composite hemostatic membrane material according to claim 2, characterized in that the purity of the type I collagen is ≥95%, and the molecular weight is about 300,000. 5. The preparation method of the collagen/chitosan micro-nano fiber composite membrane according to claim 2, characterized in that, the electrospinning conditions are: voltage 15-25kv, solution flow rate 0.01-0.3mm/min , The receiving distance is 5-25cm; the apparent viscosity of the electrospinning mother liquor is 10-120mPa ^. s, the measurement temperature is 20°C, and the rotation speed is 32r/min. 6. the preparation method of collagen/chitosan micro-nano fiber composite hemostatic membrane material as claimed in claim 2, is characterized in that, after described collagen/chitosan micro-nano fiber composite membrane is made, soak in phosphate In the buffer solution for 2 days, remove the hexafluoroisopropanol remaining in the membrane, and change the buffer solution every 4 hours during this period. 7. The preparation method of the collagen/chitosan micro-nano fiber composite hemostatic membrane material according to claim 2, characterized in that the deacetylation degree of the chitosan is 60%-80%. 8. The preparation method of the collagen/chitosan micro-nano fiber composite hemostatic membrane material according to claim 2, characterized in that, the extraction of type I collagen takes medical biological skin as a raw material, and under the condition of ultrasonic waves, The type I collagen is extracted by the acid enzyme combination method, and the type I collagen obtained in this way is more pure and has a higher extraction rate.

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