CN111921001A - Film for wound healing and preparation method thereof - Google Patents
Film for wound healing and preparation method thereof Download PDFInfo
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- CN111921001A CN111921001A CN202010839123.XA CN202010839123A CN111921001A CN 111921001 A CN111921001 A CN 111921001A CN 202010839123 A CN202010839123 A CN 202010839123A CN 111921001 A CN111921001 A CN 111921001A
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- wound healing
- film
- chitosan
- polyaspartic acid
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- 230000029663 wound healing Effects 0.000 title claims abstract description 46
- 238000002360 preparation method Methods 0.000 title claims abstract description 27
- 229920000805 Polyaspartic acid Polymers 0.000 claims abstract description 55
- 108010064470 polyaspartate Proteins 0.000 claims abstract description 55
- 229920001661 Chitosan Polymers 0.000 claims abstract description 42
- 239000003053 toxin Substances 0.000 claims abstract description 27
- 231100000765 toxin Toxicity 0.000 claims abstract description 27
- 241000219729 Lathyrus Species 0.000 claims abstract description 23
- 239000002994 raw material Substances 0.000 claims abstract description 10
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 59
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 31
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims description 30
- 238000009987 spinning Methods 0.000 claims description 19
- 239000003054 catalyst Substances 0.000 claims description 11
- 238000006116 polymerization reaction Methods 0.000 claims description 11
- 238000002156 mixing Methods 0.000 claims description 10
- 238000010041 electrostatic spinning Methods 0.000 claims description 9
- 238000000034 method Methods 0.000 claims description 9
- 239000012528 membrane Substances 0.000 claims description 8
- 239000000203 mixture Substances 0.000 claims description 7
- 238000009210 therapy by ultrasound Methods 0.000 claims description 7
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 claims description 6
- 229910000147 aluminium phosphate Inorganic materials 0.000 claims description 3
- 239000012298 atmosphere Substances 0.000 claims description 3
- 239000011261 inert gas Substances 0.000 claims description 3
- 239000010408 film Substances 0.000 abstract description 39
- 230000002439 hemostatic effect Effects 0.000 abstract description 7
- 239000010409 thin film Substances 0.000 abstract description 4
- 208000001393 Lathyrism Diseases 0.000 abstract 1
- 239000000243 solution Substances 0.000 description 61
- 238000002386 leaching Methods 0.000 description 30
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 description 20
- 229960000583 acetic acid Drugs 0.000 description 19
- 244000111261 Mucuna pruriens Species 0.000 description 15
- 235000006161 Mucuna pruriens Nutrition 0.000 description 15
- 206010052428 Wound Diseases 0.000 description 14
- 208000027418 Wounds and injury Diseases 0.000 description 14
- 230000000740 bleeding effect Effects 0.000 description 10
- 239000012362 glacial acetic acid Substances 0.000 description 10
- 230000023597 hemostasis Effects 0.000 description 10
- 239000000706 filtrate Substances 0.000 description 9
- 238000006243 chemical reaction Methods 0.000 description 8
- 238000010828 elution Methods 0.000 description 8
- 229910052751 metal Inorganic materials 0.000 description 8
- 239000002184 metal Substances 0.000 description 8
- 238000001035 drying Methods 0.000 description 7
- 210000000952 spleen Anatomy 0.000 description 7
- 238000012360 testing method Methods 0.000 description 6
- 238000005341 cation exchange Methods 0.000 description 5
- 230000000052 comparative effect Effects 0.000 description 5
- 239000000835 fiber Substances 0.000 description 5
- 239000011259 mixed solution Substances 0.000 description 5
- 239000002121 nanofiber Substances 0.000 description 5
- 229920000642 polymer Polymers 0.000 description 5
- 241000283973 Oryctolagus cuniculus Species 0.000 description 4
- LPQOADBMXVRBNX-UHFFFAOYSA-N ac1ldcw0 Chemical compound Cl.C1CN(C)CCN1C1=C(F)C=C2C(=O)C(C(O)=O)=CN3CCSC1=C32 LPQOADBMXVRBNX-UHFFFAOYSA-N 0.000 description 4
- 239000003480 eluent Substances 0.000 description 4
- 239000012065 filter cake Substances 0.000 description 4
- 238000001914 filtration Methods 0.000 description 4
- 238000004128 high performance liquid chromatography Methods 0.000 description 4
- 238000011068 loading method Methods 0.000 description 4
- 238000000520 microinjection Methods 0.000 description 4
- 239000012299 nitrogen atmosphere Substances 0.000 description 4
- 238000010898 silica gel chromatography Methods 0.000 description 4
- 238000001179 sorption measurement Methods 0.000 description 4
- 230000009286 beneficial effect Effects 0.000 description 3
- 238000013168 hemostasis test Methods 0.000 description 3
- 239000002105 nanoparticle Substances 0.000 description 3
- 238000003825 pressing Methods 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 230000009278 visceral effect Effects 0.000 description 3
- 241000196324 Embryophyta Species 0.000 description 2
- NEEQFPMRODQIKX-REOHCLBHSA-N N(3)-oxalyl-L-2,3-diaminopropionic acid Chemical compound OC(=O)[C@@H](N)CNC(=O)C(O)=O NEEQFPMRODQIKX-REOHCLBHSA-N 0.000 description 2
- 229920002472 Starch Polymers 0.000 description 2
- 208000007536 Thrombosis Diseases 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 150000002500 ions Chemical class 0.000 description 2
- 210000004379 membrane Anatomy 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 210000000056 organ Anatomy 0.000 description 2
- 230000001737 promoting effect Effects 0.000 description 2
- SQGYOTSLMSWVJD-UHFFFAOYSA-N silver(I) nitrate Inorganic materials [Ag+].[O-]N(=O)=O SQGYOTSLMSWVJD-UHFFFAOYSA-N 0.000 description 2
- 235000019698 starch Nutrition 0.000 description 2
- 239000008107 starch Substances 0.000 description 2
- 210000001519 tissue Anatomy 0.000 description 2
- 206010002091 Anaesthesia Diseases 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- 241000220485 Fabaceae Species 0.000 description 1
- 241000122904 Mucuna Species 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 210000001015 abdomen Anatomy 0.000 description 1
- 230000003187 abdominal effect Effects 0.000 description 1
- 210000003489 abdominal muscle Anatomy 0.000 description 1
- 230000037005 anaesthesia Effects 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 238000004440 column chromatography Methods 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 239000006185 dispersion Substances 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 230000005611 electricity Effects 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- PNDPGZBMCMUPRI-UHFFFAOYSA-N iodine Chemical compound II PNDPGZBMCMUPRI-UHFFFAOYSA-N 0.000 description 1
- 239000004745 nonwoven fabric Substances 0.000 description 1
- 239000003960 organic solvent Substances 0.000 description 1
- 229960001412 pentobarbital Drugs 0.000 description 1
- WEXRUCMBJFQVBZ-UHFFFAOYSA-N pentobarbital Chemical compound CCCC(C)C1(CC)C(=O)NC(=O)NC1=O WEXRUCMBJFQVBZ-UHFFFAOYSA-N 0.000 description 1
- 210000004303 peritoneum Anatomy 0.000 description 1
- 238000010298 pulverizing process Methods 0.000 description 1
- 238000005086 pumping Methods 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 230000008439 repair process Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000012279 sodium borohydride Substances 0.000 description 1
- 229910000033 sodium borohydride Inorganic materials 0.000 description 1
- 230000003068 static effect Effects 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 210000003462 vein Anatomy 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L15/00—Chemical aspects of, or use of materials for, bandages, dressings or absorbent pads
- A61L15/16—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons
- A61L15/22—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons containing macromolecular materials
- A61L15/28—Polysaccharides or their derivatives
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L15/00—Chemical aspects of, or use of materials for, bandages, dressings or absorbent pads
- A61L15/16—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons
- A61L15/22—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons containing macromolecular materials
- A61L15/26—Macromolecular compounds obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds; Derivatives thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L15/00—Chemical aspects of, or use of materials for, bandages, dressings or absorbent pads
- A61L15/16—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons
- A61L15/42—Use of materials characterised by their function or physical properties
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L15/00—Chemical aspects of, or use of materials for, bandages, dressings or absorbent pads
- A61L15/16—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons
- A61L15/42—Use of materials characterised by their function or physical properties
- A61L15/44—Medicaments
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L15/00—Chemical aspects of, or use of materials for, bandages, dressings or absorbent pads
- A61L15/16—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons
- A61L15/42—Use of materials characterised by their function or physical properties
- A61L15/62—Compostable, hydrosoluble or hydrodegradable materials
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/20—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices containing or releasing organic materials
- A61L2300/30—Compounds of undetermined constitution extracted from natural sources, e.g. Aloe Vera
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
- A61L2300/418—Agents promoting blood coagulation, blood-clotting agents, embolising agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2400/00—Materials characterised by their function or physical properties
- A61L2400/04—Materials for stopping bleeding
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- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Materials Engineering (AREA)
- Epidemiology (AREA)
- Hematology (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Medicinal Preparation (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The invention relates to the field of medical care, in particular to a film for wound healing and a preparation method thereof. The film for wound healing provided by the invention comprises the following raw materials in parts by weight: 0.2-0.5 part of lathyrus lathyris toxin, 50-60 parts of polyaspartic acid and 50-60 parts of chitosan. According to the thin film for wound healing, provided by the invention, the lathyrism, the polyaspartic acid and the chitosan in a specific ratio are matched with each other, so that the hemostatic effect of the thin film can be greatly improved, and meanwhile, the obtained thin film can be biodegraded.
Description
Technical Field
The invention relates to the field of medical care, in particular to a film for wound healing and a preparation method thereof.
Background
Mucuna pruriens is a plant of Mucuna genus of Leguminosae, and is distributed in Korean, Japan and Russian far east, and is commonly distributed in northeast, North China, Shaanxi, south Gansu and east China sea of Qinghai in China. Mucuna pruriens grows in hillsides, forest borders, roadside, meadows and the like, can live in places with the elevation of 2500 m at most, and is rough and fond of warm and humid environments. The protein content of the mucuna pruriens seeds is about 25% -28%, the starch content is about 55% -61%, and the mucuna pruriens seeds are ideal high-protein leguminous feed crops and good plant starch resources. Meanwhile, the leguma pruriens also contains legumatoxin (ODAP), and has a good hemostatic effect.
Wound healing is a way of repairing damaged tissues and organs, and during the repair process of damaged tissues and organs, a wound dressing can play a role in protecting wounds from being invaded by bacteria and accelerating wound healing. The prior art CN103088630A discloses a preparation method of a nanofiber membrane for promoting wound healing, which comprises the following steps: preparation of spinning solution: dissolving a polymer in an organic solvent, and fully stirring until the polymer is completely dissolved to obtain a polymer spinning solution; spinning conditions are as follows: placing the spinning solution into an injector of electrostatic spinning equipment, generating jet flow by the polymer spinning solution under the action of static electricity, and obtaining a film consisting of polymer electrospun nanofibers which are arranged in parallel on the receiving device; collecting the collected poly-mer in parallelSoaking thin film formed by compound electrospun nanofiber in AgNO3Taking out the solution, and washing the free Ag ions which are not adsorbed by water; taken out and soaked in NaBH4Reducing Ag ions adsorbed on the surface of the electrospun nanofiber into simple substance Ag and forming Ag nano particles in the aqueous solution to obtain the nanofiber membrane for promoting wound healing. The prepared film has an antibacterial effect by utilizing the Ag nano particles adsorbed on the fibers, and can be used for wound healing, however, the film has a poor hemostatic effect, and the Ag nano particles are not easy to degrade, so that the use of the film is greatly limited.
Disclosure of Invention
The invention aims to overcome the defects that the existing film for wound healing has poor hemostatic effect and is not easy to degrade, and further provides a film for wound healing and a preparation method thereof.
In order to achieve the purpose, the invention adopts the following technical scheme:
a film for wound healing comprises the following raw materials in parts by weight: 0.2-0.5 part of lathyrus lathyris toxin, 50-60 parts of polyaspartic acid and 50-60 parts of chitosan.
Preferably, the method comprises the following raw materials: 4 parts of lathyrus lathyris toxin, 55 parts of polyaspartic acid and 59 parts of chitosan.
Preferably, the preparation method of polyaspartic acid comprises the following steps: mixing L-aspartic acid-4-benzyl ester-N-carboxyanhydride with a catalyst, and carrying out polymerization reaction in an inert gas atmosphere to obtain the polyaspartic acid.
Preferably, the polymerization temperature is 160-200 ℃, and the polymerization time is 30-50 h.
Preferably, the method for preparing polyaspartic acid further comprises the step of adding glycerol to the mixture of L-aspartic acid-4-benzyl ester-N-carboxyanhydride and the catalyst.
Preferably, the mass ratio of the L-aspartic acid-4-benzyl ester-N-carboxyanhydride to the catalyst is 1: (0.08-0.12);
the mass ratio of the L-aspartic acid-4-benzyl ester-N-carboxyanhydride to the glycerol is 1: (0.01-0.1).
Preferably, the catalyst is Co (PMe)3)4Or phosphoric acid.
Preferably, the chitosan has a molecular weight of 10 to 20 ten thousand.
The lathyrium pruriens toxin can be extracted from lathyrium pruriens by the existing known method or can be obtained by synthesizing by the existing known method.
The preparation method of the lathyrus lathyridis toxin comprises the following steps:
1) pulverizing Mucuna pruriens, and leaching with water to obtain leaching solution;
2) filtering the leaching liquor, and concentrating the filtrate to obtain a leaching concentrated solution;
3) passing the concentrated extractive solution through cation exchange column, eluting with water as eluent, and collecting eluate;
4) concentrating the eluent, and separating and purifying the concentrated eluent by adopting a column chromatography method to obtain the lathyritoxin.
Preferably, the weight ratio of the lathyrus pruriens to the water is 1: (7-10); the leaching temperature is 50-70 ℃, and the leaching time is 0.5-3 hours.
Preferably, the eluent used in the separation and purification in the step 4) is a mixed solution of n-butanol, glacial acetic acid and water; the mass ratio of the n-butanol to the glacial acetic acid to the water is 16:8: 1.
The lathyrus lathyris toxin disclosed by the invention is beta-ODAP.
The invention also provides a preparation method of the film for wound healing, which comprises the following steps:
1) dissolving chitosan in an acetic acid solution to obtain a chitosan solution; dissolving polyaspartic acid in water to obtain a polyaspartic acid solution;
2) mixing the chitosan solution obtained in the step 1) with the polyaspartic acid solution, then adding the lathyrus radiatus toxin, and performing ultrasonic treatment to obtain a spinning solution;
3) performing electrostatic spinning on the spinning solution obtained in the step 2) to obtain the film for wound healing.
Preferably, the mass ratio of the chitosan to the acetic acid solution is (3-5): 10;
the mass ratio of the polyaspartic acid to the water is (2-3) to 10;
the mass fraction of acetic acid in the acetic acid solution is 90-96%.
The invention has the beneficial effects that:
1) the film for wound healing provided by the invention comprises the following raw materials in parts by weight: 0.2-0.5 part of lathyrus lathyris toxin, 50-60 parts of polyaspartic acid and 50-60 parts of chitosan. According to the invention, the lathyrium pruriens toxin, the polyaspartic acid and the chitosan in a specific ratio are matched with each other, so that the hemostasis effect of the film can be greatly improved, and the obtained film can be biodegraded.
2) The invention provides a film for wound healing, and further the preparation method of polyaspartic acid comprises the following steps: mixing L-aspartic acid-4-benzyl ester-N-carboxyanhydride with a catalyst, and carrying out polymerization reaction in an inert gas atmosphere to obtain the polyaspartic acid. The polyaspartic acid prepared by the preparation method is more beneficial to dispersion of the lathyria pruriens toxin and the chitosan, so that the hemostatic effect of the film is better.
3) The film for wound healing provided by the invention further comprises a step of adding glycerol into the mixture of the L-aspartic acid-4-benzyl ester-N-carboxyanhydride and the catalyst in the preparation method of the polyaspartic acid. According to the invention, the glycerol is added into the mixture of the L-aspartic acid-4-benzyl ester-N-carboxyanhydride and the catalyst, and the addition of the glycerol is beneficial to forming a cross-linked network structure by polyaspartic acid, so that the hemostatic effect of the film can be further improved through research.
4) The preparation method of the film for wound healing provided by the invention comprises the following steps: dissolving chitosan in an acetic acid solution to obtain a chitosan solution; dissolving polyaspartic acid in water to obtain a polyaspartic acid solution; mixing the chitosan solution and the polyaspartic acid solution, then adding the lathyrus lathyris toxin, and performing ultrasonic treatment to obtain a spinning solution; and (3) carrying out electrostatic spinning on the spinning solution to obtain the film for wound healing. According to the invention, the film for wound healing is prepared by an electrostatic spinning method, the distribution of ingredients such as the lathyrus toxin in the obtained film is uniform, and the film is degradable.
Detailed Description
The following examples are provided to further understand the present invention, not to limit the scope of the present invention, but to provide the best mode, not to limit the content and the protection scope of the present invention, and any product similar or similar to the present invention, which is obtained by combining the present invention with other prior art features, falls within the protection scope of the present invention.
The examples do not show the specific experimental steps or conditions, and can be performed according to the conventional experimental steps described in the literature in the field. The reagents or instruments used are not indicated by manufacturers, and are all conventional reagent products which can be obtained commercially.
Example 1
The embodiment provides a film for wound healing, which comprises the following raw materials in parts by weight: 2g of mucuna pruriens toxin, 50g of polyaspartic acid and 60g of chitosan; the molecular weight of the chitosan is 20 ten thousand.
The preparation method of the lathyrus lathyridis toxin comprises the following steps:
1) crushing 50g of mucuna pruriens, and adding water with the weight 10 times that of the mucuna pruriens for leaching to obtain leaching liquor; the leaching temperature is 70 ℃, the leaching time is 1 hour, then the leaching liquor is sieved by a 200-mesh sieve to obtain filtrate, and the filtrate is concentrated to obtain leaching concentrated solution;
2) loading the concentrated extract obtained in the step 1) on a cation exchange column (MonoS 5/50GL), adding 300ml of water for elution after sample adsorption, and collecting the eluate;
3) concentrating the eluate, separating the concentrated eluate with 200 mesh silica gel column chromatography, and eluting with eluate (mixed solution of n-butanol, glacial acetic acid and water; and the mass ratio of the n-butanol to the glacial acetic acid to the water is 16:8:1), collecting an elution fraction containing the lathyritoxin (determined by high performance liquid chromatography), concentrating, and drying to obtain the lathyritoxin.
The preparation method of the polyaspartic acid comprises the following steps: 100g L-aspartic acid-4-benzyl ester-N-carboxyanhydride, 8gCo (PMe)3)4Placing the mixture into a 500ml three-neck flask, vacuumizing, carrying out polymerization reaction for 30 hours at 200 ℃ in a nitrogen atmosphere, filtering the reaction solution after the reaction is finished, and drying a filter cake to obtain the polyaspartic acid.
The preparation method of the film for wound healing comprises the following steps:
1) dissolving 60g of chitosan in 200g of 96% acetic acid solution at 80 ℃ to obtain a chitosan solution; dissolving 50g of polyaspartic acid in 250g of water to obtain a polyaspartic acid solution;
2) mixing the chitosan solution obtained in the step 1) with a polyaspartic acid solution, then adding 2g of lathyrus radiatus toxin, and carrying out ultrasonic treatment at 30 ℃ for 1 hour to obtain a spinning solution;
3) injecting the spinning solution obtained in the step 2) into an injector with a flat head needle head with the inner diameter of 1.2mm, fixing the injector on a micro-injection pump, placing a metal receiving screen at a position 16cm away from the needle head, connecting the needle head with a high-voltage direct-current power supply of 20kV, performing electrostatic spinning at a set flow rate of 1.0ml/h, and collecting a fiber membrane on the metal receiving screen to obtain the film for wound healing.
Example 2
The embodiment provides a film for wound healing, which comprises the following raw materials in parts by weight: 2g of mucuna pruriens toxin, 50g of polyaspartic acid and 60g of chitosan; the molecular weight of the chitosan is 20 ten thousand.
The preparation method of the lathyrus lathyridis toxin comprises the following steps:
1) crushing 50g of mucuna pruriens, and adding water with the weight 10 times that of the mucuna pruriens for leaching to obtain leaching liquor; the leaching temperature is 70 ℃, the leaching time is 1 hour, then the leaching liquor is sieved by a 200-mesh sieve to obtain filtrate, and the filtrate is concentrated to obtain leaching concentrated solution;
2) loading the concentrated extract obtained in the step 1) on a cation exchange column (MonoS 5/50GL), adding 300ml of water for elution after sample adsorption, and collecting the eluate;
3) concentrating the eluate, separating the concentrated eluate with 200 mesh silica gel column chromatography, and eluting with eluate (mixed solution of n-butanol, glacial acetic acid and water; and the mass ratio of the n-butanol to the glacial acetic acid to the water is 16:8:1), collecting an elution fraction containing the lathyritoxin (determined by high performance liquid chromatography), concentrating, and drying to obtain the lathyritoxin.
The preparation method of the polyaspartic acid comprises the following steps: 100g L-aspartic acid-4-benzyl ester-N-carboxyanhydride, 8gCo (PMe)3)4Placing the mixture into a 500ml three-neck flask, adding 10g of glycerol, vacuumizing, carrying out polymerization reaction for 30 hours at 200 ℃ in a nitrogen atmosphere, filtering the reaction solution after the reaction is finished, and drying a filter cake to obtain the polyaspartic acid.
The preparation method of the film for wound healing comprises the following steps:
1) dissolving 60g of chitosan in 200g of 96% acetic acid solution at 80 ℃ to obtain a chitosan solution; dissolving 50g of polyaspartic acid in 250g of water to obtain a polyaspartic acid solution;
2) mixing the chitosan solution obtained in the step 1) with a polyaspartic acid solution, then adding 2g of lathyrus radiatus toxin, and carrying out ultrasonic treatment at 30 ℃ for 1 hour to obtain a spinning solution;
3) injecting the spinning solution obtained in the step 2) into an injector with a flat head needle head with the inner diameter of 1.2mm, fixing the injector on a micro-injection pump, placing a metal receiving screen at a position 16cm away from the needle head, connecting the needle head with a high-voltage direct-current power supply of 20kV, performing electrostatic spinning at a set flow rate of 1.0ml/h, and collecting a fiber membrane on the metal receiving screen to obtain the film for wound healing.
Example 3
The embodiment provides a film for wound healing, which comprises the following raw materials in parts by weight: 5g of lathyrus lathyris toxin, 60g of polyaspartic acid and 50g of chitosan; the molecular weight of the chitosan is 10 ten thousand.
The preparation method of the lathyrus lathyridis toxin comprises the following steps:
1) crushing 50g of mucuna pruriens, and adding water with the weight 10 times that of the mucuna pruriens for leaching to obtain leaching liquor; the leaching temperature is 70 ℃, the leaching time is 1 hour, then the leaching liquor is sieved by a 200-mesh sieve to obtain filtrate, and the filtrate is concentrated to obtain leaching concentrated solution;
2) loading the concentrated extract obtained in the step 1) on a cation exchange column (MonoS 5/50GL), adding 300ml of water for elution after sample adsorption, and collecting the eluate;
3) concentrating the eluate, separating the concentrated eluate with 200 mesh silica gel column chromatography, and eluting with eluate (mixed solution of n-butanol, glacial acetic acid and water; and the mass ratio of the n-butanol to the glacial acetic acid to the water is 16:8:1), collecting an elution fraction containing the lathyritoxin (determined by high performance liquid chromatography), concentrating, and drying to obtain the lathyritoxin.
The preparation method of the polyaspartic acid comprises the following steps: 100g L-aspartic acid-4-benzyl ester-N-carboxyanhydride and 12g phosphoric acid solution with the mass fraction of 85% are put into a 500ml three-neck flask, then 1g glycerol is added, the vacuum pumping is carried out, the polymerization reaction is carried out for 50 hours at 160 ℃ in the nitrogen atmosphere, the reaction solution is filtered after the reaction is finished, and the filter cake is dried, thus obtaining the polyaspartic acid.
The preparation method of the film for wound healing comprises the following steps:
1) dissolving 50g of chitosan in 100g of acetic acid solution with the mass fraction of 90% at 80 ℃ to obtain a chitosan solution; dissolving 60g of polyaspartic acid in 200g of water to obtain a polyaspartic acid solution;
2) mixing the chitosan solution obtained in the step 1) with a polyaspartic acid solution, then adding 5g of lathyrus radiatus toxin, and carrying out ultrasonic treatment at 30 ℃ for 1 hour to obtain a spinning solution;
3) injecting the spinning solution obtained in the step 2) into an injector with a flat head needle head with the inner diameter of 1.2mm, fixing the injector on a micro-injection pump, placing a metal receiving screen at a position 16cm away from the needle head, connecting the needle head with a high-voltage direct-current power supply of 20kV, performing electrostatic spinning at a set flow rate of 1.0ml/h, and collecting a fiber membrane on the metal receiving screen to obtain the film for wound healing.
Example 4
The embodiment provides a film for wound healing, which comprises the following raw materials in parts by weight: 4g of lathyrus lathyris toxin, 55g of polyaspartic acid and 59g of chitosan; the molecular weight of the chitosan is 15 ten thousand.
The preparation method of the lathyrus lathyridis toxin comprises the following steps:
1) crushing 50g of mucuna pruriens, and adding water with the weight 10 times that of the mucuna pruriens for leaching to obtain leaching liquor; the leaching temperature is 70 ℃, the leaching time is 1 hour, then the leaching liquor is sieved by a 200-mesh sieve to obtain filtrate, and the filtrate is concentrated to obtain leaching concentrated solution;
2) loading the concentrated extract obtained in the step 1) on a cation exchange column (MonoS 5/50GL), adding 300ml of water for elution after sample adsorption, and collecting the eluate;
3) concentrating the eluate, separating the concentrated eluate with 200 mesh silica gel column chromatography, and eluting with eluate (mixed solution of n-butanol, glacial acetic acid and water; and the mass ratio of the n-butanol to the glacial acetic acid to the water is 16:8:1), collecting an elution fraction containing the lathyritoxin (determined by high performance liquid chromatography), concentrating, and drying to obtain the lathyritoxin.
The preparation method of the polyaspartic acid comprises the following steps: 100g of 100g L-aspartic acid-4-benzyl ester-N-carboxyanhydride, 10g of Co (PMe)3)4Putting the mixture into a 500ml three-neck flask, adding 8g of glycerol, vacuumizing, carrying out polymerization reaction for 40 hours at 180 ℃ in a nitrogen atmosphere, filtering the reaction solution after the reaction is finished, and drying a filter cake to obtain the polyaspartic acid.
The preparation method of the film for wound healing comprises the following steps:
1) dissolving 59g of chitosan in 147g of acetic acid solution with the mass fraction of 93% at 80 ℃ to obtain a chitosan solution; dissolving 55g of polyaspartic acid in 180g of water to obtain a polyaspartic acid solution;
2) mixing the chitosan solution obtained in the step 1) with a polyaspartic acid solution, then adding 4g of lathyrus radiatus toxin, and carrying out ultrasonic treatment at 30 ℃ for 1 hour to obtain a spinning solution;
3) injecting the spinning solution obtained in the step 2) into an injector with a flat head needle head with the inner diameter of 1.2mm, fixing the injector on a micro-injection pump, placing a metal receiving screen at a position 16cm away from the needle head, connecting the needle head with a high-voltage direct-current power supply of 20kV, performing electrostatic spinning at a set flow rate of 1.0ml/h, and collecting a fiber membrane on the metal receiving screen to obtain the film for wound healing.
Comparative example 1
This comparative example provides a film for wound healing, which is different from example 4 in that 55 gL-aspartic acid is substituted for the polyaspartic acid of example 4.
Effect verification
Hemostasis test
Test groups: the films obtained in examples 1 to 4 and comparative example 1 were fixed to nonwoven fabrics to obtain samples for hemostasis test, i.e., example 1, example 2, example 3, example 4 and comparative example 1, respectively.
Control group: gauze hemostatic products were used as a control group in the market.
The test procedure was as follows:
(1) according to the proportion of 1ml/kg, 4% sodium pentobarbital is slowly injected into the ear vein of the rabbit for anesthesia, then a wound towel is paved on the left abdominal side of the rabbit, and the surgical site is disinfected by iodine tincture and 75% alcohol respectively.
(2) A5.0 cm longitudinal incision was made along the left inner edge, approximately 1cm to the left of the midline of the abdomen, the abdominal muscles were incised to the peritoneal layer, and the peritoneum was dissected.
(3) The spleen of the rabbit was removed with forceps and kept moist with normal saline.
(4) Blank control group: on the visceral surface of the spleen, two symmetrical wounds with the length of 1.2cm and the depth of 0.8mm are cut along the long axis of the spleen by using an operation blade, and the wound surface is actively bled. And observing the bleeding condition of the wound surface, and recording the automatic hemostasis time of the rabbit.
(5) Control group: on the visceral surface of the spleen, a wound with a length of 1.2cm and a depth of 0.8mm is cut along the long axis of the spleen by using a surgical blade, and bleeding on the wound surface is active. After 12 seconds, ordinary gauze is applied to stop bleeding, and fingers are lightly pressed for 1 minute until the bleeding is stopped. After 1 minute, observing the bleeding condition of the wound surface, if the bleeding does not occur within 2 minutes, considering that the hemostasis is successful, otherwise, removing the blood clots and the gauze of the wound surface, applying the common gauze again, slightly pressing the finger for 1 minute, and repeating the steps until the hemostasis is successful; the time to hemostasis was recorded.
(6) Test groups: on the visceral surface of the spleen, a wound with a length of 1.2cm and a depth of 0.8mm is cut along the long axis of the spleen by using a surgical blade, and bleeding on the wound surface is active. Applying a test group sample for hemostasis after 12 seconds, slightly pressing the finger for 1 minute, observing bleeding conditions of the wound surface after 1 minute with the inactive bleeding as a degree, if bleeding does not occur within 2 minutes, determining hemostasis to be successful, otherwise removing blood clots of the wound surface and the test group sample, applying the test group sample again, slightly pressing the finger for 1 minute, and repeating the steps until hemostasis is successful; the time to hemostasis was recorded. The test results are shown in table 1.
TABLE 1 hemostasis test results
| Group of | Hemostasis time (min) |
| Blank group | 40 |
| Control group | 21 |
| Example 1 sample | 12 |
| Example 2 sample | 8 |
| Example 3 sample | 6 |
| Example 4 sample | 5 |
| Comparative example 1 sample | 16 |
It should be understood that the above examples are only for clarity of illustration and are not intended to limit the embodiments. Other variations and modifications will be apparent to persons skilled in the art in light of the above description. And are neither required nor exhaustive of all embodiments. And obvious variations or modifications therefrom are within the scope of the invention.
Claims (10)
1. A film for wound healing is characterized by comprising the following raw materials in parts by weight: 0.2-0.5 part of lathyrus lathyris toxin, 50-60 parts of polyaspartic acid and 50-60 parts of chitosan.
2. The film for wound healing according to claim 1, comprising the following raw materials: 4 parts of lathyrus lathyris toxin, 55 parts of polyaspartic acid and 59 parts of chitosan.
3. The film for wound healing according to claim 1 or 2, wherein the preparation method of polyaspartic acid comprises the steps of: mixing L-aspartic acid-4-benzyl ester-N-carboxyanhydride with a catalyst, and carrying out polymerization reaction in an inert gas atmosphere to obtain the polyaspartic acid.
4. The membrane for wound healing according to any one of claims 1 to 3, wherein the polymerization temperature is 160-200 ℃ and the polymerization time is 30-50 h.
5. The film for wound healing according to any one of claims 1 to 4, wherein the method for preparing polyaspartic acid further comprises a step of adding glycerol to the mixture of L-aspartic acid-4-benzyl ester-N-carboxyanhydride and the catalyst.
6. The film for wound healing according to any one of claims 1 to 5, wherein the mass ratio of L-aspartic acid-4-benzyl ester-N-carboxyanhydride to catalyst is 1: (0.08-0.12);
the mass ratio of the L-aspartic acid-4-benzyl ester-N-carboxyanhydride to the glycerol is 1: (0.01-0.1).
7. The membrane for wound healing according to any one of claims 1 to 6, wherein the catalyst is Co (PMe)3)4Or phosphoric acid.
8. A film for use in wound healing according to any of claims 1 to 7, wherein the chitosan has a molecular weight of 10 to 20 ten thousand.
9. A method of preparing a film for wound healing according to any one of claims 1 to 8, comprising the steps of:
1) dissolving chitosan in an acetic acid solution to obtain a chitosan solution; dissolving polyaspartic acid in water to obtain a polyaspartic acid solution;
2) mixing the chitosan solution obtained in the step 1) with the polyaspartic acid solution, then adding the lathyrus radiatus toxin, and performing ultrasonic treatment to obtain a spinning solution;
3) performing electrostatic spinning on the spinning solution obtained in the step 2) to obtain the film for wound healing.
10. The method for preparing a film for wound healing according to claim 9, wherein the mass ratio of the chitosan to the acetic acid solution is (3-5): 10;
the mass ratio of the polyaspartic acid to the water is (2-3) to 10;
the mass fraction of acetic acid in the acetic acid solution is 90-96%.
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