CN117503980A - Double-layer nanofiber membrane dressing and preparation method and application thereof - Google Patents
Double-layer nanofiber membrane dressing and preparation method and application thereof Download PDFInfo
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- CN117503980A CN117503980A CN202311454438.2A CN202311454438A CN117503980A CN 117503980 A CN117503980 A CN 117503980A CN 202311454438 A CN202311454438 A CN 202311454438A CN 117503980 A CN117503980 A CN 117503980A
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- nanofiber membrane
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- polylactic acid
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- 239000012528 membrane Substances 0.000 title claims abstract description 104
- 239000002121 nanofiber Substances 0.000 title claims abstract description 103
- 238000002360 preparation method Methods 0.000 title claims abstract description 11
- 229920001661 Chitosan Polymers 0.000 claims abstract description 58
- 229920000747 poly(lactic acid) Polymers 0.000 claims abstract description 38
- 239000004626 polylactic acid Substances 0.000 claims abstract description 38
- BSYNRYMUTXBXSQ-UHFFFAOYSA-N Aspirin Chemical group CC(=O)OC1=CC=CC=C1C(O)=O BSYNRYMUTXBXSQ-UHFFFAOYSA-N 0.000 claims abstract description 30
- 229960001138 acetylsalicylic acid Drugs 0.000 claims abstract description 30
- 229920001690 polydopamine Polymers 0.000 claims abstract description 19
- CUGZWHZWSVUSBE-UHFFFAOYSA-N 2-(oxiran-2-ylmethoxy)ethanol Chemical compound OCCOCC1CO1 CUGZWHZWSVUSBE-UHFFFAOYSA-N 0.000 claims abstract description 8
- 239000000243 solution Substances 0.000 claims description 35
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 30
- 238000009987 spinning Methods 0.000 claims description 27
- 238000010791 quenching Methods 0.000 claims description 24
- 230000000171 quenching effect Effects 0.000 claims description 24
- VYFYYTLLBUKUHU-UHFFFAOYSA-N dopamine Chemical compound NCCC1=CC=C(O)C(O)=C1 VYFYYTLLBUKUHU-UHFFFAOYSA-N 0.000 claims description 22
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 21
- ZHNUHDYFZUAESO-UHFFFAOYSA-N Formamide Chemical compound NC=O ZHNUHDYFZUAESO-UHFFFAOYSA-N 0.000 claims description 18
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 claims description 18
- 238000002791 soaking Methods 0.000 claims description 14
- 239000007788 liquid Substances 0.000 claims description 12
- 229960003638 dopamine Drugs 0.000 claims description 11
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 10
- 238000000034 method Methods 0.000 claims description 10
- 238000010041 electrostatic spinning Methods 0.000 claims description 9
- 230000002439 hemostatic effect Effects 0.000 claims description 9
- 239000000463 material Substances 0.000 claims description 9
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 claims description 9
- 239000011259 mixed solution Substances 0.000 claims description 8
- 239000012046 mixed solvent Substances 0.000 claims description 7
- 229910052757 nitrogen Inorganic materials 0.000 claims description 5
- 230000003110 anti-inflammatory effect Effects 0.000 claims description 4
- 238000009777 vacuum freeze-drying Methods 0.000 claims description 4
- 230000001678 irradiating effect Effects 0.000 claims description 3
- 238000003756 stirring Methods 0.000 claims description 2
- 230000001954 sterilising effect Effects 0.000 abstract description 9
- 238000004659 sterilization and disinfection Methods 0.000 abstract description 9
- 230000015271 coagulation Effects 0.000 abstract description 6
- 238000005345 coagulation Methods 0.000 abstract description 6
- 238000001035 drying Methods 0.000 description 9
- 239000000203 mixture Substances 0.000 description 9
- 239000000835 fiber Substances 0.000 description 8
- 230000001580 bacterial effect Effects 0.000 description 6
- 230000023555 blood coagulation Effects 0.000 description 6
- 238000006243 chemical reaction Methods 0.000 description 6
- 230000001332 colony forming effect Effects 0.000 description 6
- 230000000052 comparative effect Effects 0.000 description 6
- 238000003892 spreading Methods 0.000 description 6
- 230000007480 spreading Effects 0.000 description 6
- 238000001291 vacuum drying Methods 0.000 description 6
- 102000001554 Hemoglobins Human genes 0.000 description 5
- 108010054147 Hemoglobins Proteins 0.000 description 5
- 238000002835 absorbance Methods 0.000 description 5
- 238000011156 evaluation Methods 0.000 description 5
- 238000000338 in vitro Methods 0.000 description 5
- 230000023597 hemostasis Effects 0.000 description 4
- 241000191967 Staphylococcus aureus Species 0.000 description 3
- XAGFODPZIPBFFR-UHFFFAOYSA-N aluminium Chemical compound [Al] XAGFODPZIPBFFR-UHFFFAOYSA-N 0.000 description 3
- 229910052782 aluminium Inorganic materials 0.000 description 3
- 230000000844 anti-bacterial effect Effects 0.000 description 3
- 239000007853 buffer solution Substances 0.000 description 3
- 238000004140 cleaning Methods 0.000 description 3
- 239000008367 deionised water Substances 0.000 description 3
- 229910021641 deionized water Inorganic materials 0.000 description 3
- 239000011888 foil Substances 0.000 description 3
- 238000003760 magnetic stirring Methods 0.000 description 3
- 239000002861 polymer material Substances 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical class OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 3
- 238000005406 washing Methods 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 description 2
- 206010061218 Inflammation Diseases 0.000 description 2
- 208000007536 Thrombosis Diseases 0.000 description 2
- 206010052428 Wound Diseases 0.000 description 2
- 208000027418 Wounds and injury Diseases 0.000 description 2
- 230000009471 action Effects 0.000 description 2
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- 210000003743 erythrocyte Anatomy 0.000 description 2
- 230000004054 inflammatory process Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
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- 150000003254 radicals Chemical class 0.000 description 2
- 229920002101 Chitin Polymers 0.000 description 1
- 238000001994 activation Methods 0.000 description 1
- 229940030225 antihemorrhagics Drugs 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000000601 blood cell Anatomy 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 229920002678 cellulose Polymers 0.000 description 1
- 239000001913 cellulose Substances 0.000 description 1
- 238000004132 cross linking Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- GYZLOYUZLJXAJU-UHFFFAOYSA-N diglycidyl ether Chemical compound C1OC1COCC1CO1 GYZLOYUZLJXAJU-UHFFFAOYSA-N 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 238000012377 drug delivery Methods 0.000 description 1
- 238000001523 electrospinning Methods 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
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- 150000004676 glycans Chemical class 0.000 description 1
- 230000000025 haemostatic effect Effects 0.000 description 1
- WGCNASOHLSPBMP-UHFFFAOYSA-N hydroxyacetaldehyde Natural products OCC=O WGCNASOHLSPBMP-UHFFFAOYSA-N 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 229920005615 natural polymer Polymers 0.000 description 1
- 230000010118 platelet activation Effects 0.000 description 1
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- 239000005017 polysaccharide Substances 0.000 description 1
- 238000007348 radical reaction Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000001356 surgical procedure Methods 0.000 description 1
- 230000002195 synergetic effect Effects 0.000 description 1
- 238000002145 thermally induced phase separation Methods 0.000 description 1
- 230000029663 wound healing Effects 0.000 description 1
- 230000037314 wound repair Effects 0.000 description 1
Classifications
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- 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
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- A61L15/00—Chemical aspects of, or use of materials for, bandages, dressings or absorbent pads
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- A61L15/00—Chemical aspects of, or use of materials for, bandages, dressings or absorbent pads
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- 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
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- A61L15/00—Chemical aspects of, or use of materials for, bandages, dressings or absorbent pads
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Abstract
The invention discloses a preparation method of a double-layer nanofiber membrane dressing, wherein an upper layer is an aspirin modified chitosan nanofiber membrane, a lower layer is a polydopamine coated polylactic acid nanofiber membrane, and the two layers are crosslinked through ethylene glycol glycidyl ether. The porosity of the double-layer nanofiber membrane dressing is 92.1 percent, and the specific surface area is 5.33m 2 And/g. The sterilization rate of the double-layer nanofiber membrane dressing is 100%, and the coagulation index BCI is 61.1%.
Description
Technical Field
The invention relates to a preparation method of a double-layer nanofiber membrane dressing, and belongs to the fields of functional polymer materials and biomedical materials.
Background
After massive hemostasis, inflammation and complications from wounds are an important challenge for a large number of surgical procedures. Most hemostatic materials are currently used as single hemostatic materials. Many biodegradable and biocompatible natural polymeric materials are widely used in tissue engineering scaffolds and drug delivery materials. Among these natural polymer materials, chitosan is widely used because of its excellent biocompatibility. Chitosan is a deacetylated product of chitin, and the content of chitosan is a polysaccharide polymer material inferior to cellulose in nature. Chitosan is widely used in biomedical engineering and biotechnology engineering due to its wound repair, biodegradability and biocompatibility. There have been a number of studies showing that chitosan can be used as hemostatic material (Gu B K, et al Fabrication of sonicated chitosan nanofiber mat with enlarged porosity for use as hemostatic materials.carbohydrate.Polym.2013, 97:65-73). Chitosan is adhered and aggregated with the negatively charged erythrocyte surface substances through the action of positive charges, so that blood clots are rapidly formed for hemostasis. Second, chitosan forms a three-dimensional network structure after some polymerization reaction in blood, which aggregates red blood cells by capturing them. The surface forms electronegative substances in the platelet activation process, and the substances and positively charged chitosan are aggregated under the action of electrostatic attraction force to quickly form blood clots, thereby achieving the aim of hemostasis. However, inflammation and complications after hemostasis are important causes of death. Therefore, the timely development of multifunctional hemostatic dressing with hemostatic and anti-inflammatory effects is the key point of future research.
The invention comprises the following steps:
in order to solve the technical problems, the invention aims to provide a double-layer nanofiber membrane dressing as well as a preparation method and application thereof.
The invention is realized by the following technical scheme:
a method for preparing a double-layer nanofiber membrane dressing, which comprises the following steps:
s1, preparing a chitosan nanofiber membrane by using an electrostatic spinning method;
s2, under the protection of nitrogen, irradiating the chitosan nanofiber membrane by using low-temperature plasma, soaking the chitosan nanofiber membrane in an ethanol solution of aspirin, and reacting at 60-80 ℃ to obtain an aspirin modified chitosan nanofiber membrane;
s3, adding polylactic acid into a tetrahydrofuran/formamide mixed solvent, magnetically stirring to form a quenching liquid, carrying out low-temperature quenching on the quenching liquid, and then carrying out vacuum freeze drying to obtain a polylactic acid nanofiber membrane, soaking the polylactic acid nanofiber membrane in a dopamine solution, adjusting the pH value to 8-9, and obtaining the polydopamine coated polylactic acid nanofiber membrane after soaking;
s4, paving the aspirin modified chitosan nanofiber membrane on the surface of the polydopamine coated polylactic acid nanofiber membrane in an ethylene glycol glycidyl ether/ethanol mixed solution, soaking at normal temperature, and then performing vacuum freeze drying to obtain the double-layer nanofiber membrane dressing.
As a preferred scheme, the preparation method of the chitosan nanofiber membrane comprises the following steps:
adding chitosan into a dilute acetic acid solution, and dissolving to obtain spinning solution;
carrying out electrostatic spinning operation on the spinning solution, wherein the conditions for controlling the electrostatic spinning are as follows: the spinning voltage is 15-30 kV, the distance between the spinning needle head and the collecting plate is 10-20 cm, and the spinning speed is 0.1-0.4 mL/h; the low-temperature plasma irradiation conditions are as follows: the irradiation power is 120-200W, the pressure is 200-300 Pa, and the irradiation time is 3-10 min.
Preferably, the mass concentration of the dilute acetic acid solution is 1-3%; in the spinning solution, the mass concentration of chitosan is 3-10%.
Preferably, in the quenching liquid, the mass concentration of the polylactic acid is 3-8%; in the tetrahydrofuran/formamide mixed solvent, the mass ratio of tetrahydrofuran to formamide is 1: (3-6); the temperature of the low-temperature quenching is-40 to-10 ℃ and the quenching time is 100 to 300min; the concentration of the dopamine solution is 0.1-0.3 mg/mL.
As a preferable scheme, the mass ratio of the aspirin modified chitosan nanofiber membrane to the polydopamine coated polylactic acid nanofiber membrane is (3-5): (1-3); the mass ratio of the ethylene glycol glycidyl ether to the ethanol is 1: (1-5).
A double-layered nanofiber membrane dressing obtained by the aforementioned preparation method.
Use of a bilayer nanofiber membrane dressing as hereinbefore described in haemostatic and anti-inflammatory materials.
The basic principle of the invention is as follows:
1. preparing a chitosan nanofiber membrane through electrospinning, introducing free radicals into the fiber membrane through low-temperature plasma irradiation, and grafting the medicine aspirin onto the fiber membrane through a free radical reaction to obtain the aspirin modified chitosan nanofiber membrane.
2. The polylactic acid nanofiber membrane is prepared by a thermally induced phase separation method, and dopamine is polymerized and loaded on the polylactic acid nanofiber membrane under a weak alkaline condition to obtain the polydopamine coated polylactic acid nanofiber membrane.
3. And (3) crosslinking the aspirin modified chitosan nanofiber membrane and the polydopamine coated polylactic acid nanofiber membrane through glycol glycidyl ether, and freeze-drying to obtain the double-layer nanofiber membrane dressing.
Compared with the prior art, the invention has the following beneficial effects:
1. the high porosity and large specific surface area of the chitosan and polylactic acid nanofiber membrane can be used for rapidly absorbing plasma, so that blood cells are accumulated on the surface, and blood coagulation on the wound surface is promoted.
2. The slow release of the drug-loaded aspirin and the synergistic anti-inflammatory effect of polydopamine are utilized to further promote the wound healing speed.
Drawings
Other features, objects and advantages of the present invention will become more apparent upon reading of the detailed description of non-limiting embodiments, given with reference to the accompanying drawings in which:
fig. 1 is a route pattern for the preparation of a two-layer nanofiber membrane dressing.
Detailed Description
The present invention will be described in detail with reference to specific examples. The following examples will assist those skilled in the art in further understanding the present invention, but are not intended to limit the invention in any way. It should be noted that variations and modifications could be made by those skilled in the art without departing from the inventive concept. These are all within the scope of the present invention.
Example 1
2g of chitosan is added into 30g of dilute acetic acid solution with mass concentration of 2%, and the chitosan solution is obtained by magnetic stirring and dissolution. Spinning the solution by using an electrostatic spinning device, wherein the spinning voltage is 20kV, the distance between a spinning needle head and a collecting plate is 15cm, the spinning speed is 0.2mL/h, the spinning film is torn off from aluminum foil paper, and the chitosan nanofiber film is obtained by vacuum drying. Under the protection of nitrogen, the chitosan nanofiber membrane is irradiated by low-temperature plasma, wherein the irradiation power is 150W, the pressure is 300Pa, and the irradiation time is 6min. After the irradiation is finished, the chitosan nanofiber membrane is soaked in an aspirin/ethanol mixed solution, and the reaction is carried out for 3 hours at 70 ℃. And after the reaction is finished, washing the mixture with ethanol for a plurality of times to ensure that unreacted aspirin is removed, and drying the mixture in vacuum to obtain the aspirin modified chitosan nanofiber membrane.
0.5g of polylactic acid is added into 10g of mixed solvent of tetrahydrofuran/formamide (mass ratio is 1:3), and the mixture is magnetically stirred for 2 hours at 50 ℃ to form quenching liquid. Pouring the quenching liquid into a culture dish with the diameter of 8cm, putting the culture dish into a low-temperature refrigerator with the temperature of minus 20 ℃ for quenching for 200min, and putting the culture dish into a vacuum freeze dryer for drying for 3 days after the quenching is finished to obtain the polylactic acid nanofiber membrane. 0.4g of polylactic acid nanofiber membrane is soaked in 100mL of dopamine solution with the concentration of 0.2mg/mL, the pH value is adjusted to be 8.5 by using Tris salt buffer solution, and after soaking for 30min, dopamine is deposited and polymerized on the surface of the fiber. And then cleaning the fiber membrane with deionized water for 3 times, and vacuum drying to obtain the polydopamine coated polylactic acid nanofiber membrane.
Spreading 0.1g polydopamine coated polylactic acid nanofiber membrane in a culture dish, spreading 0.4g aspirin modified chitosan nanofiber membrane on the surface of the culture dish, adding a mixed solution of ethylene glycol glycidyl ether and ethanol with the mass ratio of 1:4 into the culture dish, and soaking for 24 hours at normal temperature. Finally, the petri dish is put into a vacuum freeze dryer for drying for 3 days, and the double-layer nanofiber membrane dressing is obtained, and the preparation route is shown in figure 1.
The dressing of the double-layer nanofiber membrane prepared in example 1 has a porosity of 92.1% and a specific surface area of 5.33m 2 And/g. The bacterial colony forming units (colony forming units, CFU) on the petri dish were calculated using staphylococcus aureus (s. Aureus) as the subject, thereby calculating the bacterial sterilization rate of the dressing, and thus investigating the antibacterial performance of the dressing. The sterilization rate of the double-layer nanofiber membrane dressing prepared in example 1 was 100%. The blood coagulation rate was determined by absorbance values of the hemoglobin solution using a coagulation index (BCI) evaluation in vitro. The BCI of the double layer nanofiber membrane dressing was 61.1%.
Example 2
1.5g of chitosan is added into 30g of dilute acetic acid solution with mass concentration of 2%, and the solution is dissolved by magnetic stirring, thus obtaining chitosan solution. Spinning the solution by using an electrostatic spinning device, wherein the spinning voltage is 25kV, the distance between a spinning needle head and a collecting plate is 20cm, the spinning speed is 0.25mL/h, the spinning film is torn off from aluminum foil paper, and the chitosan nanofiber film is obtained by vacuum drying. Under the protection of nitrogen, the chitosan nanofiber membrane is irradiated by low-temperature plasma, wherein the irradiation power is 140W, the pressure is 250Pa, and the irradiation time is 5min. After the irradiation is finished, the chitosan nanofiber membrane is soaked in an aspirin/ethanol mixed solution, and the reaction is carried out for 3 hours at 70 ℃. And after the reaction is finished, washing the mixture with ethanol for a plurality of times to ensure that unreacted aspirin is removed, and drying the mixture in vacuum to obtain the aspirin modified chitosan nanofiber membrane.
0.4g of polylactic acid is added into 10g of mixed solvent of tetrahydrofuran/formamide (mass ratio is 1:3), and the mixture is magnetically stirred for 2 hours at 50 ℃ to form quenching liquid. Pouring the quenching liquid into a culture dish with the diameter of 8cm, putting the culture dish into a low-temperature refrigerator with the temperature of minus 25 ℃ for quenching for 250min, and putting the culture dish into a vacuum freeze dryer for drying for 3 days after the quenching is finished to obtain the polylactic acid nanofiber membrane. 0.4g of polylactic acid nanofiber membrane is soaked in 100mL of dopamine solution with the concentration of 0.15mg/mL, the pH value is adjusted to 8.5 by using Tris salt buffer solution, and after soaking for 30min, dopamine is deposited and polymerized on the surface of the fiber. And then cleaning the fiber membrane with deionized water for 3 times, and vacuum drying to obtain the polydopamine coated polylactic acid nanofiber membrane.
Spreading 0.2g polydopamine coated polylactic acid nanofiber membrane in a culture dish, spreading 0.3g aspirin modified chitosan nanofiber membrane on the surface of the culture dish, adding a mixed solution of ethylene glycol glycidyl ether and ethanol with the mass ratio of 1:5 into the culture dish, and soaking for 24 hours at normal temperature. Finally, the petri dish is put into a vacuum freeze dryer for drying for 3 days, and the double-layer nanofiber membrane dressing is obtained, and the preparation route is shown in figure 1.
The double-layer nanofiber membrane dressing prepared in example 1 has a porosity of 93.8% and a specific surface area of 5.69m 2 And/g. The bacterial colony forming units (colony forming units, CFU) on the petri dish were calculated using staphylococcus aureus (s. Aureus) as the subject, thereby calculating the bacterial sterilization rate of the dressing, and thus investigating the antibacterial performance of the dressing. The sterilization rate of the double-layer nanofiber membrane dressing prepared in example 1 was 100%. The blood coagulation rate was determined by absorbance values of the hemoglobin solution using a coagulation index (BCI) evaluation in vitro. The BCI of the double layer nanofiber membrane dressing was 63.6%.
Example 3
1.8g of chitosan is added into 30g of dilute acetic acid solution with mass concentration of 2%, and the solution is dissolved by magnetic stirring, thus obtaining chitosan solution. Spinning the solution by using an electrostatic spinning device, wherein the spinning voltage is 20kV, the distance between a spinning needle head and a collecting plate is 20cm, the spinning speed is 0.2mL/h, the spinning film is torn off from aluminum foil paper, and the chitosan nanofiber film is obtained by vacuum drying. And irradiating the chitosan nanofiber membrane with low-temperature plasma under the protection of nitrogen, wherein the irradiation power is 130W, the pressure is 300Pa, and the irradiation time is 4min. After the irradiation is finished, the chitosan nanofiber membrane is soaked in an aspirin/ethanol mixed solution, and the reaction is carried out for 3 hours at 70 ℃. And after the reaction is finished, washing the mixture with ethanol for a plurality of times to ensure that unreacted aspirin is removed, and drying the mixture in vacuum to obtain the aspirin modified chitosan nanofiber membrane.
0.5g of polylactic acid is added into 10g of mixed solvent of tetrahydrofuran/formamide (mass ratio is 1:5), and the mixture is magnetically stirred for 2 hours at 50 ℃ to form quenching liquid. Pouring the quenching liquid into a culture dish with the diameter of 8cm, putting the culture dish into a low-temperature refrigerator with the temperature of minus 15 ℃ for quenching for 300min, and putting the culture dish into a vacuum freeze dryer for drying for 3 days after the quenching is finished to obtain the polylactic acid nanofiber membrane. 0.4g of polylactic acid nanofiber membrane is soaked in 100mL of dopamine solution with the concentration of 0.2mg/mL, the pH value is adjusted to be 8.5 by using Tris salt buffer solution, and after soaking for 30min, dopamine is deposited and polymerized on the surface of the fiber. And then cleaning the fiber membrane with deionized water for 3 times, and vacuum drying to obtain the polydopamine coated polylactic acid nanofiber membrane.
Spreading 0.2g polydopamine coated polylactic acid nanofiber membrane in a culture dish, spreading 0.5g aspirin modified chitosan nanofiber membrane on the surface of the culture dish, adding a mixed solution of ethylene glycol glycidyl ether and ethanol with the mass ratio of 1:4 into the culture dish, and soaking for 24 hours at normal temperature. Finally, placing the culture dish into a vacuum freeze dryer for drying for 3 days to obtain the double-layer nanofiber membrane dressing.
The dressing of the double-layer nanofiber membrane prepared in example 1 has a porosity of 94.0% and a specific surface area of 5.56m 2 And/g. The bacterial colony forming units (colony forming units, CFU) on the petri dish were calculated using staphylococcus aureus (s. Aureus) as the subject, thereby calculating the bacterial sterilization rate of the dressing, and thus investigating the antibacterial performance of the dressing. The sterilization rate of the double-layer nanofiber membrane dressing prepared in example 1 was 100%. The blood coagulation rate was determined by absorbance values of the hemoglobin solution using a coagulation index (BCI) evaluation in vitro. The BCI of the double layer nanofiber membrane dressing was 63.1%.
Comparative example 1
Unlike example 1, comparative example 1 directly uses aspirin modified chitosan nanofiber membrane for subsequent hemostatic dressing. The porosity of the aspirin modified chitosan nanofiber membrane dressing prepared in comparative example 1 is 89.1%, and the specific surface area is 5.78m 2 And/g. The sterilization rate of the aspirin modified chitosan nanofiber membrane dressing prepared in example 1 is 100%. The blood coagulation rate was determined by absorbance values of the hemoglobin solution using a coagulation index (BCI) evaluation in vitro. The BCI of the aspirin modified chitosan nanofiber membrane dressing was 48.1%.
Comparative example 2
Unlike example 1, comparative example 2 directly uses polydopamine-coated polylactic acid nanofiber membrane for subsequent hemostatic dressing. The porosity of the polydopamine coated polylactic acid nanofiber membrane prepared in comparative example 2 is 90.1%, and the specific surface area is 6.25m2/g. The sterilization rate of the polydopamine coated polylactic acid nanofiber membrane dressing prepared in the example 1 is 92%. The blood coagulation rate was determined by absorbance values of the hemoglobin solution using a coagulation index (BCI) evaluation in vitro. The BCI of the aspirin modified chitosan nanofiber membrane dressing was 35.2%.
The foregoing describes specific embodiments of the present invention. It is to be understood that the invention is not limited to the particular embodiments described above, and that various changes and modifications may be made by one skilled in the art within the scope of the claims without affecting the spirit of the invention.
Claims (7)
1. The preparation method of the double-layer nanofiber membrane dressing is characterized by comprising the following steps of:
s1, preparing a chitosan nanofiber membrane by using an electrostatic spinning method;
s2, under the protection of nitrogen, irradiating the chitosan nanofiber membrane by using low-temperature plasma, soaking the chitosan nanofiber membrane in an ethanol solution of aspirin, and reacting at 60-80 ℃ to obtain an aspirin modified chitosan nanofiber membrane;
s3, adding polylactic acid into a tetrahydrofuran/formamide mixed solvent, magnetically stirring to form a quenching liquid, carrying out low-temperature quenching on the quenching liquid, and then carrying out vacuum freeze drying to obtain a polylactic acid nanofiber membrane, soaking the polylactic acid nanofiber membrane in a dopamine solution, adjusting the pH value to 8-9, and obtaining the polydopamine coated polylactic acid nanofiber membrane after soaking;
s4, paving the aspirin modified chitosan nanofiber membrane on the surface of the polydopamine coated polylactic acid nanofiber membrane in an ethylene glycol glycidyl ether/ethanol mixed solution, soaking at normal temperature, and then performing vacuum freeze drying to obtain the double-layer nanofiber membrane dressing.
2. The method for preparing the double-layer nanofiber membrane dressing according to claim 1, wherein the method for preparing the chitosan nanofiber membrane comprises the following steps:
adding chitosan into a dilute acetic acid solution, and dissolving to obtain spinning solution;
carrying out electrostatic spinning operation on the spinning solution, wherein the conditions for controlling the electrostatic spinning are as follows: the spinning voltage is 15-30 kV, the distance between the spinning needle head and the collecting plate is 10-20 cm, and the spinning speed is 0.1-0.4 mL/h; the low-temperature plasma irradiation conditions are as follows: the irradiation power is 120-200W, the pressure is 200-300 Pa, and the irradiation time is 3-10 min.
3. The method for preparing a double-layer nanofiber membrane dressing according to claim 2, wherein the mass concentration of the dilute acetic acid solution is 1-3%; in the spinning solution, the mass concentration of chitosan is 3-10%.
4. The method for preparing the double-layer nanofiber membrane dressing according to claim 1, wherein the mass concentration of polylactic acid in the quenching liquid is 3-8%; in the tetrahydrofuran/formamide mixed solvent, the mass ratio of tetrahydrofuran to formamide is 1: (3-6); the temperature of the low-temperature quenching is-40 to-10 ℃ and the quenching time is 100 to 300min; the concentration of the dopamine solution is 0.1-0.3 mg/mL.
5. The method for preparing the double-layer nanofiber membrane dressing according to claim 1, wherein the mass ratio of the aspirin modified chitosan nanofiber membrane to the polydopamine coated polylactic acid nanofiber membrane is (3-5):
(1-3); the mass ratio of the ethylene glycol glycidyl ether to the ethanol is 1: (1-5).
6. A double-layered nanofiber membrane dressing obtainable by the method of any one of claims 1 to 5.
7. Use of the bilayer nanofiber membrane dressing of claim 6 in hemostatic and anti-inflammatory materials.
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