CN111945301A - Electrostatic spinning membrane releasing nitric oxide based on near-infrared response and preparation method and application thereof - Google Patents
Electrostatic spinning membrane releasing nitric oxide based on near-infrared response and preparation method and application thereof Download PDFInfo
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- D—TEXTILES; PAPER
- D04—BRAIDING; LACE-MAKING; KNITTING; TRIMMINGS; NON-WOVEN FABRICS
- D04H—MAKING TEXTILE FABRICS, e.g. FROM FIBRES OR FILAMENTARY MATERIAL; FABRICS MADE BY SUCH PROCESSES OR APPARATUS, e.g. FELTS, NON-WOVEN FABRICS; COTTON-WOOL; WADDING ; NON-WOVEN FABRICS FROM STAPLE FIBRES, FILAMENTS OR YARNS, BONDED WITH AT LEAST ONE WEB-LIKE MATERIAL DURING THEIR CONSOLIDATION
- D04H1/00—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres
- D04H1/70—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres characterised by the method of forming fleeces or layers, e.g. reorientation of fibres
- D04H1/72—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres characterised by the method of forming fleeces or layers, e.g. reorientation of fibres the fibres being randomly arranged
- D04H1/728—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres characterised by the method of forming fleeces or layers, e.g. reorientation of fibres the fibres being randomly arranged by electro-spinning
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- D—TEXTILES; PAPER
- D01—NATURAL OR MAN-MADE THREADS OR FIBRES; SPINNING
- D01D—MECHANICAL METHODS OR APPARATUS IN THE MANUFACTURE OF ARTIFICIAL FILAMENTS, THREADS, FIBRES, BRISTLES OR RIBBONS
- D01D5/00—Formation of filaments, threads, or the like
- D01D5/0007—Electro-spinning
- D01D5/0015—Electro-spinning characterised by the initial state of the material
- D01D5/003—Electro-spinning characterised by the initial state of the material the material being a polymer solution or dispersion
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- D—TEXTILES; PAPER
- D01—NATURAL OR MAN-MADE THREADS OR FIBRES; SPINNING
- D01D—MECHANICAL METHODS OR APPARATUS IN THE MANUFACTURE OF ARTIFICIAL FILAMENTS, THREADS, FIBRES, BRISTLES OR RIBBONS
- D01D5/00—Formation of filaments, threads, or the like
- D01D5/28—Formation of filaments, threads, or the like while mixing different spinning solutions or melts during the spinning operation; Spinnerette packs therefor
- D01D5/30—Conjugate filaments; Spinnerette packs therefor
- D01D5/34—Core-skin structure; Spinnerette packs therefor
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- D01—NATURAL OR MAN-MADE THREADS OR FIBRES; SPINNING
- D01F—CHEMICAL FEATURES IN THE MANUFACTURE OF ARTIFICIAL FILAMENTS, THREADS, FIBRES, BRISTLES OR RIBBONS; APPARATUS SPECIALLY ADAPTED FOR THE MANUFACTURE OF CARBON FILAMENTS
- D01F1/00—General methods for the manufacture of artificial filaments or the like
- D01F1/02—Addition of substances to the spinning solution or to the melt
- D01F1/10—Other agents for modifying properties
- D01F1/103—Agents inhibiting growth of microorganisms
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- D—TEXTILES; PAPER
- D01—NATURAL OR MAN-MADE THREADS OR FIBRES; SPINNING
- D01F—CHEMICAL FEATURES IN THE MANUFACTURE OF ARTIFICIAL FILAMENTS, THREADS, FIBRES, BRISTLES OR RIBBONS; APPARATUS SPECIALLY ADAPTED FOR THE MANUFACTURE OF CARBON FILAMENTS
- D01F8/00—Conjugated, i.e. bi- or multicomponent, artificial filaments or the like; Manufacture thereof
- D01F8/04—Conjugated, i.e. bi- or multicomponent, artificial filaments or the like; Manufacture thereof from synthetic polymers
- D01F8/10—Conjugated, i.e. bi- or multicomponent, artificial filaments or the like; Manufacture thereof from synthetic polymers with at least one other macromolecular compound obtained by reactions only involving carbon-to-carbon unsaturated bonds as constituent
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- D—TEXTILES; PAPER
- D01—NATURAL OR MAN-MADE THREADS OR FIBRES; SPINNING
- D01F—CHEMICAL FEATURES IN THE MANUFACTURE OF ARTIFICIAL FILAMENTS, THREADS, FIBRES, BRISTLES OR RIBBONS; APPARATUS SPECIALLY ADAPTED FOR THE MANUFACTURE OF CARBON FILAMENTS
- D01F8/00—Conjugated, i.e. bi- or multicomponent, artificial filaments or the like; Manufacture thereof
- D01F8/18—Conjugated, i.e. bi- or multicomponent, artificial filaments or the like; Manufacture thereof from other substances
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- D—TEXTILES; PAPER
- D04—BRAIDING; LACE-MAKING; KNITTING; TRIMMINGS; NON-WOVEN FABRICS
- D04H—MAKING TEXTILE FABRICS, e.g. FROM FIBRES OR FILAMENTARY MATERIAL; FABRICS MADE BY SUCH PROCESSES OR APPARATUS, e.g. FELTS, NON-WOVEN FABRICS; COTTON-WOOL; WADDING ; NON-WOVEN FABRICS FROM STAPLE FIBRES, FILAMENTS OR YARNS, BONDED WITH AT LEAST ONE WEB-LIKE MATERIAL DURING THEIR CONSOLIDATION
- D04H1/00—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres
- D04H1/40—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres from fleeces or layers composed of fibres without existing or potential cohesive properties
- D04H1/42—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres from fleeces or layers composed of fibres without existing or potential cohesive properties characterised by the use of certain kinds of fibres insofar as this use has no preponderant influence on the consolidation of the fleece
- D04H1/4382—Stretched reticular film fibres; Composite fibres; Mixed fibres; Ultrafine fibres; Fibres for artificial leather
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- D—TEXTILES; PAPER
- D04—BRAIDING; LACE-MAKING; KNITTING; TRIMMINGS; NON-WOVEN FABRICS
- D04H—MAKING TEXTILE FABRICS, e.g. FROM FIBRES OR FILAMENTARY MATERIAL; FABRICS MADE BY SUCH PROCESSES OR APPARATUS, e.g. FELTS, NON-WOVEN FABRICS; COTTON-WOOL; WADDING ; NON-WOVEN FABRICS FROM STAPLE FIBRES, FILAMENTS OR YARNS, BONDED WITH AT LEAST ONE WEB-LIKE MATERIAL DURING THEIR CONSOLIDATION
- D04H3/00—Non-woven fabrics formed wholly or mainly of yarns or like filamentary material of substantial length
- D04H3/005—Synthetic yarns or filaments
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- D—TEXTILES; PAPER
- D04—BRAIDING; LACE-MAKING; KNITTING; TRIMMINGS; NON-WOVEN FABRICS
- D04H—MAKING TEXTILE FABRICS, e.g. FROM FIBRES OR FILAMENTARY MATERIAL; FABRICS MADE BY SUCH PROCESSES OR APPARATUS, e.g. FELTS, NON-WOVEN FABRICS; COTTON-WOOL; WADDING ; NON-WOVEN FABRICS FROM STAPLE FIBRES, FILAMENTS OR YARNS, BONDED WITH AT LEAST ONE WEB-LIKE MATERIAL DURING THEIR CONSOLIDATION
- D04H3/00—Non-woven fabrics formed wholly or mainly of yarns or like filamentary material of substantial length
- D04H3/02—Non-woven fabrics formed wholly or mainly of yarns or like filamentary material of substantial length characterised by the method of forming fleeces or layers, e.g. reorientation of yarns or filaments
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- D—TEXTILES; PAPER
- D10—INDEXING SCHEME ASSOCIATED WITH SUBLASSES OF SECTION D, RELATING TO TEXTILES
- D10B—INDEXING SCHEME ASSOCIATED WITH SUBLASSES OF SECTION D, RELATING TO TEXTILES
- D10B2509/00—Medical; Hygiene
- D10B2509/02—Bandages, dressings or absorbent pads
- D10B2509/022—Wound dressings
Abstract
The invention provides an electrostatic spinning film for releasing nitric oxide based on near-infrared response and a preparation method and application thereof, and relates to the technical field of medical dressings. The electrostatic spinning membrane provided by the invention is formed by interweaving core-shell nano fibers obtained by coaxial electrostatic spinning; the core-shell nanofiber comprises a shell layer and a core; the shell layer comprises polyvinyl alcohol, chitosan and Prussian blue-chitosan nanoparticles with a nitric oxide release function; the core includes polyvinyl alcohol and a bioactive molecule. The electrostatic spinning membrane capable of releasing nitric oxide based on near-infrared response provided by the invention can play a role in resisting bacteria and accelerating healing of a wound surface through the synergistic effect of the core-shell nanofiber shell layer and the core layer core and the adjustable and controllable release of nitric oxide, has good fitting property, and can be effectively applied as an antibacterial auxiliary material. The invention adopts the coaxial electrostatic spinning technology to prepare the electrostatic spinning membrane, has simple operation and low cost, and is convenient for large-scale production.
Description
Technical Field
The invention relates to the technical field of medical dressings, in particular to an electrostatic spinning film capable of releasing nitric oxide based on near-infrared response and a preparation method and application thereof.
Background
The skin, which is the largest organ of the human body, helps to prevent invasion of microorganisms such as bacteria and viruses from the outside, protects important tissues such as blood vessels and nerves in the body, and protects the body from damage, and thus, it is essential to protect the healthy integrity of the skin. However, the skin is inevitably traumatized, and if not properly treated, it may affect the health of the person or even endanger life. The skin injury is caused by many reasons, including heredity, acute wound, chronic wound, surgical wound, etc. Wound healing is a progressive, complex physiological process that is generally divided into three phases, inflammation, proliferation and remodeling. After wound infection, wounds are more difficult to heal, and diseases such as wound septicemia and the like can be caused in severe cases. Therefore, skin wounds caused by wounds, burns, chronic diseases, etc., remain a great clinical challenge worldwide. Immediately after the skin is wounded, a wound dressing should be used to prevent wound infection, relieve pain, and promote skin regeneration. The traditional wound dressing can prevent bacterial infection to a certain extent, but has no obvious promotion effect on wound healing.
Nitric Oxide (NO) isEndogenous lipophilic molecules play a key regulatory role in many physiological and pathological processes, and in particular play a critical role in fighting infection in the natural immune system. NO molecule and its by-products peroxynitrite (ONOO-) and dinitrogen trioxide (N) generated by reaction with oxidant in ambient environment2O3) Easily causes lipid peroxidation of bacterial membranes, thereby showing remarkable antibacterial performance. Vasodilation by NO is an important process in wound healing, and it promotes blood circulation and transport of nutrients to the lesion. During the proliferative phase of wound healing, NO can drive the expression of Vascular Endothelial Growth Factor (VEGF), thereby stimulating angiogenesis, and these NO-induced growth factors can also promote cell migration, adhesion and proliferation, thereby enhancing the growth of endothelial cells, fibroblasts and keratinocytes, thereby helping repair tissues and blood vessels. In the final remodeling stage, NO also plays a role in stimulating fibroblast proliferation to increase collagen formation and deposition on the wound. Patent CN201580022323.3 discloses a wound treatment membrane releasing NO, which shows better effect in promoting anti-infection wound healing, but NO is an active chemical molecule, and excessive release can cause inflammation and cause oxidative stress reaction of normal nuclear tissues, with adverse consequences. Therefore, designing a wound dressing capable of controllably releasing NO is of great significance.
Disclosure of Invention
In view of the above, the present invention aims to provide an electrospun membrane releasing nitric oxide based on near-infrared response, and a preparation method and an application thereof. The electrostatic spinning membrane capable of releasing nitric oxide based on near-infrared response provided by the invention not only can play a role in resisting bacteria and accelerating healing of a wound surface, but also can realize adjustable and controllable release of nitric oxide.
In order to achieve the above purpose, the invention provides the following technical scheme:
the invention provides an electrostatic spinning membrane for releasing nitric oxide based on near-infrared response, which is formed by interweaving core-shell nano fibers obtained by coaxial electrostatic spinning; the core-shell nanofiber comprises a shell layer and a core; the shell layer comprises polyvinyl alcohol, chitosan and Prussian blue-chitosan nanoparticles with a nitric oxide release function; the prussian blue-chitosan nano particle comprises prussian blue nano crystal doped with sodium nitroprusside and chitosan coated on the surface of the prussian blue nano crystal; the core includes polyvinyl alcohol and a bioactive molecule.
Preferably, the molecular weight of chitosan in the prussian blue-chitosan nanoparticle is 3-100 kDa; the particle size of the Prussian blue-chitosan nanoparticles is 30-200 nm; the mass content of the Prussian blue-chitosan nanoparticles in the shell layer is 0.05-2.5%.
Preferably, the polymerization degree of polyvinyl alcohol in the shell layer is 1500-2400, and the molecular weight of chitosan in the shell layer is 3-100 kDa; the mass ratio of the chitosan, the polyvinyl alcohol and the Prussian blue-chitosan nanoparticles in the shell layer is 2-5: 10-15: 0.01 to 0.5.
Preferably, the polymerization degree of polyvinyl alcohol in the core is 1500-2400, and the bioactive molecules comprise one or more of collagen, growth factors and small molecule drugs; the mass ratio of polyvinyl alcohol to bioactive molecules in the core is 10-50: 0.1 to 2.
Preferably, the mass ratio of the shell layer to the core is 2-5: 1 to 0.5; the diameter of the core-shell nanofiber is 50-500 nm.
The invention provides a preparation method of an electrostatic spinning membrane based on near-infrared response nitric oxide release, which comprises the following steps:
(1) mixing a glacial acetic acid aqueous solution of chitosan with potassium ferricyanide, sodium nitroprusside and hydrochloric acid, heating to 50-100 ℃ for coprecipitation reaction to obtain the prussian blue-chitosan nanoparticles with the nitric oxide release function;
(2) carrying out coaxial electrostatic spinning by taking a mixed aqueous solution containing the Prussian blue-chitosan nanoparticles, polyvinyl alcohol, chitosan and glacial acetic acid as a shell layer electrospinning solution and a mixed aqueous solution containing polyvinyl alcohol and bioactive molecules as a core electrospinning solution to obtain core-shell nanofibers; the core-shell nanofibers are interwoven to form the near-infrared response-based nitric oxide releasing electrostatic spinning membrane.
Preferably, in the step (1), the mass concentration of chitosan in the glacial acetic acid aqueous solution of chitosan is 0.5-20 mg/mL, and the mass concentration of glacial acetic acid is 0.5-2.5%; the mass ratio of the potassium ferricyanide to the sodium nitroprusside is 1: 1-1: 10, the mass ratio of the sum of the potassium ferricyanide and sodium nitroprusside to the chitosan is 1-5: 0.1 to 1.
Preferably, in the step (2), the mass concentration of the prussian blue-chitosan nanoparticles in the shell layer electrospinning solution is 0.5-10 mg/mL, the mass concentration of polyvinyl alcohol is 2-20%, the mass concentration of chitosan is 0.1-5%, and the mass concentration of glacial acetic acid is 0.1-2.5%; the mass concentration of polyvinyl alcohol in the core electrospinning liquid is 2-20%, and the mass concentration of bioactive molecules is 0.5-50 mg/mL;
the mass ratio of the shell layer electrospinning liquid to the core electrospinning liquid is 5-2: 1 to 0.5.
Preferably, the conditions of the coaxial electrospinning in the step (2) include: the voltage is 10-20 kV, the spinning temperature is 15-50 ℃, the relative humidity is 10-60%, the spinning needle is 5-23G, and the collection distance is 5-20 cm.
The invention provides an application of the electrostatic spinning film capable of releasing nitric oxide based on near-infrared response in the technical scheme or the electrostatic spinning film capable of releasing nitric oxide based on near-infrared response prepared by the preparation method in the technical scheme as an antibacterial dressing.
The invention provides an electrostatic spinning membrane for releasing nitric oxide based on near-infrared response, which is formed by interweaving core-shell nano fibers obtained by coaxial electrostatic spinning; the core-shell nanofiber comprises a shell layer and a core; the shell layer comprises polyvinyl alcohol, chitosan and Prussian blue-chitosan nanoparticles with a nitric oxide release function, and the Prussian blue-chitosan nanoparticles comprise Prussian blue nanocrystals doped with sodium nitroprusside and chitosan coated on the surfaces of the Prussian blue nanocrystals; the core includes polyvinyl alcohol and a bioactive molecule. In the invention, sodium nitroprusside in the shell layer of the core-shell nanofiber is used as a donor of nitric oxide, Prussian blue has excellent near-infrared photothermal conversion capability, and the formed Prussian blue-chitosan nanoparticles can realize the release of dosage control type NO at a specific target position by adjusting the irradiation position and the intensity of near-infrared laser, thereby playing the roles of resisting bacteria and promoting the healing of a wound surface, reducing the release of NO at a non-target position and reducing the toxicity and side effects possibly caused; bioactive molecules in the core of the core-shell nanofiber can improve the ability of the electrostatic spinning membrane material in accelerating wound healing. In addition, the electrostatic spinning membrane can play a supporting role, promote the intercellular interaction at the wound healing position, and stimulate the formation of new tissues through the release of bioactive molecules in the core, thereby promoting the wound healing and the reconstruction of the tissues. Therefore, the electrostatic spinning membrane capable of releasing nitric oxide based on near-infrared response provided by the invention can play a role in resisting bacteria and accelerating healing of a wound surface and realize adjustable and controllable release of nitric oxide through the synergistic effect of the core-shell nanofiber shell layer and the core; in addition, the dressing has good fitting property, can be well covered on wounds, and can be further effectively applied as an antibacterial auxiliary material.
The invention provides a preparation method of the electrostatic spinning membrane for releasing nitric oxide based on near-infrared response.
Drawings
FIG. 1 is a schematic diagram of a process for preparing the near-infrared response based nitric oxide releasing electrospun membrane according to the invention;
FIG. 2 is a nano transmission electron microscope image of Prussian blue-chitosan nanoparticles having nitric oxide releasing function prepared in example 1, wherein (a) and (b) in FIG. 2 are nano transmission electron microscope images at different magnifications, respectively;
FIG. 3 is a transmission electron microscope image of C, Fe, N, O elements in Prussian blue-chitosan nanoparticles with nitric oxide releasing function prepared in example 1;
FIG. 4 shows Prussian blue-chitosan nanoparticles with nitric oxide releasing function at different concentrations in example 1 at a power density of 0.35W/cm2808 (d) ofA temperature rise curve under the irradiation of nm near-infrared laser;
FIG. 5 is a temperature rise curve of the Prussian blue-chitosan nanoparticles with nitric oxide releasing function in example 1 at a concentration of 50 μ g/mL under 808nm near-infrared laser irradiation under different power conditions;
fig. 6 is a photo-thermal stability curve of prussian blue-chitosan nanoparticles having nitric oxide releasing function prepared in example 1;
FIG. 7 is a graph showing NO release of Prussian blue-chitosan nanoparticles having nitric oxide releasing function at a concentration of 100. mu.g/mL in example 1 under 808nm near-infrared laser irradiation under different power conditions;
FIG. 8 shows the switching power density of 0.75W/cm in example 12The 808nm near-infrared laser and a curve chart of NO release of Prussian blue-chitosan nanoparticles with the nitric oxide release function are shown;
FIG. 9 is an SEM image of core-shell nanofibers prepared in example 1, wherein (a) and (b) in FIG. 9 are SEM images at different magnifications, respectively;
FIG. 10 is a graph showing the antibacterial effect of the nucleocapsid nanofibers prepared in example 1 on Staphylococcus aureus and Escherichia coli, and in FIG. 10, (a) is a graph showing the antibacterial effect of the nucleocapsid nanofibers on Staphylococcus aureus, and (b) is a graph showing the antibacterial effect of the nucleocapsid nanofibers on Escherichia coli;
FIG. 11 is a graph showing the healing of Staphylococcus aureus infected wounds in different groups of experimental mice in example 1;
FIG. 12 is a graph of HE staining and Masson staining at skin lesions in mice infected with Staphylococcus aureus from different treatment groups of example 1.
Detailed Description
The invention provides an electrostatic spinning membrane for releasing nitric oxide based on near-infrared response, which is formed by interweaving coaxial electrostatic spinning; the core-shell nanofiber comprises a shell layer and a core; the shell layer comprises polyvinyl alcohol, chitosan and Prussian blue-chitosan nanoparticles with a nitric oxide release function, and the Prussian blue-chitosan nanoparticles comprise Prussian blue nanocrystals doped with sodium nitroprusside and chitosan coated on the surfaces of the Prussian blue nanocrystals; the core includes polyvinyl alcohol and a bioactive molecule.
In the invention, the core-shell nanofiber comprises a shell layer, wherein the shell layer comprises polyvinyl alcohol, chitosan and Prussian blue-chitosan nanoparticles with a nitric oxide release function, and the Prussian blue-chitosan nanoparticles comprise Prussian blue nanocrystals doped with sodium nitroprusside and chitosan coated on the surfaces of the Prussian blue nanocrystals. In the invention, the molecular weight of chitosan in the prussian blue-chitosan nanoparticle is preferably 3-100 kDa, and more preferably 15-50 kDa. In the invention, the particle size of the Prussian blue-chitosan nanoparticle is preferably 30-200 nm, and more preferably 50-100 nm; the mass content of the Prussian blue-chitosan nanoparticles in the shell layer is preferably 0.05-2.5%. In the invention, sodium nitroprusside in the prussian blue-chitosan nanoparticles is used as a nitric oxide donor, prussian blue has excellent near-infrared photothermal conversion capability, and prussian blue nanocrystals doped with sodium nitroprusside enable the nanoparticles to realize the release of dosage control type NO at a specific target position by adjusting the irradiation position of near-infrared laser, thereby playing the roles of resisting bacteria and promoting the healing of wound surfaces, reducing the release of NO at non-target positions and reducing the toxicity and side effects possibly caused; the chitosan in the Prussian blue-chitosan nano particles is used as a stabilizer, so that the stability and biocompatibility of the doped sodium nitroprusside-Prussian blue nano crystals are improved.
In the invention, the polymerization degree of polyvinyl alcohol in the shell layer is preferably 1500-2400, more preferably 1700-2200, and in the embodiment of the invention, the polyvinyl alcohol is 1788 type polyvinyl alcohol, 1795 type polyvinyl alcohol or 2000 type polyvinyl alcohol; the molecular weight of the chitosan in the shell layer is preferably 3-100 kDa, and more preferably 15-50 kDa. In the invention, the mass ratio of the chitosan, the polyvinyl alcohol and the Prussian blue-chitosan nanoparticles in the shell layer is preferably 2-5: 10-15: 0.01 to 0.5, more preferably 3: 10: 0.25; the chitosan in the shell layer can have good blood coagulation, hemostasis and antibacterial effects, the biocompatibility is good, the chitosan can be automatically degraded, and the polyvinyl alcohol can improve the spinnability of the chitosan nanofiber and the appearance of the chitosan nanofiber.
In the present invention, the core-shell nanofiber comprises a core comprising polyvinyl alcohol and a bioactive molecule. In the invention, the polymerization degree of the polyvinyl alcohol in the core is preferably 1500-2400, more preferably 1700-2200, in the embodiment of the invention, the polyvinyl alcohol is 1788 type polyvinyl alcohol, 1795 type polyvinyl alcohol or 2000 type polyvinyl alcohol; the bioactive molecules preferably comprise one or more of collagen, growth factors and small molecule drugs (deferoxamine, curcumin and the like) for promoting wound healing; the mass ratio of polyvinyl alcohol to bioactive molecules in the core is preferably 50-10: 2 to 0.1, more preferably 50: 1. in the invention, the bioactive molecules in the core of the core-shell nanofiber can improve the ability of the electrostatic spinning membrane material to accelerate wound healing.
In the invention, the mass ratio of the shell layer to the core is preferably 5-2: 1 to 0.5, more preferably 2: 1; the diameter of the composite core-shell nanofiber is preferably 50-500 nn, and more preferably 50-100 nm. The electrostatic spinning membrane capable of releasing nitric oxide based on near-infrared response provided by the invention has the advantages that through the synergistic effect of the core-shell nanofiber shell and the core, the effects of resisting bacteria and accelerating healing of a wound surface can be achieved, and the adjustable release of nitric oxide can be realized; in addition, the dressing has good fitting property, can be well covered on wounds, and can be further effectively applied as an antibacterial dressing.
The invention provides a preparation method of an electrostatic spinning membrane based on near-infrared response nitric oxide release, which comprises the following steps:
(1) mixing a glacial acetic acid aqueous solution of chitosan with potassium ferricyanide, sodium nitroprusside and hydrochloric acid, heating to 50-100 ℃ for coprecipitation reaction to obtain the prussian blue-chitosan nanoparticles with the nitric oxide release function;
(2) carrying out coaxial electrostatic spinning by taking a mixed aqueous solution containing the Prussian blue-chitosan nanoparticles, polyvinyl alcohol, chitosan and glacial acetic acid as a shell layer electrospinning solution and a mixed aqueous solution containing polyvinyl alcohol and bioactive molecules as a core electrospinning solution to obtain core-shell nanofibers; and the core-shell nano fibers are interwoven to form the near-infrared response nitric oxide releasing electrostatic spinning membrane.
The process for preparing the electrostatic spinning membrane based on the near infrared response to release nitric oxide is shown in figure 1.
Mixing a glacial acetic acid aqueous solution of chitosan with potassium ferricyanide, sodium nitroprusside and hydrochloric acid, heating to 50-100 ℃ for coprecipitation reaction, and obtaining the prussian blue-chitosan nanoparticle with the nitric oxide release function. In the invention, the mass concentration of chitosan in the glacial acetic acid aqueous solution of chitosan is preferably 0.5-20 mg/mL, and more preferably 5-20 mg/mL; the mass concentration of the glacial acetic acid is preferably 0.5-2.5%. In the invention, the mass ratio of potassium ferricyanide to sodium nitroprusside is preferably 1: 1-1: 10, more preferably 1: 4-1: 9; the mass ratio of the sum of the potassium ferricyanide and sodium nitroprusside to the chitosan is preferably 1-5: 0.1 to 1, more preferably 2 to 3.5: 0.1 to 0.8. In the invention, the concentration of the hydrochloric acid is preferably 0.1-0.5 mol/L, and more preferably 0.15-0.25 mol/L; the ratio of the volume of the hydrochloric acid to the volume of the glacial acetic acid aqueous solution of the chitosan is preferably 0.5-3: 1, more preferably 1: 1.
preferably, the potassium ferricyanide and the sodium nitroprusside are added into the glacial acetic acid aqueous solution of the chitosan to be mixed, and then the hydrochloric acid is slowly added. In the invention, the mixing is preferably stirring mixing, and the invention has no special requirements on the mixing speed and time and ensures that the components are uniformly mixed.
In the invention, the temperature of the coprecipitation reaction is preferably 60-80 ℃, and the time is preferably 4-24 h, and preferably 12-20 h. Because the structure of the sodium nitroprusside is very similar to that of the potassium ferricyanide for synthesizing the Prussian blue material, in the process of the coprecipitation reaction, the nitroso in the sodium nitroprusside is doped in the formed Prussian blue nano crystal, so that the Prussian blue nano crystal has the function of releasing nitric oxide; the chitosan serving as cheap aminopolysaccharide with high biocompatibility is coated on the surface of the Prussian blue nanocrystal through static electricity and complexation, so that the stability and biocompatibility of the nanomaterial are improved.
After the coprecipitation reaction, the invention preferably carries out solid-liquid separation, solid-phase washing and drying on the obtained coprecipitation reaction product in sequence to obtain the prussian blue-chitosan nano particle with the nitric oxide release function. In the invention, the solid-liquid separation method is preferably centrifugal separation, the speed of the centrifugal separation is preferably 10000-14000 r/min, and the time is preferably 8-10 min; the solid phase washing is preferably water washing, the number of the water washing is preferably at least 3, and unreacted raw materials remained on the surface of the solid phase are removed by washing; the drying is preferably freeze drying, the cold trap temperature of the freeze drying is preferably-50 ℃, the absolute pressure is preferably 3Pa, and the freeze drying time is preferably 1 day.
The Prussian blue-chitosan nanoparticles with the nitric oxide release function are preferably stored at the temperature of 4 ℃ for later use.
After obtaining the Prussian blue-chitosan nano particles with the nitric oxide release function, the invention takes the mixed aqueous solution of the Prussian blue-chitosan nano particles, polyvinyl alcohol, chitosan and glacial acetic acid as the shell layer electrospinning solution. In the invention, the mass concentration of the chitosan nanoparticles doped with sodium nitroprusside-Prussian blue in the shell layer electrospinning solution is 0.5-10 mg/mL, preferably 3-8 mg/mL; the mass concentration of the polyvinyl alcohol is preferably 2-20%, and more preferably 7-15%; the mass concentration of the chitosan is preferably 0.1-5%, and more preferably 0.4-1%; the mass concentration of the glacial acetic acid is preferably 0.1-2.5%, and more preferably 0.2-1%. The preparation method of the shell layer electrospinning solution has no special requirement, and the shell layer electrospinning solution can be obtained by uniformly mixing all the components; in the embodiment of the present invention, the shell solution is preferably prepared by the following method:
dissolving chitosan in a glacial acetic acid aqueous solution to obtain a chitosan solution; dispersing prussian blue-chitosan nanoparticles in a polyvinyl alcohol aqueous solution to obtain a polyvinyl alcohol dispersion liquid of the nanoparticles; and mixing the chitosan solution with the polyvinyl alcohol dispersion liquid of the nanoparticles to obtain the shell layer electrospinning liquid.
The core electrospinning liquid is a mixed aqueous solution containing polyvinyl alcohol and bioactive molecules, and the mass concentration of the polyvinyl alcohol in the core electrospinning liquid is preferably 2-20%, and more preferably 10-15%; the mass concentration of the bioactive molecules is preferably 0.5-50 mg/mL, and more preferably 2-20 mg/mL. The invention has no special requirements on the preparation method of the core electrospinning liquid, and can be obtained by uniformly mixing all the components.
The shell layer electrospinning solution and the core electrospinning solution are subjected to coaxial electrostatic spinning to obtain the core-shell nanofiber. In the invention, the mass ratio of the shell layer electrospinning liquid to the core electrospinning liquid is preferably 5-2: 1 to 0.5, more preferably 2 to 5: 1. in the present invention, the conditions of the coaxial electrospinning include: the voltage is preferably 10-20 kV, and more preferably 11-18 kV; the spinning temperature is preferably 15-50 ℃, more preferably 25-37 ℃, and the relative humidity is preferably 10-60%, more preferably 35-50%; the spinning needle is preferably 5-23G, and more preferably 18-23G (namely, the outer diameter of the spinning needle is 0.6mm, and the inner diameter is 0.3 mm); the collection distance is preferably 5-20 cm, and more preferably 10-16 cm. The invention has no special requirements on the specific operation method of the coaxial electrostatic spinning, and the operation method known by the technical personnel in the field can be adopted.
And after the core-shell nano fibers are obtained, interweaving the core-shell nano fibers to form the electrostatic spinning film for releasing nitric oxide based on near-infrared response. In the invention, the core-shell nanofibers are preferably accumulated on an electrospinning receiver, and then are interwoven to form an electrospinning film. The present invention does not require the electrospinning receiver to be particularly limited, and any electrospinning receiver known to those skilled in the art may be used.
The invention adopts the coaxial electrostatic spinning technology to prepare the electrostatic spinning membrane, has simple operation and low cost, and is convenient for large-scale production; and the fiber prepared by coaxial electrostatic spinning has large specific surface area, high porosity, strong ductility, better mechanical strength and toughness, and has very obvious advantages in local drug delivery compared with hydrogel or sponge.
The invention provides an application of the electrostatic spinning film capable of releasing nitric oxide based on near-infrared response in the technical scheme or the electrostatic spinning film capable of releasing nitric oxide based on near-infrared response prepared by the preparation method in the technical scheme as an antibacterial dressing. The present invention is not particularly limited to the specific embodiment of the application, and may be applied using antimicrobial dressings well known to those skilled in the art.
The following examples are provided to illustrate details of the near-infrared based nitric oxide releasing electrospun membrane of the present invention, and the preparation method and application thereof, but they should not be construed as limiting the scope of the present invention.
Example 1
(1) Preparation of prussian blue-chitosan nano-particle with nitric oxide release function
Dissolving chitosan with molecular weight of 50kDa in deionized water containing 1 wt.% of glacial acetic acid to prepare 100mL of 1mg/mL chitosan solution, adding 300mg of potassium ferricyanide and 2.5g of sodium nitroprusside into the chitosan solution under magnetic stirring, slowly adding 100mL of hydrochloric acid (0.1M), carrying out coprecipitation reaction for 16 hours under the condition of controlling the heating temperature to be 80 ℃, carrying out centrifugal separation (14000 r/min, 8 min) to collect a solid phase, washing for at least 3 times with water, removing unreacted raw materials, carrying out freeze drying for 3 days to obtain Prussian blue-chitosan nanoparticles with a nitric oxide release function, and storing at the temperature of 4 ℃.
(2) Preparation of core-shell nanofibers
Preparing a shell layer electrospinning solution: dissolving chitosan (molecular weight is 50kDa) in 1 wt% glacial acetic acid aqueous solution to prepare 3 wt% chitosan solution; dispersing prussian blue-chitosan nanoparticles with a nitric oxide release function in 10 wt% of 1788 type polyvinyl alcohol aqueous solution to obtain 5mg/mL nanoparticle polyvinyl alcohol dispersion liquid; mixing a chitosan solution and a polyvinyl alcohol dispersion liquid of nanoparticles according to a volume ratio of 1: 4, mixing to obtain a shell layer electrospinning solution;
the core electrospinning liquid comprises the following components: 1788 type polyvinyl alcohol aqueous solution containing 20mg/mL collagen, the concentration of polyvinyl alcohol in the solution is 10 wt%;
carrying out coaxial electrostatic spinning on the shell layer electrospinning solution and the core electrospinning solution, wherein the mass ratio of the shell layer electrospinning solution to the core electrospinning solution is 2: 1, spinning voltage is 11kV, collecting distance is 15cm, spinning needle is 23G, spinning temperature is 37 ℃, relative humidity is 40%, composite core-shell nanofibers are obtained, and the composite core-shell nanofibers are accumulated and collected in an electrostatic spinning receiver and are interwoven to form an electrostatic spinning film which responds to near infrared and releases nitric oxide.
Respectively characterizing the structure and the performance of the prepared Prussian blue-chitosan nanoparticles with the nitric oxide release function, the composite core-shell nanofibers and the electrostatic spinning film:
(a) characterization of prussian blue-chitosan nanoparticles (abbreviated as PB-NO NPs) having nitric oxide releasing function:
(a.1) particle size and morphology of Prussian blue-chitosan nanoparticles with nitric oxide release function
Obtaining the nanometer morphology of the nanoparticles through a transmission electron microscope: a drop of 50-fold diluted Prussian blue-chitosan nano-sample with nitric oxide release function (after washing) is placed on a supporting film of a copper mesh, after drying, the particle size, morphology and dispersion condition are observed under the voltage of 120kV by an HT7700 Transmission Electron Microscope (TEM), the transmission electron microscope picture of the Prussian blue-chitosan nano-particle with nitric oxide release function is shown as figure 2, and (a) and (b) in figure 2 are the nano transmission electron microscope pictures of the Prussian blue-chitosan nano-particle with nitric oxide release function under different magnifications.
The particle size of the prussian blue-chitosan nanoparticles with the nitric oxide release function is measured to be 200 nanometers, the PDI is 0.01-0.3, the distribution is uniform, and the shape of an electron microscope is circular.
(a.2) elemental constitution of Prussian blue-chitosan nanoparticles having nitric oxide releasing function
FIG. 3 is a transmission electron microscope image of C, Fe, N, O elements in Prussian blue-chitosan nano-particles with nitric oxide release function; C. the presence of chitosan was confirmed by element N, O, and the formation of prussian blue material was indicated by element Fe.
(a.3) photothermal Properties of Prussian blue-chitosan nanoparticles having nitric oxide releasing function
FIG. 4 is a differenceThe Prussian blue-chitosan nano-particles with the concentration and the nitric oxide release function have the power density of 0.75W/cm2Temperature rise curve under 808nm near-infrared laser irradiation. As can be seen from fig. 4, under laser irradiation, prussian blue-chitosan nanoparticles with nitric oxide release function have obvious concentration-dependent temperature rise; after irradiation for 6min, when the concentration of the nanoparticles is 25 mug/mL, the nano temperature reaches 56.2 ℃, and the temperature of water is basically unchanged, which shows that the Prussian blue-chitosan nanoparticles with the nitric oxide release function have good photo-thermal property.
FIG. 5 is a temperature rise curve of a Prussian blue-chitosan nano-particle with a nitric oxide release function at a concentration of 50 μ g/mL under 808nm near-infrared laser irradiation under different power conditions. As can be seen from FIG. 5, the laser power density with 808nm was from 0.35W/cm2Increased to 0.75W/cm2The Prussian blue-chitosan nano particles with the concentration of 50 mu g/mL and the nitric oxide release function have the temperature of 0.35W/cm2The temperature is increased by 24.3 ℃ under the power condition and is increased by 0.75W/cm2The power rise was 39.4 ℃. The result shows that the photothermal effect of the prussian blue-chitosan nano with the nitric oxide release function is positively correlated with the laser power.
The photo-thermal stability of the Prussian blue-chitosan nanoparticles with the nitric oxide release function is researched through an on/off period irradiation experiment, and the power density is 0.75W/cm when the irradiation is started2And 808nm near infrared laser irradiation, and fig. 6 is a measured photo-thermal stability curve of prussian blue-chitosan nanoparticles having a nitric oxide releasing function. As can be seen from FIG. 6, after 3 cycles, the heating and cooling performance of the Prussian blue-chitosan nanoparticles with the nitric oxide release function is kept unchanged, which shows that the material has good photo-thermal stability.
(a.4) Prussian blue-chitosan nano photoresponse NO release performance with nitric oxide release function
This example studies the near-infrared NO release behavior in vitro and monitors NO release using Griess method: adding Prussian blue-chitosan nano suspension with nitric oxide release function dispersed in 1mL of PBS into a quartz cuvetteIrradiating by 808nm near-infrared laser at different power densities; after the irradiation is finished, taking out the sample, centrifuging the sample at 18000rpm for 10min to avoid interference, and removing the precipitate; adding 100 mu L of Griess Reagent I and 100 mu L of Griess Reagent II into 100 mu L of supernatant in sequence, keeping out of the sun, and incubating for 10 minutes; the absorbance of the mixture at 540nm was then measured and quantified by the standard curve method. FIG. 7 shows that the Prussian blue-chitosan nano-particles with nitric oxide releasing function at the concentration of 50 mug/mL are in different power conditions (0, 0.5, 1, 1.5W/cm)2) 808nm near infrared laser irradiation. As can be seen from fig. 7, the nanoparticles have NO release function, and it is confirmed that sodium nitroprusside is doped in the nano system and can release NO under the irradiation of the near-infrared laser, and with the increase of irradiation power density and irradiation time, the amount of NO released by prussian blue-chitosan nanoparticles having nitric oxide release function is gradually increased, which indicates that dose-controlled NO release can be achieved by adjusting the intensity of the near-infrared laser and the irradiation time. The prussian blue-chitosan nano-particle with the nitric oxide release function can react to thermal stimulation when releasing NO, and the NO release rate of the prussian blue-chitosan nano-particle with the nitric oxide release function is in positive correlation with temperature.
This example also studies the near-infrared controllability of prussian blue-chitosan nanoparticles with nitric oxide release function on NO release: at a power density of 0.75W/cm2Under the condition of (1), using 808nm laser pulse to irradiate, researching NO release of prussian blue-chitosan nano particles with nitric oxide release function under near infrared control, firstly using laser to irradiate the nano particles for 5 minutes, and then closing the laser for 5 minutes; two additional laser irradiation cycles were performed; the amount of NO released was measured using Griess method described above over a selected time interval. FIG. 8 shows the switching power density of 0.75W/cm2808nm near infrared laser, Prussian blue-chitosan nano release NO curve chart with nitric oxide release function. As can be seen from fig. 8, when the near-infrared laser is turned on, the prussian blue-chitosan nanoparticles having a nitric oxide releasing function rapidly release NO; prussian blue-chitosan nano-particles with nitric oxide release function once near-infrared laser is turned offAlmost completely stops NO release. With the increase of the near infrared irradiation time, the NO release rate is kept relatively stable, and NO released by the Prussian blue-chitosan nano-particles with the nitric oxide release function shows extremely small fatigue signs under repeated intermittent irradiation. The prussian blue-chitosan nano NO release behavior controlled by near infrared and having the nitric oxide release function can greatly promote the release of NO at a specific site. By controlling the near-infrared laser switch and the power, the release amount of NO in the damaged skin can be controlled, and the curative effect of NO in treatment can be improved.
(b) Characterization of the core-shell nanofiber:
(b.1) morphology of core-shell nanofibers
Fig. 9 is an SEM image of the core-shell nanofibers, and (a) and (b) in fig. 9 are SEM images at different magnifications. The uniform fiber shape can be observed from FIG. 9, the diameter of the fiber is 50-100 nn, and the distribution is uniform.
(b.2) antibacterial test
PBS, staphylococcus aureus bacterial liquid, culture medium and composite core-shell nanofibers are prepared into solutions (0, 300 mug/mL, 750 mug/mL and 1.5mg/mL) with different nanofiber concentrations, and then 10-fold dilution is sequentially carried out to obtain the final concentration of 1.0x108CFU/mL of bacterial suspension. The control group was cultured in a shaker for 12h (without near infrared illumination), and the experimental group was incubated at 0.75W/cm2Irradiating with 808nm near infrared laser for 3min, and shake culturing in shaking table for 12 h. After 12 hours, the bacterial suspension of the experimental group and the control group was diluted in a double ratio and then plate-coated, and the results are shown in fig. 10 (a). The staphylococcus aureus liquid was replaced with the escherichia coli liquid, and the experiment was performed in the same manner, and the result is shown in fig. 10 (b). As can be seen from fig. 10, the nanofibers have better antibacterial properties, and particularly, after the treatment of infrared light irradiation and nanofiber, the colony counts are significantly reduced as the concentration increases.
Animal experiments are used for detecting the antibacterial and healing effects of the electrostatic spinning film prepared by the embodiment on the wound surface as the antibacterial dressing:
20 BALB/c female mice (6-8 weeks old) are taken and bred according to standard diet, after adaptive breeding for 1 week, round wounds with the diameter of 6mm are made on the backs. Smearing bacterial liquid on a mouse wound every 24 hours (smearing 20 mu L of staphylococcus aureus bacterial liquid on each mouse) and carrying out different treatments for 10 consecutive days, and establishing a wound bacterial liquid infected mouse model, which specifically comprises the following steps:
the experimental mice were divided into 4 groups of 5 mice each: a wound applied bacteria liquid non-treatment group (group one), a wound applied bacteria liquid illumination group (group two), a wound applied bacteria liquid and antibacterial dressing (prepared in the embodiment) non-illumination group (group three), and a wound applied bacteria liquid and antibacterial dressing (prepared in the embodiment) illumination group (group four); the light conditions in the experimental group were 0.75W/cm2808nm near infrared laser irradiation for 3 min.
The healing of staphylococcus aureus infected wounds of different groups of experimental mice was observed on days 1, 5 and 10 of the experiment, respectively, and the results are shown in fig. 11. As can be seen from fig. 11, after the infected wounds of the mice in the group four experimental groups were treated with near-infrared light, the scars became smaller or even disappeared, and the infected wounds were not ulcerated due to bacterial infection during the experimental procedure, while the skin layers of the other groups were incomplete and the wound surface boundaries were incomplete. These results show that the near-infrared response nitric oxide releasing electrospun membrane prepared in the example shows a good healing promotion effect in infected wounds.
The skin of 4 groups of staphylococcus aureus-infected mice with different treatments was completely removed, and placed in 4% paraformaldehyde tissue fixative for HE staining and Masson staining, respectively, with the results shown in fig. 12. As can be seen from fig. 12, compared with the group of four experimental mice before treatment, after 10 days of treatment, wound skinning appeared on the wound-bandaged part, and collagen fiber regeneration was shown, indicating that the electrostatic spinning membrane + near-infrared light group showed significantly increased collagen fiber regeneration ability, and the wound was significantly reduced compared with the other groups.
Example 2
(1) Preparation of prussian blue-chitosan nano-particle with nitric oxide release function
Dissolving chitosan with the molecular weight of 25kDa in deionized water containing 1 wt.% of glacial acetic acid to prepare 100mL of chitosan solution of 8mg/mL, adding 400mg of potassium ferricyanide and 2.0g of sodium nitroprusside into the chitosan solution under magnetic stirring, slowly adding 100mL of hydrochloric acid (0.2M), carrying out coprecipitation reaction for 24 hours under the condition of controlling the heating temperature to be 100 ℃, carrying out centrifugal separation (10000 r/min, 10 min) to collect a solid phase, washing for at least 3 times with water to remove unreacted raw materials, carrying out freeze drying for 3 days to obtain prussian blue-chitosan nanoparticles with the nitric oxide release function, and storing at the temperature of 4 ℃.
(2) Preparation of core-shell nanofibers
Preparing a shell layer electrospinning solution: dissolving chitosan (molecular weight of 25kDa) in 1 wt% glacial acetic acid aqueous solution to prepare 4 wt% chitosan solution; dispersing prussian blue-chitosan nanoparticles with a nitric oxide release function in a 2000-type polyvinyl alcohol aqueous solution with 10 wt% to obtain a polyvinyl alcohol dispersion liquid of 10mg/mL nanoparticles; mixing a chitosan solution and a polyvinyl alcohol dispersion liquid of nanoparticles according to a volume ratio of 1: 3, mixing to obtain the shell layer electrospinning solution.
The core electrospinning liquid comprises the following components: a type 2000 aqueous solution of polyvinyl alcohol containing 2mg/mL of deferoxamine, the concentration of the polyvinyl alcohol in the solution being 10% by weight;
carrying out coaxial electrostatic spinning on the shell layer electrospinning solution and the core electrospinning solution, wherein the mass ratio of the shell layer electrospinning solution to the core electrospinning solution is 3: 1, spinning voltage is 15kV, collecting distance is 20cm, a spinning needle is 20G, spinning temperature is 37 ℃, and relative humidity is 50%, so that the composite core-shell nanofiber is obtained, accumulated and collected in an electrostatic spinning receiver, and interwoven to form the near-infrared response nitric oxide releasing electrostatic spinning membrane.
Example 3
(1) Preparation of prussian blue-chitosan nano-particle with nitric oxide release function
Dissolving chitosan with the molecular weight of 15kDa in deionized water containing 1.5 wt.% of glacial acetic acid to prepare 100mL of 2mg/mL chitosan solution, adding 600mg of potassium ferricyanide and 2.5g of sodium nitroprusside into the chitosan solution under magnetic stirring, slowly adding 100mL of hydrochloric acid (0.15M), controlling the heating temperature to be 90 ℃, carrying out coprecipitation reaction for 20 hours, carrying out centrifugal separation (12000 r/min, 10 minutes) to collect a solid phase, then washing with water for at least 3 times, removing unreacted raw materials, and carrying out freeze drying for 3 days to obtain prussian blue-chitosan nanoparticles with the nitric oxide release function, and storing at the temperature of 4 ℃.
(2) Preparation of core-shell nanofibers
Preparing a shell layer electrospinning solution: dissolving chitosan (molecular weight is 15kDa) in 1.5 wt% glacial acetic acid aqueous solution to prepare 2.5 wt% chitosan solution; dispersing prussian blue-chitosan nanoparticles with a nitric oxide release function in 10 wt% of 1795 type polyvinyl alcohol aqueous solution to obtain 2mg/mL nanoparticle polyvinyl alcohol dispersion liquid; mixing a chitosan solution and a polyvinyl alcohol dispersion liquid of nanoparticles according to a volume ratio of 1: 5, mixing to obtain the shell layer electrospinning solution.
The core electrospinning liquid comprises the following components: an aqueous 1795 type polyvinyl alcohol solution containing 10mg/mL of collagen, the concentration of polyvinyl alcohol in the solution being 10% by weight.
Carrying out coaxial electrostatic spinning on the shell layer electrospinning solution and the core electrospinning solution, wherein the mass ratio of the shell layer electrospinning solution to the core electrospinning solution is 5: 1, spinning voltage is 11kV, collecting distance is 10cm, a spinning needle is 18G, spinning temperature is 25 ℃, and relative humidity is 35%, so that the composite core-shell nanofiber is obtained, accumulated and collected in an electrostatic spinning receiver, and interwoven to form the near-infrared response nitric oxide releasing electrostatic spinning membrane.
The shapes of the prussian blue-chitosan nanoparticles with nitric oxide release functions obtained in the embodiments 2 and 3 are similar to those of the embodiment 1, the electron microscope is circular, and the prussian blue-chitosan nanoparticles have good photo-thermal properties and photo-response NO release properties; the diameter of the core-shell nanofiber obtained in the embodiment 2 and the embodiment 3 is 50-100 nm, and the core-shell nanofiber has a good antibacterial property; the electrostatic spinning films obtained in the examples 2 and 3 have good antibacterial and healing effects on wound surfaces as antibacterial dressings.
The embodiment shows that the electrostatic spinning film capable of releasing nitric oxide based on near-infrared response can realize controllable release of nitric oxide, release of dosage control type NO at a specific target part is realized by adjusting the irradiation position and intensity of near-infrared laser, release of NO at a non-target part is reduced, possible toxicity and side effects are reduced, and the electrostatic spinning film has the effects of effectively resisting bacteria and accelerating healing of wound surfaces.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Claims (10)
1. An electrostatic spinning membrane for releasing nitric oxide based on near-infrared response is formed by interweaving core-shell nano fibers obtained by coaxial electrostatic spinning; the core-shell nanofiber comprises a shell layer and a core; the shell layer comprises polyvinyl alcohol, chitosan and Prussian blue-chitosan nanoparticles with a nitric oxide release function; the prussian blue-chitosan nano particle comprises prussian blue nano crystal doped with sodium nitroprusside and chitosan coated on the surface of the prussian blue nano crystal; the core includes polyvinyl alcohol and a bioactive molecule.
2. The near-infrared response nitric oxide based electrospun membrane of claim 1 wherein the molecular weight of the chitosan in the prussian blue-chitosan nanoparticle is 3-100 kDa; the particle size of the Prussian blue-chitosan nanoparticles is 30-200 nm; the mass content of the Prussian blue-chitosan nanoparticles in the shell layer is 0.05-2.5%.
3. The near-infrared response-based nitric oxide releasing electrospun membrane according to claim 1 or 2, wherein the polymerization degree of polyvinyl alcohol in the shell layer is 1500-2400, and the molecular weight of chitosan in the shell layer is 3-100 kDa; the mass ratio of the chitosan, the polyvinyl alcohol and the Prussian blue-chitosan nanoparticles in the shell layer is 2-5: 10-15: 0.01 to 0.5.
4. The near-infrared response based nitric oxide releasing electrospun membrane of claim 1 wherein the polymerization degree of polyvinyl alcohol in the core is 1500-2400, and the bioactive molecule comprises one or more of collagen, growth factor and small molecule drug; the mass ratio of polyvinyl alcohol to bioactive molecules in the core is 10-50: 0.1 to 2.
5. The near-infrared response based nitric oxide releasing electrospun membrane according to claim 1, wherein the mass ratio of the shell layer to the core is 2-5: 1 to 0.5; the diameter of the core-shell nanofiber is 50-500 nm.
6. The method for preparing the electrostatic spinning membrane for releasing nitric oxide based on near-infrared response according to any one of claims 1 to 5, characterized by comprising the following steps:
(1) mixing a glacial acetic acid aqueous solution of chitosan with potassium ferricyanide, sodium nitroprusside and hydrochloric acid, heating to 50-100 ℃ for coprecipitation reaction to obtain the prussian blue-chitosan nanoparticles with the nitric oxide release function;
(2) carrying out coaxial electrostatic spinning by taking a mixed aqueous solution containing the Prussian blue-chitosan nanoparticles, polyvinyl alcohol, chitosan and glacial acetic acid as a shell layer electrospinning solution and a mixed aqueous solution containing polyvinyl alcohol and bioactive molecules as a core electrospinning solution to obtain core-shell nanofibers; the core-shell nanofibers are interwoven to form the near-infrared response-based nitric oxide releasing electrostatic spinning membrane.
7. The preparation method according to claim 6, wherein in the step (1), the mass concentration of chitosan in the glacial acetic acid aqueous solution of chitosan is 0.5-20 mg/mL, and the mass concentration of glacial acetic acid is 0.5-2.5%; the mass ratio of the potassium ferricyanide to the sodium nitroprusside is 1: 1-1: 10, the mass ratio of the sum of the potassium ferricyanide and sodium nitroprusside to the chitosan is 1-5: 0.1 to 1.
8. The preparation method according to claim 6, wherein in the step (2), the mass concentration of the Prussian blue-chitosan nanoparticles in the shell electrospinning solution is 0.5-10 mg/mL, the mass concentration of polyvinyl alcohol is 2-20%, the mass concentration of chitosan is 0.1-5%, and the mass concentration of glacial acetic acid is 0.1-2.5%; the mass concentration of polyvinyl alcohol in the core electrospinning liquid is 2-20%, and the mass concentration of bioactive molecules is 0.5-50 mg/mL;
the mass ratio of the shell layer electrospinning liquid to the core electrospinning liquid is 5-2: 1 to 0.5.
9. The production method according to claim 6 or 8, wherein the conditions for the coaxial electrospinning in the step (2) include: the voltage is 10-20 kV, the spinning temperature is 15-50 ℃, the relative humidity is 10-60%, the spinning needle is 5-23G, and the collection distance is 5-20 cm.
10. Use of the electrostatic spinning film for releasing nitric oxide based on near-infrared response as claimed in any one of claims 1 to 5 or the electrostatic spinning film for releasing nitric oxide based on near-infrared response as prepared by the preparation method as claimed in any one of claims 6 to 9 as an antibacterial dressing.
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