CN117026517A - Electrostatic spinning nanofiber membrane, preparation method and application - Google Patents
Electrostatic spinning nanofiber membrane, preparation method and application Download PDFInfo
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- 239000012528 membrane Substances 0.000 title claims abstract description 76
- 239000002121 nanofiber Substances 0.000 title claims abstract description 57
- 238000010041 electrostatic spinning Methods 0.000 title claims abstract description 26
- 238000002360 preparation method Methods 0.000 title abstract description 6
- 239000000835 fiber Substances 0.000 claims abstract description 45
- 230000001737 promoting effect Effects 0.000 claims abstract description 4
- 238000009987 spinning Methods 0.000 claims description 29
- BYEAHWXPCBROCE-UHFFFAOYSA-N 1,1,1,3,3,3-hexafluoropropan-2-ol Chemical compound FC(F)(F)C(O)C(F)(F)F BYEAHWXPCBROCE-UHFFFAOYSA-N 0.000 claims description 15
- 238000000034 method Methods 0.000 claims description 11
- 238000003760 magnetic stirring Methods 0.000 claims description 6
- 238000000151 deposition Methods 0.000 claims description 4
- 238000002156 mixing Methods 0.000 claims description 4
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- 239000003960 organic solvent Substances 0.000 claims description 2
- 238000001132 ultrasonic dispersion Methods 0.000 claims description 2
- 229920000520 poly(3-hydroxybutyrate-co-3-hydroxyvalerate) Polymers 0.000 claims 6
- 238000001523 electrospinning Methods 0.000 claims 2
- 238000002604 ultrasonography Methods 0.000 claims 1
- 230000004069 differentiation Effects 0.000 abstract description 7
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- 238000011069 regeneration method Methods 0.000 abstract description 3
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- 239000004952 Polyamide Substances 0.000 abstract 1
- 229920002647 polyamide Polymers 0.000 abstract 1
- 229920001610 polycaprolactone Polymers 0.000 description 39
- 239000004632 polycaprolactone Substances 0.000 description 39
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- 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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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/14—Macromolecular materials
- A61L27/18—Macromolecular materials obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
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- 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
- D01D5/0007—Electro-spinning
- D01D5/0061—Electro-spinning characterised by the electro-spinning apparatus
- D01D5/0092—Electro-spinning characterised by the electro-spinning apparatus characterised by the electrical field, e.g. combined with a magnetic fields, using biased or alternating fields
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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
- D01F8/14—Conjugated, i.e. bi- or multicomponent, artificial filaments or the like; Manufacture thereof from synthetic polymers with at least one polyester as constituent
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- 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
- D04H1/43825—Composite fibres
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- A61L2400/00—Materials characterised by their function or physical properties
- A61L2400/12—Nanosized materials, e.g. nanofibres, nanoparticles, nanowires, nanotubes; Nanostructured surfaces
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Abstract
The invention relates to the technical field of high polymer materials, and discloses an electrostatic spinning nanofiber membrane, a preparation method and application thereof, wherein the nanofiber membrane is a PHBV modified PCL (polyamide fiber) fiber membrane, fiber filaments in the nanofiber membrane are orderly arranged, and the average diameter of the fiber filaments is 763.2 +/-180.9 nm. Polyhydroxybutyrate-valerate (PHBV) is combined with PCL to form the nanofiber membrane through an electrostatic spinning technology. The PHBV has good biocompatibility and degradability, and simultaneously shows piezoelectric property, so that the nanofiber membrane has piezoelectric effect, has potential of promoting differentiation of stem cells into nerve cells, and is applied to regeneration and repair of nerve defects clinically.
Description
Technical Field
The invention relates to the technical field of high polymer materials, in particular to an electrostatic spinning nanofiber membrane, a preparation method and application.
Background
Nerve injury ranges from a slight crush injury to a severe avulsion, which brings great pain to the patient. Treatment of nerve damage remains a significant clinical challenge. Current treatments mainly include direct repair of damaged nerves or nerve grafting using microsurgical techniques. However, there are problems such as lack of donor grafts, need for multiple operations, and complicated operation. Stem cells are derived from mesoderm and have self-renewal and multipotent differentiation capacity. With the development of nanotechnology, a series of biological scaffolds have been developed which mimic the natural extracellular matrix and can promote stem cell proliferation and regulate its differentiation behavior. Neural tissue engineering aims at inducing stem cell directed differentiation using functionalized biological scaffolds.
The electrostatic spinning technology is used for preparing the nanofiber membrane and has the advantages of simplicity in operation, low cost, high yield and the like. In addition, nanofiber membranes with different morphologies can be prepared by setting different spinning parameters, so that the behavior of cells can be regulated. Meanwhile, the fiber membrane prepared by the electrostatic spinning technology has higher specific surface area and porosity. The natural extracellular matrix is simulated, which is beneficial to proliferation and adhesion of cells. Polycaprolactone (PCL) is degradable semi-crystalline polyester, has stronger flexibility, and can be used as an ideal material for electrostatic spinning. Research shows that the crack bridging effect of PCL increases the toughness of the ceramic bracket and improves the mechanical property of the composite material.
Cell proliferation, migration, axon growth, etc. are all affected by endogenous electric fields. Thus, bioelectricity plays a vital role in the human body. In recent years, some researchers have utilized electrical stimulation to treat various diseases. This treatment often requires an external power source or implanted electrodes to generate a current from outside the body through the skin to function. The piezoelectric material can generate electric activity during mechanical deformation, and does not need to be connected with an external power supply or an implanted electrode, so that the human body is not damaged. It is therefore of great importance to develop a nanofiber membrane that combines PCL with piezoelectric material.
Disclosure of Invention
The invention aims to provide an electrostatic spinning nanofiber membrane, a preparation method and application thereof, wherein polyhydroxybutyrate-valerate (PHBV) is applied to the preparation of a conductive nanofiber membrane, and the electrostatic spinning nanofiber membrane has the potential of promoting differentiation of stem cells into nerve cells and is applied to regeneration and repair of nerve defects clinically.
In order to achieve the above object, in one aspect, the present invention provides an electrospun nanofiber membrane, wherein the nanofiber membrane is a PHBV modified PCL fiber membrane, fiber filaments in the nanofiber membrane are orderly arranged, and the average diameter of the fiber filaments is 763.2 ±180.9nm.
Preferably, the nanofiber membrane has a maximum tensile stress of 14.5MPa and a maximum elastic modulus of 699.18MPa.
In another aspect, the present disclosure provides a method for preparing the electrospun nanofiber membrane, including:
mixing PCL and PHBV in hexafluoroisopropanol organic solution at normal temperature, and performing magnetic stirring and ultrasonic dispersion to obtain spinning solution;
and carrying out electrostatic spinning on the spinning solution under strong voltage, and depositing the sprayed uniform fiber on a receiver to form a nanofiber membrane, wherein the receiving speed is 2400r-2500r.
Preferably, the blending of PCL and PHBV in hexafluoroisopropanol organic solution at normal temperature comprises the steps of:
PHBV is dissolved in hexafluoroisopropanol organic solvent, magnetically stirred for 5-6h, PCL is added, magnetically stirred for 11-12h and ultrasonically stirred for 0.4-0.5h.
Preferably, the dosage ratio of PHBV, PCL and hexafluoroisopropanol is 0.4-0.5g:0.01-0.02g:9-10mL.
Preferably, the PHBV, PCL and hexafluoroisopropanol are used in an amount ratio of 0.5g:0.02g:10mL.
Preferably, the voltage of the electrostatic spinning is 14-15kV, the spinning speed is 0.7-0.8mL/h, and the spinning distance is 9-10cm.
Preferably, the voltage of the electrostatic spinning is 15kV, the spinning speed is 0.8mL/h, the spinning distance is 10cm, and the receiving speed is 2500r. The X-ray pattern of the nanofiber membrane showed that the characteristic peak of beta crystals of PHBV occurred at 19.7 deg., the beta crystals were drawn from amorphous regions between orthogonal alpha-platelets, which reflected a large deformation between fibers, thus generating an electrical signal.
Preferably, the power of the ultrasonic wave is 0.4-0.5W, and the frequency is 0.9-1MHz.
In yet another aspect, the invention also provides the use of an electrospun nanofiber membrane for a biological scaffold to promote cell differentiation.
Compared with the prior art, the invention has the following beneficial effects:
through the technical scheme, polyhydroxybutyrate-valerate (PHBV) is combined with PCL, and the nanofiber membrane is formed through an electrostatic spinning technology. The PHBV has good biocompatibility and degradability, and simultaneously shows piezoelectric property, so that the nanofiber membrane has piezoelectric effect, has potential of promoting differentiation of stem cells into nerve cells, and is applied to regeneration and repair of nerve defects clinically.
The ordered nanofiber membrane has the maximum tensile stress of 14.5MPa and the maximum elastic modulus of 699.18MPa, and has good mechanical properties. Compared with the unordered PCL nanofiber membrane and PHBV/PCL nanofiber membrane, the nanofiber membrane has stronger output voltage, and the ordered spinning fibers can promote cell adhesion, survival and proliferation, guide cell migration and are beneficial to stimulating the differentiation of bone marrow mesenchymal stem cells to nerve cells.
The electrostatic spinning nanofiber membrane prepared by the invention contains 5% PHBV fiber membrane, and under ultrasonic stimulation, stem cells can be induced to differentiate into nerve cells through electric signals and ordered surface morphology, so that the electrostatic spinning nanofiber membrane can be used as a nerve repair stent for clinical application.
Additional features and advantages of the invention will be set forth in the detailed description which follows.
Drawings
The accompanying drawings are included to provide a further understanding of the invention, and are incorporated in and constitute a part of this specification, illustrate the invention and together with the description serve to explain, without limitation, the invention. In the drawings:
FIG. 1 shows a scanning electron microscope image of comparative example 3, example 1 of the present invention;
FIG. 2 shows the diameter distribution diagram of comparative example 3, example 1 of the present invention;
FIG. 3 shows XRD patterns of comparative examples 1, 2, 3 and 1 according to the present invention;
FIG. 4 shows the mechanical curves of comparative examples 2, 3 and 1 according to the invention;
fig. 5 is a graph showing the output voltage of comparative examples 1, 3 and 1 according to the present invention with time.
Detailed Description
The following describes specific embodiments of the present invention in detail with reference to the drawings. It should be understood that the detailed description and specific examples, while indicating and illustrating the invention, are not intended to limit the invention.
Comparative example 1
The electrospun PCL nanofiber membrane was prepared according to the following steps:
s1, at normal temperature, 0.02g PCL is dissolved in 10mL of hexafluoroisopropanol organic solution, magnetic stirring is carried out for 12h, ultrasonic treatment is carried out for 0.5h, and then the sprayed uniform fiber is deposited on a receiver to form a fiber membrane (spinning voltage is 15kV, spinning speed is 0.8mL/h, spinning distance is 10cm, and receiving speed is 20 r) under a strong electric field by utilizing an electrostatic spinning technology;
s2, cutting the PCL fiber membrane into a wafer, stimulating the fiber membrane by using an ultrasonic therapeutic instrument, collecting piezoelectric signals by using an oscilloscope, verifying the piezoelectric performance (the ultrasonic power is 0.5W and the frequency is 1 MHz) of the fiber membrane, and performing XRD detection, as shown in figure 3.
Comparative example 2
The electrospun PHBV nanofiber membrane is prepared according to the following steps:
s1, at normal temperature, 0.5g PHBV is dissolved in 10mL of hexafluoroisopropanol organic solution, magnetic stirring is carried out for 12h, ultrasonic treatment is carried out for 0.5h, then the sprayed uniform fiber is deposited on a receiver to form a fiber membrane by utilizing an electrostatic spinning technology under a strong electric field (spinning voltage is 15kV, spinning speed is 0.8mL/h, spinning distance is 10cm, receiving speed is 20r, ultrasonic power is 0.5W, and frequency is 1 MHz);
s2, cutting the PHBV fiber membrane into a rectangle with the length of 1 multiplied by 2cm, and placing the rectangle on a mechanical detector to detect the mechanical property of the PHBV fiber membrane, as shown in figure 2.
Comparative example 3
The electrostatic spinning PCL/PHBV nanofiber membrane is prepared according to the following steps:
s1, dissolving 0.5g PHBV in 10mL of hexafluoroisopropanol organic solution at normal temperature, magnetically stirring for 6h, adding 0.02g PCL, magnetically stirring for 12h, ultrasonically treating for 0.5h, and depositing on a receiver by using an electrostatic spinning technology under a strong electric field through sprayed uniform fibers to form a fiber membrane (spinning voltage is 15kV, spinning speed is 0.8mL/h, spinning distance is 10cm and receiving speed is 20 r);
s2, cutting the PCL/PHBV fiber film into a rectangle with the length of 1 multiplied by 2cm, and placing the rectangle on a mechanical detector to detect the mechanical property of the PCL/PHBV fiber film. The PCL/PHBV fiber membrane is cut into a wafer, the fiber membrane is stimulated by an ultrasonic therapeutic instrument, and piezoelectric signals are collected by an oscilloscope to verify the piezoelectric performance (the ultrasonic power is 0.5W and the frequency is 1 MHz). And XRD and SEM detection were performed. As shown in fig. 1-5.
Example 1
The method comprises the following steps of preparing an orderly arranged electrostatic spinning PCL/PHBV nanofiber membrane:
s1, dissolving 0.5g PHBV in 10mL of hexafluoroisopropanol organic solution at normal temperature, magnetically stirring for 6h, adding 0.02g PCL, magnetically stirring for 12h, ultrasonically treating for 0.5h, and depositing on a receiver by using an electrostatic spinning technology under a strong electric field through sprayed uniform fibers to form a fiber membrane (spinning voltage is 15kV, spinning speed is 0.8mL/h, spinning distance is 10cm, and receiving speed is 2500 r);
s2, cutting the PCL/PHBV fiber film into a rectangle with the length of 1 multiplied by 2cm, and placing the rectangle on a mechanical detector to detect the mechanical property of the PCL/PHBV fiber film. The PCL/PHBV fiber membrane is cut into a wafer, the fiber membrane is stimulated by an ultrasonic therapeutic instrument, and piezoelectric signals are collected by an oscilloscope to verify the piezoelectric performance (the ultrasonic power is 0.5W and the frequency is 1 MHz). And XRD and SEM detection were performed. As shown in fig. 1-5.
Example 2
The method comprises the following steps of preparing an orderly arranged electrostatic spinning PCL/PHBV nanofiber membrane:
s1, at normal temperature, 0.4g PHBV is dissolved in 9mL hexafluoroisopropanol organic solution, magnetic stirring is carried out for 5h, then 0.01g PCL is added, magnetic stirring is carried out for 11h, ultrasonic treatment is carried out for 0.4h, and then an electrostatic spinning technology is utilized to deposit uniform fibers sprayed out on a receiver under a strong electric field to form a fiber membrane (spinning voltage is 14kV, spinning speed is 0.7mL/h, spinning distance is 9cm and receiving speed is 2400 r). The parameters set in this example 2 are slightly different from those set in example 1, and the electrospun nanofiber membrane with good mechanical properties and output voltage can be obtained, so that the filaments in the electrospun nanofiber membrane are orderly arranged, and the average diameter of the filaments is within the range of 763.2 ±180.9nm.
Description of the Experimental results
As can be seen from the figures 1 and 2, the PCL/PHBV-20r and PCL/PHBV-2500r fibers have uniform fiber structure under an electron microscope, smooth surface, consistent and ordered arrangement direction of the PCL/PHBV-2500r fibers and average diameter smaller than that of the PCL/PHBV-20r fibers.
As can be seen from FIG. 3, XRD patterns of PCL/PHBV-2500r fiber films showed 6 alpha-orthorhombic crystal characteristic peaks of PHBV of 13.6, 17.1, 20.3, 21.7, 25.7 and 27.3, respectively. While the characteristic peak of beta-shaped crystals of PHBV appears at 19.7 deg., the beta-shaped crystals are drawn from amorphous regions between orthogonal alpha-shaped lamellae, which reflect the large deformation between fibers, thus generating an electrical signal.
As can be seen from FIG. 4, the tensile stress of the ordered nanofiber membrane (PCL/PHBV-2500 r) is stronger than that of the unordered nanofiber membrane (PCL/PHBV-20 r), and the mechanical properties are good.
As can be seen from fig. 5, the output voltage detection result of the PHBV/PCL nanofiber membrane shows that the output voltage of the ordered nanofiber membrane (PCL/PHBV-2500 r) is higher than that of the unordered nanofiber membrane (PCL/PHBV-20 r) and the PCL nanofiber membrane, and the output voltage shows good piezoelectric performance.
The preferred embodiments of the present invention have been described in detail above with reference to the accompanying drawings, but the present invention is not limited to the specific details of the above embodiments, and various simple modifications can be made to the technical solution of the present invention within the scope of the technical concept of the present invention, and all the simple modifications belong to the protection scope of the present invention. In addition, the specific features described in the above embodiments may be combined in any suitable manner, and in order to avoid unnecessary repetition, various possible combinations are not described further.
Moreover, any combination of the various embodiments of the invention can be made without departing from the spirit of the invention, which should also be considered as disclosed herein.
Claims (10)
1. An electrostatic spinning nanofiber membrane is characterized in that,
the nanofiber membrane is a PHBV modified PCL fiber membrane, fiber filaments in the nanofiber membrane are orderly arranged, and the average diameter of the fiber filaments is 763.2 +/-180.9 nm.
2. The electrospun nanofiber membrane according to claim 1, wherein the nanofiber membrane has a maximum tensile stress of 14.5MPa and a maximum elastic modulus of 699.18MPa.
3. A method of preparing an electrospun nanofiber membrane according to claim 1 or 2, comprising:
mixing PCL and PHBV in hexafluoroisopropanol organic solution at normal temperature, and performing magnetic stirring and ultrasonic dispersion to obtain spinning solution;
and carrying out electrostatic spinning on the spinning solution under strong voltage, and depositing the sprayed uniform fiber on a receiver to form a nanofiber membrane, wherein the receiving speed is 2400-2500r.
4. The method for preparing the electrospun nanofiber membrane according to claim 3, wherein the blending of PCL and PHBV in hexafluoroisopropanol organic solution at normal temperature comprises:
PHBV is dissolved in hexafluoroisopropanol organic solvent, magnetically stirred for 5-6h, PCL is added, magnetically stirred for 11-12h and ultrasonically stirred for 0.4-0.5h.
5. The method for preparing the electrospun nanofiber membrane according to claim 3 or 4, wherein the dosage ratio of PHBV, PCL and hexafluoroisopropanol is 0.4-0.5g:0.01-0.02g:9-10mL.
6. The method for preparing the electrospun nanofiber membrane according to claim 5, wherein the dosage ratio of PHBV, PCL and hexafluoroisopropanol is 0.5g:0.02g:10mL.
7. The method for preparing an electrospun nanofiber membrane according to claim 5, wherein the voltage of the electrospinning is 14-15kV, the spinning speed is 0.7-0.8mL/h, and the spinning distance is 9-10cm.
8. The method of producing electrospun nanofiber membrane according to claim 7, wherein the voltage of the electrospinning is 15kV, the spinning speed is 0.8mL/h, the spinning distance is 10cm, and the receiving speed is 2500r.
9. The method for preparing an electrospun nanofiber membrane according to claim 3, wherein the power of the ultrasound is 0.4-0.5W and the frequency is 0.9-1MHz.
10. Use of an electrospun nanofiber membrane according to claim 1 or 2 for a bioscaffold promoting cell differentiation.
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