CN115305582A - Preparation method and application of neurovascularization double-sided bionic periosteum - Google Patents
Preparation method and application of neurovascularization double-sided bionic periosteum Download PDFInfo
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- CN115305582A CN115305582A CN202210835132.0A CN202210835132A CN115305582A CN 115305582 A CN115305582 A CN 115305582A CN 202210835132 A CN202210835132 A CN 202210835132A CN 115305582 A CN115305582 A CN 115305582A
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- periosteum
- polyester
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Images
Classifications
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- D—TEXTILES; PAPER
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- D01D5/003—Electro-spinning characterised by the initial state of the material the material being a polymer solution or dispersion
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Abstract
The invention relates to a preparation method and application of a neurovascularization double-sided bionic periosteum. The template with the surface orientation structure is used as a collector to collect the polyester nano spinning fibers, and the mono-orientation nano fiber membrane and the bi-orientation nano fiber membrane are structurally realized. The microenvironment for the growth of natural periosteal cells is simulated from the structure and the function, and a new therapeutic strategy is provided for the regeneration of large-section bones.
Description
Technical Field
The invention belongs to the field of biomedical materials, and particularly relates to a preparation method and application of a neurovascularization double-sided bionic periosteum.
Background
At present, the main modes for treating the large-segment bone defect comprise bone transplantation, a tissue engineering scaffold, bone cement, biological ceramics, a bionic periosteum and the like. Wherein the bone graft may be taken from another site in the patient or from another individual. The tissue engineering scaffold is from human or animal sources. Both bone grafting and tissue engineering scaffolds may increase the incidence of complications, leading to problems of disease transmission, bacterial infection, etc. Bone cement when filled in the body may cause high pressure in the bone marrow cavity, seriously threatening life and health. Bioceramics also have the problem of how to adapt to the organism for degradation. Therefore, a bone repair material which has little harm to human body and does not generate rejection reaction as much as possible is urgently needed to be searched.
Some studies have shown that periosteal reconstruction is the primary task of bone regeneration. The abundant nerves, blood vessels and various cells in the periost play an important role in maintaining bone homeostasis and repairing bone defects. Periosteum can spontaneously induce new bone growth and also metabolize old bone tissue. Its ability to maintain bone homeostasis is primarily attributed to several aspects:
1) Periosteum provides cells with differentiation capacity for bone growth stage
Recent studies have shown that there are multipotent periosteal stem cells in the periost that are capable of self-renewal, and that there are fundamental differences in transcriptional characteristics between multipotent periosteal stem cells and other cells (skeletal stem cells, mature mesenchymal cells). Periosteal stem cells form bone directly through the intramembranous pathway, while other cells form bone through endochondral formation.
2) Periosteum provides nutrients and signaling molecules for osteogenesis
It is well known that periosteum not only contains abundant networks of nerves and blood vessels, but also provides a variety of signaling molecules for bone homeostasis. Blood vessels in the periost provide the necessary nutrients and metabolic pathways for cell growth and metabolism. Wherein the abundant blood calcium in the blood and the collagen fiber secreted by osteoblast act together to generate new mineralized tissue. Calcitonin gene-related peptide (CGRP) secreted from nerve fibers in the periosteum is also capable of opening the Wnt pathway and cyclic adenosine monophosphate (cAMP) pathway to promote bone regeneration. Most importantly, periosteum is a plurality of signal molecules in the environment, including growth factors such as bone morphogenetic protein-2 (BMP-2), osteocalcin (OCN), collagen type I (COL-I), CGRP, NGF, VEGF and the like interact, and the balance of osteogenesis and bone resorption is maintained.
3) Periosteum promotes osteogenesis under mechanical stimulation
When the bone is subjected to external mechanical force, the periosteum is in a stretching state, and the periosteum and the cortical bone are tightly connected through Sharpey fibers, so that the mechanical external force applied to the periosteum can be immediately transmitted to bone tissues. In the process, osteoblasts are rearranged along the direction of tension, the surface area is increased, and then the osteoblasts and various signal molecules and cells in the periosteal microenvironment act together to form new bone tissues.
The function and structure of periost play an important role in bone regeneration, especially the abundant neural and vascular networks in periosteal fiber layers, which are essential in regulating bone homeostasis and repairing bone defects. However, most of the current researches only start from the structure or the function, most of the researches only involve osteogenic blood vessels, and deep researches on periosteal neurogenesis are lacked. At present, research on regeneration of nerves at a bone defect part is still in an initial stage, and how to realize neurovascular network reconstruction at the bone defect part is a key point to be solved urgently at present. Therefore, the patent provides a double-sided bionic periosteum for neurovascularization to solve the problem that a neurovascular network is difficult to reconstruct.
Disclosure of Invention
In order to solve the problem, a surface micro-pattern bionic periosteum is provided, and the effects of promoting bone repair and promoting neurovascular reconstruction can be simultaneously obtained.
In order to realize the purpose, the technical scheme adopted by the invention is as follows:
in one aspect, the present invention provides a method for preparing a spun fiber membrane, comprising the steps of:
1) Solution preparation: dispersing polyester particles in an organic solvent to obtain a mixed solution with the polyester content of 0.1-30 wt%;
2) Film forming by using a solution: and (3) performing electrostatic spinning on the obtained mixed solution, and collecting spinning nano fibers by using a collector with a surface orientation structure as a collecting template to obtain the fiber membrane.
As a preferred embodiment, the polyester is a poorly water-soluble polyester compound, preferably polycaprolactone, polyglycolide, polylactide, polyglycolide, polyester-polyethylene glycol copolymer. More preferably polycaprolactone.
As a preferred example, the organic solvent is selected from trifluoroacetic acid, hexafluoroisopropanol, dithiosulfoxide, formic acid, acetic acid, ethanol, trifluoroethanol, carbon trichloride, dimethylformamide, dimethylacetamide, 1, 4-dioxane, tetrahydrofuran, ethyl acetate, acetone, pyridine and acetonitrile. More preferably hexafluoroisopropanol.
As a preferred embodiment, the mixed solution further comprises inorganic nanoparticles, and the inorganic nanoparticles can be selected from nano-hydroxyapatite, inorganic whitlockite nanoparticles (WH), and photothermal whitlockite (nd @ WH).
As a preferred embodiment, the mass content of the inorganic nanoparticles in the mixed solution is 0.1 to 15wt%.
As a preferred embodiment, the step 1) adopts stirring and ultrasonic dispersion, the stirring time is 4 to 6 hours, and the ultrasonic time is 0.5 to 1 hour.
As a preferred embodiment, the collector surface orientation structure is obtained by a laser etching technique, and the collector surface orientation structure groove cross section includes but is not limited to triangle, trapezoid, arc, square, rectangle, and polygon.
In a preferred embodiment, the collector surface orientation structure has grooves with a width of 100-500 μm, a spacing of 0-500 μm and a depth of not more than 1cm, preferably 10-1000 μm.
As a preferred embodiment, the dimensions of the orientation structure are specified as follows: the cross section of the groove is rectangular, the width of the bottom edge is 100 mu m, the depth of the groove is 100 mu m, and the interval is 100 mu m.
As a preferred embodiment, the dimensions of the orientation structure are specified as follows: the cross section of the groove is rectangular, the width of the bottom edge is 200 mu m, the depth of the groove is 100 mu m, and the interval is 200 mu m.
As a preferred embodiment, the dimensions of the orientation structure are specified as follows: the cross section of the groove is rectangular, the width of the bottom edge is 300 mu m, the depth of the groove is 100 mu m, and the interval is 300 mu m.
As a preferred embodiment, the dimensions of the orientation structure are specified as follows: the cross section of the groove is rectangular, the width of the bottom edge is 500 mu m, the depth of the groove is 100 mu m, and the interval is 500 mu m.
As a preferred embodiment, the dimensions of the orientation structure are specified as follows: the section of the groove is isosceles trapezoid, the width of the bottom edge is 100 mu m, the width of the upper bottom edge is 200 mu m, the interval is 0, and the depth of the groove is 100 mu m.
As a preferred embodiment, the dimensions of the orientation structure are specified as follows: the cross section of the groove is in the shape of an isosceles triangle, the width of the groove is 200 mu m, the interval is 0, and the depth of the groove is 100 mu m.
As a preferred embodiment, the dimensions of the orientation structure are specified as follows: the cross section of the groove is isosceles triangle, the width of the groove is 300 μm, the interval is 0, and the depth of the groove is 100 μm.
As a preferred embodiment, the dimensions of the orientation structure are specified as follows: the cross section of the groove is isosceles triangle, the width of the groove is 500 μm, the interval is 0, and the depth of the groove is 100 μm.
As a preferred embodiment, the electrostatic spinning process has a voltage of-6 to +30kV, a rotating speed of 100 to 4000rpm, and a bolus injection speed of 0.05 to 0.3mm/min.
As a preferred embodiment, the electrostatic spinning can adopt a vertical spinning machine, a horizontal spinning machine and an orientation receiving spinning machine.
As a preferred embodiment, the electrostatic spinning is a random nanofiber spinning, and the fiber membrane is composed of random nanofibers. Namely, one side of the fiber membrane is provided with an orientation structure matched with the collecting template, and the other side of the fiber membrane is in a random structure. And collecting the conventional random nanofiber filaments by adopting a surface orientation template.
As a preferred embodiment, the electrostatic spinning is oriented nanofiber spinning, and the fiber membrane is composed of oriented nanofibers. Namely, one surface of the fiber membrane is provided with an orientation structure matched with the collecting template, and the other surface of the fiber membrane is provided with a fiber orientation structure. And collecting the oriented nanofiber filaments by adopting a surface oriented template.
In a preferred embodiment, the electrostatic spinning is an alternate spinning of random nanofibers and oriented nanofibers, and the fiber membrane is formed by stacking a random nanofiber layer and an oriented nanofiber layer. The method comprises the steps of collecting oriented nano-fiber yarns by using a surface orientation template, receiving a layer of random nano-fiber yarns on the surface of the obtained membrane by using the same spinning solution, collecting the random nano-fiber yarns by using the orientation template, and receiving a layer of oriented nano-fiber yarns on the surface of the obtained membrane by using the same spinning solution.
As a preferred embodiment, the resulting fiber film is dried after demolding and the solvent is removed. The drying is preferably vacuum drying, and the vacuum environment can be achieved by various low-pressure equipment such as a vacuum drying oven, a cold trap, a freeze dryer and the like. The solvent volatilization temperature is below the melting temperature of the polymer.
In a second aspect, the invention provides a spun fiber membrane prepared by the above method.
In a third aspect, the invention provides an application of the spinning fiber membrane in preparation of a bionic bone membrane material.
The application adopts electrostatic spinning can prepare the controllable fibre of diameter, therefore can imitate the extracellular matrix microenvironment of cell growth. First, the surface-oriented micropattern can promote cell alignment and tubular formation of blood vessels. Second, the inorganic nanoparticles can promote differentiation of mesenchymal stem cells, thereby promoting bone regeneration. And Mg released in the nanoparticles 2+ Can promote the secretion of Nerve Growth Factor (NGF) and Vascular Endothelial Growth Factor (VEGF) to be up-regulated, and promote the reconstruction of neurovascular network. Finally, the photothermal response performance of Nd @ WH can also promote PPCL/Nd @ WH to be heated to the effective temperature of photothermal osteogenesis of 40 +/-0.5 ℃ under the near-infrared stimulation of 808 nm.
Drawings
FIG. 1 is an SEM photograph of the reverse structure of the biomimetic periosteum in example 4 at 50 magnification;
FIG. 2 is an SEM image of the reverse structure of the biomimetic periosteum of example 4 at a magnification of 100;
FIG. 3 is an SEM photograph of the reverse structure of the biomimetic periosteum of example 4 at 300 magnification;
FIG. 4 is an SEM photograph of the reverse structure of the biomimetic periosteum of example 6, with a magnification of 50;
FIG. 5 is an SEM photograph of the reverse structure of the biomimetic periosteum of example 6 at a magnification of 100;
FIG. 6 is an SEM photograph of the reverse structure of the biomimetic periosteum of example 6 at 500 magnification;
FIG. 7 is an SEM photograph of the front structure of the biomimetic periosteum in example 4 at a magnification of 100;
fig. 8 is a diagram illustrating the neurovascularization test and effect of the biomimetic periosteum obtained in examples 1 to 5 in the skull defect test, wherein a is a schematic diagram illustrating the test process, B is an experimental effect, and PCL is a PCL nanofiber membrane with 15% whitlockite nanoparticles (WH) added thereto, PCL/nd @ WH is a PCL nanofiber membrane with 15% photothermal whitlockite nanoparticles (nd @ WH) added thereto, PPCL/nd @ WH is a surface-patterned PCL nanofiber membrane with 15% nd @ WH added thereto, PPCL/nd @ WH (+) is a surface-patterned PCL nanofiber membrane with 15% nd @ WH added thereto, and PPCL/nd @ WH (+) is a surface-patterned PCL nanofiber membrane with 15 nd @ WH added thereto and is stimulated by an external near-infrared light source;
FIG. 9 shows the bone regeneration test and effect of the biomimetic periosteum obtained in examples 1-5 in the skull defect test, wherein A is the Micro-CT qualitative analysis result of rat skull defect test, B is the Micro-CT quantitative analysis result of rat skull defect test, and C is the H & E and Masson histological staining analysis result.
Detailed Description
In order to better understand the present invention, the following examples are further provided to illustrate the content of the present invention, but the present invention is not limited to the following examples. The following examples are not specifically described, and all reagents used are commercially available chemical reagents or industrial products.
Example 1
A preparation method of a PCL bionic periosteum comprises the following steps:
1) Solution preparation: 1g of PCL was added to 10ml of hexafluoroisopropanol, and stirred for 5 hours to completely dissolve it.
2) Construction of conventional electrospun membranes: and (3) carrying out electrostatic spinning on the spinning solution in the step 1) under the conditions of technological parameters, spinning voltage of 12kV/-2kV, receiving distance of 15cm, pushing speed of 0.08mm/min and receiving speed of 100rpm, collecting the electrostatic spinning nanofiber membrane by using a planar template, and carrying out spinning for 4h to obtain the PCL nanofiber membrane.
3) Removing the solvent: the resulting film was placed in a vacuum oven to remove the solvent.
The PCL biomimetic periosteum prepared in this example was observed by Scanning Electron Microscopy (SEM). And carrying out gold spraying treatment on the bionic periosteum, and observing the surface appearance of the bionic periosteum by an S-4800 Scanning Electron Microscope (SEM) produced by Hitachi.
Example 2
A preparation method of PCL/WH bionic periosteum comprises the following steps:
1) Solution preparation: 1.5g of WH was dispersed in 10ml of hexafluoroisopropanol, and then PCL was added and stirred for 5 hours to completely dissolve it.
2) Construction of conventional electrospun membranes: and (2) carrying out electrostatic spinning on the spinning solution in the step 1) under the conditions of technological parameters, spinning voltage of 12kV/-2kV, receiving distance of 15cm, pushing speed of 0.08mm/min and receiving speed of 100rpm, collecting the electrostatic spinning nanofiber membrane by using a planar template, and carrying out spinning for 4h to obtain the PCL/WH nanofiber membrane.
3) Removing the solvent: the resulting film was placed in a vacuum oven to remove the solvent.
The PCL/WH biomimetic periosteum prepared in this example was observed by Scanning Electron Microscopy (SEM). And carrying out gold spraying treatment on the bionic periosteum, and observing the surface appearance of the double-sided bionic periosteum by an S-4800 Scanning Electron Microscope (SEM) produced by Hitachi.
Example 3:
a preparation method of PCL/Nd @ WH bionic periosteum comprises the following steps:
1) Solution preparation: 1.5g of Nd @ WH was dispersed in 10ml of hexafluoroisopropanol, and then PCL was added and stirred for 5 hours to completely dissolve it.
2) Construction of conventional electrospun membranes: carrying out electrostatic spinning on the spinning solution in the step 1) under the conditions of technological parameters, spinning voltage of 12kV/-2kV, receiving distance of 15cm, pushing speed of 0.08mm/min and receiving speed of 100rpm, collecting the electrostatic spinning nanofiber membrane by using a planar template, and spinning for 4h to obtain the PCL/Nd @ WH nanofiber membrane.
3) Removing the solvent: the resulting film was placed in a vacuum oven to remove the solvent.
The PCL/Nd @ WH bionic periosteum prepared by the example is observed through a Scanning Electron Microscope (SEM). And carrying out gold spraying treatment on the bionic periosteum, and observing the surface appearance of the double-sided bionic periosteum by an S-4800 Scanning Electron Microscope (SEM) produced by Hitachi.
Example 4:
a preparation method of PPCL/Nd @ WH bionic periosteum comprises the following steps:
1) Solution preparation: 1.5g of Nd @ WH was dispersed in 10ml of hexafluoroisopropanol, and then PCL was added and stirred for 5 hours to completely dissolve it.
2) Constructing a reverse surface micro-pattern nanofiber membrane: carrying out electrostatic spinning on the spinning solution in the step 1) under the conditions of technological parameters, spinning voltage of 12kV/-2kV, receiving distance of 15cm, pushing speed of 0.08mm/min and receiving speed of 100rpm, and collecting the electrostatic spinning nanofiber membrane by using a template with a micro-pattern on the surface for 4 hours. The surface micro-pattern orientation structure and the size are as follows: the cross section of the groove is rectangular, the width of the bottom edge of the groove is 100 mu m, the depth of the groove is 100 mu m, and the interval is 100 mu m.
3) Construction of a front-side conventional electrospun membrane: carrying out electrostatic spinning on the spinning solution obtained in the step 1) under the conditions of technological parameters, spinning voltage of 12kV/-2kV, receiving distance of 15cm, pushing speed of 0.08mm/min and receiving speed of 100rpm, and collecting nanofibers on the demolded nanofiber membrane for 4 hours. Obtaining the double-sided nanofiber membrane.
4) Removing the solvent: the resulting film was placed in a vacuum oven to remove the solvent.
The double-sided nanofiber membrane prepared in the example is observed through a Scanning Electron Microscope (SEM), the electrostatic spinning nanofiber membrane is subjected to gold spraying treatment, and then the surface morphology of the double-sided bionic periosteum is observed through an S-4800 type Scanning Electron Microscope (SEM) produced by Hitachi, wherein the front morphology is shown in figures 1-3, and the back morphology is shown in figure 7.
Example 5:
a preparation method of PPCL/Nd @ WH (+) bionic periosteum comprises the following steps:
the specific implementation steps are the same as those in the embodiment 4, and on the basis, the defect part of the implant material is heated to about 40 ℃ by adding 808nm near-infrared stimulation, so that osteoblasts are active at the temperature, and the osteogenic capacity is enhanced.
The effect characterization of the neurovascularization double-sided bionic periosteum takes a rat skull defect model as an object, takes immunofluorescence staining, micro-CT and histological staining as means, and verifies the neurovascularization effect and the osteogenesis capacity effect of the bionic periosteum in vivo. As shown in FIGS. 8 and 9, the neuroangiogenesis and osteogenesis abilities of the PPCL/Nd @ WH group were significantly better than those of the PCL/Nd @ WH group, and it was concluded that patterning of the surface orientation structure could promote osteogenesis in rat skull defects.
Example 6
The preparation method of the double-sided bionic periosteum comprises the following steps:
1) Solution preparation: 1g of PCL was added to 10ml of hexafluoroisopropanol, and stirred for 5 hours to completely dissolve it.
2) Constructing a reverse surface micro-pattern nanofiber membrane: carrying out electrostatic spinning on the spinning solution in the step 1) under the conditions of technological parameters, spinning voltage of 12kV/-2kV, receiving distance of 15cm, pushing speed of 0.08mm/min and receiving speed of 2000rpm, and collecting the electrostatic spinning nanofiber membrane by using a template with a micro-pattern on the surface for 4 hours. The surface micro-pattern orientation structure and the size are as follows: the cross section of the groove is rectangular, the width of the bottom edge of the groove is 100 mu m, the depth of the groove is 100 mu m, and the interval is 100 mu m.
3) Construction of a front-side conventional electrospun membrane: carrying out electrostatic spinning on the spinning solution in the step 1) under the conditions of technological parameters, spinning voltage of 12kV/-2kV, receiving distance of 15cm, pushing speed of 0.08mm/min and receiving speed of 100rpm, and collecting nanofibers on the demolded nanofiber membrane for 4 hours. Obtaining the double-sided nanofiber membrane.
4) Removing the solvent: the resulting film was placed in a vacuum oven to remove the solvent.
The double-sided nanofiber membrane prepared in this example was observed by a Scanning Electron Microscope (SEM), the electrospun nanofiber membrane was subjected to gold spraying, and then the surface morphology of the double-sided biomimetic periosteum was observed by an S-4800 type Scanning Electron Microscope (SEM) produced by hitachi, with the results shown in fig. 4-6.
Example 7
A preparation method of a neurovascularization double-sided bionic periosteum comprises the following steps:
1) Solution preparation: 1.5g of WH was added to 10ml of hexafluoroisopropanol, and after ultrasonic dispersion for 30min, 1g of PCL was added to the mixed dispersion, and stirred for 5 hours to completely dissolve it.
2) Electrostatic spinning: the spinning solution is prepared by the following steps: spinning voltage: 12kV/-2kV, reception distance: 15cm, push speed: 0.08mm/min, receiving speed: and (3) carrying out electrostatic spinning at 100rpm, and collecting the electrostatic spinning nanofiber membrane by using a template with a micro-pattern on the surface for 4h.
3) Removing the solvent: the resulting film was placed in a vacuum oven to remove the solvent.
Example 8
A preparation method of a neurovascularization double-sided bionic periosteum comprises the following steps:
1) Solution preparation: 1.5g of WH was added to 10ml of hexafluoroisopropanol, and after ultrasonic dispersion for 30min, 1g of PCL was added to the mixed dispersion, and stirred for 5 hours to completely dissolve it.
2) Electrostatic spinning: the spinning solution is prepared by the following steps: spinning voltage: 12kV/-2kV, reception distance: 15cm, push speed: 0.08mm/min, receiving speed: carrying out electrostatic spinning at 2000rpm, and collecting the electrostatic spinning nanofiber membrane by using a template with a micro-pattern on the surface for 4h.
3) Removing the solvent: the resulting film was placed in a vacuum oven to remove the solvent.
Example 9
A preparation method of a neurovascularization double-sided bionic periosteum comprises the following steps:
1) Solution preparation: 1g of WH was added to 10ml of hexafluoroisopropanol, and after ultrasonic dispersion for 30min, 1g of PCL was added to the mixed dispersion, and stirred for 5 hours to completely dissolve it.
2) Electrostatic spinning: the spinning solution is prepared by the following steps: spinning voltage: 12kV/-2kV, reception distance: 15cm, push speed: 0.08mm/min, reception speed: and (3) carrying out electrostatic spinning at 100rpm, and collecting the electrostatic spinning nanofiber membrane by using a template with a micro-pattern on the surface for 4h.
3) Removing the solvent: the resulting film was placed in a vacuum oven to remove the solvent.
Example 10
A preparation method of a neurovascularization double-sided bionic periosteum comprises the following steps:
1) Solution preparation: 0.5g of WH was added to 10ml of hexafluoroisopropanol, and after ultrasonic dispersion for 30min, 1g of PCL was added to the mixed dispersion, and stirred for 5 hours to completely dissolve it.
2) Electrostatic spinning: the spinning solution is prepared by the following steps: spinning voltage: 12kV/-2kV, reception distance: 15cm, push speed: 0.08mm/min, receiving speed: and (3) carrying out electrostatic spinning at 100rpm, and collecting the electrostatic spinning nanofiber membrane by using a template with a micro-pattern on the surface for 4h.
3) Removing the solvent: the resulting film was placed in a vacuum oven to remove the solvent.
The above description is only a preferred embodiment of the present invention, and it should be noted that those skilled in the art can make various modifications and changes without departing from the inventive concept of the present invention, and these modifications and changes are within the protection scope of the present invention.
Claims (10)
1. The preparation method of the spinning fiber membrane is characterized by comprising the following steps:
1) Solution preparation: dispersing polyester particles in an organic solvent to obtain a mixed solution with the polyester content of 0.1-30 wt%;
2) Film forming by using a solution: and (3) performing electrostatic spinning on the obtained mixed solution, and collecting spinning nano fibers to obtain the fiber membrane by using a collector with a surface orientation structure as a collecting template.
2. The method for preparing a polyester-based compound according to claim 1, wherein the polyester is a poorly water-soluble polyester compound, preferably polycaprolactone, polyglycolide, polylactide, polyglycolide, polyester-polyethylene glycol copolymer.
3. The method according to claim 1, wherein the organic solvent is selected from trifluoroacetic acid, hexafluoroisopropanol, dithiosulfoxide, formic acid, acetic acid, ethanol, trifluoroethanol, carbon trichloride, dimethylformamide, dimethylacetamide, 1, 4-dioxane, tetrahydrofuran, ethyl acetate, acetone, pyridine and acetonitrile.
4. The method according to claim 1, wherein the mixed solution further contains inorganic nanoparticles, and the inorganic nanoparticles are contained in the mixed solution in an amount of 0.1 to 15wt%.
5. The method according to claim 1, wherein the mixed solution contains inorganic nanoparticles selected from the group consisting of nano-hydroxyapatite (hydroxyapatite), inorganic whitlockite nanoparticles (WH), and photo-thermal whitlockite (Nd @ WH).
6. The method of claim 1, wherein the collector surface alignment structure has grooves with a width of 100 to 500 μm, a groove interval of 0 to 500 μm, and a groove depth of 10 to 1000 μm.
7. The method of claim 1, wherein the collector surface orientation feature grooves have a cross-section selected from the group consisting of triangular, trapezoidal, curved, square, rectangular, and polygonal.
8. The method according to claim 1, wherein the electrospinning is random nanofiber spinning or oriented nanofiber spinning or alternate spinning of random nanofibers and oriented nanofibers.
9. A spun fibrous membrane, characterized in that it is obtained by a process according to any one of claims 1 to 8.
10. Use of the spun fiber membrane of claim 9 in the preparation of a biomimetic bone membrane material.
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