A fiber membrane using polydioxanone as a base material and a method for preparing the same

Abstract

The application provides a fiber membrane with poly-p-dioxanone as a base material and a preparation method thereof, and relates to the technical field of engineering tissue materials. The fiber membrane with poly-p-dioxanone as a base material comprises: an outer dense layer of PPDO material, and an inner layer of PPDO material formed by electrospinning on the outer dense layer; and the preparation method comprises the following steps: S10, PPDO is mixed with an organic solvent to prepare a PPDO solution a, the PPDO solution a is used for multiple times of film laying, and a PPDO base film is prepared through drying; S20, PPDO is mixed with an organic solvent to prepare a PPDO electrospinning solution c; and S30, electrospinning is performed on the PPDO base film by using the electrospinning solution c, so that the fiber membrane with poly-p-dioxanone as a base material is prepared; and the application can positively regulate intracranial cell behavior, shorten a degradation cycle, and accelerate the recovery process of a patient.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of engineered tissue materials, in particular a fiber membrane with poly-p-dioxanone as a base material and a preparation method thereof. BACKGROUND

[0002] Engineered tissue materials are biomaterials used in tissue engineering to construct or repair human tissues and organs. These materials can serve as scaffolds for cell growth, providing necessary structural support and biological signals to promote cell attachment, proliferation, and differentiation.

[0003] Artificial dura mater is a fiber membrane made of engineered tissue material as a substitute for human dura mater, used for repairing dura mater or spinal membrane defects caused by craniocerebral, spinal cord injury, tumor and other craniocerebral diseases, preventing serious complications such as cerebrospinal fluid leakage, intracranial infection, brain swelling, brain adhesion and scar, and restoring its integrity.

[0004] In craniocerebral surgery, according to the requirements of neurosurgery, the ideal dura mater repair material should be easy to operate, convenient for surgery, have high biocompatibility and appropriate mechanical properties, promote the formation of new tissue at the injury site during the degradation process, reduce the formation of scars, and not adhere to the surrounding tissue. To solve this problem, researchers have developed various materials for artificial dura mater, such as natural biological materials, autologous and allogeneic transplantation materials, and artificial materials. Compared with other materials, artificial materials have excellent biocompatibility, good mechanical properties, and great advantages in antibacterial, storage, manufacturing and transportation, etc. They are a very potential substitute material for dura mater. Designing new strategies for artificial dura mater repair materials and achieving their antibacterial, anti-inflammatory and rapid reconstruction of inner tissue functions has great research significance and clinical application prospects for improving the efficacy of dura mater repair and improving the postoperative recovery of patients.

[0005] The problems of the artificial dura mater substitutes in the prior art include: long repair period, poor biocompatibility, lack of ability to positively regulate cells, etc. The artificial polymer materials used for dura mater repair mainly include: polyglycolic acid (PGA), polyurethane (PU), polycaprolactone (PCL) and poly-L-lactic acid (PLLA), etc. In Japan, polyglycolic acid (PGA) patch has been approved as a clinical artificial substitute for dura mater, but the effect is not satisfactory, and a considerable part of the clinical reports show that the PGA patch has poor biocompatibility, and has adverse reactions after operation, including granulation tissue formation, meningeal infection, etc. Polycaprolactone (PCL) and poly-L-lactic acid (PLLA) are two kinds of artificial dura mater substitute materials approved by the US FDA, which have good biodegradability and biocompatibility compared with PGA, and have been proved to prevent cerebrospinal fluid leakage and combine with the surrounding tissue to repair the dura mater defect, and no obvious infection occurs in the clinical research, but the degradation period is more than half a year. In summary, the main problem of the artificial dura mater substitute made of the material for making artificial dura mater is the lack of the ability to promote tissue reconstruction, positively regulate cells to shorten the repair period, and the long degradation period of the material itself, which limits the recovery speed of the patient using the artificial dura mater and increases the burden of the patient. SUMMARY

[0006] The present application is to solve the problem of the lack of the ability to promote tissue reconstruction, positively regulate cells to shorten the repair period, and the long degradation period of the material itself in the prior art tissue engineering material substitute such as artificial dura mater, and provides two kinds of fiber membranes with poly-p-dioxanone as the base material, which can positively regulate intracranial cell behavior, promote tissue reconstruction, and quickly degrade to shorten the recovery period of the patient.

[0007] The technical scheme adopted by the present application is:

[0008] A preparation method of a fiber membrane with poly-p-dioxanone as the base material, comprising the following steps:

[0009] S10. Preparing a PPDO solution a by mixing PPDO with an organic solvent, using the PPDO solution a to perform film laying multiple times, and drying to obtain a PPDO base film;

[0010] S20. Preparing an electrospinning solution c by mixing PPDO with an organic solvent;

[0011] S30. Performing electrospinning on the PPDO base film using the electrospinning solution c to obtain a fiber membrane with poly-p-dioxanone as the base material;

[0012] Wherein, the steps S10 and S20 have no fixed sequence.

[0013] Further,

[0014] The S20 comprises: using PPDO mixed with an organic solvent to prepare a PPDO solution b, and adding a BG suspension in the PPDO solution b to prepare a PPDO electrospinning solution c.

[0015] Further,

[0016] The organic solvent used in the S10 and the S20 is hexafluoroisopropanol, and the PPDO solution a and the PPDO solution b prepared by mixing need to remove bubbles.

[0017] Further,

[0018] The S10 specifically comprises: mixing PPDO with hexafluoroisopropanol, sealing a light-proof room, stirring at room temperature, so that the PPDO is completely dissolved in the hexafluoroisopropanol, placing in a vacuum bubble removing machine to remove bubbles in the mixed solution to obtain a pure PPDO solution a; pouring the pure PPDO solution a into a glass dish in three times with intervals, and after each time of pouring, using a coater to spread to ensure the surface uniformity; after three times of film laying, placing in a constant temperature vacuum drying box for drying in a vacuum environment to prepare a PPDO base film.

[0019] Further,

[0020] The S20 specifically comprises: mixing PPDO with hexafluoroisopropanol, sealing a light-proof room, stirring at room temperature, so that the PPDO is completely dissolved in the hexafluoroisopropanol, placing in a vacuum bubble removing machine to remove bubbles in the mixed solution to obtain a pure PPDO solution b; mixing excess BG powder with deionized water, ultrasonic oscillation to mix uniformly, centrifuging to take the supernatant, then taking a certain amount of BG suspension and adding it into the pure PPDO solution b to prepare a PPDO electrospinning solution c.

[0021] Further,

[0022] The S30 specifically comprises: adding the electrospinning solution c into an electrostatic spinning liquid injector to remove bubbles; using an electrostatic spinning instrument to perform multiple electrospinning to prepare a fiber membrane with poly-p-dioxanone as a base material.

[0023] The spinning voltage is controlled at 10-30 kV, the injector advancing speed is controlled at 0.1-5 mL / h, the distance between the needle head and the receiver is controlled at 50-150 mm, and the left and right moving speed of the needle head is suitably controlled at 25-100 mm / sec, and the rotating speed of the drum receiver is controlled at 0-5000 rpm.

[0024] Further,

[0025] The PPDO concentration of the electrospinning solution c prepared in the S20 is 0.06 g / mL-0.10 g / mL.

[0026] Further,

[0027] The silicon concentration of the electrospun solution c prepared in the S20 is 0.5 mg / L to 50 mg / L.

[0028] A fiber membrane with poly-p-dioxanone as a base material, prepared by the preparation method, comprising:

[0029] An outer dense layer of the PPDO material; and

[0030] An inner layer of the PPDO material formed by electrospinning on the outer dense layer.

[0031] Another preparation method of a fiber membrane with poly-p-dioxanone as a base material, comprising the following steps:

[0032] S10. Preparing a PPDO solution a by mixing PPDO with an organic solvent, using the PPDO solution a for multiple times of film laying, and drying to obtain a PPDO base film;

[0033] S20. Preparing a PPDO solution b by mixing PPDO with an organic solvent, and adding a BG suspension to the PPDO solution b to obtain a PPDO electrospun solution c;

[0034] S22. Mixing CNPs groups with an organic solvent, and adding poloxamer to obtain an electrospun solution d;

[0035] S30. Simultaneously performing electrospinning using the electrospun solution c and the electrospun solution d on the PPDO base film to obtain a fiber membrane with poly-p-dioxanone as a base material;

[0036] The S10, S20 and S22 steps have no fixed sequence.

[0037] Further,

[0038] The organic solvent used in the S10, S20 and S22 is hexafluoroisopropanol, and the PPDO solution a and the PPDO solution b prepared by mixing need to remove bubbles.

[0039] Further,

[0040] The S10 specifically comprises: mixing PPDO with hexafluoroisopropanol, stirring in a sealed light-proof room at room temperature, so that the PPDO is completely dissolved in the hexafluoroisopropanol, and placing the mixed solution in a vacuum bubble removing machine to remove bubbles to obtain a pure PPDO solution a; pouring the pure PPDO solution a into a glass dish in three times with intervals, and coating with a coater after each time of pouring to ensure the surface uniformity; and placing the PPDO base film obtained by drying in a vacuum environment in a constant temperature vacuum drying box after three times of film laying.

[0041] Further,

[0042] S20 specifically includes: mixing PPDO with hexafluoroisopropanol, sealing in a light-proof chamber, stirring at room temperature, so that PPDO is completely dissolved in hexafluoroisopropanol, and placing in a vacuum defoaming machine to remove bubbles in the mixed solution to obtain a pure PPDO solution b; mixing excess BG powder with deionized water, ultrasonic oscillation to mix uniformly, centrifugation to take supernatant, then taking a certain amount of BG suspension and adding to the pure PPDO solution b to prepare a PPDO electrospinning solution c.

[0043] Further,

[0044] S22 specifically includes: mixing CNPs with hexafluoroisopropanol, sealing in a light-proof chamber, ultrasonic treatment at room temperature, so that the nano cerium oxide is uniformly dispersed in the hexafluoroisopropanol; adding poloxamer, sealing in a light-proof chamber, stirring and treating at room temperature, so that the poloxamer is completely dissolved in the hexafluoroisopropanol to prepare an electrospinning solution d.

[0045] Further,

[0046] S30 specifically includes: using a double-head synchronous electrospinning technology, the electrospinning solution c and the electrospinning solution d are synchronously injected using different syringes; using an electrospinning instrument to perform multiple electrospinning to prepare a fiber membrane with poly-p-dioxanone as a base material.

[0047] Further,

[0048] The preparation method of the CNPs used in S22 includes:

[0049] Ce(NO3)3·6H2O is placed in a container, and 1-octadecene, oleylamine and deionized water are added; the container containing the weighed drugs is placed in an oil bath pot, and stirring is continuously performed, so that the Ce(NO3)3·6H2O is completely dissolved in the octadecene and oleylamine to form a brownish yellow liquid; then the temperature is increased and the stirring is continuously performed for 1.5-4h to obtain a reaction liquid containing CNP nanoparticles;

[0050] The reaction liquid is taken out of the container, and after cooling to room temperature, a large amount of mixed liquid of dichloromethane and ethanol is added to disperse and settle the CNP nanoparticles, and the mixture is left to stand for 24-48h; the low-speed centrifuge is used to centrifuge and settle the liquid for 5-30min to obtain the CNPs.

[0051] Further,

[0052] The CNPs content in the electrospinning solution d prepared in S22 is 0.01%-0.1%.

[0053] A fiber membrane with poly-p-dioxanone as a base material is prepared by the above preparation method, and contains:

[0054] An outer dense layer of the PPDO material; and

[0055] An inner layer of the PPDO-poloxamer hydrogel with BG and CNP formed by electrospinning on the outer dense layer.

[0056] The present application has the following beneficial effects:

[0057] 1. The fiber membrane of the present application prevents intracranial cerebrospinal fluid penetration by using PPDO film as an outer dense layer, and improves cell compatibility and rapid degradation performance by using PPDO with good cell compatibility and degradable characteristics, thereby solving the problems of poor cell compatibility of the fiber membrane substitute and long self-degradation cycle in the prior art.

[0058] 2. The fiber membrane of the present application prevents intracranial cerebrospinal fluid penetration by using PPDO film as an outer dense layer, and prepares an inner layer by electrospinning technology with a certain concentration of BG. Since electrospinning can produce a structure similar to extracellular matrix (ECM), and PPDO has good cell compatibility and degradable characteristics, the combination of BG can promote cell growth and angiogenesis, and can be rapidly degraded, thereby solving the problems of lack of ability to promote tissue reconstruction and positively regulate cells to shorten the repair cycle in the prior art engineering tissue materials, and long self-degradation cycle.

[0059] 1. The other fiber membrane of the present application adds a combination of nano cerium oxide and poloxamer. Nano cerium oxide can provide antioxidant protection and reduce postoperative complications, while poloxamer can be used as a drug carrier to achieve local delivery of drugs in the repair area of the engineering tissue material. This combination of materials can improve the repair effect of the engineering tissue material, reduce the pain and recovery time of the patient, and has a stronger positive regulation effect on cells. BRIEF DESCRIPTION OF DRAWINGS

[0060] In order to more clearly illustrate the technical solutions in the embodiments of the present application or the prior art, the drawings needed in the description of the embodiments or the prior art will be briefly introduced. Obviously, the drawings in the following description are only some embodiments of the present application, and other drawings can be obtained by those skilled in the art without creative labor.

[0061] Figure 1 Flowchart for the preparation method of the fiber membrane of the present application;

[0062] Figure 2 SEM morphology of the fiber membrane with different PPDO concentrations in Example 1 of the present application

[0063] Figure 3 SEM images of fiber membranes for different roller rotation speeds of Example 1 of the present application;

[0064] Figure 4 SEM images of PBDs front, side, back of Example 2 of the present application;

[0065] Figure 5 SEM images of PBDs front of Example 2 of the present application;

[0066] Figure 6 Fiber diameter distribution histogram of PBDs of Example 2 of the present application;

[0067] Figure 7 Si element Mapping of PBDs front micro-morphology of Example 2 of the present application;

[0068] Figure 8 XRD results of PBDs of Example 2 of the present application;

[0069] Figure 9 DSC results of PBDs of Example 2 of the present application;

[0070] Figure 10 PBDs front of Example 2 of the present application under water droplet falling and 0.1s, 0.15s, 0.2s after contact schematic diagram;

[0071] Figure 11 PBDs back of Example 2 of the present application under water droplet falling and contact schematic diagram;

[0072] Figure 12 PBDs tensile test results of Example 2 of the present application;

[0073] Figure 13 PBDs front SEM images during degradation of Example 2 of the present application;

[0074] Figure 14 PBDs Si release line graph during degradation of Example 2 of the present application;

[0075] Figure 15 PBDs pH change line graph during degradation of Example 2 of the present application;

[0076] Figure 16 PBDs mass change line graph during degradation of Example 2 of the present application;

[0077] Figure 17 Cell live and dead staining of PBDs of Example 2 of the present application, 3-day cell DAPI and rhodamine-labeled phalloidin staining control diagram;

[0078] Figure 18 Cell CCK-8 result graph of PBDs of Example 2 of the present application;

[0079] Figure 19 Hemolysis experiment result graph of PBDs of Example 2 of the present application;

[0080] Figure 20 4h in-vivo angiogenesis experiment graph of PBDs of Example 2 of the present application;

[0081] Figure 21 Tubule connection number column graph in angiogenesis image of PBDs of Example 2 of the present application;

[0082] Figure 22 Tubule total area column graph in angiogenesis image of PBDs of Example 2 of the present application;

[0083] Figure 23 Tubule total length column graph in angiogenesis image of PBDs of Example 2 of the present application;

[0084] Figure 24 TEM micro-morphology graph of CNPs of Example 3 of the present application;

[0085] Figure 25 XRD result graph of CNPs of Example 3 of the present application;

[0086] Figure 26 XPS result graph of CNPs of Example 3 of the present application;

[0087] Figure 27 Cell CCK-8 result graph of CNPs of Example 3 of the present application;

[0088] Figure 28 Cell live and dead staining of CNPs of Example 3 of the present application, 3-day cell DAPI and rhodamine-labeled phalloidin staining control graph;

[0089] Figure 29 Frontal SEM morphology and Ce, Si element Mapping distribution schematic diagram of CBPPs of Example 4 of the present application;

[0090] Figure 30 Cell CCK-8 result graph of CBPPs of Example 4 of the present application;

[0091] Figure 31 Cell live and dead staining of CBPPs of Example 4 of the present application, 3-day cell DAPI and rhodamine-labeled phalloidin staining control graph;

[0092] Figure 32 Hemolysis experiment result graph of CBPPs of Example 4 of the present application;

[0093] Figure 33 Figure 4 shows the results of an in vivo angiogenesis experiment for CBPPs of Example 4 of the present application. DETAILED DESCRIPTION

[0094] In the description of the application, it needs to be understood that the terms "center", "longitudinal", "transverse", "length", "width", "thickness", "upper", "lower", "front", "back", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer", "clockwise", "counterclockwise", "axial", "radial", "circumferential" and the like indicate the orientation or positional relationship based on the orientation or positional relationship shown in the drawings, and are only for the convenience of describing the application and simplifying the description, and therefore cannot be understood as indicating or implying that the devices or elements referred to must have a particular orientation, be constructed and operated in a particular orientation, and therefore cannot be understood as limiting the application.

[0095] The disclosure below provides many different embodiments or examples for implementing different structures of the application. For the purpose of simplicity, the components and arrangements of the examples that are described below are shown and described in detail. It is to be understood, however, that they are merely examples and are not intended to limit the application in any way.

[0096] Poly-p-dioxanone (PPDO) is a kind of aliphatic polyester biodegradable biomedical polymer material, which has good flexibility and excellent tensile strength, and also has excellent biocompatibility, mechanical properties and degradation rate. PPDO has gradually replaced materials such as polylactic acid (PLA) and polyglycolic acid (PGA) and has become one of the main materials for surgical sutures. It is a very suitable artificial material for in vivo implants. The advantages of PPDO material are: (1) compared with the existing fiber membrane materials polycaprolactone (PCL) and poly-L-lactic acid (PLLA), PPDO material has good flexibility and excellent tensile strength and degradation period, which is suitable for medical applications that can bear tension, and avoids the need for secondary surgery removal, and the degradation product is harmless to the human body; (2) compared with the existing fiber membrane materials polyglycolic acid (PGA) and polyurethane (PU), PPDO material has good biocompatibility and mechanical properties, which can reduce the rejection reaction and inflammatory reaction with human tissues; (3) PPDO shows good stability during heat treatment, which is convenient for processing and molding, and can be chemically or physically modified to give it additional functions such as drug release and antibacterial properties.

[0097] The poly-p-dioxanone-based fiber membrane provided by the application mainly comprises: an outer dense layer of PPDO material, and an inner layer of PPDO material formed by electrospinning on the outer dense layer. The electrospinning is a special fiber manufacturing process, which uses a polymer solution or melt to perform jet spinning in a strong electric field. The principle is that under the action of an electric field, the droplet at the needle tip will change from a spherical shape to a conical shape (i.e., a "Taylor cone"), and a fiber filament is obtained by extending from the tip of the conical shape. Electrospinning can produce polymer filaments with a nanometer diameter, and in the biomedical field, nanofibers with a diameter smaller than a cell can simulate the structure and biological function of the natural extracellular matrix; and most of the tissues and organs of humans are similar in form and structure to nanofibers, which provides the possibility for nanofibers to be used for tissue and organ repair. At the same time, some electrospinning raw materials have good biocompatibility and degradability, can enter the human body as a carrier, and are easily absorbed; in addition, electrospun nanofibers also have excellent properties such as large specific surface area and porosity, therefore, electrospun nanofibers have attracted continuous attention of researchers in the biomedical field, and have been well applied in drug controlled release, wound repair, biological tissue engineering and the like.

[0098] In the preparation process of the poly-p-dioxanone-based fiber membrane of the application, first, a PPDO solution a is prepared, and a PPDO base film is prepared using the PPDO solution a; then, a PPDO electrospinning solution c is prepared; and then, electrospinning is performed on the PPDO base film using the PPDO electrospinning solution c to prepare a PPDO / BG double-layer fiber membrane. The specific preparation method of the poly-p-dioxanone-based fiber membrane is as follows:

[0099] S10. Preparing a PPDO base film: PPDO is mixed with hexafluoroisopropanol, sealed in a light-proof room, stirred at room temperature, so that the PPDO is completely dissolved in the hexafluoroisopropanol, and the mixed solution is placed in a vacuum defoaming machine to remove the bubbles in the mixed solution to obtain a pure PPDO solution a. Then, the pure PPDO solution a is poured into a glass dish in three times with intervals, and each time after pouring, a coating device is used to gently spread to ensure the uniformity of the surface. During the interval, the PPDO solution a is strictly ensured to be in a vacuum state to prevent oxidation and degradation of the PPDO. After three times of film laying, the PPDO base film is placed in a constant-temperature vacuum drying box for drying in a vacuum environment to obtain a uniform PPDO base film.

[0100] S20. Preparing a PPDO electrospinning solution c: PPDO is mixed with hexafluoroisopropanol, sealed in a light-proof room, stirred at room temperature, so that the PPDO is completely dissolved in the hexafluoroisopropanol, and the mixed solution is placed in a vacuum defoaming machine to remove the bubbles in the mixed solution to obtain the electrospinning solution c.

[0101] It should be noted that there is no fixed sequence between the S10 step and the S20 step in the present application, and the two steps can be performed simultaneously or sequentially in time.

[0102] S30. Electrospinning: The prepared PPDO electrospinning solution c is added to an electrospinning liquid injector to remove bubbles. Using an electrospinning instrument (NANON-01A MECC, Japan), a flat plate receiver is used, a conductive tin paper is attached to the flat plate receiver and the receiver is grounded, the PPDO base film is fixed on the conductive tin paper, the electrospinning needle is kept at a distance from the receiver, and the PPDO / BG double-layer fiber film is obtained after the fiber film is taken off from the conductive tin paper after the electrospinning solution is spun for multiple times.

[0103] In the fiber film of the present application, on the one hand, the PPDO film is used as an outer dense layer to prevent the penetration of intracranial cerebrospinal fluid; on the other hand, the PPDO has good cell compatibility and degradable characteristics, which improves the cell compatibility and rapid degradation performance of the fiber film, thereby solving the problems of poor cell compatibility of the fiber film substitute and long self-degradation period in the prior art.

[0104] Meanwhile, bioactive glass (Bioactive Glass, abbreviated as BG) is a kind of silicate material with biological activity, which is composed of SiO2, Na2O, CaO and P2O5 in a specific proportion. The reaction product Si ion on the surface of BG can significantly affect cell behavior, including stimulating endothelial cell angiogenesis and promoting fibroblast secretion of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF). The research in the prior art shows that the Si concentration released by the material added with bioactive glass in the body is about 1 μg / mL, which is the best for stimulating cell secretion of growth factors, and the effect will change significantly with the slight change of Si concentration.

[0105] Since PPDO has good biocompatibility and specific concentration BG has the ability to positively regulate cell behavior, and the brain relies on the precise regulation of ions to achieve its complex physiological functions, ions are crucial for the repair of dural defects, synaptic function and the stability of the entire brain environment, the present application also proposes a fiber film based on poly-p-dioxanone as a substrate, which has a main structure of a natural dura mater artificial double-layer structure film based on PPDO and BG. By finely changing the Si concentration contained in the double-layer structure film, the purpose of positively regulating intracranial cell behavior is achieved.

[0106] Another poly-p-dioxanone-based fiber membrane and a preparation method thereof are provided in the present application. The poly-p-dioxanone-based fiber membrane mainly comprises: an outer dense layer of PPDO material, and an inner layer of PPDO material with a certain concentration of BG formed by electrospinning on the outer dense layer.

[0107] In the preparation process of the poly-p-dioxanone-based fiber membrane of the present application, first, PPDO solution a is prepared, and a PPDO base film is prepared using the PPDO solution a; then, PPDO solution b is prepared, and a BG suspension is added to prepare PPDO electrospinning solution c containing biological glass; and then, electrospinning is performed on the PPDO base film using the PPDO electrospinning solution c containing biological glass to prepare a PPDO / BG double-layer artificial dura mater. The specific preparation method of the poly-p-dioxanone-based fiber membrane is as follows:

[0108] S10. Preparation of PPDO base film: PPDO is mixed with hexafluoroisopropanol, sealed in a light-proof room, stirred at room temperature, so that the PPDO is completely dissolved in the hexafluoroisopropanol, and the mixture is placed in a vacuum degassing machine to remove the bubbles in the mixture to obtain pure PPDO solution a. Then, the pure PPDO solution a is poured into a glass dish in three times with intervals, and each time after pouring, a coating device is used to gently spread it to ensure the uniformity of the surface. During the interval, the PPDO solution a is strictly ensured to be in a vacuum state to prevent oxidation and degradation of the PPDO. After three times of film laying, it is placed in a constant temperature vacuum drying box for drying in a vacuum environment to obtain a uniform PPDO base film.

[0109] S20. Preparation of PPDO electrospinning solution c containing biological glass: PPDO is mixed with hexafluoroisopropanol, sealed in a light-proof room, stirred at room temperature, so that the PPDO is completely dissolved in the hexafluoroisopropanol, and the mixture is placed in a vacuum degassing machine to remove the bubbles in the mixture to obtain pure PPDO solution b. Then, excess BG powder is mixed with deionized water, ultrasonically shaken to mix uniformly, centrifuged to take the supernatant, and then a certain amount of BG suspension is added to the pure PPDO solution b to prepare the PPDO electrospinning solution c containing biological glass.

[0110] It should be noted that there is no fixed sequence between the steps S10 and S20 in the present application, and the two steps can be performed simultaneously or sequentially in different time periods.

[0111] S30. Electrospinning: The prepared PPDO electrospinning solution containing bioglass c is added to the electrospinning liquid injector to remove bubbles. Using an electrospinning instrument (NANON-01A MECC, Japan), a flat plate receiver is used, a conductive tin paper is attached to the flat plate receiver and the receiver is grounded, the PPDO base film is fixed on the conductive tin paper, the electrospinning needle is kept at a distance from the receiver, and the PPDO / BG double-layer artificial dura mater is obtained after the electrospinning solution is spun multiple times and the fiber film is taken off from the conductive tin paper after spinning.

[0112] In another fiber membrane of the present application, on the one hand, the PPDO film is used as an outer dense layer to prevent the penetration of intracranial cerebrospinal fluid; on the other hand, the PPDO is added to a certain concentration of BG to prepare the inner layer by electrospinning technology. Since electrospinning can produce a structure similar to extracellular matrix (ECM), and PPDO has good cell compatibility and degradable properties, by combining BG, the effect of synergistically promoting cell growth and angiogenesis is achieved, and it can be quickly degraded, thereby solving the problem that the artificial dura mater substitute in the prior art lacks the ability to promote tissue reconstruction, positively regulate cells to shorten the repair cycle, and has a long self-degradation period.

[0113] In addition, the present application further adopts the scheme of adding nano cerium oxide (chemical formula CeO2, CNP for short) and poloxamer, thereby providing another fiber membrane with poly-p-dioxanone as a base material. Among them, nano cerium oxide is widely studied due to its unique physical and chemical properties, such as redox ability, catalytic activity, and biocompatibility. These properties of nano cerium oxide make it have potential application value in the field of biomedicine, especially in drug carriers and tissue engineering. For example, nano cerium oxide can be used as an antioxidant material to protect cells from oxidative stress, and also as a drug carrier to improve drug delivery efficiency and therapeutic effect. Poloxamer is a non-ionic surfactant, commonly used as a stabilizer and carrier in drug delivery systems. It can form a thermoreversible gel, which makes it very useful in controlling drug release. Poloxamer has good biocompatibility and can be degraded by the body, so it is widely used in drug delivery systems.

[0114] The following is another fiber membrane with poly-p-dioxanone as a base material and its preparation method provided by the present application. The fiber membrane with poly-p-dioxanone as a base material mainly comprises: an outer dense layer of PPDO material, and an inner layer of PPDO-poloxamer hydrogel formed by electrospinning PPDO material into BG and CNP on the outer dense layer.

[0115] Another process for preparing the fiber membrane with PPDO as the base material, first, PPDO solution a is prepared, and PPDO base film is prepared using the PPDO solution a; then, PPDO solution b is prepared, and BG suspension is added to prepare PPDO electrospinning solution c containing bioglass; and electrospinning solution d is prepared using nano cerium oxide and poloxamer; then, on the PPDO base film, electrospinning solution c and electrospinning solution d are used to perform synchronous electrospinning, and a double-layer artificial dura mater is prepared. The specific preparation method of the fiber membrane with PPDO as the base material is as follows:

[0116] S10. Preparing the PPDO base film: PPDO is mixed with hexafluoroisopropanol, sealed in a light-proof room, stirred at room temperature, so that the PPDO is completely dissolved in the hexafluoroisopropanol, and the mixed solution is placed in a vacuum degassing machine to remove the bubbles in the mixed solution to obtain pure PPDO solution a. Then, the pure PPDO solution a is poured into a glass dish in three times with intervals, and each time after pouring, a coating device is used to gently spread to ensure the uniformity of the surface. During the interval, the PPDO solution a is strictly ensured to be in a vacuum state to prevent PPDO from being oxidized and degraded. After three times of film laying, it is placed in a constant temperature vacuum drying box for drying in a vacuum environment to obtain a uniform PPDO base film.

[0117] S20. Preparing electrospinning solution c: PPDO is mixed with hexafluoroisopropanol, sealed in a light-proof room, stirred at room temperature, so that the PPDO is completely dissolved in the hexafluoroisopropanol, and the mixed solution is placed in a vacuum degassing machine to remove the bubbles in the mixed solution to obtain pure PPDO solution b. Then, the excess BG powder is mixed with deionized water, ultrasonically agitated to mix uniformly, centrifuged to take the supernatant, and then a certain amount of BG suspension is taken and added to the pure PPDO solution b to prepare the electrospinning solution c containing bioglass.

[0118] S22. Preparing electrospinning solution d: CNPs formed by CNP nanoparticles are mixed with hexafluoroisopropanol, sealed in a light-proof room, ultrasonically treated at room temperature, so that the nano cerium oxide is uniformly dispersed in the hexafluoroisopropanol. Then, poloxamer is added, sealed in a light-proof room, and stirred at room temperature to make the poloxamer completely dissolved in the hexafluoroisopropanol to prepare the electrospinning solution d.

[0119] It should be pointed out that in the present application, there is no definite order between S10 for preparing the PPDO base film, S20 for preparing the electrospinning solution c, and S22 for preparing the electrospinning solution d, and the three steps can be performed simultaneously or sequentially in time periods.

[0120] S30. Electrospinning: Using a double-head synchronous electrospinning technology, electrospinning solution c and electrospinning solution d are propelled by different syringes, a drum receiver is used, a conductive tin paper is attached to the drum receiver and the receiver is grounded, the electrospinning needle maintains a distance from the receiver, electrospinning solution c and electrospinning solution d are spun synchronously, after spinning is completed, the fiber membrane is removed from the conductive tin paper, and an electrospun double-layer artificial dura mater containing BG and CNP is obtained.

[0121] In another poly-p-dioxanone-based fiber membrane of the present application, the combination of nano cerium oxide and poloxamer brings some unique advantages. Among them, nano cerium oxide can provide antioxidant protection and reduce postoperative complications, while poloxamer can be used as a drug carrier to achieve local delivery of drugs in the dura mater repair area. This combination of materials can improve the effect of dura mater repair and reduce the pain and recovery time of patients.

[0122] It should be noted that the poly-p-dioxanone-based fiber membrane of the present application can be used not only for artificial dura mater repair, but also as a tissue engineering scaffold and artificial tissue membrane for other parts.

[0123] To specifically illustrate the beneficial effects of the present application, specific examples and corresponding experimental result analyses are provided below.

[0124] Example 1

[0125] This example provides a PPDO fiber membrane (including multiple groups) prepared by electrospinning and a preparation method thereof. The poly-p-dioxanone-based fiber membrane mainly comprises: an outer dense layer of PPDO material, and an inner layer of PPDO material formed by electrospinning on the outer dense layer. The preparation process of the poly-p-dioxanone-based fiber membrane is as follows:

[0126] S10. Preparation of PPDO base film:

[0127] In one or more embodiments, 1.8 g of PPDO is mixed with 30 mL of hexafluoroisopropanol, sealed, and stirred at room temperature for 4 h, and in one or more other embodiments, 4-8 h, so that the PPDO is completely dissolved in the hexafluoroisopropanol. The mixed solution is placed in a vacuum defoaming machine for 0.5 h to remove bubbles in the mixed solution to obtain 30 mL of a pure PPDO solution a with a concentration of 6 g / mL. The pure PPDO solution a is poured into a glass dish with a radius of 10 cm in three 10 mL portions with a 0.5 h interval, and after each pouring, the surface is gently spread with a spreader to ensure uniformity. In one or more other embodiments, the PPDO solution a can be poured with an interval of 0.5-1.5 h, and during the interval, the PPDO solution a is strictly kept in a vacuum state to prevent oxidative degradation of the PPDO. After three times of film coating, the glass dish is placed in a 37°C constant temperature vacuum drying oven, and dried for 36 h in a vacuum environment to obtain a uniform PPDO base film, and in one or more other embodiments, the drying can be performed for 24-72 h, and the purpose is to obtain a uniform PPDO base film after drying.

[0128] S20. Preparation of a PPDO electrospinning solution c:

[0129] In one or more embodiments, 0.6 g, 0.7 g, 0.8 g, 0.9 g, 1.0 g, and 1.1 g of PPDO are mixed with 10 mL of hexafluoroisopropanol, respectively, sealed, and stirred at room temperature for 4 h, and in one or more other embodiments, 4-8 h, so that the PPDO is completely dissolved in the hexafluoroisopropanol. The mixed solution is placed in a vacuum defoaming machine for 0.5 h to remove bubbles in the mixed solution to obtain 10 mL of a PPDO electrospinning solution c with a concentration of 0.06 g / mL, 0.07 g / mL, 0.08 g / mL, 0.09 g / mL, 0.1 g / mL, and 0.11 g / mL, respectively.

[0130] S30. Preparation of an oriented PPDO electrospinning fiber film:

[0131] The prepared electrospinning solution c was added into a 5 mL electrospinning liquid injector with a diameter of 12.5 mm, and bubbles were removed to ensure uniform solution ejection during electrospinning. Using an electrospinning instrument (NANON-01A MECC, Japan), the voltage was adjusted to 18 kV, the injector advancing speed was 5 mL / h, a drum receiver was used, conductive tin paper was attached to the drum receiver and the receiver was grounded, the electrospinning needle was 50 mm away from the receiver, the left and right moving speed of the needle was 100 mm / sec, 10 mL of electrospinning solution c was spun each time, and after spinning, the fiber membrane was removed from the conductive tin paper to obtain a plurality of groups of PPDO electrospinning fiber membranes with different orientation degrees and fiber diameters. It should be noted that in one or more other embodiments, the voltage is controlled in the range of 10-30 kV, the injector advancing speed is controlled in the range of 0.1-5 mL / h, the electrospinning needle is controlled in the range of 50-150 mm away from the receiver, and the left and right moving speed of the needle is controlled in the range of 25-100 mm / sec, and the receiver rotating speed is controlled in the range of 0-5000 rpm.

[0132] Example 1 benefit analysis

[0133] See Figure 2 , Figure 3 In order to test the benefits of the present application, the surface morphology structure of the PPDO double-layer fiber membrane of Example 1 was detected. Among them, the morphology and microstructure of the front surface of the dry sample of Example 1 were tested. By cutting the fibers, gold spraying treatment was performed for cross-section imaging. After obtaining the SEM image, the fiber diameter of each sample was counted.

[0134] The front surface structure of the PPDO electrospinning fiber membrane in Example 1 was observed by scanning electron microscope, as shown in Figure 2 , Figure 3 When the PPDO concentration of the electrospinning solution c is in the range of 0.06 g / mL-0.1 g / mL, and the rotating speed of the electrospinning instrument drum is in the range of 900 rpm-1200 rpm, a relatively uniform and dense structure can be obtained in the inner layer of electrospinning.

[0135] Example 2

[0136] This embodiment provides a fiber membrane with poly-p-dioxanone as a base material (including six groups) and a preparation method thereof. The fiber membrane with poly-p-dioxanone as a base material mainly comprises: an outer dense layer of PPDO material, and an inner layer of PPDO material with a certain concentration of BG formed by electrospinning on the outer dense layer. The preparation process of the fiber membrane with poly-p-dioxanone as a base material is as follows:

[0137] S10. Preparation of PPDO base film:

[0138] Mix 1.8 g PPDO with 30 mL hexafluoroisopropanol, seal and stir at room temperature for 4 h, in one or more other embodiments, it can also be stirred for 4-8 h, the purpose is to make PPDO completely dissolved in hexafluoroisopropanol, and then place it in a vacuum degassing machine for 0.5 h to remove bubbles in the mixed solution to obtain 30 mL of pure PPDO solution a with a concentration of 6 g / mL. Pour 10 mL of pure PPDO solution a at a time into a glass dish with a radius of 10 cm, with an interval of 0.5 h, and after each pouring, use a coater to gently spread it to ensure uniformity of the surface. In one or more other embodiments, it can also be poured with an interval of 0.5-1.5 h, and during the interval, strictly ensure that the PPDO solution a is in a vacuum state to prevent PPDO oxidative degradation. After three times of film laying, place it in a 37°C constant temperature vacuum drying oven and dry it in a vacuum environment for 36 h, in one or more other embodiments, it can also be dried for 24-72 h, the purpose is to obtain a uniform PPDO base film after drying.

[0139] S20. Preparation of PPDO electrospinning solution c containing bioglass:

[0140] Mix 0.6 g PPDO with 10 mL hexafluoroisopropanol, respectively, seal and stir at room temperature for 4 h to make PPDO completely dissolved in hexafluoroisopropanol, in one or more other embodiments, it can also be stirred for 2-8 h, because the electrospinning solution is also stirred again after the addition of BG powder, so the first stirring time can be appropriately shortened. Then place it in a vacuum degassing machine for 0.5 h to remove bubbles in the mixed solution to obtain 10 mL of PPDO solution b with a concentration of 0.06 g / mL, respectively. Mix excess BG powder with deionized water, ultrasonic oscillation for 1 h to make it uniformly mixed, centrifuge at 2000 rpm for 5 min, in one or more other embodiments, it can also be centrifuged at 2000-4000 rpm, and then take the supernatant and use icp-oes to quantitatively determine the silicon concentration in the BG suspension. Then, take the quantified BG suspension and add it to the PPDO solution b to make the silicon concentration in the mixed electrospinning solution c be 0.5 mg / L, 1 mg / L, 1.5 mg / L, 2 mg / L, 10 mg / L, and 50 mg / L, respectively, to obtain six groups of electrospinning solutions c.

[0141] S30. Electrospinning:

[0142] The prepared six groups of electrospinning solution c were added into 5 mL electrospinning liquid syringe with a diameter of 12.5 mm respectively, and the bubbles were removed. Using electrospinning instrument (NANON-01A MECC, Japan), the voltage was adjusted to 14 kV, the syringe advancing speed was 3 mL / h, the flat plate receiver was used, the conductive tin paper was attached to the flat plate receiver and the receiver was grounded, the PPDO base film in step S10 was fixed on the conductive tin paper, the distance between the electrospinning needle and the receiver was 120 mm, the left and right moving speed of the needle was 50 mm / sec, 10 mL electrospinning solution c was spun each time, and the fiber membrane was taken off from the conductive tin paper after spinning. Six groups of electrospinning solution c were used to obtain six groups of PPDO / BG double-layer artificial dura mater with different BG contents, which were named as PBD-1, PBD-2, PBD-3, PBD-4, PBD-5 and PBD-6 respectively for comparative experiments. It should be pointed out that in one or more other embodiments, the voltage is controlled in the range of 10-30 kV, the syringe advancing speed is controlled in the range of 0.1-5 mL / h, the distance between the electrospinning needle and the receiver is controlled in the range of 50-150 mm, and the left and right moving speed of the needle is controlled in the range of 25-100 mm / sec, and the receiver rotating speed is controlled in the range of 0-5000 rpm.

[0143] Beneficial effect analysis of example 2

[0144] Please refer to Figures 4-23 In order to test the beneficial effects of the present application, the physicochemical properties, mechanical properties, water absorption rate and in vitro degradation performance of the PPDO / BG double-layer artificial dura mater of example 2 were tested, and the influence of the BG content of the dura mater on cell proliferation, cell morphology and cell compatibility of the material was tested.

[0145] (1) Surface morphology structure analysis

[0146] (1) Surface morphology structure analysis

[0147] The morphology and microstructure of the front and back surfaces and the cross section of the dried samples of example 2 were tested, and the element distribution and relative abundance on the surface of the samples of example 2 were tested by EDS. The cross section imaging was carried out by cutting the fibers and gold spraying treatment. After obtaining the SEM images, the fiber diameter of each sample was counted.

[0148] Among them, the front and back surface and side surface structure observed by scanning electron microscope is as follows Figure 4SEM images of the PBDs show that the front side of the PBDs has a classic isotropic electrospinning structure, with a uniform distribution of the mesh at different magnifications and no obvious beading, indicating that the PPDO / BG double-layer material (PBD) has the properties of traditional electrospinning films, and can particularly mimic the extracellular matrix ECM. The back side SEM images show that the fused layer has a dense and uniform structure without pores, indicating that the preparation process can ensure the uniformity of the back side. The side SEM images show that the dense fused layer and the electrospinning layer are tightly combined without cracking, and the double-layer structure stably exists. At the same time, as shown in Figure 5 、 Figure 6 the fiber diameter distribution obtained by Image J shows that the fiber diameter of the spinning layer gradually decreases with the addition of the bioglass, and no obvious uneven structure appears in all samples. In addition, the results of Mapping in Figure 7 show that the bioglass BG with a specific Si content is uniformly distributed in the front side of the PBD spinning layer, and the Si content shown in Mapping increases with the increase of the Si content, which is consistent with the expected experimental results.

[0149] (2) XRD analysis

[0150] The dried PPDO raw material and PBDs dura mater were subjected to XRD testing. The XRD spectrum of the PBDs is shown in Figure 8 . The experimental results show that the PBDs have two characteristic peaks of the PPDO raw material and the PBDs near 2θ = 21° and 2θ = 23°, which are the diffraction peaks of their similar crystal structures, and the half-peak width becomes wider and the relative peak height becomes lower with the increase of the bioglass content. The results show that the crystallinity of the PBDs with different Si contents decreases. This is because the bioglass is usually a non-crystalline or low-crystallinity material. When it is mixed into the polymer, especially when the fiber film is prepared by the electrospinning process, the non-crystalline nature of the bioglass can affect the crystallinity of the overall material. The non-crystalline nature of the bioglass can inhibit the crystallization of the polymer molecules, making the overall material more non-crystalline. At the same time, after adding the BG, the physical angle reduces the freedom of the long and short chains of ppdo, thereby changing the crystallization properties.

[0151] (3) DSC analysis

[0152] DSC was used to characterize the thermodynamic properties of the PBDs. The melting temperature (Tm) and melting enthalpy (ΔHm) of the PBDs were determined, and the melting enthalpy (ΔHm) of 100% crystalline PPDO was 141.18 J / g. The crystallinity was calculated according to the following formula:

[0153]

[0154] ​​DSC results of PBDs are shown in FIG. 1, the melting temperature (Tm) and melting enthalpy (ΔHm) of all samples are almost the same. Figure 9 The relative crystallinity of PBDs is calculated and shown in FIG. 2, the results show that the relative crystallinity of PBDs decreases with the addition of bio-glass, which is basically consistent with the XRD analysis results.

[0155] (II) Water contact angle analysis

[0156] The water contact angle of PBDs was measured by optical contact angle measurement instrument. The water contact angle results are shown in FIG. 3 and FIG. 4, the experimental results show that the front surface of PBD is super-hydrophilic, and the water droplet is completely absorbed within 0.2s after falling. The back surface of PBD is dense and hydrophilic, and the hydrophilic angle is 73°. The results are consistent with the properties of PPDO material itself. The hydrophilicity can support cell adhesion, growth and differentiation, and reduce inflammation and foreign body reaction caused by the material in the brain tissue. Figure 10 Figure 11 (III) Mechanical property analysis

[0157] The tensile properties of PBDs were tested. The maximum tensile deformation displacement and the maximum tensile stress of the sample were measured by an electronic universal testing machine. As shown in FIG. 5, the results show that all sample groups meet the minimum maximum tensile stress (5Mpa) requirement of artificial dura mater. There is no obvious difference in the maximum tensile deformation displacement and the maximum tensile stress in PBD-1~PBD-5 groups, but there is a significant downward trend in PBD-6 group. This is because bio-glass is usually a relatively hard and brittle material, while PPDO has relatively high toughness. The tensile strength of pure PPDO fiber membrane can reach 20Mpa. When the addition of bio-glass reaches a certain amount, the rigidity of the material increases, and the overall toughness of the material decreases, resulting in a step change in PBD-6 compared with other groups.

[0158] (IV) In vitro degradation analysis Figure 12 (1) In vitro degradation surface morphology analysis

[0159] PBS (Biosharp, US) buffer was used as the simulated body fluid, and different concentrations of PBDs samples were weighed and immersed in the degradation solution. The samples were incubated at constant temperature and shaken for 1, 4, 7, 14, 21 and 28 days, and the solution was changed every 3 days. There were 3 control groups at each time node. The samples and degradation solution at 6 time nodes were taken out for characterization.

[0160] During the in vitro degradation period, the morphology changes of PBD-1~PBD-6 groups are shown in FIG. 6 and FIG. 7.

[0161]

[0162] Figure 13 ​​​​The experimental results show that the degradation trends of PBD-1~PBD-6 groups are roughly the same within the first 4 days, and no significant morphological changes are observed. Under the observation of high-magnification and low-magnification scanning electron microscopy (SEM), the fiber structure presents clear, uniform and linear electrospinning characteristics. At the end of the second week, the surface structure of PBD-1~PBD-6 groups begins to collapse, and the fibrous structure also begins to show slight bending. With the continuous development of the degradation process, by the fourth week, compared with the previous three periods, the destruction of the surface structure is further intensified, and most of the fibrous structure has failed to maintain its original linear form. Under high-magnification SEM, the observed structure presents a state similar to "melting", and the deformation of the fibrous structure is extremely significant, which reflects the gradual collapse of the structure of the material in the 28-day degradation process, and the characteristics of a shorter degradation period.

[0163] (2) In vitro degradation weight loss rate, pH change, Si release analysis

[0164] The pH of the degradation solution at each time point in vitro degradation was determined, and the Si concentration in the degradation solution at a specific time point was determined using a spectrometer. The mass of the sample before the experiment is denoted as . After the in vitro degradation sample is taken out, it is placed in a vacuum drying box, and the mass after drying to a constant weight is denoted as . The degradation rate is calculated by the formula:

[0165]

[0166] The weight loss rate, pH change, and Si release of PBDs during 4 weeks of PBS degradation are shown in Figure 14 , Figure 15 , Figure 16 . The experimental results show that within the first week, the weight loss rate of PBDs is nearly zero. Within the second week, the weight loss rate of all PBDs samples experiences a stepwise increase, generally exceeding 5%. After four weeks of degradation, the weight loss rate of PBDs reaches a peak, with PBD-6 group showing the most significant performance (9.9%). The in vitro degradation weight loss of PBDs is mainly due to the hydrolysis of PPDO and the dissolution of low molecular weight components. The observed differences in weight loss rate among the PBDs groups can be attributed to the differences in fiber diameter during electrospinning. The fiber diameter of PBD-1~PBD-6 groups gradually decreases, and the thinning of the fiber diameter and the large pore structure produced by electrospinning increase the contact area between water molecules and the fiber structure, thereby accelerating the degradation rate.

[0167] Regarding pH changes, the trends across the sample groups were generally consistent. On the first day of degradation, the release of bioglass, which was not encapsulated within the fibers but rather free within the PBD fiber structure, caused a brief increase in pH, with the PBD-6 group showing a pH peak exceeding 8 during this phase. As the degradation process continued, the pH degradation trends across the groups gradually converged, stabilizing between 6.8 and 7.0 within four weeks.

[0168] The result of silicon release is as follows Figure 14 As shown, on the first day of the experiment, the silicon concentration briefly increased due to the release of free bioglass, and then remained stable for 4 to 7 days. Starting on day 14, the partial hydrolysis of PBDs led to the release of bioglass encapsulated within the PPDO fibers, causing the silicon concentration in the degradation solution to peak on day 21. However, due to the slightly acidic environment created by the PPDO degradation product 2-hydroxyethoxyacetic acid, the pH of the degradation solution did not increase significantly. By the fourth week, with the further collapse of the PBD surface structure, the internally encapsulated bioglass was released, leading to an increase in the overall silicon release. These experimental results are largely consistent with the trends in structural changes, pH changes, and weight loss observed by SEM, collectively verifying the systematic regularity of PBD degradation behavior.

[0169] (v) Biocompatibility analysis

[0170] L929 cells (mouse fibroblasts) were selected for in vitro experiments and cultured in MEM medium until sufficient numbers and healthy morphology were achieved. PBD samples were immersed in the medium, placed in an incubator, and centrifuged to obtain the sample extract. Cells were seeded in 96-well plates, and after cell adhesion, the sample extract was replaced. Cell viability was assessed on days 1, 3, and 5 of incubation. CCK-8 reagent was bound to the cells, and the cells were incubated in the dark, with absorbance measured at 450 nm. Cell viability was also assessed using a live / dead staining kit. Cells were seeded in 48-well plates, and after cell adhesion, the sample extract was replaced and the cells were incubated for 3 days. The sample extract was then removed and the cells were washed. Cells were then fixed with paraformaldehyde and washed. Subsequently, cells were permeabilized with Triton X-100 solution and washed. TRITC-labeled phalloidin staining solution was added to each well, and the cells were incubated at room temperature in the dark before removing the staining solution and washing. Finally, the cell nuclei were stained with DAPI solution, incubated at room temperature in the dark, and then the staining solution was removed and the cells were washed. Cell morphology was observed and photographed using an inverted microscope.

[0171] (1) Cell viability

[0172] The cytocompatibility of PBDs was evaluated using the L929 cell line. For example... Figure 17As shown in Fig. 1, the cell viability of PBD-1, PBD-2, PBD-3, PBD-4 groups was more than 100% within 1, 3, 5 days of culture, and was significantly higher than that of the control group on the 5th day, indicating that they could promote the proliferation of L929 cell line. However, the cell viability of PBD-5 and PBD-6 groups was much lower than that of other groups within the culture period, with the cell viability of PBD-5 being less than 60% and that of PBD-6 being less than 20%, indicating that the two groups of materials had obvious cytotoxicity. This is because the PBDs leaching solution releases ions such as silicon ions, calcium ions, and phosphate ions under the culture conditions with L929 cells, and these ions have no obvious effect on cells or can positively promote cells at low doses. However, when the ion concentration exceeds a certain threshold, it will affect the stability of the intracellular and extracellular environment, leading to cell damage or death. Secondly, the degradation of 45S5 bioglass also leads to the release of alkaline substances, and a high alkaline environment may have a negative regulation on the survival and normal function of cells, affecting the stability of the cell membrane and the normal function of ion channels.

[0173] (2) Live / Dead

[0174] In order to further confirm the cytotoxicity of the samples, the cell activity was detected by Live / Dead experiment. As shown in Fig. 2, Figure 17 Figure 18 The experimental results show that, except for the PBD-6 group, the cell density of the other groups increases with the increase of culture days. The cell behavior of the PBD-1 group is similar to that of the control group within the culture period. On the 3rd and 5th days, the cell density of the PBD-2, PBD-3, PBD-4 groups is greater than that of the control group. However, the cell density of the PBD-5 group is low, and the number of living cells is less than that of the control group. The cell density of the PBD-6 group changes little from the 1st day of culture to the 3rd and 5th days. The Live / Dead staining results are basically consistent with the CCK-8 test results.

[0175] (3) Cell morphology

[0176] DAPI and phalloidin solution were used to detect the effect of the samples on cell morphology, and further evaluate the cell compatibility. As shown in Fig. 3, Figure 17 Figure 18 The experimental results show that the cells of PBD-1, PBD-2, PBD-3, PBD-4 groups remain spindle-shaped and star-shaped, and the structure of cytoplasm and nucleus is clear after staining. However, the cells of PBD-5 and PBD-6 groups are in a diffuse round or oval shape, the cytoplasm staining is not clear, and the nucleus is atrophic, indicating that PBD-5 and PBD-6 have high cytotoxicity, which is consistent with the results of CCK-8 and Live / Dead staining.

[0177] (6) Blood compatibility analysis​​

[0178] Rabbit blood was centrifuged to separate red blood cells and washed and diluted. Subsequently, PBDs were combined with the red blood cell suspension, and the mixture was incubated by shaking, centrifuged, and the supernatant was placed in a 96-well plate, and the absorbance of the supernatant was measured at 540 nm. The absorbance of red blood cells treated with 0.1% Triton X-100 and PBS was the positive control group and the negative control group , and the absorbance of the material treated with PBS was the material control group , and each sample was repeated three times. The formula for calculating the hemolysis rate of the sample (Hemolysis rate) is:

[0179]

[0180] As shown in Figure 19 , the experimental results show that the supernatant obtained by treating the red blood cell suspension with the PBDs sample solution and centrifugation is transparent in color, but the quantitative absorbance analysis data shows that the hemolysis rate of the PBD-1~PBD-4 group material is less than 5%, and the hemolysis rate of the PBD-5~PBD-6 group is higher than 5%, and there is no statistical difference between the PBDs material groups, and there is a significant statistical difference compared with the positive control group Triton X-100, which shows that the PBD-1~PBD-4 group material has blood safety.

[0181] (Seven) In vitro blood vessel formation

[0182] Due to the poor cell compatibility of PBD-5 and PBD-6 groups, they are not added to the experimental group in subsequent experiments. HUVECs (human umbilical vein endothelial cells) are used to evaluate the blood vessel formation ability of PBDs. HUVECs are resuscitated, passaged and induced to form blood vessels using ECM medium (Endothelial Cell Medium) and endothelial cell growth factor ECGS (Endothelial cell growth supplement).

[0183] After rewarming, HUVEC cell suspension was transferred to centrifuge tubes and ECM complete medium was added. The cells were centrifuged, medium was added, and the suspension was continuously pipetted to obtain a uniformly distributed cell suspension. The cell suspension was seeded into culture flasks and placed in a CO2 incubator, with fresh medium replaced every 48 hours. When cell confluence reached 70%, passage was performed. The medium was removed from the flasks, the cells were washed, and trypsin was added and the flasks were agitated. When the cells became rounded and white mist appeared on the flask walls, complete medium was added to stop trypsin digestion. The cell suspension was then centrifuged, medium was added, and the cells were pipetted until uniformly suspended before being seeded into culture flasks and placed in an incubator. Additionally, a matrix gel was evenly spread across the bottom of a 48-well culture plate and placed in an incubator for further incubation. Once the HUVEC cells have reached the required level in the incubator, remove them, centrifuge them, and add the sample extract solution soaked in ECM (the preparation method of the extract solution is the same as above). Count the cells in the processed cell suspension. After counting, add 1.5 × 10⁶ cells to each well of a culture plate covered with matrix gel and incubate them in a cell culture incubator. Observe the cell morphology and take pictures using an inverted microscope. Repeat each sample three times.

[0184] like Figures 20-22 As shown in the figure, the angiogenesis experiment results indicate that, compared with the control group, the extracts of PBD-1, PBD-2, PBD-3, and PBD-4 groups significantly promoted in vitro angiogenesis of HUVECs. Furthermore, with increasing Si concentration, the length of the vessels, the number of connections, the number of formed vessels, and the area of ​​the formed vessels all increased. This is because, when Si ions in the bioglass are present in appropriate amounts, they can stimulate vascular endothelial cells to secrete growth factors, such as vascular endothelial growth factor (VEGF), thereby promoting the proliferation and migration of vascular endothelial cells.

[0185] The experimental results above indicate that groups PBD-1 to PBD-4 showed better performance. Specifically, when the silicon concentration in electrospinning solution c was 0.5–2 mg / L, the resulting PBDs met the requirements for artificial dura mater in terms of microstructure, crystallinity, tensile strength, and hydrophilicity. Furthermore, they exhibited a shorter degradation cycle, good compatibility with cells and blood, and no significant cytotoxicity, while also promoting cell proliferation and migration. This combination of a shorter degradation cycle and the ability to promote cell and tissue remodeling allows patients using these PBDs to recover more quickly.

[0186] Example 3

[0187] This embodiment provides six types of cerium oxide nanoparticles (CNPs) and their preparation methods, demonstrating the process of selecting suitable nanoparticles to prepare another type of fiber membrane based on poly(p-dioxanone) as a substrate according to this invention. The preparation process of the six types of cerium oxide nanoparticles is based on existing technology, using a modified thermal decomposition method to synthesize cerium oxide nanoparticles of different particle sizes and valence states. The specific process is as follows:

[0188] Take 10 mmol of cerium (III) nitrate hexahydrate (Ce(NO3)3·6H2O, 4.342 g) in a three-necked round-bottom flask, and add 150 mmol of 1-octadecene (39.455 g), 30 mmol of oleylamine (8.025 g). Then, place the three-necked flask containing the weighed chemicals in an oil bath, and continue stirring at 80°C and 200 rpm for 30 min, so that the cerium (III) nitrate hexahydrate is completely dissolved in the octadecene and oleylamine to form a brownish-yellow liquid. Subsequently, increase the temperature to 160°C and continue stirring for 2 h to obtain CNP-1 group nanoparticles. In one or more other embodiments, the stirring time after temperature increase is controlled to be 1.5-4 h.

[0189] Take the same 10 mmol of cerium (III) nitrate hexahydrate (Ce(NO3)3·6H2O, 4.342 g), 150 mmol of 1-octadecene (39.455 g), 30 mmol of oleylamine (8.025 g), and add 400 mmol of deionized water (7.2 g), repeat the steps of CNP-1 group to obtain CNP-2 group nanoparticles.

[0190] Reduce the proportion of oleylamine to 15 mmol (4.025 g) and 10 mmol (2.675 g) respectively, repeat the steps of CNP-2 group to obtain CNP-3 and CNP-4 group nanoparticles.

[0191] Repeat the steps of CNP-2 group, and prepare the process throughout the nitrogen as a protective gas to repeat its steps to obtain CNP-5 group nanoparticles.

[0192] Repeat the steps of CNP-5, and after stirring at 80°C under nitrogen protection for 30 min, increase the temperature to 190°C and continue stirring for 2 h to obtain CNP-6 group nanoparticles.

[0193] Take the reaction liquid of each group from the three-necked flask, and after cooling to room temperature, pour it into a beaker. Add a large amount of dichloromethane and ethanol 1:5 mixture to disperse and settle the nanoparticles, and stand for 24 h. Centrifuge the settled liquid at 4000 rpm for 5 min to obtain CNPs pellets. In one or more other embodiments, the standing time is controlled to be 24-48 h, and the centrifugation time of the settled liquid is controlled to be 5-30 min. Then, use anhydrous ethanol as a washing agent and dispersant, disperse the CNPs pellets and then centrifuge, wash away the residual dichloromethane and other impurities, and repeat 5 times until the supernatant after centrifugation becomes transparent. Finally, disperse the CNPs pellets in anhydrous ethanol using an ultrasonic instrument to ensure uniform dispersion for storage.

[0194] Example 3 analysis

[0195] See Figures 24-28 , in order to test another preparation method of CNPs suitable for the poly-p-dioxycyclohexanone-based fiber membrane of the present application, the physicochemical properties, micro-morphology, and cell compatibility of CNPs were experimented.

[0196] (I) Analysis of particle size and physicochemical properties of CNPs

[0197] As Figure 24 , the TEM results showed that the particle sizes of the 6 groups of CNPs were 3.4 ± 0.3 nm, 3.4 ± 0.8 nm, 7.2 ± 0.1 nm, 10 ± 0.2 nm, 14 ± 1.6 nm, and 8.7 ± 0.7 nm, respectively, and the particle size distribution of each group was uniform. Figure 25 , the XRD results showed that the characteristic peaks of CNPs appeared at 28.46° (111), 33.10° (200), 47.40° (220), 56.22° (311), 59.01° (222), and 69.69° (400), which was consistent with the results of existing research, proving the successful synthesis of CNPs. In addition, the XPS results showed that the escape energy of CNPs powder tested at C 1s (284.6 eV) was at 570.25 eV, 580.2 eV, 585.7 eV, 588.6 eV, 598.5 eV, and 604.5 eV, which was Ce4+, and the escape energy was at 583.1 eV, 587.1 eV, 601.6 eV, and 606.4 eV, which was Ce3+. The oxidation state ratio of CNPs was obtained by peak fitting of XPS data using avantage software. Figure 26 , the results showed that the proportion of Ce3+ in the 6 groups of CNPs was 33.73%, 56.65%, 35.00%, 53.21%, 22.42%, and 18.91%, respectively.

[0198] (II) Analysis of cell compatibility of CNPs

[0199] Different particle sizes and valence states of CNPs were mixed with MEM medium to keep the CNPs content in the medium at 0.1%, and were co-cultured with L929 cells. As Figure 27CCK-8 results showed that the cell proliferation rates of all experimental groups were greater than 95% after co-culturing for 1, 3, and 5 days. It was indicated that CNPs in each group at a content of 0.1% had no cytotoxicity. After incubation for 1 day, the CNP-5 group had the best effect on promoting cell proliferation, followed by the CNP-2 group and the CNP-6 group. However, as the incubation time reached 3 days and 5 days, the CNP-4 group, the CNP-5 group, and the CNP-6 group showed a phenomenon that the cell proliferation rate was lower than that of the control group, and at this time, the CNP-2 group had the best effect on promoting cell proliferation. It was indicated that after co-culturing with cells for 1 day, the cells had a relatively strong toxic side effect on the CNP-4 group, the CNP-5 group, and the CNP-6 group. With the increase of the culture time, the proliferation rates of the three groups decreased sharply and were lower than that of the control group. The reason might be that the Ce3+ content of the CNP-5 group and the CNP-6 group was relatively low, and the ability to regulate the active oxygen content in the cell environment was weak. The Ce3+ content of the CNP-4 group was relatively high, but because the particle size was large (14 nm ± 1.6 nm), the exchange of cells with external substances was affected when co-cultured with cells, thereby causing the cell proliferation rate to decrease significantly after the strong toxic side effect appeared on the first day.

[0200] In addition, as shown in Figure 28 , the cell proliferation ability of CNPs was further evaluated by a Live / Dead test. A large number of living cells were observed in all groups within 1, 3, and 5 days. After incubation for 3 days and 5 days, it was observed that the proportion of living cells in the CNP-2 and CNP-3 groups was much higher than that in the control group and other experimental groups, and the proportion of living cells in the CNP-4 to CNP-6 groups was significantly lower than that in the control group. This was basically consistent with the results of the CCK-8 experiment, which again indicated that CNPs with a particle size of about 3.4 ± 0.8 nm and a relatively high Ce3+ content were more conducive to promoting cell proliferation.

[0201] At the same time, as shown in Figure 28 , the morphology of L929 cells in all groups was normal, most of which were spindle-shaped, a small amount of triangular and circular, and the pseudopods were long and the density was high. It was indicated that CNPs at a suitable concentration had no effect on the cell morphology, and appropriate addition of the material could promote the proliferation of cells and play a positive regulatory role in the repair of tissues in specific areas.

[0202] Based on the above experiments, it was known that the CNPs prepared by the CNP-2 group were more suitable for use in the artificial dura mater material of the present application.

[0203] Example 4

[0204] The embodiment provides a fiber membrane with poly-p-dioxanone as a base material (divided into four groups) and a preparation method thereof. The fiber membrane with poly-p-dioxanone as a base material mainly comprises: an outer dense layer of PPDO material, and an inner layer of PPDO-poloxamer hydrogel with BG and CNP participating in PPDO formed by electrospinning on the outer dense layer. The fiber membrane with poly-p-dioxanone as a base material is prepared as follows:

[0205] S10. Preparing a PPDO base film: 1.8g of PPDO is mixed with 30mL of hexafluoroisopropanol, sealed, stirred in a dark room at room temperature for 4h, and in one or more other embodiments, can be stirred for 4-8h, so that the PPDO is completely dissolved in the hexafluoroisopropanol, and then placed in a vacuum defoaming machine for 0.5h to remove bubbles in the mixed solution to obtain 30mL of a pure PPDO solution a with a concentration of 6g / mL. The pure PPDO solution a is poured into a glass dish with a radius of 10cm in three times of 10mL each time with an interval of 0.5h, and after each time of pouring, a coater is used to gently spread to ensure uniformity of the surface. In one or more other embodiments, it can also be poured with an interval of 0.5-1.5h, and during the interval, the PPDO solution a is strictly ensured to be in a vacuum state to prevent oxidation and degradation of the PPDO. After three times of film laying, it is placed in a 37℃ constant temperature vacuum drying box, and dried in a vacuum environment for 36h, and in one or more other embodiments, can be dried for 24-72h, so that a uniform PPDO base film is obtained after drying.

[0206] S20. Preparing a PPDO electrospinning solution c containing bioglass: 2.4g of PPDO is mixed with 30mL of hexafluoroisopropanol, respectively, sealed, stirred in a dark room at room temperature for 4h, so that the PPDO is completely dissolved in the hexafluoroisopropanol, and in one or more other embodiments, can be stirred for 2-8h, because the electrospinning solution is prepared, and after the BG powder is added, it is stirred again, so the first stirring time can be appropriately shortened. Then, it is placed in a vacuum defoaming machine for 0.5h to remove bubbles in the mixed solution to obtain 30mL of a PPDO solution b with a concentration of 0.08g / mL, respectively. Excess 45s5-BG powder is mixed with deionized water, ultrasonically oscillated for 1h to make it uniformly mixed, centrifuged at 2000rpm for 5min, and in one or more other embodiments, can be centrifuged at 2000-4000rpm, and the supernatant is taken, and the silicon concentration in the BG suspension is determined by icp-oes. The quantitative BG suspension is added to the pure PPDO solution b so that the silicon concentration in the mixed electrospinning solution c is 1.5mg / L, respectively, to obtain the electrospinning solution c.

[0207] S22. Preparation of electrospinning solution d: 0.001 g, 0.01 g, 0.05 g, 0.1 g CNPs of nano cerium oxide group 2 (CNP-2) in Example 3 were mixed with 10 mL of hexafluoroisopropanol respectively, and sealed in a dark room for ultrasonic treatment for 2 h. In one or more other embodiments, the ultrasonic treatment time is controlled to be 1-4 h, with the purpose of uniformly dispersing the nano cerium oxide in the hexafluoroisopropanol. 3 g of poloxamer was added to each group respectively, and sealed in a dark room for stirring for 2 h. In one or more other embodiments, the stirring time is controlled to be 2-8 h, with the purpose of completely dissolving the poloxamer in the hexafluoroisopropanol, to obtain four groups of mixed electrospinning solution d. The CNP content of the electrospinning solution d in each group is 0.01%, 0.1%, 0.5%, and 1% respectively.

[0208] S30. Electrospinning: Using an electrospinning instrument (NANON-01A MECC, Japan), a double-head synchronous electrospinning technique was adopted, the voltage was adjusted to 18 kV, the syringe advancing speed of the electrospinning solution c was 3 mL / h, and the syringe advancing speed of the electrospinning solution d was 1 mL / h. A drum receiver was used, a conductive tin paper was attached to the drum receiver and the receiver was grounded, the distance between the electrospinning needle and the receiver was 50 mm, the left and right moving speed of the needle was 25 mm / sec, and the drum rotating speed was set to 800 rpm. 30 mL of the electrospinning solution c and 10 mL of the electrospinning solution d were electrospun each time, and after the electrospinning was completed, the fiber membrane was taken off from the conductive tin paper. Four groups of CBPPs containing BG and CNP of PPDO-poloxamer hydrogel were obtained by using the four groups of mixed electrospinning solution d with different CNP content, and were named as CBPP-1, CBPP-2, CBPP-3 and CBPP-4 respectively, which were used for comparative experiments. It should be pointed out that in one or more other embodiments, the voltage is controlled to be in the range of 15-25 kV, the syringe advancing speed of the electrospinning solution c is controlled to be in the range of 1-5 mL / h, the syringe advancing speed of the electrospinning solution d is controlled to be in the range of 0.1-2 mL / h, the distance between the electrospinning needle and the receiver is controlled to be in the range of 25-75 mm, the left and right moving speed of the needle is controlled to be in the range of 25-100 mm / sec, and the drum rotating speed is set to be in the range of 200-800 rpm.

[0209] Beneficial effect analysis of Example 4

[0210] Please refer to Figures 29-33 In order to test the beneficial effect of the present application, the surface morphology structure of the CBPP containing BG and CNP of PPDO-poloxamer hydrogel, and the influence of CNPs and poloxamer on cell proliferation, cell morphology and cell compatibility of the material were tested.

[0211] (1) Surface morphology structure analysis

[0212] The detection process refers to Example 2. The hydrogel fiber membrane was observed by scanning electron microscope, as shown in Figure 29 The surface of the electrospun double-layer material (CBPP) containing BG and CNP of the PPDO-poloxamer hydrogel was a combination of isotropic electrospun structure and hydrogel film structure, the poloxamer hydrogel was uniformly distributed in the pore size of the electrospun fiber membrane, there was no beading of the electrospun, the overall structure was dense, indicating that the preparation process was perfect. The results of Mapping showed that the specific Si element content of the bioactive glass BG was uniformly distributed in the CBPP spun fiber hydrogel layer, and the Ce element increased in the Mapping as the CNP content increased, which was consistent with the expected experimental results.

[0213] (B) Biocompatibility analysis

[0214] The experimental process refers to Example 2.

[0215] Cell activity

[0216] The cell compatibility of CBPP was evaluated using L929 cell line. The results are shown in Figure 30 The cell viability of CBPP-1, CBPP-2, CBPP-3 groups was more than 100% within 1 day and 3 days of culture, among which the CBPP-1 and CBPP-2 groups were much greater than the control group. At 5 days, the CBPP-1 and CBPP-2 groups decreased to almost the same level as the control group, while the CBPP-3 and CBPP-4 groups were much lower than the control group. The overall results showed that the BG bioactive glass and CNP nanoparticles could significantly promote cell proliferation within a reasonable concentration range, but excessive addition of CNP nanoparticles had cytotoxicity.

[0217] Live / Dead

[0218] As shown in Figure 31 , the experimental results showed that the cell density of all groups increased with the increase of culture days. At 5 days, the cell density of CBPP-1 and CBPP-2 groups was much greater than that of the control group, and the cell behavior of the remaining groups was similar to that of the control group within the culture period. The CBPP-4 group had part of the cells spheroid from the first day of culture, and the cell density changed little at 1 day and 3 days. The Live / Dead staining results were basically consistent with the CCK-8 test results, proving that appropriate BG-CNP could promote the proliferation of L929 cells.

[0219] Cell morphology

[0220] DAPI and phalloidin solution were used to detect the effect of the sample on cell morphology, further evaluating its cell compatibility. As shown in Figure 31As shown, cells in the CBPP-1 and CBPP-2 groups maintained a spindle shape, and the cytoplasm and nucleus showed clear structures after staining. With increasing CNP content, the proportion of diffusely round and oval cells increased, cytoplasmic staining became less clear, and nuclei shrank. This was particularly evident in the CBPP-4 group, demonstrating its high cytotoxicity, which is largely consistent with the results of CCK-8 and Live / Dead staining.

[0221] (vi) Blood compatibility analysis

[0222] The experimental procedure is as described in Example 2. Figure 32 As shown, the experimental results indicate that the supernatant obtained by centrifuging red blood cell suspension treated with CBPPs sample solution was transparent. However, quantitative absorbance analysis showed that the hemolysis rate of CBPP-1 to CBPP-4 groups was less than 5%, and there was no statistical difference between the CBPPs material groups. Compared with the positive control group Triton X-100, there was a significant statistical difference. The results indicate that the CBPP-1 to CBPP-4 groups have blood safety.

[0223] (vii) In vitro angiogenesis

[0224] The experimental procedure is as described in Example 2. Figure 33 As shown in the figure, the angiogenesis experiment results indicate that, compared with the control group, the extracts of CBPP-1, CBPP-2, and CBPP-3 groups significantly promoted in vitro angiogenesis of HUVECs. Furthermore, with increasing CNP concentration, the length of vessels, the number of connections, the number of formed vessels, and the vessel area first increased and then decreased. When the concentration reached a certain level, the CBPP-4 group could no longer promote in vitro angiogenesis of HUVECs. This is because CNP's ability to regulate oxidative stress and scavenge free radicals allows the CBPP-1, CBPP-2, and CBPP-3 groups to significantly positively regulate cells compared to the control group. However, the cytotoxicity caused by excessively high concentrations negatively impacted cell proliferation and differentiation.

[0225] Based on the above experimental results, it can be seen that the CBPP-1~CBPP-2 groups have better effects. That is, when the CNP nanoparticle content in the electrospinning solution d is 0.01~0.1%, the obtained CBPPs can be well compatible with cells and blood, do not produce obvious cytotoxicity, and have a stronger positive regulatory effect on cells.

[0226] The above shows and describes the basic principles, main features and advantages of the present application. Those skilled in the art should understand that the present application is not limited to the above-mentioned embodiments, and the above-mentioned embodiments and descriptions in the specification are only preferred examples of the present application and are not intended to limit the present application. Various changes and improvements can be made to the present application without departing from the spirit and scope of the present application, and these changes and improvements all fall within the scope of the claimed present application. The scope of protection of the present application is defined by the appended claims and their equivalents.

Claims

1. A method for preparing a fiber membrane using poly(p-dioxanone) as a substrate, characterized in that, Includes the following steps: S10. Prepare PPDO solution a by mixing PPDO with an organic solvent, use the PPDO solution a to lay a film multiple times, and dry it to obtain a PPDO substrate film; S20. Prepare PPDO solution b by mixing PPDO with an organic solvent, and add bioglass suspension to PPDO solution b to obtain PPDO electrospinning solution c; S22. Electrospinning solution d is prepared by mixing nano-cerium oxide particles with an organic solvent and adding poloxamer; S30. Electrospinning is performed simultaneously on a PPDO substrate membrane using electrospinning solution c and electrospinning solution d to prepare a fiber membrane with polydioxanone as the substrate. There is no fixed order between steps S10, S20 and S22; The preparation method of the cerium oxide nanoparticle clusters used in S22 includes: Cerium(III) nitrate hexahydrate (Ce(NO3)3·6H2O) was placed in a container, and 1-octadecene, oleylamine, and deionized water were added. The container containing the weighed reagents was placed in an oil bath and stirred continuously until the cerium(III) nitrate hexahydrate was completely dissolved in octadecene and oleylamine, forming a brownish-yellow liquid. Then the temperature was increased and the mixture was stirred continuously for 1.5 to 4 hours to obtain a reaction solution containing cerium oxide nanoparticles. Remove the reaction solution from the container, cool it to room temperature, and then add a large amount of a mixture of dichloromethane and ethanol to disperse and precipitate the cerium oxide nanoparticles. Let it stand for 24-48 hours. Centrifuge the precipitate for 5-30 minutes using a low-speed centrifuge to obtain nano-cerium oxide particle clusters. The molar ratio of cerium(III) nitrate hexahydrate (Ce(NO3)3·6H2O), 1-octadecene, oleylamine and deionized water is 1:15:3:40 or 2:30:3:

80.

2. The method for preparing a fiber membrane based on poly(p-dioxanone) as described in claim 1, characterized in that, The organic solvent used in S10, S20 and S22 is hexafluoroisopropanol, and both the PPDO solution a and PPDO solution b prepared by mixing need to have air bubbles removed.

3. The method for preparing a fiber membrane based on poly(p-dioxanone) as described in claim 2, characterized in that: S10 specifically includes: mixing PPDO with hexafluoroisopropanol, sealing and stirring at room temperature in a light-proof environment to completely dissolve PPDO in hexafluoroisopropanol, and placing the mixture in a vacuum degassing machine to remove air bubbles to obtain a pure PPDO solution a; pouring the pure PPDO solution a into a glass dish three times at intervals, and spreading it with a coater after each pour to ensure a uniform surface; placing the three layers into a constant temperature vacuum drying oven for vacuum drying to obtain a PPDO substrate film.

4. The method for preparing a fiber membrane based on poly(p-dioxanone) as described in claim 2, characterized in that: S20 specifically includes: mixing PPDO with hexafluoroisopropanol, sealing and stirring at room temperature in a light-proof environment to completely dissolve PPDO in hexafluoroisopropanol, and placing the mixture in a vacuum defoamer to remove air bubbles to obtain a pure PPDO solution b; mixing excess BG powder with deionized water, ultrasonically vibrating to ensure uniform mixing, centrifuging to collect the supernatant, and then adding a quantitative amount of BG suspension to the pure PPDO solution b to obtain a PPDO electrospinning solution c.

5. The method for preparing a fiber membrane based on poly(p-dioxanone) as described in claim 2, characterized in that: S22 specifically includes: mixing CNPs groups with hexafluoroisopropanol, sealing and protecting from light, and ultrasonically treating at room temperature to make the nano-cerium oxide uniformly dispersed in hexafluoroisopropanol; adding poloxamer, sealing and protecting from light, and stirring at room temperature to make the poloxamer completely dissolve in hexafluoroisopropanol to obtain electrospinning solution d.

6. The method for preparing a fiber membrane based on poly(p-dioxanone) as described in claim 1, characterized in that: Specifically, S30 includes: employing a dual-head synchronous electrospinning technique, wherein the electrospinning solution c and electrospinning solution d are injected synchronously using different syringes; and performing multiple electrospinning operations using an electrospinning instrument to produce a fiber membrane based on poly(p-dioxanone).

7. The method for preparing a fiber membrane based on poly(p-dioxanone) as described in any one of claims 1-6, characterized in that, The content of nano-cerium oxide particle clusters in the electrospinning solution d prepared in S22 is 0.0001 g / mL to 0.01 g / mL.

8. A fiber membrane based on poly(p-dioxanone) as a substrate, characterized in that, Prepared by the preparation method according to any one of claims 1-7, characterized in that it comprises: The outer dense layer of PPDO material; and An inner layer of PPDO-poloxam hydrogel incorporating bioglass and cerium oxide is formed by electrospinning on the outer dense layer.

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