Nanofiber electrospun membrane as well as preparation method and application thereof

Abstract

The invention provides a nanofiber electrospun membrane as well as a preparation method and application thereof, and belongs to the technical field of tooth implantation repair materials. The novel GBR membrane is prepared by taking Col, BC, Mg-MOF and P28 as raw materials and using an electrostatic spinning technology, and a P28 sustained-release system taking MOFs as a carrier is constructed in the membrane, so that the defects of the clinical GBR membrane are overcome, the bone defect repairing effect of the GBR operation is improved, and a new thought is expected to be provided for implant repairing of tooth deficiency.

Description

Technical Field

[0001] The present invention relates to the technical field of dental implant restoration materials, and particularly to a nanofiber electrospun membrane, a preparation method thereof, and an application thereof. Background Art

[0002] Tooth defect and tooth loss are common diseases in clinical diagnosis and treatment of stomatology. At present, dental implant technology has become a conventional restoration treatment method for tooth loss. How to broaden the indications of dental implantation and improve the restoration effect of dental implants is still an urgent problem to be solved in clinical work and scientific research.

[0003] Guided Tissue Regeneration (GBR) uses the mechanical barrier effect of the GBR membrane to block the surrounding tissues with a faster growth rate, so that they cannot preferentially contact the bone defect area, providing a relatively closed tissue growth space for the healing of the bone defect, enabling the cells with regenerative ability in the bone defect area to proliferate and differentiate to the maximum extent, and realizing the reparative regeneration of the bone defect area. For the problem of bone defect in dental implantation, GBR has currently been proven to be an effective method and is widely used in clinical practice. The key link in GBR is the application of the GBR membrane.

[0004] GBR membranes are divided into absorbable membranes and non-absorbable membranes. Currently, absorbable membranes are more commonly used clinically, among which the domestic Hyaff oral repair membrane and the imported Bio-Gide biofilm are the most widely used. The Bio-Gide biofilm belongs to a non-crosslinked absorbable collagen membrane made from pigskin. Due to problems such as the lack of osteoinductive activity, too fast degradation rate, and weak mechanical strength in itself, it is generally used in combination with bone filling materials clinically, such as Bio-Oss bone powder material and Bio-Oss Collagen material. Since Bio-Oss bone powder does not contain active ingredients such as bone growth factors and lacks the active induction of bone, it cannot maintain a high osteogenic activity for a long time and has poor effects in repairing large bone defects. Summary of the Invention

[0005] In view of this, the purpose of the present invention is to provide a nanofiber electrospun membrane, a preparation method thereof, and an application thereof. The nanofiber electrospun membrane prepared by the present invention can construct a P28 sustained-release system with MOFs as a carrier in the membrane, aiming to make up for the deficiencies of the GBR membrane clinically and improve the repair effect of GBR on bone defects.

[0006] To achieve the above-mentioned invention purpose, the present invention provides the following technical solutions:

[0007] The present invention provides a preparation method of a nanofiber electrospun membrane, comprising the following steps:

[0008] Mix the P28 active polypeptide, Mg-MOF and water, and then incubate and dry them in sequence to obtain a powder;

[0009] Mix the powder with a chitosan solution to obtain Mg-MOF@P28;

[0010] Mix sulfadiazine and a 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid / diaminetetraacetic acid solution to obtain a sulfadiazine solution;

[0011] Mix hydroxylamine, a 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid / diaminetetraacetic acid solution and an RGD solution (arginine-glycine-aspartic acid solution), and then conduct the first incubation to obtain an incubation product;

[0012] Mix the incubation product, the sulfadiazine solution, MAL-PEG-COOH and a 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid / diaminetetraacetic acid solution, and conduct the second incubation to obtain SD-PEG-COOH;

[0013] Mix the SD-PEG-COOH, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride, N-hydroxysuccinimide and water to conduct an activation reaction to obtain an SSDD solution;

[0014] Mix the Mg-MOF@P28 with the SSDD solution to conduct a host-guest reaction to obtain a Mg-MOF@P28@SDSSD solid;

[0015] Mix collagen, bacterial cellulose and a solvent to obtain a shell spinning solution;

[0016] Mix the Mg-MOF@P28@SDSSD with water to obtain a core spinning solution;

[0017] Conduct electrospinning on the shell spinning solution and the core spinning solution to obtain the nanofiber electrospun membrane.

[0018] Preferably, the mass ratio of the P28 active polypeptide to Mg-MOF is 2.9-3.1:8.9-9.1.

[0019] Preferably, the mass ratio of the P28 active polypeptide in the powder to chitosan in the chitosan solution is 2.9-3.1:2.

[0020] Preferably, the mass ratio of SD-PEG-COOH used for preparing the SSDD solution to Mg-MOF@P28 is 0.95-1.04:1.

[0021] Preferably, the mass ratio of collagen to bacterial cellulose is 0.745-0.754:0.745-0.754.

[0022] Preferably, the mass fraction of the shell spinning solution is 15%.

[0023] Preferably, the concentration of the core spinning solution is 10 μg / mL.

[0024] Preferably, the parameters of the electrospinning include: the voltage is 20 kV, the receiving distance is 20 cm, the injection rate of the shell spinning solution is 2.0 mL / h, and the injection rate of the core spinning solution is 0.4 mL / h.

[0025] The present invention also provides a nanofiber electrospun membrane prepared by the preparation method described in the above technical solution.

[0026] The present invention also provides the application of the nanofiber electrospun membrane described in the above technical solution in the preparation of dental implant restoration materials.

[0027] The present invention provides a method for preparing a nanofiber electrospun membrane, comprising the following steps: mixing P28 active polypeptide, Mg-MOF and water, incubating and drying in sequence to obtain a powder; mixing the powder with a chitosan solution to obtain Mg-MOF@P28; mixing sulfadiazine and a 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid / diaminetetraacetic acid solution to obtain a sulfadiazine solution; mixing hydroxylamine, a 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid / diaminetetraacetic acid solution and an RGD solution, and performing a first incubation to obtain an incubation product; mixing the incubation product, the sulfadiazine solution, MAL-PEG-COOH and a 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid / diaminetetraacetic acid solution, and performing a second incubation to obtain SD-PEG-COOH; mixing the SD-PEG-COOH, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride, N-hydroxysuccinimide and water, and performing an activation reaction to obtain an SSDD solution; mixing the Mg-MOF@P28 with the SSDD solution to perform a host-guest reaction to obtain a Mg-MOF@P28@SDSSD solid; mixing collagen, bacterial cellulose and a solvent to obtain a shell spinning solution; mixing the Mg-MOF@P28@SDSSD with water to obtain a core spinning solution; performing electrospinning on the shell spinning solution and the core spinning solution to obtain the nanofiber electrospun membrane.

[0028] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0029] In the present invention, collagen (Collagen, Col) is the main component of the extracellular matrix. The Col membrane prepared therefrom has good biocompatibility, low antigenicity and low cytotoxicity, and can better guide bone regeneration. However, it has deficiencies in mechanical and mechanical properties, is easily torn and degraded. Since the actual application environment of the GBR membrane is a wet body fluid environment, it is necessary to improve the mechanical properties of the thin film in the wet state; bacterial cellulose (Bacterial Cellulose, BC) is a biopolymer produced by bacteria and fungi, and has unique physical and chemical properties and mechanical properties, including high crystallinity, strong water-holding capacity, excellent tensile strength and Young's modulus. It has been proven that it can be used as a reinforcing agent for nanocomposites. Preparing a nanofiber membrane by electrospinning technology with Col combined with BC can make up for the deficiencies of the Col membrane; bone morphogenetic protein (Bone Morphogenetic Protein, BMPs) is a member of the transforming growth factor-β superfamily and is also the growth factor that appears most frequently in the field of bone tissue engineering in recent years. It is generally considered to be a positive regulator of new bone formation. Among them, BMP-2 has the strongest osteogenic effect. P28 is a small molecule active polypeptide containing 28 amino acids designed by analyzing the "core domain" in the natural BMP-2 molecular structure that truly exerts osteogenic induction activity. P28 can promote the expression of osteogenesis-related gene mRNA of bone marrow stromal stem cells (BMSCs) and increase the activity of alkaline phosphatase, significantly stimulate osteoporotic bone regeneration, and can also effectively promote the proliferation, aggregation and osteogenic differentiation of MC3T3-E1 cells. It is a growth factor with strong osteogenic induction activity that can be used on a large scale in bone tissue engineering. Metal-organic frameworks (Metal-Organic Frameworks, MOFs) are metal nanoparticles constructed by bridging metal ions or metal clusters with organic ligands. Due to their unique structural diversity, large surface area, adjustable pore size, affinity for various functionalization methods and biodegradability, they can be used as excellent nanocarrier materials in the fields of tissue engineering and regeneration. MOFs can play an antibacterial role through metal ion release, oxidative stress or non-oxidative mechanisms, and are not easily caused bacterial drug resistance. MOFs can not only play an antibacterial and sterilization role, but also be used as a sustained-release performance carrier to load P28 peptide to prevent its premature degradation and continuously exert osteogenic activity. The present invention uses Col, BC, Mg-MOF and P28 as raw materials and prepares a novel GBR membrane by electrospinning technology, and constructs a P28 sustained-release system with MOFs as the carrier in the membrane, aiming to make up for the deficiencies of the GBR membrane in clinical practice, improve the repair effect of the GBR technique on bone defects, and hope to provide new ideas for the implant restoration of tooth loss.

[0030] Moreover, the nanofiber electrospun membrane (electrospun nanofiber membrane) of the present invention is a type of membranous material prepared by electrospinning technology, which has the characteristics of simple preparation process, suitable mechanical properties, large specific surface area, high drug loading capacity, and good air permeability. Moreover, the fiber diameter, porosity, interconnected structure, and biodegradation rate of the nanofiber electrospun membrane can be adjusted to meet various functions required by the GBR membrane. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] Figure 1 It is a schematic diagram for the preparation of the Col / BC / Mg-MOF@P28@SDSSD membrane and a schematic diagram of its action on osteoblasts;

[0032] Figure 2 It is a preparation flow chart of Col / BC / Mg-MOF@P28@SDSSD and its application flow chart in the repair of critical bone defects;

[0033] Figure 3 It is a comparison chart of DMEM medium, traditional GBR membrane extract, and the extract of the CBMPS nanofiber membrane in Example 1 for culturing osteoblasts;

[0034] Figure 4 It is a CCK-8 detection comparison chart of blank control, traditional GBR membrane, and the CBMPS nanofiber membrane in Example 1 at different detection days. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0035] The present invention provides a method for preparing a nanofiber electrospun membrane, which includes the following steps:

[0036] Mix P28 bioactive polypeptide, Mg-MOF, and water, and then incubate and dry them in sequence to obtain a powder;

[0037] Mix the powder with a chitosan solution to obtain Mg-MOF@P28;

[0038] Mix sulfadiazine and 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid / diaminetetraacetic acid solution to obtain a sulfadiazine solution;

[0039] Mix hydroxylamine, 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid / diaminetetraacetic acid solution, and RGD solution, and then perform the first incubation to obtain an incubation product;

[0040] Mix the incubation product, the sulfadiazine solution, MAL-PEG-COOH, and 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid / diaminetetraacetic acid solution, and perform the second incubation to obtain SD-PEG-COOH;

[0041] Mix the SD-PEG-COOH, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride, N-hydroxysuccinimide and water for an activation reaction to obtain an SSDD solution;

[0042] Mix the Mg-MOF@P28 with the SSDD solution for a host-guest reaction to obtain a Mg-MOF@P28@SDSSD solid;

[0043] Mix collagen, bacterial cellulose and a solvent to obtain a shell spinning solution;

[0044] Mix the Mg-MOF@P28@SDSSD with water to obtain a core spinning solution;

[0045] Electrospin the shell spinning solution and the core spinning solution to obtain the nanofiber electrospun membrane.

[0046] In the present invention, unless otherwise specified, the raw materials used are all commercially available products in the art.

[0047] In the present invention, the P28 active polypeptide, Mg-MOF and water are mixed and then incubated and dried in sequence to obtain a powder

[0048] In the present invention, the mass ratio of the P28 active polypeptide to the Mg-MOF is preferably 2.9-3.1:8.9-9.1, and more preferably 1:3.

[0049] In the present invention, the mass ratio of the P28 active polypeptide to water is preferably 2.9-3.1:1, and more preferably 3:1.

[0050] In the present invention, the Mg-MOF is preferably prepared by a method including the following steps:

[0051] Weigh 0.7115-0.7124 g of Mg(NO 3 )·6H 2 O and 0.1665-0.1674 g of 2,5-dihydroxyterephthalic acid, dissolve them in 67.5 mL of N,N-dimethylformamide, 4.5 mL of deionized water, and 4.5 mL of ethanol in a 100 mL beaker, ultrasonically dissolve for 30 min, then transfer to a high-pressure reaction kettle, react at 398 K for 24 h, after the reaction is completed, cool to room temperature and pour out the pale yellow supernatant to obtain a dark yellow Mg-MOF sample, transfer the sample to methanol and shake for three days to displace the DMF and H 2 O in the framework, replace methanol 6 times during this period, then place the sample in a vacuum at 373 K for 12 h to obtain the Mg-MOF.

[0052] In the present invention, the incubation temperature is preferably 4 °C and the time is preferably 24 h.

[0053] In the present invention, the drying is preferably freeze-drying, the temperature of the freeze-drying is preferably -55 °C, the time is preferably 24 h, and the freeze-drying is preferably carried out in a negative pressure freeze-dryer.

[0054] In the present invention, the P28 active polypeptide is preferably first dissolved in water, the Mg-MOF is added to the obtained solution, and then ultrasonic dispersion is carried out to make it uniform. The obtained mixed solution is incubated in a 4 °C refrigerator for 24 h and then put into a -55 °C negative pressure freeze-dryer for freezing and drying for 24 h.

[0055] After obtaining the powder, the present invention mixes the powder with a chitosan solution to obtain Mg-MOF@P28. The Mg-MOF@P28 is a metal-organic framework-small molecule active peptide composite structure formed with P28 as the ligand and Mg-MOF as the skeleton. The chitosan in the chitosan solution is linked to Mg-MOF@P28 in a molecular form.

[0056] In the present invention, the mass ratio of the P28 active polypeptide in the powder to the chitosan in the chitosan solution is preferably 2.9 - 3.1:2, and more preferably 3:2.

[0057] In the present invention, the mixing is preferably stirring.

[0058] The present invention mixes sulfadiazine (SD) and 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid / diaminetetraacetic acid solution (HEPES / EDTA) to obtain a sulfadiazine solution (SD solution).

[0059] In the present invention, the concentration of the HEPES / EDTA is preferably 0.5 M, and the pH value is preferably 8.0.

[0060] In the present invention, the dosage ratio of the SD to the HEPES / EDTA is preferably 1 mg:500 μL.

[0061] The present invention mixes hydroxylamine, 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid / diaminetetraacetic acid solution and RGD solution and then carries out the first incubation to obtain an incubation product.

[0062] In the present invention, the 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid / diaminetetraacetic acid solution is preferably the same as the above scheme and will not be elaborated here.

[0063] In the present invention, the volume ratio of the hydroxylamine to the 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid / diaminetetraacetic acid solution is preferably 2:5.

[0064] In the present invention, the temperature of the first incubation is preferably 37 °C, and the time is preferably 1.5 h.

[0065] After obtaining the incubation product, the present invention mixes the incubation product, sulfadiazine solution, MAL-PEG-COOH (carboxyl-polyethylene glycol-maleimide), and 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid / diaminetetraacetic acid solution for a second incubation to obtain SD-PEG-COOH, which is a PEG functionalized with SD.

[0066] In the present invention, the temperature of the second incubation is preferably 4°C, and the time is preferably 12 h.

[0067] After the second incubation is completed, the present invention preferably washes the obtained second incubation product three times with PBS to obtain the SD-PEG-COOH.

[0068] In the present invention, the PBS preferably contains 0.5 M EDTA.

[0069] After obtaining SD-PEG-COOH, the present invention mixes the SD-PEG-COOH, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC), N-hydroxysuccinimide (NHS), and water for an activation reaction to obtain an SSDD solution.

[0070] In the present invention, the mass ratio of SD-PEG-COOH, EDC, and NHS is preferably 0.95 - 1.04:0.95 - 1.04:0.95 - 1.04, more preferably 1:1:1.

[0071] In the present invention, the temperature of the activation reaction is preferably 4°C, and the time is preferably 10 min.

[0072] After obtaining the SSDD solution, the present invention mixes the Mg-MOF@P28 with the SSDD solution for a host-guest reaction to obtain a Mg-MOF@P28@SDSSD solid.

[0073] In the present invention, the mass ratio of SD-PEG-COOH to Mg-MOF@P28 used in preparing the SSDD solution is preferably 0.95 - 1.04:1, more preferably 1:1.

[0074] In the present invention, the temperature of the host-guest reaction is preferably 4°C, and the time is preferably 12 h. During the host-guest reaction, SSDD is connected to chitosan on Mg-MOF@P28, thereby obtaining Mg-MOF@P28@SDSSD. Mg-MOF@P28@SDSSD is a composite material formed with Mg-MOF as the metal-organic framework, P28 as the ligand, and SSDD as the load connected by chitosan. @ represents that the substance is loaded on Mg-MOF.

[0075] After the host-guest reaction is completed, the present invention preferably purifies the obtained reaction solution by dialysis (MWCO = 3500) three times, and rinses it with PBS (pH = 7.4) to obtain the Mg-MOF@P28@SDSSD solid.

[0076] The present invention mixes collagen (Col), bacterial cellulose (BC) and a solvent to obtain a shell spinning solution.

[0077] In the present invention, the mass ratio of the collagen to the bacterial cellulose is preferably 0.745 - 0.754:0.745 - 0.754.

[0078] In the present invention, the solvent is preferably trifluoroethanol.

[0079] In the present invention, the mass fraction of the shell spinning solution is preferably 15%.

[0080] In the present invention, the mixing is preferably carried out by stirring with a magnetic stirrer. The present invention has no special limitation on the specific manner of stirring with the magnetic stirrer, and parameters well-known to those skilled in the art can be adopted.

[0081] The present invention mixes the Mg-MOF@P28@SDSSD with water to obtain a core spinning solution.

[0082] In the present invention, the concentration of the core spinning solution is preferably 10 μg / mL.

[0083] In the present invention, the water preferably further contains a protein stabilizer. The protein stabilizer is preferably BSA, and the concentration of the protein stabilizer in the water is preferably 1 mg / mL.

[0084] After obtaining the shell spinning solution and the core spinning solution, the present invention electrospins the shell spinning solution and the core spinning solution to obtain the nanofiber electrospun membrane.

[0085] In the present invention, the parameters of the electrospinning preferably include: the voltage is 20 kV, the receiving distance is 20 cm, the injection rate of the shell spinning solution is 2.0 mL / h, and the injection rate of the core spinning solution is 0.4 mL / h

[0086] The present invention connects the shell spinning solution and the core spinning solution to the shell and core needles respectively.

[0087] The present invention also provides a nanofiber electrospun membrane (CBMPS or Col / BC / Mg-MOF@P28@SDSSD membrane) prepared by the preparation method described in the above technical solution.

[0088] The present invention also provides an application of the nanofiber electrospun membrane described in the above technical solution in the preparation of dental implant restoration materials.

[0089] The technical solutions of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all of the embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts belong to the scope of protection of the present invention.

[0090] Example 1

[0091] Figure 1 It is a schematic diagram of the preparation principle of the Col / BC / Mg-MOF@P28@SDSSD membrane and the schematic diagram of its action on osteoblasts. The preparation process is as follows:

[0092] Synthesis of Mg-MOF

[0093] Weigh 0.712 g of Mg(NO 3 )·6H 2 O and 0.167 g of 2,5-dihydroxyterephthalic acid, dissolve them in 67.5 mL of N,N-dimethylformamide, 4.5 mL of deionized water, and 4.5 mL of ethanol in a 100 mL beaker; after ultrasonic dissolution for 30 min, transfer it to a high-pressure reaction kettle; react at 398 K for 24 h, and after the reaction is completed, cool it to room temperature and pour out the pale yellow supernatant to obtain a dark yellow Mg-MOF sample; transfer the sample to methanol and shake it for three days to displace DMF and H 2 O in the framework. Replace methanol 6 times during this period, and then place the sample in a vacuum dryer at 373 K for 12 h to obtain Mg-MOF solid.

[0094] Preparation of Mg-MOF@P28@SDSSD

[0095] 1. Preparation of Mg-MOF@P28

[0096] Weigh 3 mg of P28 active polypeptide, dissolve it thoroughly with 1 mL of deionized water, add 9 mg of Mg-MOF to the solution, disperse it evenly by ultrasonic treatment, place the mixed solution in a refrigerator at 4 °C for incubation for 24 h, then put it into a -55 °C negative pressure freeze dryer for freezing and drying for 24 h, and then mix 1 mL of chitosan solution with a concentration of 2 mg / mL with the freeze-dried powder and stir it evenly to obtain Mg-MOF@P28.

[0097] 2. Preparation of SD-PEG-COOH

[0098] Dissolve SD (1 mg) in 500 μL of 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid / diaminetetraacetic acid (HEPES / EDTA) solution (0.5 M, pH 8.0) to obtain an SD solution. Hydroxylamine (200 μL) was mixed with HEPES / EDTA (500 μL), and then added to the RGD solution. After incubation at 37 °C for 90 min, 300 mg of the incubation product and MAL-PEG-COOH were dissolved in 200 μL of 0.5 M HEPES / EDTA, and then added to the prepared SD solution, followed by incubation overnight at 4 °C. The resulting material was washed three times with PBS (containing 0.5 M EDTA) to obtain SD-PEG-COOH.

[0099] 2. Preparation of Mg-MOF@P28@SDSSD

[0100] Weigh SD-PEG-COOH (1 mg) and dissolve it in deionized water. React it with 1 mg of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) and 1 mg of N-hydroxysuccinimide (NHS) at 4 °C for 10 min to obtain an SSDD solution. Add Mg-MOF@P28 (1 mg) to the obtained SSDD solution and react at 4 °C for 12 h. The reaction solution was purified by dialysis three times (MWCO = 3500), and after rinsing with PBS (500 μL, pH = 7.4), Mg-MOF@P28@SDSSD solid was obtained.

[0101] Preparation of CBMPS membrane

[0102] Preparation of CBMPS nanofiber membrane

[0103] Weigh 0.75 g of Col and 0.75 g of BC respectively and dissolve them in 8.5 g of trifluoroethanol solution. Stir with a magnetic stirrer overnight to obtain a homogeneous spinning solution with a mass fraction of 15% as the shell spinning solution. Dissolve Mg-MOF@P28@SDSSD in deionized water (using 1 mg / mL BSA as a protein stabilizer) to obtain a 10 μg / mL Mg-MOF@P28@SDSSD aqueous solution as the core spinning solution. Connect the two spinning solutions to the shell and core needles respectively, and perform electrospinning at a voltage of 20 kV, a receiving distance of 20 cm, and an injection rate of 2.0 mL / h for the shell layer and 0.4 mL / h for the core layer to obtain a CBMPS nanofiber membrane for storage and standby.

[0104] Figure 2 It is the preparation flow chart of Col / BC / Mg-MOF@P28@SDSSD and its application flow chart in critical bone defect repair.

[0105] DMEM medium, the leaching solution of the traditional GBR membrane, and the leaching solution of the CBMPS nanofiber membrane of this example were used to culture osteoblasts, and the results are shown in Figure 3 , Figure 3 where A in Figure 3 is DMEM medium, that is, the blank control, B is the leaching solution of the traditional GBR membrane, and C is the leaching solution of the CBMPS nanofiber membrane of this example. The scale bars in

[0106] are all 500 μm. It can be seen that there are no obvious differences among the three groups.

[0107] CCK-8 assay was performed on the blank control, the traditional GBR membrane, and the CBMPS nanofiber membrane of this example. Figure 4 is the comparison chart of OD values at different detection days. It can be seen that as time gradually increases, the cell viability is normal.

[0108] The above are only the preferred embodiments of the present invention and do not impose any formal limitations on the present invention. It should be noted that for those of ordinary skill in the art of this technology, without departing from the principle of the present invention, several improvements and refinements can still be made, and these improvements and refinements should also be regarded as the protection scope of the present invention.

Claims

1. A method for preparing a nanofiber electrospun membrane, characterized in that: The following steps are involved: The P28 active polypeptide, Mg-MOF and water are mixed, incubated and dried in sequence to obtain a powder; The powder is mixed with a chitosan solution to obtain Mg-MOF@P28; mixing sulfadiazine and 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid / diaminetetraacetic acid solution to obtain a sulfadiazine solution; The hydroxylamine, 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid / diaminetetraacetic acid solution and RGD solution are mixed and then first incubated to obtain an incubation product; The incubation product, sulfadiazine solution, MAL-PEG-COOH and 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid / diaminetetraacetic acid solution are mixed for a second incubation to obtain SD-PEG-COOH; The SD-PEG-COOH, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride, N-hydroxysuccinimide and water are mixed for activation reaction to obtain a SDSSD solution; The Mg-MOF@P28 and SDSSD solution are mixed to perform a host-guest reaction to obtain a Mg-MOF@P28@SDSSD solid; Mixing collagen, bacterial cellulose and a solvent to obtain a shell spinning solution; Mixing the Mg-MOF@P28@SDSSD with water to obtain a core layer spinning solution; The shell layer spinning solution and the core layer spinning solution are subjected to electrostatic spinning to obtain the nanofiber electrospun membrane.

2. The preparation method according to claim 1, characterized in that: The mass ratio of the P28 active polypeptide to Mg-MOF is 2.9-3.1:8.9-9.

1.

3. The preparation method according to claim 1 or 2, characterized in that: The mass ratio of the P28 active polypeptide in the powder to the chitosan in the chitosan solution is 2.9-3.1:

2.

4. The preparation method according to claim 1, characterized in that: The mass ratio of SD-PEG-COOH to Mg-MOF@P28 used in preparing the SDSSD solution is 0.95-1.04:

1.

5. The preparation method according to claim 1, characterized in that: The mass ratio of the collagen to the bacterial cellulose is 0.745-0.754:0.745-0.

754.

6. The preparation method according to claim 1 or 5, characterized in that: The mass fraction of the shell layer spinning solution is 15%.

7. The preparation method according to claim 1, characterized in that: The concentration of the core layer spinning solution is 10 μg / mL.

8. The preparation method according to claim 1, characterized in that: The electrospinning parameters include: voltage of 20 kV, receiving distance of 20 cm, injection rate of the shell layer spinning solution of 2.0 mL / h, and injection rate of the core layer spinning solution of 0.4 mL / h.

9. The nanofiber electrospun membrane prepared by the preparation method according to any one of claims 1 to 8.

10. Use of the nanofiber electrospun membrane according to claim 9 in preparing implant restoration materials for missing teeth.

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