Multifunctional bio-fiber membrane and preparation method and application thereof

Abstract

The application discloses a multifunctional biological fiber membrane and a preparation method and application thereof, and belongs to the technical field of biomedical materials. The multifunctional biological fiber membrane is of a double-layer structure, the outer layer is dense, the fiber surface is smooth, the fibers are cross-linked with each other, and the pores are reduced and closed; the inner layer is porous, has a grid-shaped aperture, and the fibers are straight. The outer layer is made of spinning solution A, the spinning solution A comprises a high polymer material A, a photo initiator, a functional component and an organic solvent; and the inner layer is made of spinning solution B, the spinning solution B comprises a high polymer material B, a functional component and an organic solvent. The multifunctional biological fiber membrane is of a double-layer structure, the outer layer is dense, the fiber surface is smooth, the fibers are cross-linked with each other, and the pores are reduced and closed; the inner layer is porous, has a grid-shaped aperture, and the fibers are straight, the dense barrier structure of the outer layer can effectively block the invasion of epithelial cells, the loaded antibacterial component can effectively resist bacteria, and the inner layer has a grid-shaped aperture and loads a bone formation promoting component, so that the bone regeneration can be promoted.

Description

Technical Field

[0001] This invention relates to the field of biomedical materials technology, specifically to a multifunctional biofiber membrane, its preparation method, and its applications. Background Technology

[0002] Periodontal disease, periapical periodontitis, and trauma often lead to varying degrees of alveolar bone defects, severely impacting subsequent restorative treatment. Guided bone regeneration (GBR) is currently the most commonly used bone defect repair technique in clinical practice, and the barrier membrane plays a crucial role in GBR. Currently, the most commonly used membranes in clinical practice are commercially available absorbable collagen membranes, such as Bio-Gide and Hyo collagen membranes. However, the rapid degradation of collagen membranes causes them to lose their ability to maintain osteogenic space. More importantly, collagen membranes lack osteoinductive activity and antibacterial capabilities, posing a risk of infection under bacterial influence and affecting surgical prognosis. Therefore, constructing a GBR barrier membrane with both antibacterial and osteogenic functions is particularly important. Osteogenic and antibacterial functions are directions for the development of biomaterials. This fibrous membrane is expected to overcome the performance limitations of traditional collagen membrane materials, providing new ideas for the research and development of bone regeneration materials.

[0003] The mechanical properties, surface morphology, and degradation rate of fiber composite membrane scaffold structures based on electrospinning and in-situ crosslinking technologies can be adjusted by the type and ratio of polymers and the parameters of electrospinning. This is simple, easy to implement, and can be mass-produced. It is widely used in tissue engineering and regenerative medicine, filtration, sensors and other fields. How to prepare functional GBR membranes through electrospinning and in-situ crosslinking technologies has become a technical problem that urgently needs to be solved by those skilled in the art. Summary of the Invention

[0004] The purpose of this invention is to overcome the shortcomings of the existing technology and provide a multifunctional bio-fiber membrane, its preparation method and application. The dense barrier structure of the outer layer of the multifunctional bio-fiber membrane can effectively block the invasion of epithelial cells, and the loaded antibacterial and osteogenic components can effectively prevent bacteria and induce osteogenic growth.

[0005] To achieve the above objectives, the technical solution adopted by the present invention is as follows:

[0006] A multifunctional bio-cellulose membrane, wherein the multifunctional bio-cellulose membrane has a double-layer structure, the outer layer is dense with smooth fiber surfaces, the fibers are cross-linked and the pores are reduced and closed; the inner layer is loose and porous with a mesh-like pore size and straight fibers.

[0007] The outer layer is made of spinning solution A, which includes polymer material A, photoinitiator, functional components and organic solvent;

[0008] The inner layer is made of spinning solution B, which includes polymer material B, functional components and organic solvents.

[0009] The multifunctional bio-fiber membrane of this invention has a bilayer structure. The outer layer is dense with a smooth fiber surface, and the fibers are cross-linked, resulting in narrowed and closed pores. The inner layer is loose and porous with a mesh-like pore size and straight fibers. The dense barrier structure of the outer layer can effectively block the invasion of epithelial cells, and the loaded antibacterial agents provide effective antibacterial protection. The mesh-like pore size of the inner layer allows it to load osteogenic components, promoting bone regeneration. The fibrous membrane is designed with layered pores from the inside out, forming a fibrous membrane with a composite function of "antibacterial-osteogenic-barrier". In addition, the multifunctional bio-fiber membrane of this invention can serve as a structural framework to load various active ingredients such as drugs, proteins, DNA, and nanoparticles, modifying their functions. The release of active ingredients can be controlled according to the degree of cross-linking. The amino groups within the outer layer structure can be further chemically modified to optimize the function. This composite fibrous membrane has a simple preparation technology, stable composition, controllable structure, reliable performance, and adjustable function, showing broad prospects for biomedical applications.

[0010] In a preferred embodiment of the present invention, the mass concentration of polymeric material A in the spinning solution A is 0.1% to 50%; and / or

[0011] In the spinning solution A, the mass ratio of the polymer material A to the functional component is 1000:(0.1~200);

[0012] In the spinning solution A, the mass ratio of the polymer material A to the photoinitiator is 1000:(0.1~200).

[0013] In a preferred embodiment of the present invention, the mass concentration of polymer material B in the spinning solution B is 0.1-50%; and / or

[0014] In the spinning solution B, the mass ratio of the polymer material B to the functional component is 1000:(0.1~200).

[0015] As a preferred embodiment of the present invention, the degree of crosslinking of the outer layer is 1 to 100%; and / or

[0016] The degree of crosslinking of the inner layer is 1 to 100%.

[0017] As a preferred embodiment of the present invention, polymeric material A and polymeric material B are each independently at least one of the following: alkenylated collagen, alkenylated gelatin, methacrylic anhydride gelatin, alkenylated chitosan, alkenylated cellulose, alkenylated silk fibroin, alkenylated polylactic acid-glycolic acid copolymer, alkenylated polycaprolactone, alkenylated polylactide, alkenylated polylactic acid, alkenylated polyglycolic acid, alkenylated polyβ-hydroxybutyl ester, alkenylated polyglycolic acid, alkenylated polyanhydride, alkenylated polyethylene glycol, alkenylated polyphosphate ester, alkenylated carrageenan, alkenylated polyvinylpyrrolidone, alkenylated polystyrene, alkenylated polyvinyl alcohol, alkenylated polyphosphononitrile, alkenylated hydrogenated styrene-butadiene block copolymer, and poly(ethylene glycol) methyl ether-block-poly(lactide-co-glycolic acid).

[0018] In a preferred embodiment of the present invention, the organic solvent is at least one selected from hexafluoroisopropanol, chloroform, tetrahydrofuran, acetone, methyl acetate, N,N-dimethylformamide, methyl isobutyl ketone, N,N-dimethylacetamide, dimethyl sulfoxide, dichloromethane, methyl cyanide, and ethanol; and / or

[0019] The photoinitiator is at least one of 2-hydroxy-2-methyl-1-[4-(2-hydroxyethoxy)phenyl]-1-propanone, diphenyl-(2,4,6-trimethylbenzoyl)phosphine oxide, phenylbis(2,4,6-trimethylbenzoyl)phosphine oxide, and 2-hydroxy-2-methyl-1-phenyl-1-propanone.

[0020] In a preferred embodiment of the present invention, the functional component is at least one of bone-promoting components and antibacterial components;

[0021] The osteogenic component is at least one of hydroxyapatite, calcium phosphate, β-tricalcium phosphate, osteopontin-derived peptide, bone morphogenetic protein 2, osteogenic growth peptide, magnesium nanoparticles, gold nanoparticles, iron nanoparticles, niobium nanoparticles, and magnetic nanoparticles.

[0022] The antibacterial component is at least one of antibiotics, metal nanoparticles, and antimicrobial peptides.

[0023] This invention also provides a method for preparing a multifunctional bio-cellulose membrane, comprising the following steps:

[0024] Spinning solution A is loaded into an injection device and connected to an electrostatic positive high voltage. The flow rate of spinning solution A is controlled by an injection pump, and fiber A is collected by a take-up device.

[0025] Using fiber A as the receiving device, the spinning solution B is loaded into the injection device and connected to an electrostatic positive high voltage. The flow rate of the spinning solution B is controlled by the injection pump, and the fiber membrane is collected.

[0026] The fiber membrane was immersed in a solution and cross-linked under ultraviolet light to obtain a multifunctional bio-fiber membrane.

[0027] This invention also provides a method for preparing a multifunctional bio-cellulose membrane, comprising the following steps:

[0028] The spinning solution B is loaded into the injection device and connected to a positive electrostatic voltage. The flow rate of the spinning solution B is controlled by the injection pump, and the fiber B is collected by the take-up device.

[0029] Using fiber B as the receiving device, the spinning solution A is loaded into the injection device and connected to an electrostatic positive high voltage. The flow rate of the spinning solution A is controlled by the injection pump, and the fiber membrane is collected.

[0030] The fiber membrane was immersed in a solution and cross-linked under ultraviolet light to obtain a multifunctional bio-fiber membrane.

[0031] The electrostatic positive high voltage ranges from 5 to 60 kV, the distance between the wire output and take-up devices is 5 to 100 cm, and the injection rate is 0.1 to 10 mL / h.

[0032] The receiving device can be: ① a circular flatbed winding device; ② a square roller winding device, wherein the roller rotation speed is 10-3000 r / min. During roller winding, after each layer is spun, the collected fibers are rotated 0-90° and reattached to the roller surface to continue spinning, ultimately forming a multi-layered and multi-directional oriented three-dimensional structure.

[0033] The present invention also provides the application of the aforementioned multifunctional bio-cellulose membrane as a GBR membrane or a GTR membrane.

[0034] The beneficial effects of this invention are as follows: The multifunctional bio-fiber membrane of this invention has a bilayer structure. The outer layer is dense with a smooth fiber surface, and the fibers are cross-linked, resulting in reduced and closed pores. The inner layer is loose and porous with a mesh-like pore size and straight fibers. The dense barrier structure of the outer layer can effectively block the invasion of epithelial cells, and the loaded antibacterial agents are effective in inhibiting bacterial growth. The mesh-like pore size of the inner layer allows it to load osteogenic components, promoting bone regeneration. The fibrous membrane is designed with stratified pores from the inside out, forming a fibrous membrane with a composite function of "antibacterial-osteogenic-barrier". In addition, the multifunctional bio-fiber membrane of this invention can serve as a structural framework to load various other active ingredients such as drugs, proteins, DNA, and nanoparticles, modifying their functions. The release of active ingredients can be controlled according to the degree of cross-linking. The amino groups in the outer layer structure can be further chemically modified to optimize the function. The preparation technology of this composite fibrous membrane is simple, the composition is stable, the structure is controllable, the performance is reliable, and the function is adjustable, showing broad prospects for biomedical applications. Attached Figure Description

[0035] Figure 1The fiber membrane (outer layer) formed by spinning solution A.

[0036] Figure 2 The fiber membrane (inner layer) formed by spinning solution B.

[0037] Figure 3 The present invention relates to a multifunctional bio-cellulose membrane.

[0038] Figure 4 This is a test diagram for Test Example 1 of the present invention.

[0039] Figure 5 This is a test diagram for Test Example 2 of the present invention.

[0040] Figure 6 This is a test diagram for test example 3 of the present invention.

[0041] Figure 7 This is a test diagram for test example 4 of the present invention. Detailed Implementation

[0042] To make the objectives, technical solutions, and advantages of the embodiments of this application clearer, the technical solutions in the embodiments of this application will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of this application, not all embodiments. Based on the embodiments in this application, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of this application.

[0043] In this application, the technical features described in an open-ended manner include both closed technical solutions consisting of the listed features and open technical solutions that include the listed features.

[0044] In this application, numerical ranges are referred to as continuous unless otherwise specified, and include the minimum and maximum values ​​of the range, as well as every value between the minimum and maximum values. Furthermore, when the range refers to integers, it includes every integer between the minimum and maximum values ​​of the range. Additionally, when multiple ranges are provided to describe a feature or characteristic, the ranges may be merged. In other words, unless otherwise specified, all ranges disclosed herein should be understood to include any and all subranges to which they are incorporated.

[0045] In this application, there are no particular restrictions on the specific dispersion and mixing methods.

[0046] Unless otherwise specified, all components, raw materials, or instruments used in the embodiments and comparative examples of this invention are commercially available, and the same type of components and raw materials are used in each parallel experiment.

[0047] Example 1

[0048] A method for preparing a multifunctional bio-cellulose membrane includes the following steps:

[0049] (1) Poly(ethylene glycol) methyl ether-block-poly(lactide-co-lactide), methacrylic anhydride gelatin, antibacterial agent, 2-hydroxy-2-methyl-1-[4-(2-hydroxyethoxy)phenyl]-1-propanone were added to hexafluoroisopropanol and stirred at 800 rpm for 6 h to obtain spinning solution A;

[0050] In the spinning solution A, the mass percentage of poly(ethylene glycol) methyl ether-block-poly(lactide-co-glycolic acid) is 9%, the mass percentage of methacrylic anhydride gelatin is 6%, the mass percentage of antibacterial agent is 2%, and the mass percentage of 2-hydroxy-2-methyl-1-[4-(2-hydroxyethoxy)phenyl]-1-propanone is 1.5%.

[0051] (2) Spinning solution A was loaded into a 5ml syringe (23G needle). A roller collector was used to collect the oriented fiber membrane at 3000rpm. The distance between the exit needle and the collector was 15cm. A high voltage of 20KV was applied, and the solution was ejected at a rate of 6ml / h. After wetting the fiber membrane, it was irradiated with UV light for 30min to induce crosslinking. The prepared fiber membrane was dried in a vacuum oven at 25℃ for at least 2 days until the solvent was completely evaporated. Its surface morphology was observed using a scanning electron microscope. Figure 1 It can be seen that after ultraviolet light crosslinking, the fibers aggregate and crosslink, the pores are closed, and the degree of crosslinking reaches 100%.

[0052] (3) Poly(ethylene glycol) methyl ether-block-poly(lactide-co-lactide) and metal nanoparticles were added to hexafluoroisopropanol and stirred at 800 rpm for 6 h to obtain spinning solution B;

[0053] In the spinning solution B, the mass percentage of poly(ethylene glycol) methyl ether-block-poly(lactide-co-glycolic acid) is 20%, and the mass percentage of metal nanoparticles is 2%.

[0054] (4) Using the fiber membrane from step (2) as the receiver, the spinning solution B was loaded into a 5ml syringe (23G needle). A roller collector was used to collect the oriented fiber membranes at a rotation speed of 3000rpm. The distance between the needle and the collector was 15cm. A high voltage of 20KV was applied, and the solution was ejected at a rate of 6ml / h. The solution was dried in a vacuum oven at 25℃ for at least 2 days until the solvent was completely evaporated. The surface morphology was observed using a scanning electron microscope. Figure 2It can be seen that the inner layer was successfully prepared. The surface of the inner layer fiber is smooth, and the visible nanoparticles are uniformly attached to the fiber structure, with a crosslinking degree of 1%. (5) After wetting the membrane prepared in step (4), it was irradiated with ultraviolet light for 30 min to perform crosslinking. The prepared fiber membrane was dried in a vacuum oven at 25°C for more than 2 days until the solvent was completely evaporated. Its cross-sectional morphology was observed by scanning electron microscopy. Figure 3 It can be seen that the bilayer membrane is tightly connected; the cross-linked outer layer is dense, while the inner layer is loose and porous.

[0055] Example 2

[0056] A method for preparing a multifunctional bio-cellulose membrane includes the following steps:

[0057] (1) Poly(ethylene glycol) methyl ether-block-poly(lactide-co-lactide), methacrylic anhydride gelatin, antibacterial agent, 2-hydroxy-2-methyl-1-[4-(2-hydroxyethoxy)phenyl]-1-propanone were added to hexafluoroisopropanol and stirred at 800 rpm for 6 h to obtain spinning solution A;

[0058] In the spinning solution A, the mass percentage of poly(ethylene glycol) methyl ether-block-poly(lactide-co-glycolic acid) is 6%, the mass percentage of methacrylic anhydride gelatin is 4%, the mass percentage of antibacterial agent is 1%, and the mass percentage of 2-hydroxy-2-methyl-1-[4-(2-hydroxyethoxy)phenyl]-1-propanone is 1%.

[0059] (2) Spinning solution A was loaded into a 5ml syringe (23G needle). A roller collector was used to collect the oriented fiber membrane at 3000rpm. The distance between the exit needle and the collector was 15cm. A high voltage of 20KV was applied, and the solution was ejected at a rate of 6ml / h. After wetting the fiber membrane, it was irradiated with ultraviolet light for 30min to induce crosslinking. The prepared fiber membrane was dried in a vacuum oven at 25℃ for at least 2 days until the solvent was completely evaporated and the degree of crosslinking was 100%.

[0060] (3) Poly(ethylene glycol) methyl ether-block-poly(lactide-co-lactide) and metal nanoparticles were added to hexafluoroisopropanol and stirred at 800 rpm for 6 h to obtain spinning solution B;

[0061] In the spinning solution B, the mass percentage of poly(ethylene glycol) methyl ether-block-poly(lactide-co-glycolic acid) is 10%, and the mass percentage of metal nanoparticles is 1%.

[0062] (4) Using the fiber membrane from step (2) as the receiver, the spinning solution B is loaded into a 5ml syringe (23G needle). The collector for the oriented fiber membrane is a roller collector, with a rotation speed of 3000rpm. The distance between the needle and the collector is 15cm. A high voltage of 20KV is applied, and the solution is ejected at a rate of 6ml / h. The solution is dried in a vacuum oven at 25℃ for more than 2 days until the solvent is completely evaporated.

[0063] (5) After wetting the membrane prepared in step (4), crosslink it by irradiating it with ultraviolet light for 30 min. The prepared fiber membrane was dried in a vacuum oven at 25°C for more than 2 days until the solvent was completely evaporated. The cross-sectional morphology was observed by scanning electron microscopy. The double membrane was tightly connected; the outer layer after crosslinking was dense, and the inner layer was loose and porous.

[0064] Example 3

[0065] A method for preparing a multifunctional bio-cellulose membrane includes the following steps:

[0066] (1) Poly(ethylene glycol) methyl ether-block-poly(lactide-co-lactide), methacrylic anhydride gelatin, antibacterial agent, 2-hydroxy-2-methyl-1-[4-(2-hydroxyethoxy)phenyl]-1-propanone were added to hexafluoroisopropanol and stirred at 800 rpm for 6 h to obtain spinning solution A;

[0067] In the spinning solution A, the mass percentage of poly(ethylene glycol) methyl ether-block-poly(lactide-co-glycolic acid) is 12%, the mass percentage of methacrylic anhydride gelatin is 8%, the mass percentage of antibacterial agent is 2.2%, and the mass percentage of 2-hydroxy-2-methyl-1-[4-(2-hydroxyethoxy)phenyl]-1-propanone is 1.8%.

[0068] (2) Spinning solution A was loaded into a 5ml syringe (23G needle). A roller collector was used to collect the oriented fiber membrane at 3000rpm. The distance between the exit needle and the collector was 15cm. A high voltage of 20KV was applied, and the solution was ejected at a rate of 6ml / h. After wetting the fiber membrane, it was irradiated with UV light for 30min to induce crosslinking. The prepared fiber membrane was dried in a vacuum oven at 25℃ for at least 2 days until the solvent was completely evaporated. The surface morphology was observed using a scanning electron microscope. The degree of crosslinking reached 100%.

[0069] (3) Poly(ethylene glycol) methyl ether-block-poly(lactide-co-lactide) and metal nanoparticles were added to hexafluoroisopropanol and stirred at 800 rpm for 6 h to obtain spinning solution B;

[0070] In the spinning solution B, the mass percentage of poly(ethylene glycol) methyl ether-block-poly(lactide-co-glycolic acid) is 30%, and the mass percentage of metal nanoparticles is 2.2%.

[0071] (4) Using the fiber membrane from step (2) as the receiver, the spinning solution B is loaded into a 5ml syringe (23G needle). The collector for the oriented fiber membrane is a roller collector, with a rotation speed of 3000rpm. The distance between the needle and the collector is 15cm. A high voltage of 20KV is applied, and the solution is ejected at a rate of 6ml / h. The solution is dried in a vacuum oven at 25℃ for more than 2 days until the solvent is completely evaporated.

[0072] (5) After wetting the membrane prepared in step (4), crosslink it by irradiating it with ultraviolet light for 30 min. The prepared fiber membrane was dried in a vacuum oven at 25°C for more than 2 days until the solvent was completely evaporated. The cross-sectional morphology was observed by scanning electron microscopy. The double membrane was tightly connected; the outer layer after crosslinking was dense, and the inner layer was loose and porous.

[0073] Comparative Example 1

[0074] A method for preparing a multifunctional bio-cellulose membrane includes the following steps:

[0075] (1) Poly(ethylene glycol) methyl ether-block-poly(lactide-co-lactide), methacrylic anhydride gelatin, and 2-hydroxy-2-methyl-1-[4-(2-hydroxyethoxy)phenyl]-1-propanone were added to hexafluoroisopropanol and stirred at 800 rpm for 6 h to obtain spinning solution A.

[0076] In the spinning solution A, the mass percentage of poly(ethylene glycol) methyl ether-block-poly(lactide-co-glycolic acid) is 9%, the mass percentage of methacrylic anhydride gelatin is 6%, and the mass percentage of 2-hydroxy-2-methyl-1-[4-(2-hydroxyethoxy)phenyl]-1-propanone is 1.5%.

[0077] (2) Spinning solution A was loaded into a 5ml syringe (23G needle). A roller collector was used to collect the oriented fiber membrane at 3000rpm. The distance between the exit needle and the collector was 15cm. A high voltage of 20KV was applied, and the solution was ejected at a rate of 6ml / h. After wetting the fiber membrane, it was irradiated with UV light for 30min to induce crosslinking. The prepared fiber membrane was dried in a vacuum oven at 25℃ for at least 2 days until the solvent was completely evaporated.

[0078] (3) Poly(ethylene glycol) methyl ether-block-poly(lactide-co-lactide) was added to hexafluoroisopropanol and stirred at 800 rpm for 6 h to obtain spinning solution B;

[0079] The spinning solution B contains 20% poly(ethylene glycol) methyl ether-block-poly(lactide-co-glycolic acid) by mass.

[0080] (4) Using the fiber membrane from step (2) as the receiver, the spinning solution B is loaded into a 5ml syringe (23G needle). The collector for the oriented fiber membrane is a roller collector, with a rotation speed of 3000rpm. The distance between the needle and the collector is 15cm. A high voltage of 20KV is applied, and the solution is ejected at a rate of 6ml / h. The solution is dried in a vacuum oven at 25℃ for more than 2 days until the solvent is completely evaporated.

[0081] (5) After wetting the membrane prepared in step (4), irradiate it with ultraviolet light for 30 min to crosslink it. The prepared fiber membrane is dried in a vacuum oven at 25°C for more than 2 days until the solvent is completely evaporated.

[0082] Test Example 1

[0083] The fibrous membrane prepared in Example 1 and the commercially available collagen membrane were cut to the same size. After SD rats were anesthetized, the subcutaneous tissue was separated using mosquito forceps, and the fibrous membrane and the commercially available collagen membrane were implanted into the subcutaneous fascia layer and fixed with sutures. The incision was then sutured. One week later, paraffin sections were prepared, stained with hematoxylin and eosin (HE), and observed and images were acquired using an optical microscope. The results are as follows: Figure 4 As shown, cells can be seen entering the entire collagen membrane, while the dense outer layer of the composite fiber membrane can effectively act as a barrier to block cells, and the loose and porous internal structure is conducive to cell entry, proliferation and differentiation.

[0084] Test Example 2

[0085] Staphylococcus aureus and Escherichia coli were mixed at a ratio of 10... 7 CFU / mL concentrations were inoculated onto agar plates, and the functionalized component (PGLG / GM / PL@Nb) prepared in Example 1 and the non-functionalized component bilayer membrane (PGLG / GM) of Comparative Example 1 were placed on them. The plates were incubated at 37°C for 24 h, and the antibacterial activity was evaluated based on the size of the inhibition zone. The results are as follows: Figure 5 As shown, fiber membranes loaded with antibacterial components can effectively inhibit bacterial proliferation.

[0086] Test Example 3

[0087] Bone marrow mesenchymal stem cells were seeded on the inner surface of the functional component-loaded fiber membrane (PGLG / GM / PL@Nb) prepared in Example 1, the non-functional component-loaded fiber membrane (PGLG / GM) in Comparative Example 1, and the HA-loaded fiber membrane (PGLG / GM / PL@HA). PGLG / GM / PL@HA served as a positive control. The osteogenic induction solution was changed on the second day, and Western blot was performed after 7 days of induction to assess the expression of osteogenic proteins Col-I, OSX, Runx2, and OPN. The results are as follows: Figure 6 As shown, the osteogenic protein expression in the osteogenic component fibrous membrane (PGLG / GM / PL@Nb) was higher than that in other groups, thus promoting osteogenic protein expression.

[0088] Test Example 4

[0089] In Example 1, a fiber membrane carrying the functional component (PGLG / GM / PL@Nb) was prepared, and in Comparative Example 1, a fiber membrane without the functional component (PGLG / GM) was prepared. 3 mL of rabbit blood was centrifuged at 1500 rpm for 15 min, and the membrane was gently washed three times with PBS to obtain red blood cells. The obtained red blood cells were diluted with PBS to an appropriate concentration and mixed thoroughly, then added to EP tubes for each group. The fiber membrane for each group was placed in an EP tube containing 1 mL of the above solution. PBS was used as the negative control, and deionized water (DW) was used as the positive control. After incubating at 37°C for 2 h, the support was removed, and the supernatant from each group was collected. The OD value at 540 nm was measured using a spectrophotometer. After measuring the absorbance, the hemolysis rate was calculated using the following formula: Hemolysis rate (%) = (OD0.05) / (540 nm) * ... 实验组 -OD 阴性对照组 ) / (OD 阳性对照组 -OD 阴性对照组 x 100%. The result is as follows: Figure 7 As shown, the hemolysis rate of the material group was no more than 2%, which was not statistically different from that of the negative group.

[0090] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit the scope of protection of the present invention. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the essence and scope of the technical solutions of the present invention.

Claims

1. A multifunctional bio-cellulose membrane, characterized in that, The multifunctional bio-cellulose membrane has a bilayer structure. The outer layer is dense with a smooth fiber surface and cross-linked fibers, resulting in reduced and closed pores. The inner layer is loose and porous with a mesh-like pore size and straight fibers. The outer layer is made of spinning solution A, which includes polymer material A, photoinitiator, functional components and organic solvent; The inner layer is made of spinning solution B, which includes polymer material B, functional components and organic solvents; The polymer material A is methacrylic anhydride gelatin and poly(ethylene glycol) methyl ether-block-poly(lactide-co-lactide), and the polymer material B is poly(ethylene glycol) methyl ether-block-poly(lactide-co-lactide). The functional component is at least one of bone-promoting components and antibacterial components; The osteogenic component is at least one of hydroxyapatite, calcium phosphate, β-tricalcium phosphate, osteopontin-derived peptide, bone morphogenetic protein 2, osteogenic growth peptide, magnesium nanoparticles, gold nanoparticles, iron nanoparticles, niobium nanoparticles, and magnetic nanoparticles. The antibacterial component is at least one of antibiotics, metal nanoparticles, and antimicrobial peptides.

2. The multifunctional bio-cellulose membrane according to claim 1, characterized in that, The mass concentration of polymer material A in the spinning solution A is 0.1%~50%; and / or In the spinning solution A, the mass ratio of the polymer material A to the functional component is 1000:(0.1~200). In the spinning solution A, the mass ratio of the polymer material A to the photoinitiator is 1000:(0.1~200).

3. The multifunctional bio-cellulose membrane according to claim 1, characterized in that, The mass concentration of polymer B in the spinning solution B is 0.1%~50%; and / or In the spinning solution B, the mass ratio of the polymer material B to the functional component is 1000:(0.1~200).

4. The multifunctional bio-cellulose membrane according to claim 1, characterized in that, The degree of crosslinking of the outer layer is 1% to 100%; and / or The degree of crosslinking of the inner layer is 1% to 100%.

5. The multifunctional bio-cellulose membrane according to claim 1, characterized in that, The organic solvent is at least one selected from hexafluoroisopropanol, chloroform, tetrahydrofuran, acetone, methyl acetate, N,N-dimethylformamide, methyl isobutyl ketone, N,N-dimethylacetamide, dimethyl sulfoxide, dichloromethane, methyl cyanide, and ethanol; and / or The photoinitiator is at least one of 2-hydroxy-2-methyl-1-[4-(2-hydroxyethoxy)phenyl]-1-propanone, diphenyl-(2,4,6-trimethylbenzoyl)phosphine oxide, phenylbis(2,4,6-trimethylbenzoyl)phosphine oxide, and 2-hydroxy-2-methyl-1-phenyl-1-propanone.

6. The method for preparing the multifunctional bio-cellulose membrane according to claim 1, characterized in that, Includes the following steps: Spinning solution A is loaded into an injection device and connected to an electrostatic positive high voltage. The flow rate of spinning solution A is controlled by an injection pump, and fiber A is collected by a take-up device. Using fiber A as the receiving device, the spinning solution B is loaded into the injection device and connected to an electrostatic positive high voltage. The flow rate of the spinning solution B is controlled by the injection pump, and the fiber membrane is collected. The fiber membrane was immersed in a solution and cross-linked under ultraviolet light to obtain a multifunctional bio-fiber membrane.

7. The method for preparing the multifunctional bio-cellulose membrane according to claim 1, characterized in that, Includes the following steps: The spinning solution B is loaded into the injection device and connected to a positive electrostatic voltage. The flow rate of the spinning solution B is controlled by the injection pump, and the fiber B is collected by the take-up device. Using fiber B as the receiving device, the spinning solution A is loaded into the injection device and connected to an electrostatic positive high voltage. The flow rate of the spinning solution A is controlled by the injection pump, and the fiber membrane is collected. The fiber membrane was immersed in a solution and cross-linked under ultraviolet light to obtain a multifunctional bio-fiber membrane.

8. The application of the multifunctional bio-cellulose membrane according to any one of claims 1 to 6 as a GBR membrane or GTR membrane.

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