Barrier membrane with photocatalytic antibacterial activity and preparation method and application thereof

Abstract

The invention discloses a barrier membrane with photocatalytic antibacterial activity and a preparation method and application thereof, and belongs to the technical field of biological materials. The preparation method of the barrier membrane comprises the following steps: S1, dissolving aminated gelatin, oxidized sodium alginate and mesoporous bioactive glass in water, and mixing to obtain a hydrogel pre-polymerized solution; s2, polycaprolactone, aminated gelatin and the Bi / BiOCl heterojunction are dissolved, mixed and subjected to film forming, and a first fiber film is obtained; and S3, pouring the hydrogel pre-polymerized liquid on the first fiber membrane to obtain the barrier membrane. The barrier film provided by the invention has a photocatalytic antibacterial function, an osteogenesis function, mechanical properties, biocompatibility and adhesion properties, can meet the bidirectional environmental requirements of bones and soft tissues in a bone reconstruction process, and solves the problems of an existing single-layer film in the aspects of antibacterial-osteogenesis function separation, insufficient mechanical properties, dependence on external fixation and the like.

Description

Technical Field

[0001] The present invention belongs to the technical field of biomaterials, and particularly relates to a barrier membrane with photocatalytic antibacterial activity, a preparation method thereof, and an application thereof. Background Art

[0002] The Guided Bone Regeneration (GBR) technology has achieved excellent results in promoting periodontal repair. The GBR technology covers a barrier membrane in the bone defect area to block the invasion of epithelial cells and fibroblasts, ensuring the smooth progress of the bone regeneration process. The barrier membrane is a key component of this technology. The types of barrier membranes can be divided into two major categories: bioabsorbable barrier membranes and non-absorbable barrier membranes. Bioabsorbable barrier membranes include collagen membranes, chitosan membranes, gelatin membranes, and other synthetic polymer membranes, which have good biocompatibility, but usually have disadvantages such as poor antibacterial performance and insufficient mechanical strength; non-absorbable barrier membranes, such as polytetrafluoroethylene, titanium-reinforced e-PTFE membranes, and dense PTFE membranes, although having good mechanical properties and durability, cannot be absorbed by organisms, may lead to incomplete postoperative recovery, and require additional removal steps.

[0003] In recent years, the development of multi-functional hierarchical barrier membranes has received great attention in the field of biomaterials. Multi-functional hierarchical membranes usually include a dense layer and a loose porous layer. The dense layer can effectively prevent the invasion of connective tissues and bacterial infections, and the loose porous layer helps to promote the proliferation and differentiation of osteoblasts, further accelerating bone repair. The most commonly used in current clinical applications is the collagen membrane, which relies on the porous skeleton composed of collagen fibers to achieve the shielding effect on soft tissue cells. However, the collagen membrane has disadvantages such as poor antibacterial performance, poor durability, and susceptibility to microbial contamination. Especially in the oral cavity, which is a microbial-rich environment, the deficiencies of the collagen membrane are becoming increasingly prominent. In addition, in traditional GBR surgeries, membrane nails or membrane needles are used to fix the collagen membrane to prevent its displacement, but this may also cause damage to important surrounding structures and affect the treatment effect. Therefore, how to improve the antibacterial activity and mechanical properties of the barrier membrane and reduce the dependence on external fixation devices has become one of the key challenges in the current technological development. Summary of the Invention

[0004] In order to solve the problems existing in the above-mentioned prior art, the primary object of the present invention is to provide a barrier membrane.

[0005] Another object of the present invention is to provide the application of the above barrier membrane in guided bone regeneration.

[0006] To achieve the above objects, the present invention provides the following technical solutions:

[0007] A barrier membrane, which is prepared by the following preparation method:

[0008] S1. Dissolve amino gelatin, oxidized sodium alginate, and mesoporous bioactive glass in water and mix them to obtain a hydrogel prepolymer solution.

[0009] S2. Dissolve and mix polycaprolactone, amino gelatin, and Bi / BiOCl heterojunction, and obtain a first fiber membrane by electrospinning.

[0010] S3. Pour the hydrogel prepolymer solution onto the first fiber membrane to obtain the barrier membrane.

[0011] The present invention adopts a double-layer structure design and prepares a barrier membrane containing a hydrogel layer and a fiber membrane layer through an electrospinning and hydrogel composite technology, endowing it with photocatalytic antibacterial function, osteogenic function, mechanical properties, biocompatibility, and adhesion properties to meet the two-way environmental requirements of bones and soft tissues during the bone reconstruction process.

[0012] The hydrogel layer includes amino gelatin, oxidized sodium alginate, and mesoporous bioactive glass. The mesoporous bioactive glass not only provides an alkaline environment to accelerate the formation of imine bonds but also can act as a cross-linking agent to rapidly release Ca through mesoporous channels. 2+ Ca 2+ and carboxyl groups in the hydrogel layer form physically cross-linked polymer chains through chelation. This highly reversible cross-linking improves the viscosity and cohesion of the hydrogel. In addition, the three-dimensional pore structure of the mesoporous bioactive glass can enhance the interlocking effect with tissues through physical adsorption, further enhancing the interfacial adhesion between the hydrogel and tissues. Oxidized sodium alginate contains aldehyde groups, and the aldehyde groups can cross-link with amino groups on the surface of the alveolar bone through Schiff base reaction. Hydrophilic groups in the hydrogel such as (-COOH, -OH, -CONH, and -NH2) can form hydrogen bond interactions with amino groups on the surface of the alveolar bone, thereby generating strong tissue adhesion. The combined action of the three reduces the dependence of the barrier membrane on external fixation devices.

[0013] The fiber layer includes polycaprolactone, amino gelatin, and Bi / BiOCl heterojunction. The fiber layer generates highly reactive oxygen species (ROS) through the near-infrared light response characteristics of the Bi / BiOCl heterojunction, achieving broad-spectrum antibacterial against Staphylococcus aureus and Escherichia coli and effectively inhibiting the occurrence of infections. The fiber layer endows the barrier membrane with excellent mechanical strength and can long-term block the penetration of soft tissue cells to maintain the osteogenic space.

[0014] Preferably, the mass ratio of amino gelatin, oxidized sodium alginate, and mesoporous bioactive glass in the hydrogel prepolymer solution is (1 - 4):(1 - 4):(0.01 - 1).

[0015] More preferably, the mass ratio of the amidated gelatin, oxidized sodium alginate, and mesoporous bioactive glass in the hydrogel prepolymer solution is (2.5 to 3.5):(1.5 to 2.5):(0.5 to 0.7).

[0016] Preferably, the mass ratio of polycaprolactone, amidated gelatin, and Bi / BiOCl heterojunction in the first fiber membrane is (7 to 10):(0.1 to 3):(0.01 to 0.1).

[0017] More preferably, the mass ratio of polycaprolactone, amidated gelatin, and Bi / BiOCl heterojunction in the first fiber membrane is (7 to 10):(0.5 to 1.5):(0.04 to 0.06).

[0018] Specifically, in step S1, the mesoporous bioactive glass is first dispersed in the oxidized sodium alginate solution and then mixed with the amidated gelatin solution.

[0019] Specifically, the particle size distribution of the mesoporous bioactive glass is 100 to 200 nm.

[0020] Preferably, the main components of the mesoporous bioactive glass include SiO2 and CaO.

[0021] The mesoporous bioactive glass has excellent osteogenic activity. The mesoporous bioactive glass introduced into the hydrogel layer in the present invention promotes the alkaline phosphatase (ALP) activity and calcium nodule deposition of osteoblasts by slowly releasing calcium ions, further accelerating the repair of the bone defect area, thereby endowing the barrier membrane with osteogenic function.

[0022] Specifically, the mesoporous bioactive glass is prepared by a template method.

[0023] More specifically, the template is one of cetyltrimethylammonium bromide (CTAB), P123, and tetradecyltrimethylammonium bromide.

[0024] Specifically, the preparation method of the mesoporous bioactive glass includes the following steps: mixing a template agent with a calcium source and a silicon source, and calcining after reaction to obtain the mesoporous bioactive glass.

[0025] Specifically, the mesoporous pore diameter of the mesoporous bioactive glass is 2 to 50 nm.

[0026] Preferably, the concentration of the amidated gelatin in the hydrogel prepolymer solution is 50 to 200 mg / mL.

[0027] More preferably, the concentration of the amidated gelatin in the hydrogel prepolymer solution is 100 to 200 mg / mL.

[0028] Specifically, the amidated gelatin is obtained by reacting gelatin with an amino group supplier.

[0029] More specifically, the preparation method of the amidated gelatin includes: dissolving gelatin in a phosphate buffer solution, adding an amino group supplier, and obtaining amidated gelatin after the reaction.

[0030] More specifically, the mass ratio of gelatin to the amino group supplier is 1:1 to 1.5.

[0031] More specifically, the Bloom value of the gelatin is 100 to 250 g.

[0032] More specifically, the amino group supplier is at least one of 2-chloroethylamine hydrochloride, ethylenediamine, and propylenediamine.

[0033] Preferably, the amino group supplier is 2-chloroethylamine hydrochloride.

[0034] Preferably, the concentration of oxidized sodium alginate in the hydrogel prepolymer solution is 50 to 200 mg / mL.

[0035] More preferably, the concentration of oxidized sodium alginate in the hydrogel prepolymer solution is 75 to 100 mg / mL.

[0036] Preferably, the concentration of mesoporous bioactive glass in the hydrogel prepolymer solution is 0.5 to 50 mg / mL.

[0037] More preferably, the concentration of mesoporous bioactive glass in the hydrogel prepolymer solution is 30 to 50 mg / mL.

[0038] Specifically, in the step S2, the solvent used for dissolution is at least one of 1,1,1,3,3,3-hexafluoro-2-propanol, 2,2-difluoroethanol, and 2,2,2-trifluoroethanol.

[0039] Specifically, in the step S2, electrospinning is used to prepare the first fiber membrane.

[0040] Specifically, the injection rate of the electrospinning is 0.8 to 1.2 mL / h.

[0041] Specifically, the receiving distance of the electrospinning is 15 to 20 cm.

[0042] Specifically, the time of the electrospinning is 4 to 12 h.

[0043] Specifically, the diameter of the fibers in the first fiber membrane is 1 to 3 μm.

[0044] Specifically, in the step S3, 0.1 to 3 g of the hydrogel prepolymer solution is poured onto the 0.5 to 6 cm 2 first fiber membrane.

[0045] Specifically, the thickness of the barrier membrane is 1 to 3 mm.

[0046] Specifically, the barrier membrane comprises a hydrogel layer and a fiber membrane layer.

[0047] More specifically, the hydrogel layer comprises aminated gelatin, oxidized sodium alginate and mesoporous bioactive glass.

[0048] More specifically, the fiber membrane layer comprises aminated gelatin, polycaprolactone (PCL) and Bi / BiOCl heterojunction.

[0049] The present invention also protects the application of the above barrier membrane in guiding bone regeneration.

[0050] Compared with the prior art, the present invention has the following beneficial effects:

[0051] (1) The present invention adopts a double-layer structure design, and through an electrospinning and hydrogel composite technology, a barrier membrane containing a hydrogel layer and a fiber membrane layer is prepared. By introducing mesoporous bioactive glass into the hydrogel layer, the adhesion performance of the barrier membrane is improved, the dependence on the membrane fixing device is reduced, and the barrier membrane is endowed with osteogenic function; the fiber layer improves the tensile stress of the fiber membrane by introducing gelatin, meeting the requirements of the oral environment for mechanical properties. Secondly, the fiber membrane layer realizes broad-spectrum antibacterial against bacteria through the visible light response characteristics of the Bi / BiOCl heterojunction, effectively inhibits the occurrence of infection, and reduces the single-treatment cost.

[0052] (2) The barrier membrane provided by the present invention has photocatalytic antibacterial function, osteogenic function, mechanical properties, biocompatibility and adhesion performance, can meet the two-way environmental requirements of bones and soft tissues during the bone reconstruction process, solves the technical problems of the existing single-layer membrane in the separation of antibacterial-osteogenic functions, insufficient mechanical properties and dependence on external fixation, etc., and is particularly suitable for the guided bone regeneration (GBR) technology in oral bone defect repair, and has broad application prospects.

[0053] (3) The preparation method of the present invention is simple, can be customized according to the surgical needs, has high feasibility and comprehensive functions. BRIEF DESCRIPTION OF THE DRAWINGS

[0054] Figure 1 It is a flow chart of the preparation method of the barrier membrane of the present invention.

[0055] Figure 2 It is a shear lap stress-strain curve graph of the hydrogel layer in Example 1.

[0056] Figure 3 It is a tensile stress-strain curve graph of the first fiber membrane in Example 1.

[0057] Figure 4The near-infrared response antibacterial effect diagram of the first fiber membrane in Example 1.

[0058] Figure 5 The cell viability and death staining result diagram of the hydrogel layer and the first fiber membrane in Example 1.

[0059] Figure 6 The SEM diagram of the prepared mesoporous bioactive glass nanoparticles.

[0060] Figure 7 The TEM diagram of the prepared mesoporous bioactive glass nanoparticles. Detailed implementation manners

[0061] The present invention will be further elaborated below in conjunction with embodiments. These embodiments are only used to illustrate the present invention and not to limit the scope of the present invention. For the experimental methods without specific conditions in the following embodiments, they are usually carried out according to the conventional conditions in the art or according to the conditions recommended by the manufacturer; the raw materials, reagents, etc. used, unless otherwise specified, are all raw materials and reagents that can be obtained from commercial channels such as conventional markets. Any non-substantial changes and substitutions made by those skilled in the art based on the present invention fall within the scope claimed by the present invention.

[0062] The preparation method of the Bi / BiOCl heterojunction can refer to steps (1) to (6) in Example 1 of CN114748450A.

[0063] The preparation method of the mesoporous bioactive glass nanoparticles includes the following steps:

[0064] Place 2.80 g of cetyltrimethylammonium bromide (CTAB) in deionized water, and stir it at a constant temperature in a 35 °C water bath until CTAB is completely dissolved; gradually add 40 mL of ethyl acetate, 1 M ammonia water, 14.4 mL of tetraethyl orthosilicate, and 9.12 g of calcium nitrate tetrahydrate, and continue to stir and react for 4 h. Then, collect the white precipitate by centrifugation and wash it; next, dry the collected sediment overnight and grind it with a mortar; finally, heat the ground sediment in a high-temperature furnace to 700 °C and keep it warm to obtain the mesoporous bioactive glass nanoparticles.

[0065] The particle size distribution of the prepared mesoporous bioactive glass is 100 - 200 nm; the mesoporous pore size is 2 - 50 nm.

[0066] The preparation method of the amino-functionalized gelatin includes the following steps:

[0067] Dissolve 9 g of gelatin in 90 mL of phosphate buffer solution at 40 °C, and then add 10.5 g of 2-chloroethylamine hydrochloride to the above solution. After adding 1 M aqueous sodium hydroxide solution, continuously stir at 40 °C for 24 h. Neutralize the solution with 1 M hydrochloric acid, adjust the pH value to 7.0 - 7.2, dialyze in deionized water, and then lyophilize to obtain amino-gelatin.

[0068] The preparation method of oxidized sodium alginate includes the following steps:

[0069] Dissolve 4 g of sodium alginate in 400 mL of deionized water, and continuously stir to obtain a sodium alginate solution; take 4.32 g of sodium periodate and dissolve it in deionized water, then dropwise add it to sodium alginate, and stir overnight in the dark; then add 6 mL of ethylene glycol and stir to terminate the oxidation reaction; then transfer the solution to a dialysis bag and dialyze in deionized water for 3 d, and lyophilize to obtain white foamy oxidized sodium alginate.

[0070] Polycaprolactone: Manufacturer: Shanghai Aladdin Biochemical Technology Co., Ltd.; Product number: P579406, Molecular weight: 80,000 - 100,000.

[0071] Sodium alginate: Manufacturer: Shanghai Aladdin Biochemical Technology Co., Ltd.; Product number S278630: Molecular weight structural unit: Theoretical value 198.11, Average true value 222.00.

[0072] Gelatin: Manufacturer: Shanghai Aladdin Biochemical Technology Co., Ltd.; Product number: G108398; Bloom value: 100 g.

[0073] Example 1

[0074] S0. Dissolve 300 mg of amino-gelatin (Gel-NH2) in 1 mL of deionized water to obtain an amino-gelatin solution with a concentration of 300 mg / mL.

[0075] S1.1 Dissolve 200 mg of oxidized sodium alginate (OSA) in 1 mL of deionized water to obtain an oxidized sodium alginate solution with a concentration of 200 mg / mL. Subsequently, ultrasonically disperse 60 mg of mesoporous bioactive glass nanoparticles evenly in the oxidized sodium alginate solution for 30 min to obtain a first mixture, and the concentration of mesoporous bioactive glass in the oxidized sodium alginate solution is 60 mg / mL.

[0076] S1.2 Drop 1 mL of the amino-gelatin solution into the first mixture to obtain a hydrogel prepolymer solution; the concentration of amino-gelatin in the hydrogel prepolymer solution is 150 mg / mL; the concentration of oxidized sodium alginate in the hydrogel prepolymer solution is 100 mg / mL; the concentration of mesoporous bioactive glass in the hydrogel prepolymer solution is 30 mg / mL.

[0077] S2. Dissolve 900 mg of polycaprolactone (PCL), 100 mg of amino-functionalized gelatin, and 5 mg of Bi / BiOCl heterojunction in 10 mL of 1,1,1,3,3,3-hexafluoro-2-propanol, stir for 12 h to obtain a mixed solution, and prepare a first fiber membrane using electrospinning technology; the diameter of the fibers is 2 μm;

[0078] The settings for electrospinning are as follows: the rotational speed of the receiving mandrel is 140 rpm, the working voltage is 13 ± 0.5 kV, the injection speed is 1 mL / h, the receiving distance is 15 cm, and the time is 10 h.

[0079] S3. Pour 1 g of the hydrogel prepolymer solution onto the first fiber membrane of 3 cm 2 to obtain the barrier membrane. The thickness of the barrier membrane is 2 mm.

[0080] The raw material ratios used in the examples and comparative examples of the present invention are shown in Table 1, and the preparation method refers to Example 1.

[0081]

[0082] Testing method:

[0083] (1) Shear lap test

[0084] Prove the adhesion strength between the hydrogel and biological tissue through a lap shear test. Cut fresh pig skin into strips with a length of 30 mm and a width of 10 mm, apply the pre-gel solution evenly between two pieces of pig skin tissue, and the contact area of the hydrogel with the two pieces of pig skin is 10 mm × 10 mm. Use an electronic universal testing machine to conduct a lap shear test at a rate of 5 mm / min.

[0085] As can be seen from Figure 2 taking the hydrogel layer prepared in Example 1 as the experimental group and the hydrogel without mesoporous active bioactive glass nanoparticles prepared in Comparative Example 1 as the control group, when the concentration of mesoporous active bioactive glass nanoparticles in the hydrogel prepolymer solution is 30 mg / mL, the adhesion strength of the hydrogel to wet pig skin can reach 68.12 ± 3.05 kPa, while the adhesion strength of Comparative Example 1 is 21.45 ± 1.90 kPa.

[0086] (2) Mechanical property test of the first fiber membrane

[0087] Take the first fiber membrane prepared in Example 1 as the experimental group and the polycaprolactone (PCL) fiber membrane prepared by the method described in Reference Example 1 as the control group, and use an electronic universal testing machine to test the mechanical properties of the elastomer. For the tensile test, prepare rectangular samples (length: 30 mm; width: 9 mm) from the first fiber membrane and at 10 mm·min -1The displacement speed is stretched. The test results are as Figure 3 shown.

[0088] It can be seen from Figure 3 that compared with the pure PCL film without the addition of amino-gelatin, with the addition of amino-gelatin, the stress that the first fiber film can withstand increases, which can meet the requirements of the oral environment for mechanical properties.

[0089] (3) Antibacterial experiment

[0090] The first fiber film prepared in Example 1 was used as the experimental group for antibacterial effect testing. The plate counting method was used to determine the in vitro antibacterial effect of the first fiber film prepared in Example 1 against Escherichia coli under 808 nm near-infrared irradiation (+NIR) / non-irradiation (-NIR). The fiber film and the bacterial suspension (800 μL, 10 6 CFU / mL) were placed in a 48-well plate together. Subsequently, the samples were irradiated with near-infrared laser (808 nm, 1.0 W·cm -2 , 10 min). Then, the bacterial suspension was diluted 200 times, and 50 μL of the diluted bacterial suspension was spread on an agar plate and cultured at 37 °C for 18 to 24 h. The typical images of the culture plates were recorded using a digital camera. It can be seen from Figure 4 that in the blank experimental group and the experimental group, the fiber film added with Bi / BiOCl heterojunction showed a certain antibacterial effect on Escherichia coli, and the antibacterial rate was about 82.8%.

[0091] (4) Live / dead cell tests of the hydrogel and the first fiber film

[0092] Live / dead cell test of the hydrogel and bone marrow mesenchymal stem cells (BMSCs). Before cell seeding, the hydrogel was disinfected with 75% ethanol for 12 h and washed about 4 times with phosphate buffer to remove the residual ethanol. Then the treated hydrogel was placed in a 48-well plate and soaked in 500 μL of low-glucose medium containing 10% fetal bovine serum for 12 h. The cell density was adjusted to 1×10 4 cell / mL, and its suspension was inoculated into the extract of the hydrogel. After 3 days of co-culture, the cells were stained with AM (live cell dye) and PI (dead cell dye). Observation was carried out using an inverted microscope.

[0093] Live / dead cell test of the first fiber film and L929 fibroblasts. Before cell seeding, the fiber film was disinfected with 75% ethanol for 12 h and washed about 4 times with phosphate buffer to remove the residual ethanol. Then the treated fiber film was placed in a 48-well plate and soaked in 500 μL of low-glucose medium containing 10% fetal bovine serum for 12 h. The cell density was adjusted to 1×10 4cells / mL, and inoculate its suspension into the extract of the fiber membrane. After 3 days of co-culture, stain the cells with AM (live cell dye) and PI (dead cell dye). Observe using an inverted microscope.

[0094] The results of the live / dead cell test of the hydrogel layer and the first fiber membrane of Example 1 are as Figure 5 shown. It can be seen from Figure 5 that in the live / dead cell staining assay, no significant difference was observed among the blank control, the first fiber membrane, and the hydrogel layer. This indicates that the barrier membrane provided by the present invention exhibits excellent biocompatibility when used as a barrier material for guiding bone regeneration.

[0095] (5) Scanning electron microscopy of mesoporous bioactive glass (MGBNs)

[0096] The microtopography of MBGNs was tested by field emission scanning electron microscopy. Before the test, ultrasonically disperse MBGNs in absolute ethanol (50 μg / mL), take the supernatant and drop it on a silicon wafer to air-dry, and observe its morphology. Figure 6 The scanning electron microscopy image shows the spherical shape of the nanoparticles and the highly rough mesoporous morphology, with uniform particles and a particle size of about 110 nm.

[0097] (6) Transmission electron microscopy of mesoporous bioactive glass (MGBNs)

[0098] Observe the morphology of MBGNs by transmission electron microscopy. Ultrasonically disperse MBGNs in absolute ethanol, take the supernatant and drop it on a copper grid to air-dry, and observe its morphology at different magnifications. Figure 7 The transmission electron microscopy image shows the mesoporous structure of the mesoporous bioactive glass nanoparticles, with regular pore structures.

[0099] Obviously, the above embodiments of the present invention are merely examples for clearly illustrating the present invention, rather than limitations on the implementation manners of the present invention. For those of ordinary skill in the art, other different forms of changes or modifications can be made based on the above description. It is not necessary and impossible to enumerate all the implementation manners here. Any modifications, equivalent replacements, and improvements made within the spirit and principle of the present invention shall be included within the protection scope of the claims of the present invention.

Claims

1. A barrier membrane, characterized in that, The barrier membrane is prepared by the following preparation method: S1. Dissolve amino gelatin, oxidized sodium alginate and mesoporous bioactive glass in water and mix to obtain a hydrogel prepolymer solution; S2. Dissolve and mix polycaprolactone, amino gelatin and Bi / BiOCl heterojunction, and obtain a first fiber membrane by electrospinning; S3. Pour the hydrogel prepolymer solution onto the first fiber membrane to obtain the barrier membrane.

2. The barrier film according to claim 1, wherein In the first fiber membrane, the mass ratio of polycaprolactone, amino gelatin and Bi / BiOCl heterojunction is (7-10):(0.1-3):(0.01-0.1).

3. The barrier film according to claim 1, wherein The concentration of amino gelatin in the hydrogel prepolymer solution is 50-200 mg / mL.

4. The barrier film according to claim 1, characterized in that The concentration of oxidized sodium alginate in the hydrogel prepolymer solution is 50-200 mg / mL.

5. The barrier film according to claim 1, characterized in that, The concentration of mesoporous bioactive glass in the hydrogel prepolymer solution is 0.5-50 mg / mL.

6. The barrier film according to claim 1, wherein The amino gelatin is obtained by reacting gelatin with an amino group supplying agent.

7. The barrier film according to claim 1, wherein, In step S2, the first fiber membrane is prepared by electrospinning.

8. The barrier film according to claim 1, characterized in that, The diameter of the fibers in the first fiber membrane is 1-3 μm.

9. The barrier film according to claim 1, wherein The thickness of the barrier membrane is 1-3 mm.

10. Use of the barrier membrane according to any one of claims 1-9 in guided bone regeneration.

Images