Oral cavity restoration membrane for guiding tissue regeneration and preparation method thereof

Abstract

The invention relates to the technical field of oral medical materials, in particular to a tissue regeneration-guided oral repair membrane and a preparation method thereof. Comprising an outer layer and an inner layer, wherein the outer layer is a porous scaffold layer formed by a degradable polymer A, and the polymer A is selected from at least one of polylactic acid, polyglycolic acid, polycaprolactone or a copolymer thereof; the inner layer is loaded with a biocompatible hydrogel layer of a bioactive factor, and the bioactive factor is selected from bone morphogenetic protein, platelet-derived growth factor, vascular endothelial growth factor or a combination thereof. Comprising the following steps: preparing an outer layer solution; forming an outer-layer stent; preparing an inner layer precursor solution; gradient compounding and interface construction; performing crosslinking and curing; and post-processing. The invention provides a novel repairing film with intelligent degradation adaptability and precise biological factor controlled release capability.

Description

Technical Field

[0001] The present invention relates to the technical field of oral medical materials, and in particular to an oral repair membrane for guiding tissue regeneration and a preparation method thereof. Background Art

[0002] Guided tissue regeneration (GTR) is a core treatment method in the field of oral and maxillofacial restoration. It isolates soft tissue through a physical barrier, providing space for the selective regeneration of bone tissue and periodontal tissue. As the carrier of this technology, oral repair membranes must have mechanical barrier function, biocompatibility, and the ability to actively promote tissue regeneration. Currently, commonly used repair membrane materials in clinical practice include natural polymers (such as collagen) and synthetic polymers (such as polylactic acid). The core of their design lies in balancing the degradation rate and the matching of tissue regeneration cycles, while integrating bioactive factors to regulate cell behavior.

[0003] Existing oral repair membranes still have the following key limitations: it is difficult to achieve precise control of the degradation rate of a single-layer membrane structure. If the degradation is too fast, the barrier function will be lost, leading to soft tissue invasion; if the degradation is too slow, the growth of new tissue will be hindered, and even a foreign body reaction will be triggered. Growth factors directly loaded on the membrane material are prone to burst release (Burst Release), and cannot be released on demand during different healing stages such as the inflammatory phase, proliferation phase, and remodeling phase. Changes in the local microenvironment (such as pH, enzyme activity) will accelerate factor inactivation. Traditional membrane materials either focus on mechanical strength or on biological activity. It is difficult to form a synergistic interface between the porous scaffold (supporting cell migration / vascularization) and the factor sustained-release carrier (maintaining long-term pro-regeneration signals). In particular, there is a lack of gradient degradation design to dynamically adapt to the tissue regeneration process. Summary of the Invention

[0004] The above technical problems seriously restrict the regeneration effect of complex oral defects (such as alveolar bone defects and periodontal bone defects). The present invention provides a new repair membrane that has both intelligent degradation adaptability and precise biological factor controlled release capabilities.

[0005] The technical solution adopted by the present invention is: an oral repair membrane that guides tissue regeneration, comprising:

[0006] Outer layer: a porous scaffold layer composed of a degradable polymer A, wherein the polymer A is selected from at least one of polylactic acid, polyglycolic acid, polycaprolactone or a copolymer thereof;

[0007] Inner layer: a biocompatible hydrogel layer loaded with bioactive factors, wherein the bioactive factors are selected from bone morphogenetic protein, platelet-derived growth factor, vascular endothelial growth factor or a combination thereof;

[0008] The outer layer and the inner layer form an integral structure through interface bonding, and the degradation rate of the outer layer is greater than the degradation rate of the inner layer, forming a gradient degradation characteristic. The inner layer hydrogel has the characteristic of controlling the release of bioactive factors in response to the local microenvironment.

[0009] As a further improvement of the present invention, the interface bonding mode is selected from: the synergistic effect of physical crosslinking and chemical crosslinking; introducing an amphiphilic polymer B as a bonding layer at the interface, and the polymer B has good compatibility with both polymer A and the hydrogel.

[0010] As a further improvement of the present invention, the gradient degradation characteristics are as follows: the initial degradation time of the outer layer is 7-14 days earlier than that of the inner layer, and the complete degradation time of the outer layer is 20-50% shorter than that of the inner layer.

[0011] As a further improvement of the present invention, the inner layer hydrogel responds to the local microenvironment to control the release of bioactive factors, which means that: during the inflammatory period, the hydrogel network structure swells and partially dissociates, achieving the initial rapid release of bioactive factors; during the tissue regeneration period, the hydrogel achieves sustained and controllable release of bioactive factors through enzyme-sensitive bonds and specific pH-responsive groups.

[0012] As a further improvement of the present invention, the porosity of the outer porous scaffold layer is 70%-90%, and the average pore size is 100-300 μm, so as to facilitate fibroblast migration and blood vessel ingrowth, while providing an initial mechanical barrier function.

[0013] As a further improvement of the present invention, the inner layer hydrogel is selected from at least one of gelatin, hyaluronic acid, sodium alginate, chitosan or modified derivatives and composites thereof.

[0014] The bioactive factors are pre-compounded with carrier materials through microencapsulation technology and then loaded into the hydrogel to enhance their stability and control release effect.

[0015] A method for preparing an oral repair membrane for guiding tissue regeneration comprises the following steps:

[0016] S1, preparing the outer layer solution: dissolving the degradable polymer A in the organic solvent C to form a spinning solution and a casting solution;

[0017] S2, forming an outer layer scaffold: treating the solution obtained in S1 by electrospinning and freeze-drying technology to form an outer layer scaffold with a porous structure;

[0018] S3, preparing the inner layer precursor solution: dissolving and dispersing the biocompatible hydrogel matrix material D in the aqueous solution, adding the bioactive factors, and mixing uniformly;

[0019] S4, gradient composite and interface construction: the inner layer precursor liquid obtained in S3 is evenly coated and injected onto one side surface of the outer layer scaffold obtained in S2;

[0020] S5, cross-linking and curing: Under specific (T1) and humidity (H1) conditions, a physical interaction is first induced at the interface between the outer scaffold and the inner precursor solution. Under specific conditions (specific temperature T2, addition of cross-linking agent E), cross-linking between the molecular chains of the outer polymer A and chemical / physical cross-linking curing of the inner hydrogel are triggered. The cross-linking agent E achieves effective chemical bonding between the polymer A and the hydrogel matrix D at the interface while minimizing damage to the bioactive factors.

[0021] S6, post-processing: cleaning, drying, and sterilization to obtain the oral restoration membrane.

[0022] As a further improvement of the present invention, the specific temperature T1 is 4-25°C, the humidity H1 is 60%-90% RH; the specific temperature T2 is 25-37°C; the crosslinking agent E is selected from one of genipin, glutaraldehyde, EDC / NHS, and a photoinitiator, and its concentration in the interface area is higher than that in the main body area of ​​the membrane.

[0023] As a further improvement of the present invention, the bioactive factors are added to the inner layer precursor solution in the form of being loaded on a carrier material.

[0024] The beneficial effects of the present invention are as follows: (1) By designing a gradient structure with a rapidly degradable outer layer and a slowly degradable inner layer, the present invention enables the repair membrane to perform differentiated functions at different healing stages: in the early stage, the outer layer maintains mechanical barrier strength and effectively isolates soft tissue; in the middle stage, the outer layer gradually degrades to form biological channels, promoting cell migration and vascularization; and in the late stage, the inner layer continues to provide spatial support and is slowly absorbed. This design significantly reduces the risk of barrier failure caused by too rapid degradation and the foreign body reaction caused by too slow degradation.

[0025] (2) The inner hydrogel of the present invention achieves precise and timely release of growth factors based on changes in the local microenvironment (such as pH value and enzyme activity): during the inflammatory phase, the hydrogel network dissociates and rapidly releases anti-inflammatory and pro-angiogenic factors, accelerating the improvement of the wound microenvironment; during the regeneration phase, responsive chemical bonds control the release of osteogenic factors, continuously stimulating tissue regeneration. Combined with microencapsulation carrier technology, this effectively overcomes the defects of sudden release and inactivation of growth factors in traditional repair membranes.

[0026] (3) The present invention utilizes a step-by-step interface strengthening process that combines physical pre-crosslinking with chemical bonding to achieve dual optimization of high interlayer bonding strength and protection of biological factor activity under low-temperature and humidity-controlled conditions. The introduction of an amphiphilic interface layer further enhances compatibility with different material properties, ensuring that the repair membrane maintains its structural integrity over the long term in a wet oral environment, addressing the difficulty in synergizing mechanical properties and biological functions of traditional membrane materials. DETAILED DESCRIPTION

[0027] In order to make the technical problems, technical solutions and beneficial effects to be solved by this application more clearly understood, this application is further described in detail below in conjunction with the embodiments. It should be understood that the embodiments described herein are only used to explain this application and are not intended to limit this application.

[0028] The present invention provides an oral repair membrane for guiding tissue regeneration, comprising:

[0029] Outer layer: a porous scaffold layer composed of a degradable polymer A, wherein the polymer A is selected from at least one of polylactic acid, polyglycolic acid, polycaprolactone or a copolymer thereof;

[0030] Inner layer: a biocompatible hydrogel layer loaded with bioactive factors, wherein the bioactive factors are selected from bone morphogenetic protein, platelet-derived growth factor, vascular endothelial growth factor or a combination thereof;

[0031] Among them, the outer layer and the inner layer form an integral structure through interface bonding, and the degradation rate of the outer layer is greater than that of the inner layer, forming a gradient degradation characteristic. The inner layer hydrogel has the characteristic of controlling the release of bioactive factors in response to the local microenvironment.

[0032] In the present invention, the interface bonding method is selected from: the synergistic effect of physical crosslinking and chemical crosslinking; an amphiphilic polymer B is introduced at the interface as a bonding layer, and the polymer B has good compatibility with both the polymer A and the hydrogel.

[0033] The gradient degradation characteristics of the present invention are as follows: the initial degradation time of the outer layer is 7-14 days earlier than that of the inner layer, and the complete degradation time of the outer layer is 20-50% shorter than that of the inner layer.

[0034] In the present invention, the inner layer hydrogel responds to the local microenvironment to control the release of bioactive factors, which means that: during the inflammatory period, the hydrogel network structure swells and partially dissociates, achieving the initial rapid release of bioactive factors; during the tissue regeneration period, the hydrogel achieves sustained and controllable release of bioactive factors through enzyme-sensitive bonds and specific pH-responsive groups.

[0035] The porosity of the outer porous scaffold layer of the present invention is 70%-90%, and the average pore size is 100-300 μm, so as to facilitate fibroblast migration and blood vessel ingrowth, and provide an initial mechanical barrier function.

[0036] The inner layer hydrogel in the present invention is selected from at least one of gelatin, hyaluronic acid, sodium alginate, chitosan or modified derivatives and composites thereof.

[0037] In the present invention, the bioactive factors are pre-compounded with the carrier material through microencapsulation technology and then loaded into the hydrogel to enhance their stability and control release effect.

[0038] A method for preparing an oral repair membrane for guiding tissue regeneration comprises the following steps:

[0039] S1, preparing the outer layer solution: dissolving the degradable polymer A in the organic solvent C to form a spinning solution and a casting solution;

[0040] S2, forming an outer layer scaffold: treating the solution obtained in S1 by electrospinning and freeze-drying technology to form an outer layer scaffold with a porous structure;

[0041] S3, preparing the inner layer precursor solution: dissolving and dispersing the biocompatible hydrogel matrix material D in the aqueous solution, adding the bioactive factors, and mixing uniformly;

[0042] S4, gradient composite and interface construction: the inner layer precursor liquid obtained in S3 is evenly coated and injected onto one side surface of the outer layer scaffold obtained in S2;

[0043] S5, cross-linking and curing: Under specific (T1) and humidity (H1) conditions, a physical interaction is first induced at the interface between the outer scaffold and the inner precursor solution. Under specific conditions (specific temperature T2, addition of cross-linking agent E), cross-linking between the molecular chains of the outer polymer A and chemical / physical cross-linking curing of the inner hydrogel are triggered. The cross-linking agent E achieves effective chemical bonding between the polymer A and the hydrogel matrix D at the interface while minimizing damage to the bioactive factors.

[0044] S6, post-processing: cleaning, drying, and sterilization to obtain the oral restoration membrane.

[0045] In the present invention, the specific temperature T1 is 4-25°C, the humidity H1 is 60%-90% RH; the specific temperature T2 is 25-37°C; the crosslinking agent E is selected from one of genipin, glutaraldehyde, EDC / NHS, and a photoinitiator, and its concentration in the interface area is higher than that in the main membrane area.

[0046] In the present invention, the bioactive factors are added to the inner layer precursor solution in the form of being loaded on a carrier material.

[0047] Example 1:

[0048] Material

[0049] Outer layer polymer A: poly(lactic acid-co-glycolic acid) (PLGA, LA:GA=75:25, molecular weight 100,000, Jinan Daigang Biological);

[0050] Inner hydrogel D: gelatin (Type A, 300 Bloom, Sigma) + sodium alginate (viscosity 350 mPa·s, Qingdao Mingyue Seaweed);

[0051] Bioactive factors: recombinant human bone morphogenetic protein-2 (rhBMP-2, PeproTech);

[0052] Cross-linking agent E: genipin (purity 98%, Shanghai Yuanye Biotechnology);

[0053] Organic solvent C: hexafluoroisopropanol (HFIP, 99.5%, Aladdin).

[0054] Preparation steps

[0055] S1, outer layer solution preparation:

[0056] 1.2 g of PLGA was dissolved in 10 mL of HFIP and magnetically stirred for 6 h (25° C.) to form a uniform spinning solution.

[0057] S2, outer support molding:

[0058] Electrospinning was performed (voltage 18 kV, propulsion rate 1.2 mL / h, receiving distance 15 cm), and the nanofiber membrane was collected and dried under vacuum at 60° C. for 24 hours to remove the solvent. The porosity was measured to be 85% and the average pore size was 180 μm.

[0059] S3, preparation of inner layer precursor solution:

[0060] 5% gelatin and 2% sodium alginate were dissolved in PBS (pH 7.4) and stirred at 55° C.; 20 μg rhBMP-2 (pre-encapsulated in heparin-chitosan microspheres) was added and mixed in an ice bath.

[0061] S4, gradient composite:

[0062] 1 mL of the inner layer precursor solution was evenly coated on one side of the PLGA scaffold (coating thickness 0.5 mm).

[0063] S5, step-by-step cross-linking and curing:

[0064] Physical pre-crosslinking: incubate at 4°C and 85% RH for 2 hours to induce hydrogen bonding between gelatin chains and PLGA fibers;

[0065] Chemical cross-linking: spray 0.5% genipin solution (focus on strengthening the interface area) and react at 37°C for 1 hour to trigger gelatin-sodium alginate covalent cross-linking.

[0066] S6, post-treatment: rinse with sterile PBS three times, freeze-dry, and sterilize with γ rays (25 kGy) to obtain a PLGA / gelatin-sodium alginate double-layer repair membrane.

[0067] Example 2:

[0068] Material

[0069] Outer layer polymer A: polycaprolactone (PCL, molecular weight 80,000, Sigma-Aldrich);

[0070] Amphiphilic polymer B: Pluronic F127 (polyethylene glycol-polypropylene glycol block copolymer, Sigma-Aldrich);

[0071] Inner hydrogel D: oxidized hyaluronic acid (HA-ALD, molecular weight 200,000, Jinan Lujian Biological) + carboxymethyl chitosan (CMCS, deacetylation degree ≥90%, Dalian Meilun Biological);

[0072] Bioactive factors: vascular endothelial growth factor (VEGF165, Sino Biological);

[0073] Cross-linker E: EDC / NHS (1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride / N-hydroxysuccinimide, molar ratio 2:1, Sigma);

[0074] Organic solvent C: dichloromethane (99.8%, Sinopharm Group).

[0075] Preparation steps

[0076] S1, outer layer solution preparation:

[0077] 1.0 g of PCL was dissolved in 10 mL of dichloromethane and stirred in a 40°C water bath for 4 hours until completely dissolved to form a casting solution.

[0078] S2, outer layer stent molding and interface layer construction:

[0079] The solution was cast into a polytetrafluoroethylene mold (10 cm×10 cm); immediately placed in a -80°C ultra-low temperature freezer for 48 hours; and transferred to a freeze dryer (-50°C, 0.1 mbar) for drying for 24 hours to obtain a porous scaffold (porosity 82%, average pore size 250 μm).

[0080] A 5% Pluronic F127 ethanol solution was evenly sprayed on the surface of the PCL stent; it was allowed to evaporate at room temperature for 30 minutes to form an amphiphilic binding layer with a thickness of about 10 μm.

[0081] S3, preparation of inner layer precursor solution:

[0082] 3% HA-ALD and 2% CMCS were dissolved in PBS (pH 7.4) and stirred at 37° C. for 2 hours. 15 μg VEGF165 (previously complexed with chondroitin sulfate carrier) was added and the mixture was mixed in an ice bath in the dark.

[0083] S4, gradient composite:

[0084] 1.5 mL of the inner layer precursor solution was applied to the surface of the PCL scaffold treated with F127 (thickness 0.8 mm).

[0085] S5, step-by-step cross-linking and curing:

[0086] Physical pre-crosslinking: standing at 15°C and 75% RH for 3 hours to promote hydrogen bonding between the CMCS amino groups and the PEO segments of F127;

[0087] Chemical cross-linking: A 5 mM EDC / NHS solution was precisely delivered to the interface region using a microinjector (injection volume: 0.1 mL / cm 2 ) and reacted at 25°C for 2 hours to trigger the formation of an amide bond between the aldehyde group of HA-ALD and the amino group of CMCS.

[0088] S6, post-processing:

[0089] The unreacted cross-linking agent was removed by rinsing with ultrapure water, freeze-dried, and sterilized with ethylene oxide to obtain a PCL / HA-CMCS double-layer repair membrane.

[0090] Example 3:

[0091] Material

[0092] Outer layer polymer A: poly (L-lactic acid) (PLLA, molecular weight 150,000, Keran Biotechnology);

[0093] Amphiphilic polymer B: polyethylene glycol-polylactic acid block copolymer (PEG-PLGA, PEG:PLGA = 20:80, molecular weight 20,000, Jinan Daigang Biological);

[0094] Inner hydrogel D: type I collagen (bovine source, 4 mg / mL, Beijing Solebao);

[0095] Bioactive factors: platelet-derived growth factor-BB (PDGF-BB, ProSpec);

[0096] Cross-linker E: riboflavin (photoinitiator, 98%, Aladdin);

[0097] Organic solvent C: chloroform (99%, Sinopharm Group).

[0098] Preparation steps

[0099] S1, outer layer solution preparation:

[0100] 1.5 g of PLLA was dissolved in 12 mL of chloroform and stirred at 50°C for 3 hours until transparent.

[0101] S2, outer layer stent molding and rear interface layer processing:

[0102] Electrospinning: voltage 20 kV, propulsion rate 1.5 mL / h, receiving distance 18 cm; vacuum drying at 60° C. for 12 hours to obtain a fiber membrane (porosity 80%, average pore size 200 μm).

[0103] 8% PEG-PLGA tetrahydrofuran solution was spin-coated on the surface of the PLLA film (rotation speed 1500 rpm, time 30 s); evaporated at 40° C. to form an amphiphilic transition layer (thickness ≈ 15 μm).

[0104] S3, preparation of inner layer precursor solution:

[0105] 4% collagen solution (dissolved in 0.1 M acetic acid, pre-cooled at 4°C); add 25 μg PDGF-BB (pre-loaded on polylactic-co-glycolic acid microspheres) and mix well in an ice bath.

[0106] S4, gradient composite:

[0107] 1.2 mL of collagen precursor solution was injected into the surface of the PEG-PLGA treated scaffold (thickness 0.6 mm).

[0108] S5, photo-crosslinking and curing:

[0109] Physical pre-crosslinking: incubate at 10°C and 80% RH for 1.5 hours to promote hydrophobic interaction between collagen and PEG-PLGA;

[0110] Photochemical crosslinking: spraying 0.2% riboflavin solution (containing 0.1% triethanolamine); 365nm ultraviolet light (intensity 10mW / cm 2 ) for 8 minutes to trigger cross-linking between collagen molecules.

[0111] S6, post-processing:

[0112] The membrane was rinsed with PBS three times, freeze-dried, and sterilized by electron beam (15 kGy) to obtain a PLA / collagen bilayer repair membrane.

[0113] As can be seen from the above examples 1-3, the present invention demonstrates the wide applicability of the gradient double-layer structure design through three typical material combinations (synthetic polymer / natural polysaccharide, hydrophobic polyester / modified polysaccharide, polylactic acid / collagen). Different systems have successfully achieved: the high porosity structure (70%-90%) and rapid degradation characteristics of the outer layer scaffold; the microenvironment-responsive controlled release function of the inner layer hydrogel; and the stable integration of heterogeneous materials by the interface binding layer (physical-chemical synergy or amphiphilic polymer B). It has the following advantages: (1) Degradation timing matches the healing cycle: the outer layer (degradation in 7-35 days) provides a mechanical barrier first, and the inner layer (degradation in >42 days) provides continuous spatial support; (2) Intelligent adaptation of factor release: rapid release of angiogenic factors (VEGF / PDGF) during the inflammatory phase and sustained release of osteogenic factors (BMP) during the regeneration phase; (3) Breakthrough in interface stability: the interlayer bonding strength is significantly higher than that of traditional membranes (peeling force >0.78MPa), and there is no delamination under wet conditions.

[0114] In summary, the oral repair membrane that guides tissue regeneration and the preparation method thereof of the present invention have good biocompatibility, gradient degradation characteristics and controlled release function of bioactive factors, can effectively guide tissue regeneration in the oral environment, solves many problems of traditional oral repair membranes in mechanical properties, biological functions and interface bonding, etc., provides a more effective solution for oral tissue repair, and has broad application prospects in the field of oral medicine.

[0115] The above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit the same. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or make equivalent replacements for some of the technical features therein. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. An oral repair membrane for guiding tissue regeneration, characterized in that: include: Outer layer: a porous scaffold layer composed of a degradable polymer A, wherein the polymer A is selected from at least one of polylactic acid, polyglycolic acid, polycaprolactone or a copolymer thereof; Inner layer: a biocompatible hydrogel layer loaded with bioactive factors, wherein the bioactive factors are selected from bone morphogenetic protein, platelet-derived growth factor, vascular endothelial growth factor or a combination thereof; The outer layer and the inner layer form an integral structure through interface bonding, and the degradation rate of the outer layer is greater than the degradation rate of the inner layer, forming a gradient degradation characteristic. The inner layer hydrogel has the characteristic of controlling the release of bioactive factors in response to the local microenvironment.

2. The oral repair membrane for guiding tissue regeneration according to claim 1, characterized in that: The interface bonding mode is selected from: the synergistic effect of physical cross-linking and chemical cross-linking; introducing an amphiphilic polymer B as a bonding layer at the interface, and the polymer B has good compatibility with both polymer A and the hydrogel.

3. The oral repair membrane for guiding tissue regeneration according to claim 1, characterized in that: The gradient degradation characteristics are as follows: the initial degradation time of the outer layer is 7-14 days earlier than that of the inner layer, and the complete degradation time of the outer layer is 20-50% shorter than that of the inner layer.

4. The oral repair membrane for guiding tissue regeneration according to claim 1, characterized in that: The inner layer hydrogel responds to the local microenvironment to control the release of bioactive factors, which means that during the inflammatory period, the hydrogel network structure swells and partially dissociates, achieving an initial rapid release of the bioactive factors; during the tissue regeneration period, the hydrogel achieves a sustained and controllable release of the bioactive factors through enzyme-sensitive bonds and specific pH-responsive groups.

5. The oral repair membrane for guiding tissue regeneration according to claim 1, characterized in that: The porosity of the outer porous scaffold layer is 70%-90%, and the average pore size is 100-300 μm, so as to facilitate fibroblast migration and blood vessel growth, and provide an initial mechanical barrier function.

6. The oral repair membrane for guiding tissue regeneration according to claim 1, characterized in that: The inner layer hydrogel is selected from at least one of gelatin, hyaluronic acid, sodium alginate, chitosan or modified derivatives and composites thereof.

7. The oral repair membrane for guiding tissue regeneration according to claim 1, characterized in that: The bioactive factors are pre-compounded with carrier materials through microencapsulation technology and then loaded into the hydrogel to enhance their stability and control release effect.

8. A method for preparing an oral repair membrane that guides tissue regeneration, characterized in that: The following steps are involved: S1, preparing the outer layer solution: dissolving the degradable polymer A in the organic solvent C to form a spinning solution and a casting solution; S2, forming an outer layer scaffold: treating the solution obtained in S1 by electrospinning and freeze-drying technology to form an outer layer scaffold with a porous structure; S3, preparing the inner layer precursor solution: dissolving and dispersing the biocompatible hydrogel matrix material D in the aqueous solution, adding the bioactive factors, and mixing uniformly; S4, gradient composite and interface construction: the inner layer precursor liquid obtained in S3 is evenly coated and injected onto one side surface of the outer layer scaffold obtained in S2; S5, cross-linking and curing: Under specific (T1) and humidity (H1) conditions, a physical interaction is first induced at the interface between the outer scaffold and the inner precursor solution. Under specific conditions (specific temperature T2, addition of cross-linking agent E), cross-linking between the molecular chains of the outer polymer A and chemical / physical cross-linking curing of the inner hydrogel are triggered. The cross-linking agent E achieves effective chemical bonding between the polymer A and the hydrogel matrix D at the interface while minimizing damage to the bioactive factors. S6, post-processing: cleaning, drying, and sterilization to obtain the oral restoration membrane.

9. The method for preparing an oral repair membrane for guiding tissue regeneration according to claim 8, characterized in that: The specific temperature T1 is 4-25°C, the humidity H1 is 60%-90% RH; the specific temperature T2 is 25-37°C; the crosslinking agent E is selected from one of genipin, glutaraldehyde, EDC / NHS, and a photoinitiator, and its concentration in the interface area is higher than that in the main membrane area.

10. The method for preparing an oral repair membrane for guiding tissue regeneration according to claim 8, characterized in that: The bioactive factors are added to the inner layer precursor solution in the form of being loaded on a carrier material.