Multifunctional active nerve scaffold and preparation method thereof

Abstract

The invention discloses a multifunctional active nerve scaffold and a preparation method thereof, and belongs to the technical field of tissue engineering nerve scaffolds. The nerve scaffold is composed of a gel layer, a response layer and a cell layer. Wherein the gel layer has conductivity similar to that of nervous tissue and program-controlled deformation and tissue wet adhesion capability under the triggering of water or physiological liquid; the response layer has specific surface topological morphology and power generation capacity under the remote control of external fields such as light, ultrasound and magnetism, and can regulate and control migration and proliferation of cells in the cell layer and release of bioactive substances such as growth factors and exosomes based on topological morphology mediated biomechanical signals and bioelectrical signals generated under the remote control of the external fields. Based on the functions, the multifunctional active nerve scaffold can be used for tissue regeneration and functional reconstruction of nerve injury (such as brain, spinal cord, optic nerve and peripheral nerve). The invention also provides a preparation method of the multifunctional active nerve scaffold.

Description

Technical Field

[0001] The present invention belongs to the technical field of tissue engineering and regenerative medicine, and particularly relates to a multifunctional active nerve scaffold and a preparation method thereof. Background Art

[0002] Nerve damage is extremely common, severely impacting patients' health and quality of life. Autologous transplantation remains the gold standard for clinical treatment, but it can not only cause loss of nerve function at the donor site but also face mismatches such as mismatched nerve size between the donor and implanted sites. There is an urgent clinical need to develop new strategies to address nerve defects.

[0003] To this end, tissue-engineered neural scaffolds designed to promote in situ nerve regeneration have garnered widespread attention in recent years. By selecting biocompatible building materials, designing specific surface topologies, and loading and delivering specific drugs or bioactive factors, tissue-engineered neural scaffolds can regulate the behavioral functions of endogenous cells (Schwann cells and neurons), such as adhesion, migration, proliferation, and axonal elongation, thereby promoting tissue regeneration and functional reconstruction of damaged nerves. Currently, tissue-engineered neural scaffold products based on decellularized tissue and materials such as chitosan have been developed for the clinical treatment of nerve injury. However, due to the insufficient number of endogenous cells involved in tissue repair at the injury site and the inadequate bioregulatory capabilities of existing strategies, existing tissue-engineered neural scaffolds face significant challenges in repairing damaged nerves, hindering their large-scale clinical application or translation. Therefore, the novel strategy of cell-loaded active neural scaffolds has the potential to compensate for the lack of endogenous cells and achieve physiological-like bioregulatory effects through paracrine effects, providing a new research direction for nerve injury repair. However, existing active neural scaffolds face prominent challenges such as loss of loaded cells and a gradual decline in paracrine function. Compared to autologous transplantation, their effectiveness in repairing nerve tissue regeneration and restoring function still lags significantly. Therefore, minimizing the loss of loaded cells and enhancing their paracrine function are key to improving the effectiveness of active neural scaffolds in repairing nerve damage. Summary of the Invention

[0004] The purpose of the present invention is to overcome the shortcomings of the above-mentioned prior art and provide a multifunctional active nerve scaffold and a preparation method to solve the problems of active nerve scaffolds in the prior art such as loss of load cells and gradual decrease in paracrine capacity during use.

[0005] In order to achieve the above object, the present invention adopts the following technical solutions: A multifunctional active neural scaffold comprises a stacked base layer and a conductive layer; a topological response layer is embedded on the surface of the conductive layer, and a cell layer is grown on the response layer; The base layer deforms to allow the conductive layer with nerve tissue-like conductivity to adhere to the nerves. The response layer is used to convert external field energy into electrical energy. The electrical energy combines with the topological form of the response layer to generate bioelectric signals. The bioelectric signals are used to regulate cell migration, proliferation and release of bioactive substances in the cell layer.

[0006] A further improvement of the present invention is: Preferably, the thickness of the base layer is 10 μm-5 mm; the thickness of the conductive layer is 10 μm-5 mm; and the thickness of the response layer is 20 μm-1 cm.

[0007] Preferably, the deformed shape of the nerve stent is a curved membrane, a semi-closed tube or a closed tube, with a curvature radius of 200 μm-10 cm.

[0008] Preferably, the topological response layer and the conductive layer are arranged alternately and spaced apart; The spacing between the alternately arranged response layers is 20 μm-1 cm, and the spacing between the alternately arranged conductive layers is 20 μm-1 cm.

[0009] Preferably, the surface morphology of the response layer is a planar structure or a three-dimensional structure, the planar structure is at least one of a rectangle, an irregular curved surface, a circle or an ellipse; the three-dimensional structure is at least one of a microgroove, a microcone, a microcolumn, a truncated cone, spinning, a rotating parabola, and a hyperbola.

[0010] Preferably, the material of the substrate is one or more of the following materials: starch, cellulose, lignin, chitin, chitosan, alginate, hyaluronic acid, collagen, gelatin, silk fibroin, albumin, soy protein, polypeptide, decellularized matrix, polyglutamic acid, poly (3-hydroxybutyrate), polylysine, polyglycolic acid, polylactic acid, polycaprolactone, polyethylene glycol, polyvinyl alcohol, polydioxanone, polyphosphate, polyamino acid, polyanhydride, polycarbonate, polyphosphazene, polyorthoester or copolymers or derivatives containing the above units.

[0011] Preferably, the conductive layer is made of one or more of Class I materials and Class II materials, and the Class I material consists of Class III materials and Class IV materials.

[0012] The three types of materials are any one or more of the following materials: collagen, chitin, chitosan, gelatin, elastin-like polypeptide, agarose, cellulose, polyvinyl alcohol, dextran, hyaluronic acid, sodium hyaluronate, fibrin, polypeptide, protein, DNA, starch, polyhydroxyethyl methacrylate, polyethylene glycol, alginic acid, sodium alginate, poly-L-lysine, poly-L-glutamic acid, hydroxypropyl methylcellulose, hydroxyethyl cellulose, carboxymethyl cellulose, poly-L-glutamic acid, polyhistidine, polyaspartic acid, or copolymers or derivatives thereof; The second type of material is any one or more of the following materials: polyaniline, polypyrrole, polyacrylic acid gel, polyacrylamide gel, polyacrylonitrile gel, chitosan gel, PSS-PEDOT composite gel, composite gel combining metal ions and polymers, composite gel combining ionic liquids and polymers, and composite gel materials based on the above materials; The four types of materials are any one or more of the following materials: metal materials, inorganic non-metallic materials, and two-dimensional materials.

[0013] Preferably, the material of the response layer is at least one of piezoelectric material, piezoelectric ion gel, magnetocaloric material composite pyroelectric material, magnetostrictive material composite piezoelectric material, photodeformation material composite piezoelectric material, photovoltaic material, upconversion material composite photovoltaic material, photothermal material composite pyroelectric material or photocooling material composite pyroelectric material.

[0014] Preferably, the cells are at least one of Schwann cells, mesenchymal stem cells, neural crest stem cells, neural progenitor cells, PC12 cells, gene-overexpressed Schwann cells, gene-overexpressed mesenchymal stem cells, gene-overexpressed neural crest stem cells, gene-overexpressed neural progenitor cells, and Schwann cell-like cells derived from mesenchymal stem cells.

[0015] A method for preparing the multifunctional active nerve scaffold comprises the following steps: preparing the response layer; Cross-linking to form a conductive layer below the responsive layer; cross-linking to form a base layer below the conductive layer; Cells are seeded on the response layer to form the multifunctional active neural scaffold.

[0016] Compared with the prior art, the present invention has the following beneficial effects: The present invention provides a multifunctional active neural scaffold, which is composed of a basal layer, a conductive layer, a response layer, and a cell layer. The basal layer deforms when triggered by water or physiological fluids, the conductive layer has a conductivity similar to that of neural tissue and has tissue wet adhesion, which can promote neural information interaction and can conform to the target nerve without suturing. The response layer has a specific surface topology and the ability to generate electricity under external field remote control such as light, ultrasound, and magnetism. Based on the biomechanical signals mediated by the topology and the bioelectric signals generated under external field remote control, it can passively and wirelessly regulate the migration and proliferation of cells in the cell layer, as well as the release of bioactive substances such as growth factors and exosomes, promote the proliferation of loaded cells, and enhance the bioregulatory function of loaded cells. This is of great significance for solving the problems of nerve injury regeneration and functional reconstruction.

[0017] The present invention also discloses a method for preparing a multifunctional active neural scaffold. This method first prepares a topologically responsive layer in a mold, cross-links the layer below the responsive layer to form a conductive layer, and then cross-links the layer below the conductive layer to form a base layer. Cells are then seeded on the responsive layer to form the multifunctional active neural scaffold. This multifunctional active neural scaffold can be used for tissue regeneration and functional reconstruction of damaged nerves (such as those in the brain, spinal cord, optic nerve, and peripheral nerves). BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 A schematic structural diagram of a multifunctional active nerve scaffold provided in an embodiment of the present invention; Figure 2 This is a physical picture of the multifunctional active nerve scaffold shown in Example 1 of the present invention; Among them, 1. Basal layer; 2. Conductive layer; 3. Response layer; 4. Cell layer; 5. Nerve. DETAILED DESCRIPTION

[0019] The present invention is described in further detail below with reference to the accompanying drawings: To facilitate understanding of the features and effects of the present invention by those skilled in the art, the following provides a general description and definition of the terms and expressions used in the specification and claims. Unless otherwise indicated, all technical and scientific terms used herein have the ordinary meanings as understood by those skilled in the art regarding the present invention. In the event of conflict, the definitions in this specification shall prevail.

[0020] In this document, unless otherwise specified, “include,” “including,” “contains,” “has” or similar terms cover the meanings of “consisting of” and “mainly consisting of,” for example, “A includes a” covers the meanings of “A includes a and other” and “A only includes a.”

[0021] Below in conjunction with specific embodiment, further set forth the present invention.Should be understood that these embodiments are only used to illustrate the present invention and are not used in limiting the scope of the present invention.In addition, should be understood that after reading the content taught by the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms fall equally within the scope limited by the appended claims of the application.

[0022] The following examples were prepared using conventional instruments and equipment in the art. Experimental methods in the following examples, where specific conditions are not specified, were generally performed under conventional conditions or according to the conditions recommended by the manufacturer. The various raw materials used in the following examples, unless otherwise specified, were conventional commercially available products, with specifications conventional in the art. In the present specification and the following examples, unless otherwise specified, "%" indicates percentage by weight, "part" indicates parts by weight, and "ratio" indicates weight ratio.

[0023] In response to the challenges faced in repairing nerve damage, the first aspect of the present invention discloses a multifunctional active nerve scaffold, which is a multifunctional scaffold combining a base layer 1, a conductive layer 2, a response layer 3 and a cell layer; the conductive layer 2 is stacked on the base layer 1, and the surface of the conductive layer 2 is embedded with a topological response layer 3, and the response layer 3 has a cell layer 4 grown thereon; the base layer 1 deforms to make the conductive layer 2 adhere to the nerve, and the response layer 3 is used to convert external field energy into electrical energy, and the electrical energy generates bioelectric signals in combination with the topological form of the response layer 3, and the bioelectric signals are used to regulate cell migration, migration and release of bioactive substances.

[0024] The base layer 1 and the conductive layer 2 of the present invention together form a gel layer, which has the electrical conductivity of nerve tissue and the ability to programmable deformation and tissue wet adhesion under the triggering of water or physiological fluid; the neural stent of the present invention is ultimately used to repair Figure 1 The broken nerve 5 shown is used at the interface of two nerves 5, so it needs to be wrapped around the broken nerve. The base layer 1 is on the outermost side of the entire material. The material selected has strong deformation ability, can match the shape of the broken nerve, completely wrap the broken nerve, and can change with the change of the nerve shape; the conductive layer 2 of the present invention has the conductivity of nerve tissue, and restores the neural electrical signal communication at both ends of the damaged nerve through the conductivity of the gel, thereby guiding nerve regeneration; and the material selected for the conductive layer 2 can adhere to the nerve surface due to electrostatic or chemical effects, tissue wet adhesion ability, and enhance the bonding ability of the entire material with the nerve surface; the response layer 3 has a specific surface topology and the ability to generate electricity under remote control of external field energy such as light, ultrasound, and magnetism, and can regulate cell migration, proliferation, and the release of bioactive substances such as growth factors and exosomes based on the biomechanical signals mediated by the topology and the bioelectric signals generated under external field remote control, so as to repair the broken and damaged nerves.

[0025] The response layer 3 of the present invention is embedded in the conductive layer 2, so that the conductive layer 2 not covered by the response layer can directly contact the nerve, thereby improving the adhesion between the active nerve scaffold and the nerve.

[0026] In some embodiments, the deformed shape of the nerve stent includes but is not limited to a curved membrane, a semi-closed tube or a closed tube, with a curvature radius of 200 μm-10 cm, so as to wrap the nerve.

[0027] In some embodiments, the thickness of the gel layer is 20 μm-1 cm; the thickness of the response layer 3 is 20 μm-1 cm; further, the thickness of the base layer 1 in the gel layer is 10 μm-5 mm, and the thickness of the conductive layer 2 in the gel layer is 10 μm-5 mm.

[0028] In some embodiments, the response layers embedded in the conductive layer 2 are arranged in an alternating topology, and are alternately arranged with the conductive layer 2 . The minimum alternating unit group is two response layers 3 and one conductive layer 2 .

[0029] Preferably, the spacing between the alternately arranged response layers is 20 μm-1 cm, and the spacing between the alternately arranged gel layers is 20 μm-1 cm.

[0030] In some embodiments, the surface morphology of the response layer 3 is a planar structure or a three-dimensional structure, and the planar structure includes but is not limited to at least one of a rectangle, an irregular curved surface, a circle or an ellipse; the three-dimensional structure includes but is not limited to at least one of a microgroove, a microcone, a microcolumn, a truncated cone, spinning, a rotating parabola, and a hyperbola.

[0031] In some embodiments, the three-dimensional structure has a width of 10 nm-500 μm, a height of 10 nm-500 μm, and a pitch of 10 nm-500 μm.

[0032] In some embodiments, the material of the base layer 1 includes one or more of the following groups: starch, cellulose, lignin, chitin, chitosan, alginate, hyaluronic acid, collagen, gelatin, silk fibroin, albumin, soy protein, polypeptide, decellularized matrix, polyglutamic acid, poly (3-hydroxybutyrate), polylysine, polyglycolic acid, polylactic acid, polycaprolactone, polyethylene glycol, polyvinyl alcohol, polydioxanone, polyphosphate, polyamino acid, polyanhydride, polycarbonate, polyphosphazene, polyorthoester or copolymers or derivatives containing the above units.

[0033] In some embodiments, the conductive layer 2 is made of one or more of Category 1 materials and Category 2 materials, and Category 1 materials are composed of Category 3 materials and Category 4 materials.

[0034] The three types of materials are any one of the following materials: collagen, chitin, chitosan, gelatin, elastin-like polypeptide, agarose, cellulose, polyvinyl alcohol, dextran, hyaluronic acid, sodium hyaluronate, fibrin, polypeptide, protein, DNA, starch, polyhydroxyethyl methacrylate, polyethylene glycol, alginic acid, sodium alginate, poly-L-lysine, poly-L-glutamic acid, hydroxypropyl methylcellulose, hydroxyethyl cellulose, carboxymethyl cellulose, poly-L-glutamic acid, polyhistidine, polyaspartic acid or copolymers or derivatives containing the above units.

[0035] The second type of material is any one of the following materials: polyaniline, polypyrrole, polyacrylic acid gel, polyacrylamide gel, polyacrylonitrile gel, chitosan gel, PSS-PEDOT composite gel, Na + , K + , Ca 2+ Composite gels combining metal ions and polymers, composite gels combining choline and other ionic liquids and polymers, and composite gel materials based on the above materials.

[0036] The four types of materials are conductive fillers, which include but are not limited to conductive metal materials (such as Au, Ag, Cu, etc.), conductive inorganic non-metallic materials (such as carbon black, graphite, carbon fiber, carbon nanotubes, graphene, etc.), conductive two-dimensional materials (such as Mxene, phosphorus material series, etc.), etc.

[0037] Of the above materials, three types of materials have adhesive properties but lack electrical conductivity. Therefore, they require the addition of four types of conductive fillers. These fillers are dispersed within the first-stage gel material to impart conductivity. The amount of conductive fillers added is adjusted to ensure that the first-stage material possesses both adhesive properties and a certain degree of electrical conductivity. The second-stage material can be used directly, exhibiting both adhesiveness and electrical conductivity. In some embodiments, the first and second-stage materials can be used together.

[0038] It should be noted that the above-mentioned type 1 material can form chemical cross-links with nerve tissue and therefore has adhesion; the second type of material is a material that is conductive in itself but only has electrostatic interaction with nerve tissue and adheres to the nerve through electrostatic action.

[0039] In some embodiments, for a specific neural scaffold material, the conductive layer 2 and base layer 1 are composed of gels of different materials. Some materials can be used to form the conductive layer 2 in a specific combination and the base layer 1 in another combination. However, to ensure deformation, it is best to avoid having the conductive layer 2 and base layer 1 of a scaffold composed of the same material.

[0040] In some embodiments, the material of the response layer 3 is at least one of piezoelectric material, piezoelectric ion gel, magnetocaloric material composite pyroelectric material, magnetostrictive material composite piezoelectric material, photodeformation material composite piezoelectric material, photovoltaic material, upconversion material composite photovoltaic material, photothermal or photocooling material composite pyroelectric material.

[0041] In some embodiments, the piezoelectric material is at least one of the following materials: poly (vinylidene fluoride), vinylidene fluoride-trifluoroethylene copolymer [P (VDF-TrFE)], vinylidene fluoride-chlorofluoroethylene copolymer [P (VDF-CFE)], vinylidene fluoride-chlorinated trifluoroethylene copolymer [P (VDF-CTFE)], vinylidene fluoride-hexafluoropropylene copolymer [P (VDF-HFP)], vinylidene fluoride-trifluoroethylene-chlorofluoroethylene terpolymer [P (VDF-TrFE-CFE)], vinylidene fluoride-trifluoroethylene-chlorinated trifluoroethylene terpolymer [P (VDF-TrFE-CTFE)] and vinylidene fluoride-trifluoroethylene-hexafluoropropylene terpolymer [P (VDF-TrFE-HFP)], nylon with an odd number of carbon atoms, polyacrylonitrile, polyimide, polyurethane Ferroelectric polymers represented by vinyl dicyanide and its copolymers, polyurea, polyphenyl cyanoether, polyvinyl chloride, polyvinyl acetate, polypropylene, polytetrafluoroethylene, polylactic acid, etc.; inorganic oxide ferroelectrics represented by lead titanate, lead zirconate titanate, barium titanate, potassium niobate, lithium niobate, lithium tantalate, bismuth titanate, strontium ruthenium, bismuth ferrite, etc.; hydrogen bond ferroelectrics represented by potassium dihydrogen phosphate and triammonium sulfate; molecular-based ferroelectric compounds represented by Roshi salt; organic small molecule ferroelectrics represented by thiourea, cretone acid, benzimidazole, β-glycine, γ-glycine, 2,2,3,3,4,4-hexafluoro-1,5-pentanediol (HFPD), etc.; metal-organic hybrid ferroelectrics; bismuth layered perovskite structure ferroelectrics; tungsten bronze type ferroelectrics; perovskite type organometallic halide ferroelectrics; and composite materials based on the above materials.

[0042] In some embodiments, the piezoelectric ion gel is at least one of the following materials: polyacrylic acid gel, polyacrylamide gel, polyacrylonitrile gel, chitosan gel, PSS-PEDOT composite gel, Na + , K + , Ca 2+ Composite gels combining metal ions and polymers, composite gels combining choline and other ionic liquids and polymers, and composite gel materials based on the above materials.

[0043] In some embodiments, the magnetocaloric material composite pyroelectric material includes any combination of magnetocaloric material and pyroelectric material; wherein the magnetocaloric material is at least one of the following materials: ferromagnetic metals and alloys represented by iron, cobalt, nickel, gadolinium, nickel, Ni-Co alloy, Ni-Co-Cr alloy, Fe-Ni alloy, Fe-Al alloy, Fe-Co-V alloy, and Nd-Fe-B alloy; ferromagnetic metals and alloys represented by ferroferric oxide, cobalt ferrite, zinc ferrite, Ni-Co alloy, and Ni-Fe-B alloy; , ferromagnetic ferrite materials represented by Ni-Co-Cu ferrite materials, rare earth magnetic materials represented by Tb-Dy-Fe and composite materials based on the above materials; the pyroelectric material is at least one of the following materials: poly (vinylidene fluoride), vinylidene fluoride-trifluoroethylene copolymer [P(VDF-TrFE)], vinylidene fluoride-chlorofluoroethylene copolymer [P(VDF-CFE)], vinylidene fluoride-chlorinated trifluoroethylene copolymer [P(VDF-CTFE)], vinylidene fluoride-hexafluoropropylene copolymer [P(VDF-HFP)], vinylidene fluoride-trifluoroethylene-chlorofluoroethylene terpolymer [P(VDF-TrFE-CFE)], vinylidene fluoride-trifluoroethylene-chlorinated trifluoroethylene terpolymer [P(VDF-TrFE-CTFE)] and vinylidene fluoride-trifluoroethylene-hexafluoropropylene terpolymer [P(VDF-TrFE-HFP) ], ferroelectric polymers represented by nylon with an odd number of carbon atoms, polyacrylonitrile, polyimide, polyvinylidene dicyanate and copolymers, polyurea, polyphenylene cyanoether, polyvinyl chloride, polyvinyl acetate, polypropylene, polytetrafluoroethylene, polylactic acid, etc., inorganic oxide ferroelectrics represented by lead titanate, lead titanate zirconate, barium titanate, potassium niobate, lithium niobate, lithium tantalate, bismuth titanate, strontium ruthenium, bismuth ferrite, etc., potassium dihydrogen phosphate, triammonium sulfate, etc. Hydrogen bond ferroelectrics, molecular-based ferroelectric compounds represented by Roshi salt, organic small molecule ferroelectrics represented by thiourea, cretone acid, benzimidazole, β-glycine, γ-glycine, 2,2,3,3,4,4-hexafluoro-1,5-pentanediol (HFPD), etc., metal-organic hybrid ferroelectrics, bismuth layered perovskite structure ferroelectrics, tungsten bronze type ferroelectrics, perovskite type organic metal halide ferroelectrics and composite materials based on the above materials.

[0044] In some embodiments, the magnetostrictive composite piezoelectric material is a combination of any one of the following magnetostrictive materials and the pyroelectric material; wherein the magnetostrictive material is at least one of the following materials: magnetostrictive metals and alloys represented by iron, cobalt, nickel, gadolinium, nickel, Ni-Co alloy, Ni-Co-Cr alloy, Fe-Ni alloy, Fe-Al alloy, Fe-Co-V alloy, and Nd-Fe-B alloy; ferrite magnetostrictive materials represented by ferroferric oxide, cobalt ferrite, zinc ferrite, Ni-Co, and Ni-Co-Cu ferrite materials; rare earth giant magnetostrictive materials represented by Tb-Dy-Fe; and composite materials based on the above materials. The pyroelectric material is any one of the above pyroelectric materials.

[0045] In some embodiments, the photodeformable material composite piezoelectric material includes any combination of a photodeformable material and a piezoelectric material; wherein the photodeformable material is at least one of a photoisomerization material or a ferroelectric inorganic photodeformable material, and the photoisomerization material includes at least one of azobenzene and its derivatives, or spiropyran and its derivatives; the ferroelectric inorganic photodeformable material includes at least one of lead titanate, barium titanate, potassium niobate, lithium niobate, lithium tantalate, bismuth layered perovskite ferroelectric, tungsten bronze ferroelectric, bismuth ferrite, potassium dihydrogen phosphate, triammonium sulfate, Roche salt, and perovskite-type organometallic halide ferroelectric. The piezoelectric material is any of the above piezoelectric materials.

[0046] In some embodiments, the photovoltaic material is at least one of inorganic thin-film photovoltaic materials represented by single-crystal silicon, amorphous silicon, copper indium gallium selenide thin films, and cadmium telluride thin films, dye-sensitized photovoltaic materials represented by titanium dioxide and composites, perovskite photovoltaic materials based on perovskite-type organic metal halides, and organic photovoltaic materials represented by polyacetylene, polythiophene, polyaniline, polypyrrole and derivatives and copolymers.

[0047] In some embodiments, the up-conversion composite photovoltaic material is any combination of the following up-conversion materials and the piezoelectric material; wherein the up-conversion material includes yttrium oxide, yttrium oxysulfide, lanthanum fluoride, sodium yttrium fluoride, sodium gadolinium fluoride, and Yb 3+ Doped ZBLANP (ZrF4-BaF2-LaF3-AlF3-NaF-PbF2), Yb 3+ Doped KGd(WO4)2 crystal, Yb 3+ Doped yttrium lithium fluoride (YLF) crystal, Yb 3+ Doped KPb2Cl5 crystal, Yb 3+ Doped Y3Al5O 12 (YAG) crystal, Tm 3+ Doped ZBLANP, Tm3+ Doped KGd(WO4)2 crystal, Tm 3+ Doped YLF crystal, Tm 3+ Doped KPb2Cl5 crystal, Tm 3+ Rare earth ion doped heavy metal glass or crystal material with anti-Stokes effect represented by doped YAG crystal and composite materials based on the above materials. The piezoelectric material is any one of the above piezoelectric materials.

[0048] In some embodiments, the photothermal or photocold material composite pyroelectric material is a combination of any one of the following photothermal or photocold materials and the pyroelectric material; wherein the photothermal material is at least one of the following materials: carbon black, carbon nanotubes, graphene, black phosphorus, polydopamine, gold nanoparticles, gold nanorods, gallium-indium alloy liquid metal, black titanium dioxide, supramolecular metal organic framework (MOF) material, molybdenum disulfide (MoS2), transition metal carbide nitride and carbonitride two-dimensional material (MXene), small molecule photothermal agent represented by fluorine boron complex (BF2) and composite materials based on the above materials; the photocold material is at least one of the following materials: Yb 3+ Doped ZBLANP (ZrF4-BaF2-LaF3-AlF3-NaF-PbF2), Yb 3+ Doped KGd(WO4)2 crystal, Yb 3+ Doped yttrium lithium fluoride (YLF) crystal, Yb 3+ Doped KPb2Cl5 crystal, Yb 3+ Doped Y3Al5O 12 (YAG) crystal, Tm 3+ Doped ZBLANP, Tm 3+ Doped KGd(WO4)2 crystal, Tm 3+ Doped YLF crystal, Tm 3+ Doped KPb2Cl5 crystal, Tm 3+ Rare earth ion doped heavy metal glass or crystal material with anti-Stokes effect represented by doped YAG crystal and composite materials based on the above materials. The pyroelectric material is any one of the above pyroelectric materials.

[0049] In some embodiments, the cells constituting the cell layer 4 include at least one of: Schwann cells, mesenchymal stem cells, neural crest stem cells, neural progenitor cells, PC12 cells, gene-overexpressed Schwann cells, gene-overexpressed mesenchymal stem cells, gene-overexpressed neural crest stem cells, gene-overexpressed neural progenitor cells, and Schwann cell-like cells derived from mesenchymal stem cells.

[0050] The second aspect of the present invention provides a method for preparing the neural scaffold, comprising the following steps: S1, preparing the response layer 3; S2, cross-linking below the response layer to form a conductive layer 2; S3, cross-linking below the conductive layer to form a base layer 1; S4, seeding cells on the response layer to form a cell layer 4, thereby obtaining the multifunctional active neural scaffold.

[0051] In some embodiments, in S1, after the response layer 3 is prepared by coating, spin coating or casting, it is cut by a mold to obtain the response layer 3 of a set form; In some embodiments, in S2, a conductive layer is formed by cross-linking below the response layer 3 through ultraviolet, ionic or thermal cross-linking methods, forming a structure in which the conductive layer 2 and the response layer 3 are alternately arranged.

[0052] In some embodiments, in S2, the responsive layer 3 is cross-linked below the conductive layer 2 by ultraviolet, ionic or thermal cross-linking methods, and the responsive layer 3 and the conductive layer 2 together constitute a gel layer.

[0053] In some embodiments, in S3, cells are seeded on the response layer 3 to form the multifunctional active neural scaffold; the seeding cell density is 10 3 ~10 6 cells / cm2 The following is further described with reference to specific embodiments.

[0054] Example 1 A multifunctional active nerve scaffold, the structure of which is as follows Figure 1 As shown in the actual picture Figure 2 As shown, the multifunctional active neural scaffold consists of a gel layer, an ultrasound-triggered response layer, and a cell layer.

[0055] In this example, the stent is curled into a closed tube with a radius of curvature of 1 mm. The base layer of the gel layer is 50 μm thick and made of chitosan hydrogel. The conductive layer is 100 μm thick and made of sodium alginate choline composite gel. The response layer has a surface morphology of a microgroove array (microgroove width: 200 nm, height: 200 nm, spacing: 200 nm), is 100 μm thick, and is made of polydopamine and poly(vinylidene fluoride-trifluoroethylene) copolymer. The response layer and gel layer are arranged alternately, with six and five layers, respectively, spaced 500 μm apart. The response layers and conductive layers are spaced 500 μm apart. The cell layer is seeded with rat Schwann cells.

[0056] The preparation method of the multifunctional active nerve scaffold comprises the following steps: 1) Preparation of response layer: Using dimethyl sulfoxide as a solvent, a mixed solution containing 1 w / w% polydopamine and 10 w / v% poly(vinylidene fluoride-trifluoroethylene) was prepared; the mixed solution was cast on the surface of a silicon wafer with a surface microgroove array, dried at 80 degrees Celsius, and the dried film material was peeled off from the microgroove array silicon wafer template to obtain a response layer with the surface morphology of the microgroove array.

[0057] 2) Gel layer preparation: The responsive layer was nested within a Teflon mold. A precursor solution of a sodium alginate-choline composite gel containing the crosslinker ethylene glycol dimethacrylate and the initiator ammonium persulfate was poured beneath the responsive layer. A conductive layer alternating with the responsive layer was constructed by thermal polymerization at 70 degrees Celsius. A chitosan solution was poured beneath the conductive layer to form a base layer, resulting in a structure with alternating gel and responsive layers. A self-curling structure was formed by swelling equilibrium in a calcium chloride solution. The resulting gel and responsive layers were sterilized by irradiation with a cobalt source at a dose of 15 kGy for 30 minutes.

[0058] 3) Cell seeding The cell suspension of rat Schwann cells was obtained by trypsin digestion and placed on the surface of the response layer at 10 6 The cells were seeded at a seeding density of 10 cells / cm2.

[0059] Example 2 A multifunctional active nerve scaffold, the structure of which is as follows Figure 1 As shown, the multifunctional active neural scaffold consists of a gel layer, a magnetic field-triggered response layer, and a cell layer.

[0060] In this example, the scaffold is curled into a semi-closed tube with a curvature radius of 1 cm. The base layer of the gel layer is 200 μm thick and made of polyethylene glycol; the conductive layer is 100 μm thick and made of polyacrylamide gel; the response layer has a microgroove array surface (microgroove width: 10 μm, height: 10 μm, spacing: 20 μm), is 100 μm thick, and is made of iron-doped vinylidene fluoride-trifluoroethylene copolymer. The response layer and gel layer are arranged alternately, with 6 and 5 layers, respectively, spaced 500 μm apart between the response layers and 400 μm apart between the conductive layers. The cells seeded are neural progenitor cells.

[0061] The preparation method of the multifunctional active nerve scaffold comprises the following steps: 1) Preparation of response layer: Using dimethyl sulfoxide as a solvent, a vinylidene fluoride-trifluoroethylene copolymer solution with a concentration of 10 w / v% was prepared; the vinylidene fluoride-trifluoroethylene copolymer solution was cast on the surface of a silicon wafer with a surface microgroove array, and dried at 80 degrees Celsius. The dried film material was peeled off from the microgroove array silicon wafer template to obtain a response layer with the surface morphology of the microgroove array.

[0062] 2) Gel layer preparation: The responsive layer was nested within a Teflon mold. A precursor solution of polyacrylamide gel containing the initiator ammonium persulfate and the crosslinker N,N'-methylenebisacrylamide was poured beneath the responsive layer. Thermal polymerization at 60 degrees Celsius created a conductive layer alternating with the responsive layer. A polyethylene glycol solution was poured beneath the conductive layer to form a base layer, creating a structure alternating between the gel layer and the responsive layer. A self-curling structure was formed by swelling equilibrium in a sodium dodecyl sulfate solution. The resulting gel layer and responsive layer were sterilized by irradiation with a cobalt source at a dose of 15 kGy for 30 minutes.

[0063] 3) Cell seeding The neural progenitor cell suspension was obtained by trypsin digestion and was placed on the surface of the sterilized gel layer and response layer at 10 6 The cells were seeded at a seeding density of 10 cells / cm2.

[0064] Example 3 A multifunctional active nerve scaffold, the structure of which is as follows Figure 1 As shown, the multifunctional active neural scaffold consists of a gel layer, a light-triggered response layer, and a cell layer.

[0065] In this embodiment, the scaffold is curled into a closed tube with a radius of curvature of 8 mm. The base layer of the gel layer is 200 μm thick and made of lignin; the conductive layer is 200 μm thick and made of poly(3,4-ethylenedioxythiophene):polystyrenesulfonic acid) (PEDOT:PSS). The response layer has a microcone array structure (microcone width: 10 μm, height: 10 μm, spacing: 20 μm), is 500 μm thick, and is made of poly-3-hexylthiophene. The response layer and gel layer are arranged alternately, with 8 and 7 layers, respectively, spaced 200 μm apart. The cells seeded in the cell layer are neural crest cells.

[0066] The preparation method of the multifunctional active nerve scaffold comprises the following steps: 1) Preparation of response layer: Using 1,2-dichlorobenzene as the solvent, a P3HT solution with a concentration of 30 mg / mL was prepared; the poly-3-hexylthiophene solution was cast on the surface of a silicon wafer with a surface microcone array, spin-coated at 60 degrees Celsius, and dried. The dried film material was peeled off from the microcone array silicon wafer template to obtain a response layer with the surface morphology of the microcone array.

[0067] 2) Gel layer preparation: The responsive layer was nested within a Teflon mold. A premixed solution of PEDOT:PSS and 4-dodecylbenzenesulfonic acid was poured beneath the responsive layer, followed by physical crosslinking at room temperature to construct a conductive layer alternating with the responsive layer. A lignin solution was poured beneath the conductive layer to form a base layer, forming a structure alternating between the gel layer and the responsive layer. The resulting gel layer and responsive layer formed a self-curling structure through swelling equilibrium in the aqueous solution. The resulting gel layer and responsive layer were sterilized by irradiation with a cobalt source at a dose of 15 kGy for 30 minutes.

[0068] 3) Cell seeding Neural crest cells were obtained by trypsin digestion and placed on the surface of the sterilized gel layer and response layer at 10 3 The cells were seeded at a seeding density of 10 cells / cm2.

[0069] Example 4 A multifunctional active nerve scaffold, the structure of which is as follows Figure 1 As shown, the multifunctional active neural scaffold consists of a gel layer, a light-triggered response layer, and a cell layer.

[0070] In this embodiment, the scaffold is a curved membrane with a curvature radius of 5 cm; the base layer in the gel layer is 100 μm thick and is made of polyacrylic acid; the conductive layer is 200 μm thick and is made of a sodium alginate choline composite gel with liquid metal added; the surface morphology of the response layer is a flat, unstructured surface with a thickness of 20 μm and is made of poly-3-hexylthiophene; the response layers and gel layers are alternately arranged in 4 and 3 numbers, respectively, with the response layers spaced 400 μm apart and the gel layers spaced 200 μm apart; the inoculated cells are PC12 cells.

[0071] The preparation method of the multifunctional active nerve scaffold comprises the following steps: 1) Preparation of response layer: Using 1,2-dichlorobenzene as a solvent, a mixture solution containing 30 mg / mL poly-3-hexylthiophene and 0.1 mg / mL sodium gadolinium fluoride was prepared; the mixed solution was poured onto the surface of a flat silicon wafer and spin-coated at 60 degrees Celsius. After drying, the planar film material was peeled off from the flat silicon wafer template to obtain a response layer with a planar morphology.

[0072] 2) Gel layer preparation: A planar responsive layer was nested within a Teflon mold. A precursor material for a liquid metal sodium alginate-choline composite gel was poured beneath the responsive layer. CaCl₂ ions were added for crosslinking, creating a conductive layer alternating with the responsive layer. A polyacrylic acid solution was poured beneath the conductive layer to form a base layer, creating a structure with alternating gel and responsive layers. The resulting structure formed a self-curling structure through swelling equilibrium in aqueous solution. The resulting gel and responsive layers were sterilized by irradiation with a cobalt source at a dose of 15 kGy for 30 minutes.

[0073] 3) Cell seeding PC12 cells were extracted and placed on the surface of the sterilized gel layer and response layer at 10 3 The cells were seeded at a seeding density of 10 cells / cm2.

[0074] Example 5 A multifunctional active nerve scaffold, the structure of which is as follows Figure 1 As shown, the multifunctional active neural scaffold consists of a gel layer, a light-triggered response layer, and a cell layer.

[0075] In this embodiment, the stent is curled into a closed tube with a curvature radius of 200 μm; the base layer in the gel layer is 300 μm thick and is made of carboxymethyl cellulose; the conductive layer is 200 μm thick and is made of a composite gel of sodium alginate and choline; the surface morphology of the response layer is a microcolumn array surface (microcolumn width: 5 μm, height: 5 μm, spacing: 5 μm), with a thickness of 50 μm, and its material is a composite of polydopamine and poly(vinylidene fluoride-trifluoroethylene) copolymer; the number of response layers and gel layers alternating is 10 and 9 respectively, the response layers are spaced 300 μm apart, and the gel layers are spaced 200 μm apart; the inoculated cells are adipose stem cells.

[0076] The preparation method of the multifunctional active nerve scaffold comprises the following steps: 1) Preparation of response layer: Using dimethyl sulfoxide as a solvent, a mixed solution containing 0.1 w / v% polydopamine and 10 w / v% poly(vinylidene fluoride-trifluoroethylene) was prepared; the mixed solution was cast on the surface of a silicon wafer with a surface micropillar array, dried at 80 degrees Celsius, and the dried film material was peeled off from the micropillar array silicon wafer template to produce a response layer with the surface morphology of the micropillar array.

[0077] 2) Gel layer preparation: The responsive layer was nested within a Teflon mold. A precursor material of a sodium alginate and choline composite gel was poured beneath the responsive layer. CaCl₂ was added for ion crosslinking to create a conductive layer alternating with the responsive layer. A mixed solution of acrylic acid, acrylamide, and carboxymethyl cellulose was poured beneath the conductive layer to form a base layer, creating a structure with alternating gel layers and responsive layers. The resulting structure formed a self-curling structure through swelling equilibrium in a high pH solution. The resulting gel and responsive layers were sterilized by irradiation with a cobalt source at a dose of 15 kGy for 30 minutes.

[0078] 3) Cell seeding Adipose-derived stem cells were extracted and placed on the surface of the sterilized gel layer and response layer at a temperature of 10 3 The cells were seeded at a seeding density of 10 cells / cm2.

[0079] Example 6 A multifunctional active nerve scaffold, the structure of which is as follows Figure 1 As shown, the multifunctional active neural scaffold consists of a gel layer, a magnetic field-triggered response layer, and a cell layer.

[0080] In this embodiment, the stent is curled into a closed tube with a curvature radius of 2 cm; the base layer in the gel layer is 300 μm thick and is made of hydroxybutyl starch; the conductive layer is 200 μm thick and is made of a composite gel of sodium alginate and choline; the surface morphology of the response layer is a microcone array surface (microcone width: 50 nm, height: 50 nm, spacing: 50 nm), with a thickness of 200 μm, and its material is a composite of polyvinylidene fluoride and ferroferric oxide; the response layer and the gel layer are alternately arranged in 8 and 7 numbers, respectively, with a spacing of 200 μm between the response layers and 200 μm between the gel layers; the inoculated cells are adipose stem cells.

[0081] The preparation method of the multifunctional active nerve scaffold comprises the following steps: 1) Preparation of response layer: Using dimethyl sulfoxide as a solvent, a mixed solution containing 10 w / v% polyvinylidene fluoride and 5 mg / mL ferrosoferric oxide nanoparticles was prepared; the mixed solution was cast on the surface of a silicon wafer with a surface microcone array, dried at 80 degrees Celsius, and the dried film material was peeled off from the microcone array silicon wafer template to obtain a layer with a microcone array surface morphology response.

[0082] 2) Gel layer preparation: The responsive layer was nested within a Teflon mold. A precursor material of a sodium alginate and choline composite gel was poured beneath the responsive layer and above the responsive units within the mold. CaCl2 was added for ionic crosslinking to create a gel layer alternating with the responsive layer. A mixed solution of hydroxybutyl starch and N-isopropylacrylamide was poured beneath the gel layer to form an alternating structure of gel and responsive layers. The resulting gel layer and responsive layer formed a self-curling structure through swelling equilibrium in aqueous solution. The resulting gel layer and responsive layer were sterilized by irradiation with a cobalt source at a dose of 15 kGy for 30 minutes.

[0083] 3) Cell seeding Adipose-derived stem cells were extracted and placed on the surface of the sterilized gel layer and response layer at a temperature of 10 4 The cells were seeded at a seeding density of 10 cells / cm2.

[0084] Example 7 A multifunctional active nerve scaffold, the structure of which is as follows Figure 1 As shown, the multifunctional active neural scaffold consists of a gel layer, an ultrasound-triggered response layer, and a cell layer.

[0085] In this embodiment, the stent is curled into a closed tube with a curvature radius of 6 mm; the base layer in the gel layer is 300 μm thick and is made of hyaluronic acid; the gel is 200 μm thick and is made of a composite gel of sodium alginate and choline; the surface morphology of the response layer is a microcolumn array surface (microcolumn width: 50 nm, height: 50 nm, spacing: 50 nm), with a thickness of 100 μm, and its material is a composite of polymethacrylic acid and choline; the response layer and the gel layer are alternately arranged in 8 and 7 numbers, respectively, with a spacing of 500 μm between the response layers and 500 μm between the gel layers; the inoculated cells are Schwann cell-like cells derived from mesenchymal stem cells.

[0086] The preparation method of the multifunctional active nerve scaffold comprises the following steps: 1) Preparation of response layer: An aqueous solution containing 10 w / v% methacrylic acid, 3 w / v% choline chloride, and 0.1 w / v% 2959 photoinitiator was prepared; the mixed solution was cast on the surface of a silicon wafer with a micropillar array. The mixture was reacted under UV cross-linking instrument irradiation for 10 minutes. The dried film material was peeled off from the micropillar array silicon wafer template to obtain a response layer with the micropillar array surface morphology.

[0087] 2) Gel layer preparation: The responsive layer was nested within a Teflon mold. A precursor material of a sodium alginate and choline composite gel was poured beneath the responsive layer and above the responsive units within the mold. CaCl2 was added for ion crosslinking to create a conductive layer alternating with the responsive layer. A mixed solution of hyaluronic acid and chondroitin sulfate was poured beneath the conductive layer to form a structure alternating between the gel layer and the responsive layer. The resulting gel layer and responsive layer formed a self-curling structure through swelling equilibrium in aqueous solution. The resulting gel layer and responsive layer were sterilized by irradiation with a cobalt source at a dose of 15 kGy for 30 minutes.

[0088] 3) Cell seeding The Schwann cell-like cells derived from mesenchymal stem cells were extracted and placed on the surface of the sterilized gel layer and response layer at 10 4 The cells were seeded at a seeding density of 10 cells / cm2.

[0089] Example 8 A multifunctional active nerve scaffold, the structure of which is as follows Figure 1 As shown, the multifunctional active neural scaffold consists of a gel layer, a magnetic field-triggered response layer, and a cell layer.

[0090] In this embodiment, the stent is curled into a closed tube with a curvature radius of 300 μm; the base layer in the gel layer is 10 μm thick and is made of hydroxybutyl starch; the conductive layer is 10 μm thick and is made of a composite gel of sodium alginate and choline; the surface morphology of the response layer is a microcone array surface (microcone width: 50 nm, height: 50 nm, spacing: 50 nm), with a thickness of 20 μm, and its material is a composite of polyvinylidene fluoride and ferroferric oxide; the response layer and the gel layer are alternately arranged in 8 and 7 numbers respectively, with a 20 μm interval between the response layers and a 20 μm interval between the gel layers; the inoculated cells are adipose stem cells.

[0091] The preparation method of the multifunctional active nerve scaffold comprises the following steps: 1) Preparation of response layer: Using dimethyl sulfoxide as a solvent, a mixed solution containing 10 w / v% polyvinylidene fluoride and 5 mg / mL ferrosoferric oxide nanoparticles was prepared; the mixed solution was cast on the surface of a silicon wafer with a surface microcone array, dried at 80 degrees Celsius, and the dried film material was peeled off from the microcone array silicon wafer template to obtain a layer with a microcone array surface morphology response.

[0092] 2) Gel layer preparation: The responsive layer was nested within a Teflon mold. A precursor material of a sodium alginate and choline composite gel was poured beneath the responsive layer and above the responsive units within the mold. CaCl2 was added for ionic crosslinking to create a gel layer alternating with the responsive layer. A mixed solution of hydroxybutyl starch and N-isopropylacrylamide was poured beneath the gel layer to form an alternating structure of gel and responsive layers. The resulting gel layer and responsive layer formed a self-curling structure through swelling equilibrium in aqueous solution. The resulting gel layer and responsive layer were sterilized by irradiation with a cobalt source at a dose of 15 kGy for 30 minutes.

[0093] 3) Cell seeding Adipose-derived stem cells were extracted and placed on the surface of the sterilized gel layer and response layer at a temperature of 10 4 The cells were seeded at a seeding density of 10 cells / cm2.

[0094] Example 9 A multifunctional active nerve scaffold, the structure of which is as follows Figure 1 As shown, the multifunctional active neural scaffold consists of a gel layer, an ultrasound-triggered response layer, and a cell layer.

[0095] In this embodiment, the stent is curled into a closed tube with a curvature radius of 10 cm; the base layer in the gel layer is 5 mm thick and is made of hyaluronic acid; the gel is 1 cm thick and is made of a composite gel of sodium alginate and choline; the surface morphology of the response layer is a microcolumn array surface (microcolumn width: 50 nm, height: 50 nm, spacing: 50 nm), with a thickness of 1 cm, and is made of a composite of polymethacrylic acid and choline; the response layer and the gel layer are alternately arranged in 8 and 7 numbers, respectively, with a spacing of 1 cm between the response layers and 1 cm between the gel layers; the inoculated cells are Schwann cell-like cells derived from mesenchymal stem cells.

[0096] The preparation method of the multifunctional active nerve scaffold comprises the following steps: 1) Preparation of response layer: An aqueous solution containing 10 w / v% methacrylic acid, 3 w / v% choline chloride, and 0.1 w / v% 2959 photoinitiator was prepared; the mixed solution was cast on the surface of a silicon wafer with a micropillar array. The mixture was reacted under UV cross-linking instrument for 10 minutes. The dried film material was peeled off from the micropillar array silicon wafer template to obtain a response layer with the micropillar array surface morphology.

[0097] 2) Gel layer preparation: The responsive layer was nested within a Teflon mold. A precursor material of a sodium alginate and choline composite gel was poured beneath the responsive layer and above the responsive units within the mold. CaCl2 was added for ion crosslinking to create a conductive layer alternating with the responsive layer. A mixed solution of hyaluronic acid and chondroitin sulfate was poured beneath the conductive layer to form a structure alternating between the gel layer and the responsive layer. The resulting gel layer and responsive layer formed a self-curling structure through swelling equilibrium in aqueous solution. The resulting gel layer and responsive layer were sterilized by irradiation with a cobalt source at a dose of 15 kGy for 30 minutes.

[0098] 3) Cell seeding The Schwann cell-like cells derived from mesenchymal stem cells were extracted and placed on the surface of the sterilized gel layer and response layer at 10 4 The cells were seeded at a seeding density of 10 cells / cm2.

[0099] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. A multifunctional active nerve scaffold, characterized in that: It comprises a stacked base layer and a conductive layer; a topological response layer is embedded on the surface of the conductive layer, and a cell layer is grown on the response layer; The base layer deforms to allow the conductive layer with nerve tissue-like conductivity to adhere to the nerves. The response layer is used to convert external field energy into electrical energy. The electrical energy combines with the topological form of the response layer to generate bioelectric signals. The bioelectric signals are used to regulate cell migration, proliferation and release of bioactive substances in the cell layer.

2. A multifunctional active nerve scaffold according to claim 1, characterized in that: The thickness of the base layer is 10 μm-5 mm; the thickness of the conductive layer is 10 μm-5 mm; and the thickness of the response layer is 20 μm-1 cm.

3. The multifunctional active nerve scaffold according to claim 1, characterized in that: The deformed shape of the nerve stent is a curved membrane, a semi-closed tube or a closed tube, and the curvature radius is 200 μm-10 cm.

4. The multifunctional active nerve scaffold according to claim 1, characterized in that: The topological response layers and the conductive layers are alternately arranged; The spacing between the alternately arranged response layers is 20 μm-1 cm, and the spacing between the alternately arranged conductive layers is 20 μm-1 cm.

5. The multifunctional active nerve scaffold according to claim 1, characterized in that: The surface morphology of the response layer is a planar structure or a three-dimensional structure, wherein the planar structure is at least one of a rectangle, an irregular curved surface, a circle or an ellipse; and the three-dimensional structure is at least one of a microgroove, a microcone, a microcolumn, a truncated cone, a spinning, a rotating parabola, and a hyperboloid.

6. The multifunctional active nerve scaffold according to claim 1, characterized in that: The material of the substrate is one or more of the following materials: starch, cellulose, lignin, chitin, chitosan, alginate, hyaluronic acid, collagen, gelatin, silk fibroin, albumin, soy protein, polypeptide, decellularized matrix, polyglutamic acid, poly (3-hydroxybutyrate), polylysine, polyglycolic acid, polylactic acid, polycaprolactone, polyethylene glycol, polyvinyl alcohol, polydioxanone, polyphosphate, polyamino acid, polyanhydride, polycarbonate, polyphosphazene, polyorthoester or copolymers or derivatives containing the above units.

7. The multifunctional active nerve scaffold according to claim 1, characterized in that: The conductive layer is made of one or more of Class I materials and Class II materials, and the Class I material is composed of Class III materials and Class IV materials. The three types of materials are any one or more of the following materials: collagen, chitin, chitosan, gelatin, elastin-like polypeptide, agarose, cellulose, polyvinyl alcohol, dextran, hyaluronic acid, sodium hyaluronate, fibrin, polypeptide, protein, DNA, starch, polyhydroxyethyl methacrylate, polyethylene glycol, alginic acid, sodium alginate, poly-L-lysine, poly-L-glutamic acid, hydroxypropyl methylcellulose, hydroxyethyl cellulose, carboxymethyl cellulose, poly-L-glutamic acid, polyhistidine, polyaspartic acid, or copolymers or derivatives thereof; The second type of material is any one or more of the following materials: polyaniline, polypyrrole, polyacrylic acid gel, polyacrylamide gel, polyacrylonitrile gel, chitosan gel, PSS-PEDOT composite gel, composite gel combining metal ions and polymers, composite gel combining ionic liquids and polymers, and composite gel materials based on the above materials; The four types of materials are any one or more of the following materials: metal materials, inorganic non-metallic materials, and two-dimensional materials.

8. The multifunctional active nerve scaffold according to claim 1, characterized in that: The material of the response layer is at least one of piezoelectric material, piezoelectric ion gel, magnetocaloric material composite pyroelectric material, magnetostrictive material composite piezoelectric material, photodeformation material composite piezoelectric material, photovoltaic material, upconversion material composite photovoltaic material, photothermal material composite pyroelectric material or photocooling material composite pyroelectric material.

9. The multifunctional active nerve scaffold according to claim 1, characterized in that: The cells are at least one of Schwann cells, mesenchymal stem cells, neural crest stem cells, neural progenitor cells, PC12 cells, gene-overexpressed Schwann cells, gene-overexpressed mesenchymal stem cells, gene-overexpressed neural crest stem cells, gene-overexpressed neural progenitor cells, and Schwann cell-like cells derived from mesenchymal stem cells.

10. A method for preparing the multifunctional active nerve scaffold according to claim 1, characterized in that: The following steps are involved: preparing the response layer; Cross-linking to form a conductive layer below the responsive layer; cross-linking to form a base layer below the conductive layer; Cells are seeded on the response layer to form the multifunctional active neural scaffold.

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