Anti-inflammatory-regenerative porous nerve conduit based on hot-melt template technology and preparation method and application of anti-inflammatory-regenerative porous nerve conduit

Abstract

The invention discloses an anti-inflammatory-regenerative porous nerve conduit based on a hot-melt template technology and a preparation method and application thereof. The preparation method comprises the following steps: (1) preparing a plurality of gelatin cylinders and a cylindrical pouring mold with a fine needle array at the bottom; 2) preparing nano manganese dioxide and a SilMA solution loaded with manganese dioxide by a hydrothermal method; (3) respectively inserting the gelatin cylinders into the needle-shaped array of a mold, pouring a composite SilMA solution, carrying out UV curing and freezing demolding, and removing the internal gelatin cylinders in combination with a hot melting technology to obtain a porous catheter; and 4) carrying out in-situ auto-polymerization on the surface of the conduit to form a dopamine coating, and dipping and adsorbing a nerve growth promoting drug to obtain the required anti-inflammatory-regenerative porous nerve conduit material. The nerve conduit can promote neural stem cells, internal ingrowth and neural differentiation, meanwhile, neuroinflammation is prevented, and a brand new treatment strategy is provided for nerve injury.

Description

Technical Field

[0001] The invention relates to a preparation technology of a nerve damage repair material, and in particular to an anti-inflammatory-regenerative porous nerve conduit based on a hot melt template technology, and a preparation method and application thereof. Background Art

[0002] The nervous system is composed of the brain, spinal cord, and peripheral nerves, which work together to transmit and process information inside and outside the body. The brain, as the core of the nervous system, is responsible for higher-level neural activities such as thinking, judgment, and memory; the spinal cord is like an information highway, connecting the brain and peripheral nerves, transmitting motor commands and sensory information; the peripheral nerves are like a huge network, distributed in every corner of the body, sensing external stimuli and transmitting them to the brain. Among them, the peripheral nerves are very fragile and are often damaged in daily life due to traffic accidents, surgery, and other reasons. Once the nerves are damaged, it will affect the brain's ability to communicate with muscles and organs. Mild nerve damage has the possibility of self-repair, while those with severe nerve damage have a lower possibility of self-repair.

[0003] At present, the means used clinically to treat nerve damage mainly include drug therapy, electrical stimulation therapy, etc., but their actual effects are not significant. In recent years, with the rapid development of tissue engineering technology, materials for nerve repair have gradually been developed. However, the nerve repair scenario is complex, and there are problems such as neuroinflammation, lack of neural stem cells, and poor directional differentiation. Most of the current nerve repair materials cannot fully meet the above-mentioned actual clinical scenario needs. Therefore, the development of a material that has anti-inflammatory, supplements stem cell sources, promotes cell adhesion and migration, and induces directional differentiation of cells into neurons is of great significance for nerve injury repair.

[0004] In response to the above problems, the present invention proposes an anti-inflammatory-regenerative porous nerve conduit based on hot melt template technology and a preparation method thereof. Through a combination of processes such as casting, hot melt, and dipping, components / structures such as conduits, anti-inflammatory materials, adhesion-promoting coatings, and neural differentiation-promoting materials are introduced into the nerve repair material, providing a new and efficient treatment strategy for nerve injury repair. Summary of the invention

[0005] The purpose of the present invention is to provide an anti-inflammatory-regenerative porous nerve conduit based on hot melt template technology and its preparation method and application in view of the problems that traditional nerve damage repair materials are not effective. The nerve conduit can promote neural stem cells, internal growth, and neurogenic differentiation, while preventing neuroinflammation, providing a new treatment strategy for nerve damage.

[0006] The present invention is implemented by the following technical solution, which specifically includes the following steps:

[0007] A method for preparing an anti-inflammatory-regenerative porous nerve conduit based on a hot melt template technology comprises the following steps:

[0008] 1) preparing a number of physically gelled gelatin cylinders, and designing a cylindrical casting mold with an array of fine needles at the bottom;

[0009] 2) preparing nano manganese dioxide and a methacrylylated silk fibroin (SilMA) solution loaded with the manganese dioxide by a hydrothermal method, and cooling the SilMA solution loaded with manganese dioxide to room temperature;

[0010] 3) inserting the gelatin cylinders obtained in step 1) into the corresponding fine needles of the cylindrical casting mold, and casting the SilMA solution loaded with manganese dioxide, UV curing, freezing demolding, and combining hot melt technology to remove the gelatin cylinders on the fine needles to obtain a porous catheter;

[0011] 4) In situ self-polymerization of dopamine coating is carried out on the surface of the porous conduit, and the anti-inflammatory-regenerative porous nerve conduit material is obtained after impregnation and adsorption of nerve growth-promoting drugs. After culturing the material with neural stem cells in vitro, the material can be implanted into the nerve injury area for application.

[0012] Furthermore, in step 1), during the preparation of each physically gelled gelatin cylinder, the gelatin solution concentration is 5-10wt%, the dissolution temperature is 40-60°C, and the gelatin cylinder is obtained by casting and cooling process, and the size of the gelatin cylinder is: 0.5-1mm in diameter and 0.5-2cm in length;

[0013] The inner diameter of the cylindrical casting mold is 1-3 mm, the length is 0.5-2 cm, the diameter of the fine needles is 0.1-0.2 mm, the length is 0.1-0.3 cm, and the number is 3-10.

[0014] Furthermore, in step 2), during the hydrothermal process for preparing nano manganese dioxide, a mixed solution of sodium dodecylbenzene sulfonate and manganese chloride is prepared, a potassium permanganate solution is added dropwise to the mixed solution under stirring conditions, and then high-speed centrifugal washing is performed to obtain manganese dioxide nanoparticles with a particle size of 20-30 nm; wherein the concentration of sodium dodecylbenzene sulfonate is 1-2 g / L, the concentration of manganese chloride is 100-200 g / L, the concentration of potassium permanganate is 15-80 g / L, the reaction temperature is 80-100° C., the stirring rate is 800-1000 r / min, and the reaction time is 3-8 h.

[0015] Furthermore, in step 2), the SilMA solution loaded with manganese dioxide contains a photoinitiator, the SilMA substitution degree is 30-60%, the concentration is 10-20wt%, the photoinitiator is one or more of LAP, Irgacure, and photoinitiator 2959, the concentration is 0.25-0.5wt%, the concentration of the loaded manganese dioxide is 0.05-0.25wt%, and the solvent is PBS.

[0016] Furthermore, in step 3), during the UV curing process, the UV wavelength is 405 nm, the power is 20-30 W, and the time is 5-12 min;

[0017] During the freezing demoulding process, freezing is performed at 0 to -80°C for 5 to 60 minutes;

[0018] During the thermal dissolution process, the sample is immersed in a deionized water solution at 35-50° C., with an oscillation condition of 30-80 r / min for 30 min-10 h.

[0019] Further, in step 4), during the in-situ self-polymerization process, the dopamine hydrochloride solution is 2-8 mg / ml, the solvent is Tris-HCl solution, the pH value is 8.5, and the polymerization time is 4-12 h;

[0020] During the immersion adsorption process, the nerve growth promoting drug is one or more of nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), neurotrophic factor 3 (NT-3), neurotrophic factor 4 / 5 (NT-4 / 5), and neurotrophic factor 6 (NT-6), with a concentration of 100-500 ng / ml, an immersion time of 30 min-2 h, and a temperature of 4-8°C.

[0021] An anti-inflammatory and regenerative porous nerve conduit based on hot melt template technology is obtained by using any of the above-mentioned preparation methods.

[0022] A nerve damage repair material comprises the nerve guide.

[0023] Furthermore, the neural stem cell culture process is as follows: neural stem cells of generation P2-P5 are collected and cultured at 1-3×10 7 The cells are resuspended in a neural basal medium at a concentration of 100 / mL, and the anti-inflammatory-regenerative porous nerve conduit is immersed in the cell-containing medium, and then cultured continuously for 14-28 days in a constant temperature incubator at 37°C and 5% CO2, and the medium is changed every other day. When the nerve cells are observed to grow in the nerve conduit and differentiate into neurofilaments, the culture is terminated. Compared with the prior art, the present invention has the following advantages:

[0024] 1) The present invention uses gelatin, nano manganese dioxide, nerve growth promoting drugs, dopamine hydrochloride and the like as raw materials, and combines casting method, hot melt method, nanocomposite, in-situ UV light curing, in-situ self-polymerization, impregnation adsorption and other combined processes to prepare an anti-inflammatory-regenerative porous nerve conduit material based on hot melt template technology. The above raw material selection and process combination are original to the present invention.

[0025] 2) The present invention uses the casting method + hot melt method to successfully introduce an axial spatial conduit structure into the nerve conduit for cell migration and growth, which is beneficial to the repair of nerve damage. First, a physically gelled gelatin cylinder is prepared, fixed in a customized cylindrical casting mold with a fine needle array at the bottom, and then the gelatin cylinder is further wrapped in a cooled SilMA solution loaded with manganese dioxide by a casting method. After the SilMA solution loaded with manganese dioxide is gelled and formed, the hot melt method is further used to heat all the physically gelled gelatin cylinders to dissolve. After dissolution, the internal gelatin solution diffuses out of the SilMA gel matrix under oscillation conditions to form the required axial pore structure, which can be used as the main channel for cells to grow in the axial direction in the future, accelerating the repair of nerve damage. The above-mentioned axial pore structure is mainly obtained by controlling the characteristics of gelatin cooling and gelling, and heating and melting, that is, the pore structure is formed by using the physically gelled gelatin cylinder as a hot melt template. The pore structure obtained by this method is accurate in size and will not damage the SilMA matrix loaded with manganese dioxide. It is also simple to operate and has high practical value.

[0026] 3) The nerve conduit in the present invention has a composite structure of nano manganese dioxide particles + axial pipe structure + polydopamine adhesion coating from the inside to the outside. When the nerve conduit is in use, the above structures can work together to play specific roles: first, the polydopamine coating structure of the outer layer has the function of adsorbing proteins, etc., which can promote the efficient adhesion of subsequent stem cells on the surface of the nerve conduit; and the axial pipe structure provides an axial internal 3D growth space for the nerve conduit, and cells can quickly grow into the interior of the nerve conduit, and the cell growth direction has a directional (axial) orientation, and the two ends of the damaged nerve can be efficiently connected later; the nano manganese dioxide particles composited inside the SilMA matrix loaded with manganese dioxide can slowly release manganese ions to inhibit neuroinflammation and provide a non-inflammatory microenvironment for nerve damage repair. The design synergy of the above structures provides the possibility for nerve repair.

[0027] 4) The nerve conduit material in the present invention has the functions of high biocompatibility, degradability, anti-inflammatory, cell adhesion / migration, and neural differentiation, meeting the complex regeneration and repair needs of nerve damage. First of all, the raw materials selected in the present invention are all biocompatible and easily degradable, which ensures that the whole material has high biocompatibility, and completely degrades during the process of nerve damage repair, without the need for secondary surgery; secondly, the internal composite nano manganese dioxide particles can effectively prevent and treat neuroinflammation by releasing manganese ions, providing a good microenvironment guarantee for nerve repair; and the axial channel structure inside the nerve conduit and the polydopamine coating structure on the surface simultaneously ensure the adhesion of cells on the surface and the axial migration effect, solving the problem of lack of nerve damage repair cells, and providing a large number of cell sources for nerve damage repair; after supplementing the cell source, the nerve growth promoting drug loaded in the SilMA matrix loaded with manganese dioxide further induces its neural differentiation. It is through the organic synergy between the above functions of the nerve conduit that the typical problems in the process of nerve injury are solved.

[0028] 5) In the present invention, nerve conduit materials of different sizes can be prepared through the above-mentioned combined process and raw materials, which can meet the needs of different patients and different diseases. Nerve conduit materials of different sizes can be prepared by customizing molds of specific sizes to connect / repair nerve damage areas of different patients. The above-mentioned technology is convenient and universal. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 The figure is a schematic diagram of the preparation process of the anti-inflammatory-regenerative porous nerve conduit material based on the hot melt template technology of the present invention. DETAILED DESCRIPTION

[0030] The present invention is further described below with reference to specific examples and accompanying drawings.

[0031] Comparative Example 1:

[0032] 1) preparing nano manganese dioxide with a particle size of 20-30 nm by a hydrothermal method (preparing a mixed solution of sodium dodecylbenzene sulfonate and manganese chloride, adding potassium permanganate solution dropwise to the above solution under high-speed stirring, reacting at 80° C. for 4 h under stirring at 800 r / min, and then centrifuging and washing at high speed to obtain manganese dioxide nanoparticles, wherein the concentration of sodium dodecylbenzene sulfonate is 1 g / L, the concentration of manganese chloride is 150 g / L, and the concentration of potassium permanganate is 30 g / L) and a SilMA solution (SilMA substitution degree is 60%, concentration is 10 wt%, the photoinitiator is LAP, concentration is 0.25 wt%, and the solvent is PBS) loaded with manganese dioxide (concentration is 0.1 wt%), and cooling the SilMA mixed solution to room temperature;

[0033] 3) The composite SilMA solution was poured into the mold, UV-cured (UV wavelength of 405 nm, power of 20 W, time of 6 min), frozen demoulding (freezing at -80 ° C for 5 min), and the internal gelatin cylinder was removed by hot melt technology (immersion in 50 ° C deionized water solution, 50 r / min oscillation condition, time of 2 h) to obtain a porous catheter;

[0034] 4) In situ self-polymerization of dopamine coating on the surface of the catheter (dopamine hydrochloride solution is 6 mg / ml, solvent is Tris-HCl solution, pH value is 8.5, polymerization time is 8 h), and NGF is absorbed by immersion (concentration is 300 ng / ml, time is 30 min, temperature is 4°C) to obtain the desired anti-inflammatory-regenerative nerve catheter material, which is then cultured in vitro with neural stem cells (P2 neural stem cells are collected and 1×10 7 The cells were resuspended in a neural basal medium at a concentration of 100 / mL, and the anti-inflammatory-regenerative porous nerve conduit was immersed in the medium containing the cells, and then cultured continuously for 14 days in a constant temperature incubator at 37°C and 5% CO2. The medium was changed every other day. After the nerve cells were observed to grow in the conduit and differentiate into neurofilaments, the culture was terminated and the conduit could be implanted in the nerve injury area for application.

[0035] The extract prepared from the above-mentioned nerve conduit material was co-cultured with neural stem cells for 7 days for CCK-8 detection, and the results showed that the cell survival rate was about 88.3%; after the nerve conduit material was directly co-cultured with neural stem cells for 14 days, the surface of the nerve conduit was covered with neuronal cells, and after being implanted into the seminal nerve injury model for 8 weeks, no neuroinflammation occurred, but tissue sections showed that the damaged nerves were not completely repaired.

[0036] Embodiment 1:

[0037] 1) Prepare 7wt% gelatin solution by dissolving at 60℃, and obtain the required gelatin cylinder by combining casting method and cooling process. The size of the obtained gelatin cylinder is: 1mm in diameter and 2cm in length. At the same time, design a cylindrical casting mold with a fine needle array at the bottom (the inner diameter of the cylindrical casting mold is 3mm and the length is 2cm. The diameter of the fine needle is 0.1mm and the length is 0.3cm. The number is 4 according to the diameter of the mold);

[0038] 2) preparing nano manganese dioxide with a particle size of 20-30 nm by a hydrothermal method (preparing a mixed solution of sodium dodecylbenzene sulfonate and manganese chloride, adding potassium permanganate solution dropwise to the above solution at 800 r / min and 80° C., reacting for 4 hours, and then washing by high-speed centrifugation to obtain the desired manganese dioxide nanoparticles, wherein the concentration of sodium dodecylbenzene sulfonate is 1 g / L, the concentration of manganese chloride is 150 g / L, and the concentration of potassium permanganate is 30 g / L) and a SilMA solution loaded with manganese dioxide (concentration is 0.1 wt%) (SilMA substitution degree is 60%, concentration is 10 wt%, photoinitiator is LAP, concentration is 0.25 wt%, solvent is PBS), and cooling the SilMA solution loaded with manganese dioxide to room temperature;

[0039] 3) The above four gelatin cylinders were respectively inserted into the needle array of the mold (arranged in a square), and the SilMA solution loaded with manganese dioxide was poured, UV-cured (UV wavelength of 405nm, power of 20W, time of 6min), frozen demolding (freezing at -80℃ for 5min), and combined with hot melt technology (immersion in 50℃ deionized water solution, 50r / min oscillation condition, time of 2h) to remove all gelatin cylinders inside the mold to obtain porous catheters;

[0040] 4) In situ self-polymerization of dopamine coating on the surface of the porous conduit (dopamine hydrochloride solution of 6 mg / ml, solvent of Tris-HCl solution, pH value of 8.5, polymerization time of 8 h), impregnation and adsorption of NGF (concentration of 300 ng / ml, time of 30 min, temperature of 4°C) to obtain an anti-inflammatory-regenerative porous nerve conduit material, which was cultured in vitro with neural stem cells (P2 neural stem cells were collected and 1×10 7 The cells were resuspended in a neural basal medium at a concentration of 100 / mL, and the anti-inflammatory-regenerative porous nerve conduit was immersed in the medium containing the cells, and then cultured continuously for 14 days in a constant temperature incubator at 37°C and 5% CO2. The medium was changed every other day. After the nerve cells were observed to grow in the conduit and differentiate into neurofilaments, the culture was terminated and the conduit could be implanted in the nerve injury area for application.

[0041] Compared with Comparative Example 1, the embodiment adds a process of preparing the axial pore structure by hot melt method, and constructs a pore structure for the nerve conduit. The extract prepared from the above nerve conduit material was co-cultured with neural stem cells for 7 days for CCK-8 detection, and the results showed that the cell survival rate was about 85.6%; after the nerve conduit material was directly co-cultured with neural stem cells for 14 days, the surface of the nerve conduit and the pore structure were full of neuronal cells, and after being implanted into the myelin nerve injury model for 8 weeks, no neuroinflammation occurred, and the tissue sections showed that the damaged nerve repair effect was good.

[0042] Embodiment 2:

[0043] 1) Prepare 7wt% gelatin solution by dissolving at 60℃, and obtain the required gelatin cylinder by combining casting method and cooling process. The size of the obtained gelatin cylinder is: 1mm in diameter and 2cm in length. At the same time, design a cylindrical casting mold with a fine needle array at the bottom (the inner diameter of the cylindrical casting mold is 3mm and the length is 2cm. The diameter of the fine needle is 0.1mm and the length is 0.3cm. The number is 4 according to the diameter of the mold);

[0044] 2) preparing a SilMA solution (SilMA substitution degree is 60%, concentration is 10wt%, photoinitiator is LAP, concentration is 0.25wt%, solvent is PBS), and cooling the SilMA mixed solution to room temperature;

[0045] 3) The above four gelatin cylinders were respectively inserted into the needle array of the mold (arranged in a square), and SilMA solution was poured, UV-cured (UV wavelength of 405 nm, power of 20 W, time of 6 min), frozen demolding (freezing at -80 ° C for 5 min), and combined with hot melt technology (immersion in 50 ° C deionized water solution, 50 r / min oscillation condition, time of 2 h) to remove all gelatin cylinders inside the mold to obtain a porous catheter;

[0046] 4) In situ self-polymerization of dopamine coating on the surface of the porous conduit (dopamine hydrochloride solution of 6 mg / ml, solvent of Tris-HCl solution, pH value of 8.5, polymerization time of 8 h), impregnation and adsorption of NGF (concentration of 300 ng / ml, time of 30 min, temperature of 4°C) to obtain a porous nerve conduit material, which was cultured in vitro with neural stem cells (P2 neural stem cells were collected and 1×10 7 The cells were resuspended in a neural basal medium at a concentration of 100 / mL, and the anti-inflammatory-regenerative porous nerve conduit was immersed in the medium containing the cells, and then cultured continuously for 14 days in a constant temperature incubator at 37°C and 5% CO2. The medium was changed every other day. After the nerve cells were observed to grow in the conduit and differentiate into neurofilaments, the culture was terminated and the conduit could be implanted in the nerve injury area for application.

[0047] Compared with Example 1, this example reduces the addition of nano manganese dioxide. The extract prepared from the above-mentioned nerve conduit material was co-cultured with neural stem cells for 7 days for CCK-8 detection, and the results showed that the cell survival rate was about 91.4%; after the nerve conduit material was directly co-cultured with neural stem cells for 14 days, the surface and pore structure of the nerve conduit were full of neuronal cells, and after being implanted into the myelin nerve injury model for 8 weeks, the tissue section showed that local inflammation occurred and the damaged nerve was not completely repaired.

[0048] Embodiment 3:

[0049] 1) Prepare 7wt% gelatin solution by dissolving at 60℃, and obtain the required gelatin cylinder by combining casting method and cooling process. The size of the obtained gelatin cylinder is: 1mm in diameter and 2cm in length. At the same time, design a cylindrical casting mold with a fine needle array at the bottom (the inner diameter of the cylindrical casting mold is 3mm and the length is 2cm. The diameter of the fine needle is 0.1mm and the length is 0.3cm. The number is 4 according to the diameter of the mold);

[0050] 2) preparing nano manganese dioxide with a particle size of 20-30 nm by a hydrothermal method (preparing a mixed solution of sodium dodecylbenzene sulfonate and manganese chloride, adding potassium permanganate solution dropwise to the above solution at 800 r / min and 80° C., reacting for 4 hours, and then washing by high-speed centrifugation to obtain the desired manganese dioxide nanoparticles, wherein the concentration of sodium dodecylbenzene sulfonate is 1 g / L, the concentration of manganese chloride is 150 g / L, and the concentration of potassium permanganate is 30 g / L) and a SilMA solution loaded with manganese dioxide (concentration is 0.1 wt%) (SilMA substitution degree is 60%, concentration is 10 wt%, photoinitiator is LAP, concentration is 0.25 wt%, solvent is PBS), and cooling the SilMA solution loaded with manganese dioxide to room temperature;

[0051] 3) The above four gelatin cylinders were respectively inserted into the needle array of the mold (arranged in a square), and the SilMA solution loaded with manganese dioxide was poured, UV-cured (UV wavelength of 405nm, power of 20W, time of 6min), frozen demolding (freezing at -80℃ for 5min), and combined with hot melt technology (immersion in 50℃ deionized water solution, 50r / min oscillation condition, time of 2h) to remove all gelatin cylinders inside the mold to obtain porous catheters;

[0052] 4) The porous conduit was immersed in NGF (concentration of 300 ng / ml, time of 30 min, temperature of 4°C) to obtain a nerve conduit material, which was then cultured with neural stem cells in vitro (P2 neural stem cells were collected and 1×10 7 The cells were resuspended in a neural basal medium at a concentration of 100 / mL, and the anti-inflammatory-regenerative porous nerve conduit was immersed in the medium containing the cells, and then cultured continuously for 14 days in a constant temperature incubator at 37°C and 5% CO2. The medium was changed every other day. After the nerve cells were observed to grow in the conduit and differentiate into neurofilaments, the culture was terminated and the conduit could be implanted in the nerve injury area for application.

[0053] Compared with Example 1, the design of the polydopamine coating is reduced in this example. The extract of the above-mentioned nerve conduit material was prepared, and the extract was co-cultured with neural stem cells for 7 days for CCK-8 detection. The results showed that the cell survival rate was about 86.4%; after the nerve conduit material was directly co-cultured with neural stem cells for 14 days, the surface of the nerve conduit and some areas in the pore structure were still not covered with cells. After implanting it into the myelin nerve injury model for 8 weeks, no neuroinflammation occurred, but the tissue section showed that the damaged nerve was not completely repaired, and the effect was between Examples 1 and 2.

Claims

1. A method for preparing an anti-inflammatory-regenerative porous nerve conduit based on hot melt template technology, characterized in that: The following steps are involved: 1) preparing a number of physically gelled gelatin cylinders, and designing a cylindrical casting mold with an array of fine needles at the bottom; 2) preparing nano manganese dioxide and a methacrylylated silk fibroin (SilMA) solution loaded with the manganese dioxide by a hydrothermal method, and cooling the SilMA solution loaded with manganese dioxide to room temperature; 3) inserting the gelatin cylinders obtained in step 1) into the corresponding fine needles of the cylindrical casting mold, and casting the SilMA solution loaded with manganese dioxide, UV curing, freezing demolding, and combining hot melt technology to remove the gelatin cylinders on the fine needles to obtain a porous catheter; 4) In situ self-polymerizing a dopamine coating on the surface of the porous conduit, and impregnating and adsorbing a nerve growth-promoting drug to obtain an anti-inflammatory-regenerative porous nerve conduit material.

2. The method for preparing an anti-inflammatory-regenerative porous nerve conduit based on hot melt template technology according to claim 1, characterized in that: In step 1), during the preparation of each physically gelled gelatin cylinder, the gelatin solution concentration is 5-10wt%, the dissolution temperature is 40-60°C, and the gelatin cylinder is obtained by casting and cooling process. The size of the gelatin cylinder is: 0.5-1mm in diameter and 0.5-2cm in length; The inner diameter of the cylindrical casting mold is 1-3 mm, the length is 0.5-2 cm, the diameter of the fine needles is 0.1-0.2 mm, the length is 0.1-0.3 cm, and the number is 3-10.

3. The method for preparing an anti-inflammatory-regenerative porous nerve conduit based on hot melt template technology according to claim 1, characterized in that: In step 2), during the hydrothermal method for preparing nano manganese dioxide, a mixed solution of sodium dodecylbenzene sulfonate and manganese chloride is prepared, a potassium permanganate solution is added dropwise to the mixed solution under stirring conditions, and then high-speed centrifugal washing is performed to obtain manganese dioxide nanoparticles with a particle size of 20-30 nm; The concentration of sodium dodecyl sulfate is 1-2 g / L, the concentration of manganese chloride is 100-200 g / L, the concentration of potassium permanganate is 15-80 g / L, the reaction temperature is 80-100° C., the stirring rate is 800-1000 r / min, and the reaction time is 3-8 h.

4. The method for preparing an anti-inflammatory-regenerative porous nerve conduit based on hot melt template technology according to claim 1, characterized in that: In step 2), the SilMA solution loaded with manganese dioxide contains a photoinitiator, the SilMA substitution degree is 30-60%, the concentration is 10-20wt%, the photoinitiator is one or more of LAP, Irgacure, and photoinitiator 2959, the concentration is 0.25-0.5wt%, the loaded manganese dioxide concentration is 0.05-0.25wt%, and the solvent is PBS.

5. The method for preparing an anti-inflammatory-regenerative porous nerve conduit based on hot melt template technology according to claim 1, characterized in that: In step 3), during the UV curing process, the UV wavelength is 405nm, the power is 20-30W, and the time is 5-12min; During the freezing demoulding process, freezing is performed at 0 to -80°C for 5 to 60 minutes; During the thermal dissolution process, the sample is immersed in a deionized water solution at 35-50° C., with an oscillation condition of 30-80 r / min for 30 min-10 h.

6. The method for preparing an anti-inflammatory-regenerative porous nerve conduit based on hot melt template technology according to claim 1, characterized in that: In step 4), during the in-situ self-polymerization process, the dopamine hydrochloride solution is 2-8 mg / ml, the solvent is Tris-HCl solution, the pH value is 8.5, and the polymerization time is 4-12 h; During the immersion adsorption process, the nerve growth promoting drug is one or more of nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), neurotrophic factor 3 (NT-3), neurotrophic factor 4 / 5 (NT-4 / 5), and neurotrophic factor 6 (NT-6), with a concentration of 100-500 ng / ml, an immersion time of 30 min-2 h, and a temperature of 4-8°C.

7. An anti-inflammatory and regenerative porous nerve conduit based on hot melt template technology, characterized in that: The method is adopted as claimed in any one of claims 1 to 6.

8. A nerve damage repair material, characterized in that: Comprising the nerve guide as claimed in claim 7.

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