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CN-116983040-B - Multilayer composite capacitance membrane, preparation method and application thereof in nerve repair stent

CN116983040BCN 116983040 BCN116983040 BCN 116983040BCN-116983040-B

Abstract

The invention belongs to the technical field of nerve grafts, and discloses a multilayer composite capacitance membrane, a preparation method and application thereof in a nerve repair stent. The support consists of three layers, namely an A layer, a B layer and an A layer. The layer A is a two-sided interlayer and comprises a mixture of chitosan (or silk fibroin or acellular matrix), conductive polymer PEDOT: PSS and crosslinking agent polyethylene glycol diglycidyl ether (PEGDE). The B layer is an intermediate hydrogel interlayer, and comprises methacryloylated gelatin (Gel-MA). And (3) forming viscoelastic hydrogel bonding on the layers A and B after ultraviolet irradiation, curing and air-drying, and curling to form the composite capacitor bracket with the multilayer structure with the through middle cavity. The formed bracket has good electric conduction performance, mechanical performance and biological safety, can obviously and effectively induce the proliferation, migration and differentiation of RSC96 schwann cells and nerve line cells PC12, and provides good physical stable support and favorable microenvironment for repairing peripheral nerve injury.

Inventors

  • YANG YUMIN
  • LI TAO
  • GE YIFAN
  • JIANG YUHUI
  • ZHANG LUZHONG
  • TANG XIAOXUAN
  • Bian Taotao

Assignees

  • 南通大学

Dates

Publication Date
20260508
Application Date
20230818

Claims (10)

  1. 1. The preparation method of the multilayer composite capacitor film is characterized by comprising the following steps of: S1, adding a biological material, a conductive polymer and a cross-linking agent into deionized water, stirring in a water bath at 40-50 ℃ for 2-4 hours, adding acetic acid, continuing stirring in the water bath at 40-50 ℃ for 2-4 hours, carrying out ultrasonic treatment for 30-45 minutes, standing for 2-3 hours, and removing residual bubbles to obtain a mixed solution, wherein the biological material is one of chitosan, silk fibroin and acellular matrix, the conductive polymer is PEDOT PSS, and the cross-linking agent is polyethylene glycol diglycidyl ether; S2, uniformly wetting the mixed solution on an adhesive glass slide, placing the glass slide into an ultra-clean fume hood, air-drying for 8-12 hours, taking down a film layer, sequentially cleaning with NaOH and deionized water, and air-drying to obtain a film A; s3, dissolving the methacryloyl gelatin in the photoinitiated LAP solution to obtain a layer B solution, putting a layer A film after the layer B solution is coated on the layer A film, and curing by ultraviolet irradiation to obtain the multilayer composite capacitor film.
  2. 2. The preparation method of claim 1, wherein the biological material is chitosan, and the molecular weight is 200-300 kDa.
  3. 3. The preparation method according to claim 2, wherein in the step S1, the amount ratio of the biological material, the conductive polymer, the crosslinking agent, the deionized water and the acetic acid is (0.6-1.0) g, (0.02-0.06) g, (0.1-0.2) g, (10-20) mL and (0.1-0.2) mL.
  4. 4. The method according to claim 1, wherein the methacrylated gelatin has an amino substitution degree of 30-90+ -5% and a molecular weight of 100-200 Kda.
  5. 5. The method according to claim 4, wherein the concentration of the methacryloylated gelatin is 5 to 10%.
  6. 6. The preparation method of the film according to claim 1, wherein the film is taken out and washed by NaOH and deionized water in sequence, specifically, the film is soaked in 5-10% NaOH for 1-2 hours to remove residual acetic acid, and after the film is taken out, the film is soaked in deionized water, water is replaced every 30-60 minutes until the pH of a soaking solution is the same as the pH of the deionized water, and the residual NaOH and acetic acid are removed.
  7. 7. A multilayer composite capacitive film, characterized in that it is prepared by the preparation method of any one of claims 1 to 6, and is composed of a composite conductive biomaterial outer layer and a hydrogel interlayer.
  8. 8. The multilayer composite capacitive film of claim 7 wherein the composite conductive biomaterial outer layer is formed from chitosan and conductive polymer PEDOT: PSS chemically cross-linked by polyethylene glycol diglycidyl ether (PEGDE), and the hydrogel interlayer is methacryloylated gelatin.
  9. 9. Use of a multilayer composite capacitive membrane prepared by the preparation method of any one of claims 1 to 6 or a multilayer composite capacitive membrane of claim 7 or 8 in the preparation of a multilayer composite capacitive scaffold for nerve damage repair.
  10. 10. The multilayer composite capacitor bracket is characterized by being prepared from a multilayer composite capacitor film prepared by the preparation method of any one of claims 1-6 or a multilayer composite capacitor film of claim 7 or 8, wherein the preparation method comprises the steps of winding the multilayer composite capacitor film on a stainless steel round bar, coating methacrylic acid gelatin on a side opening, and curing by irradiation of an ultraviolet lamp to form the multilayer composite capacitor bracket with a through middle cavity.

Description

Multilayer composite capacitance membrane, preparation method and application thereof in nerve repair stent Technical Field The invention belongs to the technical field of nerve grafts, and relates to a multilayer composite capacitance membrane, a preparation method and application thereof in a nerve repair stent. Background Peripheral Nerve Injury (PNI) is a worldwide clinical problem, mainly due to transient or lifelong neurological dysfunction due to traction, cutting, firearm, compression, ischemia, etc. Damage to the nervous system typically results in transection of the nerve, disruption of communication between the neuronal cells and their supporting cells, and disruption of the blood nerve barrier. Unlike the central nerve, the peripheral nerve exhibits a certain regeneration ability after injury. While the inherent regeneration capacity of the peripheral nerve helps to relieve the surgical pressure on the peripheral nerve injury, the process of spontaneous regeneration capacity of the peripheral nerve is very limited and insufficient to support spontaneous regeneration after a long severe trauma such as a nerve gap after nerve cutting, to achieve complete functional recovery. Given the vast number of cases of peripheral nerve injury, surgical repair of peripheral nerve injury remains an urgent issue. Up to now, the nerve transplantation with the most definite curative effect is the autologous nerve transplantation, which is still used as a 'gold standard' for evaluating the effectiveness of various nerve grafts, but the defects of limited nerve sources, difficult matching of tissue structures and sizes, long-term denervation of a transplantation donor area, different transplantation curative effects and the like limit the wide development of the autologous nerve transplantation. Because of the need for long-term immunosuppressant application, the success rate of transplantation is low, and allogeneic nerve transplantation is also limited. These limitations necessitate the study of the application of the nerve substitutes. To overcome the drawbacks of autologous nerve grafting, artificial nerve catheterization is increasingly considered an alternative treatment. Nerve conduits (Nerve guide conduits, NGCs) are tissue engineering tubular structures made of natural and/or synthetic biopolymers with mechanical and biochemical properties required for nerve regeneration. NGCs can overcome the limitations of nerve grafting and suturing methods. NGCs act as bridges between damaged nerve endings and provide structural and nutritional support for both ends, supporting invasion of surrounding tissues and regeneration of axons along the catheter. Electrical signals have been shown to provide key bioactive cues, promote neurite extension and accelerate nerve function recovery. Current studies have shown that alternating or direct current electric fields promote growth and functional recovery of rodent sciatic nerve, while sustained electrical stimulation not only stimulates calcium activity and neurite growth in DRG neurons, but also promotes proliferation of schwann cells and up-regulation of neurotrophic factors. Among the numerous tissue engineering catheters, chitosan (or silk fibroin or acellular matrix) catheters have been used for peripheral nerve injury repair due to their biocompatibility, non-toxicity, hydrophilicity, degradability and high processability, but because of the poor conductivity of natural biomaterials, they cannot perfectly simulate the electrical signal oscillation transmission rhythm of autologous nerves, have poor signal response to injured downstream nerve cells, and cannot promote specific cell event sequences in time to influence the regeneration process. The conductive polymer, such as PEDOT: PSS, is used as a novel intelligent biological material, and has great potential in the biomedical application directions of tissue regeneration, drug delivery, nerve interfaces and the like in recent years because the conductive polymer can directly transmit electric stimulation and electrochemical signals to cells to regulate cell fate. Disclosure of Invention In view of the above, the present invention aims to provide a multilayer composite capacitor membrane, a preparation method and an application thereof in nerve repair scaffold, wherein the multilayer composite capacitor membrane provides good nutrition permeability and conductivity besides sufficient mechanical structural support, and has better promotion effect on peripheral nerve regeneration repair. The technical scheme provided by the invention is as follows: a preparation method of a multilayer composite capacitance film comprises the following steps: S1, adding a biological material, a conductive polymer and a cross-linking agent into deionized water, stirring in a water bath at 40-50 ℃ for 2-4 hours, adding acetic acid, continuing stirring in the water bath at 40-50 ℃ for 2-4 hours, carrying out ultrasonic treatment for 30-45 m