CN111744057B - Preparation method of integrally-formed magnetic induction nerve conduit - Google Patents
Preparation method of integrally-formed magnetic induction nerve conduit Download PDFInfo
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Abstract
本发明公开了一种一体化成型磁感应神经导管的制备方法，其包括以下步骤：利用PDMS模具制备一管道，在管道中放置一根圆柱状模具，并使圆柱状模具的外侧壁与管道的内侧壁之间留有间隙，使用微流控的方法在间隙内分别注入磁性纳米颗粒层和神经生长因子层，将磁性纳米颗粒层和神经生长因子层进行冻干，冻干后将圆柱状模具从神经生长因子层的内腔抽出，使用微流控的方法沿着神经生长因子层的内腔侧壁注入神经干细胞层，拆除PDMS模具得到神经导管。其目的是为了提供一种一体化成型磁感应神经导管的制备方法，通过该方法制得的神经导管同时具有磁感应颗粒、神经生长因子以及神经干细胞，能够很好地促进神经缺损的修复。
The invention discloses a method for preparing an integrated magnetic induction nerve conduit, which comprises the following steps: using a PDMS mold to prepare a pipe, placing a cylindrical mold in the pipe, and aligning the outer side wall of the cylindrical mold with the inner side of the pipe There is a gap between the walls, and the magnetic nanoparticle layer and the nerve growth factor layer are injected into the gap by a microfluidic method, and the magnetic nanoparticle layer and the nerve growth factor layer are freeze-dried. The lumen of the nerve growth factor layer was drawn out, and the neural stem cell layer was injected along the inner lumen of the nerve growth factor layer using a microfluidic method, and the PDMS mold was removed to obtain a nerve conduit. The purpose is to provide a method for preparing an integrated magnetic induction nerve conduit. The nerve conduit prepared by the method has magnetic induction particles, nerve growth factors and neural stem cells at the same time, which can well promote the repair of nerve defects.
Description
Technical Field
The invention relates to the field of medical instruments, in particular to a nerve conduit for repairing nerve defects.
Background
The treatment of nerve defects caused by serious peripheral nerve injury is always a clinical problem. The treatment currently varies according to the distance of the defect. When the defect distance is short, the broken end suturing method is often adopted directly. When the defect distance is longer, alternative treatment is required. At present, autologous nerve transplantation is the "gold standard", but has the problems of donor site damage, functional impairment and the like. The method adopting the tissue engineering nerve conduit has greatly progressed in recent years, and partial research shows that the treatment effect of the nerve conduit can approach autologous nerve transplantation. Studies have shown that the following three methods are helpful in the repair of neurological defects: (1) the magnetic induction particles are added into the nerve conduit to play a magnetic induction effect, so that the nerve repair and regeneration promoting effect of the nerve conduit can be effectively promoted; (2) factors such as Nerve Growth Factor (NGF) can effectively promote the repair of peripheral nerve injury; (3) the nerve stem cells are added into the nerve conduit to promote the repair of peripheral nerve injury. However, the conventional nerve conduit cannot combine the above three methods.
Disclosure of Invention
The invention aims to solve the technical problem of providing a preparation method of an integrally formed magnetic induction nerve conduit, wherein the nerve conduit prepared by the method simultaneously has magnetic induction particles, nerve growth factors and nerve stem cells, and can well promote the repair of nerve defects.
The invention discloses a preparation method of an integrally formed magnetic induction nerve conduit, which comprises the following steps:
a pipe is prepared by using the PDMS mold,
placing a cylindrical mold in the pipeline, and leaving a gap between the outer side wall of the cylindrical mold and the inner side wall of the pipeline,
respectively injecting a magnetic nanoparticle layer and a nerve growth factor layer into the gap by a microfluidic method, wherein the magnetic nanoparticle layer is injected along the inner side wall of the pipeline, the nerve growth factor layer is injected along the outer side wall of the cylindrical mold,
freeze-drying the magnetic nanoparticle layer and the nerve growth factor layer, extracting the cylindrical mold from the inner cavity of the nerve growth factor layer after freeze-drying,
injecting a neural stem cell layer along the inner cavity side wall of the nerve growth factor layer by using a microfluidic method,
and (4) removing the PDMS mold to obtain the nerve conduit.
The invention relates to a preparation method of an integrally formed magnetic induction nerve conduit, wherein a magnetic nanoparticle layer comprises magnetic nanoparticles and a material coated outside the magnetic nanoparticles, and the material coated outside the magnetic nanoparticles is at least one of PLGA, chitosan and GelMA.
The invention discloses a preparation method of an integrated magnetic induction nerve conduit, wherein a nerve growth factor layer is a solution containing nerve growth factors, BSA (bovine serum albumin) and a water-soluble material, the nerve growth factors are at least one of HIF-1 alpha and rapamycin, and the water-soluble material is at least one of PVA (polyvinyl acetate), silk fibroin and type I collagen.
The invention discloses a preparation method of an integrated forming magnetic induction nerve conduit, wherein a nerve stem cell layer is hydrogel containing nerve stem cells, and the hydrogel is at least one of PEG, gelatin and collagen.
The invention discloses a preparation method of an integrally-molded magnetic induction nerve conduit, wherein when a pipeline is prepared by using a PDMS mold, two PDMS templates are adopted, a chemical etching method is used for manufacturing a strip-shaped groove on each PDMS template, then the two PDMS templates are subjected to oxygen beating in a plasma cleaning machine for 30 seconds to destroy the silicon-oxygen bond of the PDMS templates, then the two PDMS templates are tightly bonded under the condition that the two strip-shaped grooves correspond to each other to obtain the PDMS mold, and the pipeline is composed of the two strip-shaped grooves.
The invention discloses a preparation method of an integrated forming magnetic induction nerve conduit, wherein when a cylindrical mold is placed in a pipeline, the cylindrical mold and the pipeline are coaxially arranged.
The preparation method of the integrally formed magnetic induction nerve conduit is different from the prior art in that the nerve conduit is manufactured by a micro-fluidic control method and comprises an outer layer, a middle layer and an inner layer, wherein the outer layer is a magnetic nanoparticle layer, the middle layer is a nerve growth factor layer, the inner layer is a nerve stem cell layer, and the magnetic nanoparticles, the nerve growth factor and the nerve stem cells in the nerve conduit can promote the repair of nerve defects, so the effect is better.
The invention will be further explained with reference to the drawings.
Drawings
FIG. 1 is a schematic view of a method for manufacturing an integrally formed magnetically inductive nerve conduit according to the present invention;
fig. 2 is a schematic cross-sectional view of a nerve conduit prepared using the present invention.
Detailed Description
As shown in fig. 1, the method for preparing the integrally formed magnetic induction nerve conduit of the present invention comprises the following steps:
a pipe is prepared using the PDMS mold 4,
placing a cylindrical mold in the pipeline, and leaving a gap between the outer side wall of the cylindrical mold and the inner side wall of the pipeline,
respectively injecting a magnetic nanoparticle layer 5 and a nerve growth factor layer 6 into the gap by using a microfluidic method, wherein the magnetic nanoparticle layer 5 is injected along the inner side wall of the pipeline, the nerve growth factor layer 6 is injected along the outer side wall of the cylindrical mould,
freeze-drying the magnetic nanoparticle layer 5 and the nerve growth factor layer 6, extracting the cylindrical mold from the inner cavity of the nerve growth factor layer 6 after freeze-drying,
injecting the neural stem cell layer 7 along the inner cavity side wall of the nerve growth factor layer 6 by using a microfluidic method,
and (4) removing the PDMS mold 4 to obtain the nerve conduit.
The invention discloses a preparation method of an integrally formed magnetic induction nerve conduit, wherein a magnetic nanoparticle layer 5 comprises magnetic nanoparticles 501 and a material coated outside the magnetic nanoparticles 501, and the material coated outside the magnetic nanoparticles 501 is at least one of PLGA, chitosan and GelMA.
The invention discloses a preparation method of an integrated magnetic induction nerve conduit, wherein a nerve growth factor layer 6 is a solution containing nerve growth factors, BSA (bovine serum albumin) and a water-soluble material, the nerve growth factors are at least one of HIF-1 alpha and rapamycin, and the water-soluble material is at least one of PVA (polyvinyl acetate), silk fibroin and type I collagen.
The invention discloses a preparation method of an integrated forming magnetic induction nerve conduit, wherein a nerve stem cell layer 7 is hydrogel containing nerve stem cells, and the hydrogel is at least one of PEG, gelatin and collagen.
As shown in fig. 1 in conjunction with fig. 2, the magnetic nanoparticle layer 5 is injected into the gap between the pipe and the cylindrical mold using the first injector 1, and the magnetic nanoparticle layer 5 becomes the outer layer of the nerve conduit since the magnetic nanoparticle layer 5 is injected along the inner sidewall of the pipe; injecting the nerve growth factor layer 6 into the gap between the pipeline and the cylindrical mold by using the second injector 2, wherein the nerve growth factor layer 6 becomes the middle layer of the nerve conduit because the nerve growth factor layer 6 is injected along the outer side wall of the cylindrical mold; the nerve stem cell layer 7 is injected along the lumen sidewall of the nerve growth factor layer 6 using the third syringe 3, and thus the nerve stem cell layer 7 becomes an inner layer of the nerve conduit.
The invention relates to a preparation method of an integrally molded magnetic induction nerve conduit, wherein when a pipeline is prepared by using a PDMS mold 4, two PDMS templates are adopted, a chemical etching method is used for manufacturing a strip-shaped groove on each PDMS template, then the two PDMS templates are subjected to oxygen beating in a plasma cleaning machine for 30 seconds to destroy the silicon-oxygen bond of the PDMS templates, then the two PDMS templates are tightly bonded under the condition that the two strip-shaped grooves correspond to each other to obtain the PDMS mold 4, and the two strip-shaped grooves form the pipeline.
The invention discloses a preparation method of an integrated forming magnetic induction nerve conduit, wherein when a cylindrical mold is placed in a pipeline, the cylindrical mold and the pipeline are coaxially arranged.
As shown in fig. 2, the nerve conduit is manufactured by a micro-fluidic control method, and comprises an outer layer, a middle layer and an inner layer, wherein the outer layer is a magnetic nanoparticle layer 5, the middle layer is a nerve growth factor layer 6, and the inner layer is a nerve stem cell layer 7, and the magnetic nanoparticles 501, the nerve growth factors and the nerve stem cells in the nerve conduit can promote the repair of nerve defects, so that the effect is better.
The above-mentioned embodiments are merely illustrative of the preferred embodiments of the present invention, and do not limit the scope of the present invention, and various modifications and improvements of the technical solution of the present invention by those skilled in the art should fall within the protection scope defined by the claims of the present invention without departing from the spirit of the present invention.
Claims (3)
1. The preparation method of the integrally formed magnetic induction nerve conduit is characterized by comprising the following steps of:
a pipe is prepared by using the PDMS mold,
placing a cylindrical mold in the pipeline, and leaving a gap between the outer side wall of the cylindrical mold and the inner side wall of the pipeline,
respectively injecting a magnetic nanoparticle layer and a nerve growth factor layer in the gap by using a microfluidic method, wherein the magnetic nanoparticle layer is injected along the inner side wall of the pipeline, the nerve growth factor layer is injected along the outer side wall of the cylindrical mold, the magnetic nanoparticle layer comprises magnetic nanoparticles and a material wrapping the magnetic nanoparticles, the material wrapping the magnetic nanoparticles is at least one of PLGA, chitosan and GelMA, the nerve growth factor layer is a solution containing nerve growth factor, BSA and a water-soluble material, the nerve growth factor is at least one of HIF-1 alpha and rapamycin, and the water-soluble material is at least one of PVA, silk fibroin and type I collagen,
freeze-drying the magnetic nanoparticle layer and the nerve growth factor layer, extracting the cylindrical mold from the inner cavity of the nerve growth factor layer after freeze-drying,
injecting a neural stem cell layer along the inner cavity side wall of the nerve growth factor layer by using a microfluidic method, wherein the neural stem cell layer is hydrogel containing neural stem cells, and the hydrogel is at least one of PEG, gelatin and collagen,
and (4) removing the PDMS mold to obtain the nerve conduit.
2. The method for preparing an integrally formed magnetically inductive nerve conduit as claimed in claim 1, wherein: when the pipeline is prepared by using the PDMS mold, two PDMS templates are adopted, a strip-shaped groove is formed in each PDMS template by using a chemical etching method, then the two PDMS templates are subjected to oxygen beating in a plasma cleaning machine for 30 seconds to destroy a silicon-oxygen bond of the PDMS templates, the two PDMS templates are tightly bonded under the condition that the two strip-shaped grooves correspond to each other, the PDMS mold is obtained, and the pipeline is composed of the two strip-shaped grooves.
3. The method for preparing an integrally formed magnetically inductive nerve conduit as claimed in claim 2, wherein: when a cylindrical mold is placed in the pipeline, the cylindrical mold is arranged coaxially with the pipeline.
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| CN115382022B (en) * | 2022-05-12 | 2023-08-22 | 大连工业大学 | Continuous preparation method of nerve conduit material based on sodium alginate and krill protein |
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| CN102048595B (en) * | 2010-12-21 | 2013-05-15 | 东华大学 | Degradable nerve guide with highly oriented tube-in-tube structure and preparation method thereof |
| CN102343114A (en) * | 2011-10-24 | 2012-02-08 | 南通大学 | Tissue engineering nerve graft and application thereof |
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| CN1843307A (en) * | 2005-04-07 | 2006-10-11 | 首都医科大学北京神经科学研究所 | Double-layer artificial nerve catheter and preparation method thereof |
| CN101439205A (en) * | 2008-12-29 | 2009-05-27 | 上海市第六人民医院 | Nerve catheter as well as preparation method and application thereof |
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