JP2011201793A - Fiber-like structure and method for manufacturing fiber-like structure - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a fiber-like structure capable of introducing viruses selectively and efficiently to a specific part, and a method for manufacturing the fiber-like structure.SOLUTION: The fiber-like structure is characterized by being bonded with a heparin having binding affinity for viruses. The method for manufacturing the fiber-like structure includes a first step of subjecting fibers to heparin binding treatment, a second step of adding the viruses to the fibers bonded with the heparin in the first step, and a third step of washing the fibers added with the viruses in the second step to form the fiber-like structure bonded with the viruses via the heparin.

Description

  The present invention relates to a fibrous structure and a fibrous structure manufacturing method.

  Conventionally, gene introduction methods for expressing a gene in a target cell, tissue, organ, etc. using a virus incorporating the gene have been developed.

  For example, a method in which a virus incorporating a gene is injected into a blood vessel and transferred to a target organ by blood flow, or a method in which a virus is directly injected into a target organ with a syringe or the like has been developed.

  Here, Non-Patent Document 1 discloses a method for preparing highly purified adeno-associated virus by adding a crude purified sample of adeno-associated virus to a column to which heparin is bound.

E Burova and E Ioffe, “Chromatographic purification of recombinant adenovial and adeno-associated virtual vectors: methods and implications” 5p.

  However, in the conventional gene transfer method, although the site of virus injection can be controlled to some extent, the injected virus diffuses in the body in a short time, so that it is a non-selective and inefficient gene transfer method.

  That is, even if a highly purified virus is prepared as described in Non-Patent Document 1, the virus is stably localized at a high concentration without being diluted at a specific site in the body, and the virus is efficiently introduced. There was no way.

  The present invention has been made in view of the above problems, and an object of the present invention is to provide a fibrous structure capable of selectively and efficiently introducing a virus into a specific site, and a method for producing the fibrous structure. And

  In order to achieve such an object, the fibrous structure of the present invention is characterized in that heparin having binding affinity with a virus is bound to the fiber.

  The fibrous structure of the present invention is characterized in that, in the fibrous structure described above, the virus is an adeno-associated virus including a gene transfer vector.

  Further, the fibrous structure of the present invention is the fibrous structure described above, wherein the fibers are polylactic acid, polyglycolic acid, polylactone, polyvinyl alcohol, nylon, other chemical fibers, silk, cotton, collagen, gelatin , Other natural fibers, magnetic nanowires, and other metal fibers.

  Further, the fibrous structure of the present invention is characterized in that in the fibrous structure described above, the fiber has biodegradability.

  The fibrous structure of the present invention is characterized in that in the fibrous structure described above, the fibers are formed into a thread shape, a net shape, a cloth shape, a spherical shape, or a sponge shape.

  The fibrous structure of the present invention is the fibrous structure described above, wherein laminin, other heparin-binding adhesion molecule, chemokine, or other heparin-binding liquid factor is bound to the heparin. It is characterized by that.

  Moreover, the fibrous structure of the present invention is characterized in that, in the fibrous structure described above, the diameter of the fiber is 0.1 to 5000 μm.

  Moreover, the fibrous structure manufacturing method of the present invention includes a first step of applying a heparin-binding treatment to a fiber, a second step of adding a virus to the fiber to which heparin is bonded in the first step, and the second step. And a third step of producing a fibrous structure in which the virus is bound via the heparin by washing the fiber to which the virus has been added in the step.

  According to the fibrous structure of the present invention, since heparin having binding affinity with the virus is bound to the fiber, the virus can be selectively and efficiently introduced at a specific site. . Specifically, the virus binds to the fibrous structure via heparin, and stable binding can be maintained in the tissue for a long time without the virus being destroyed, and the cells in contact with the fibrous structure are selectively Since it is infected with a virus, it becomes possible to introduce the virus selectively and efficiently at a specific site in the body. In addition to the body (in vivo), the virus infects specific sites (specific cells in contact with the fibrous structure) in cell culture systems such as culture dishes (in vitro), so select specific sites. It becomes possible to express the target gene.

  In addition, according to the fibrous structure of the present invention, since the virus is an adeno-associated virus containing a gene introduction vector, the gene is specific to cells that are susceptible to infection with an adeno-associated virus, such as nerve cells (neurons). And the transgene can be expressed stably for a long period of time.

  Further, according to the fibrous structure of the present invention, the fibers are polylactic acid, polyglycolic acid, polylactone, polyvinyl alcohol, nylon, other chemical fibers, silk, cotton, collagen, gelatin, other natural fibers, or Since magnetic nanowires and other metal fibers are used, various fibers can be applied for virus introduction according to the purpose.

  Further, according to the fibrous structure of the present invention, since the fiber is biodegradable, there is an effect that the working time in the body can be freely controlled.

  Further, according to the fibrous structure of the present invention, the fibers are formed into a thread shape, a net shape, a cloth shape, a spherical shape, or a sponge shape. By providing it, etc., there is an effect that the virus can be efficiently introduced.

  Further, according to the fibrous structure of the present invention, laminin, other heparin-binding adhesion molecules, or chemokine, other heparin-binding humoral factors are bound to heparin. It is possible to increase the chances of contact between cells and viruses by attracting or activating specific cells, or by attaching cells with adhesion factors such as laminin, etc., and increasing the efficiency of virus infection. Play.

  Moreover, according to the fibrous structure of the present invention, since the diameter of the fiber is 0.1 to 5000 μm, it is possible to efficiently introduce a virus using a fiber having a size suitable for cell contact. , Has the effect.

  Moreover, according to the fibrous structure manufacturing method of the present invention, the first step of performing heparin binding treatment on the fibers, the second step of adding virus to the fibers to which heparin is bound in the first step, and the second step And a third step of generating a fibrous structure in which the virus is bound via heparin by washing the fiber to which the virus has been added in step 1, so that the virus is selectively and efficiently introduced at a specific site. The fibrous structure which can be manufactured is produced.

FIG. 1 is a diagram showing a structure of a fibrous structure according to an embodiment of the present invention as an example. FIG. 2 is a flowchart showing an example of a method for manufacturing a fibrous structure. FIG. 3 shows an example of a fibrous structure injection control system that moves and localizes the fibrous structure by magnetic force. FIG. 4 is a photograph of a GFP / AAV2-wire complex placed on 293 cells. FIG. 5 is a fluorescence micrograph showing the fluorescence of GFP expressed by 293 cells using a FITC filter. FIG. 6 is a fluorescence micrograph showing the fluorescence of GFP expressed by rat hippocampal neurons on the wire of the GFP / AAV2-wire complex.

  Embodiments of a fibrous structure according to the present invention will be described below in detail with reference to the drawings. Note that the present invention is not limited to the embodiments. In particular, in the following embodiment, an example in which the fibrous structure of the present invention is clinically applied to improve nerve function or the like may be described. However, the present embodiment is not limited to this, and a new It can be used for basic research such as new drug delivery systems, new drug screening and brain function analysis.

[Outline of Embodiment of the Present Invention]
The outline of the embodiment of the present invention will be described below with reference to FIG. FIG. 1 is a diagram showing a structure of a fibrous structure according to an embodiment of the present invention as an example.

  As shown in FIG. 1, in the fibrous structure of the present invention, heparin having a binding affinity with a virus is bound to a fiber (for example, a diameter of 0.1 to 5000 μm) such as a synthetic fiber or a natural fiber. It is characterized by. Here, the virus having binding affinity for heparin may be, for example, an adeno-associated virus (hereinafter referred to as “AAV virus”) including a gene transfer vector. Since the AAV virus has a heparin binding site, the virus can be reversibly and firmly fixed to the fibrous structure. The virus is not limited to this, and may be a retrovirus, Sendai virus, adenovirus, or the like. The vector for gene transfer can be freely designed according to the gene to be expressed such as a foreign gene expected to have a therapeutic effect. For example, a chemokine that attracts a specific cell or a specific cell can be selected. Examples include vectors encoding genes such as nutrient factors to be grown (GDNF, HGF, etc.), extracellular matrix, laminin, fibronectin, NCAM, nectin, cadherin, chemokine, cytokine and the like. Heparin may be a heparin-like molecule.

  Here, the “fiber” is an elongated structure such as a resin, and is a chemical fiber such as polylactic acid (PLA), polyglycolic acid (PGA), polylactone, polyvinyl alcohol (PVA), nylon or the like. It may be natural fibers such as silk, cotton, collagen, gelatin, or metal fibers such as magnetic nanowires. Examples of the material of the metal fiber include metals such as iron, gold, copper, lead, nickel, and platinum. Further, the fiber may be biodegradable, for example, a material or thread that dissolves in the body as a structure (for example, polyvinyl alcohol (PVA), polylactic acid fiber, polyglycolic acid fiber, collagen fiber, gelatin). By using it, the one that can freely control the action time in the body may be used.

  Further, the fiber may be formed into a thread shape, a net shape, a cloth shape, a spherical shape, or a sponge shape. For example, the fibrous structure may be one that is formed into a yarn shape by drawing and twisting the fibers, or may be one that is formed into a net shape by weaving the fibers in a lattice shape, and the fibers may be looms, knitting machines, etc. May be molded into a cloth shape such as woven fabric, knitted fabric, lace, and non-woven fabric, or may be molded into a spherical or sponge-like structure using polylactone, polylactic acid, gelatin beads, polystyrene, or the like.

  In addition, on the surface of the fibrous structure, for example, proteins (adhesion molecules, nutrient factors, antibodies, growth factors, etc.), hormones, peptides, drugs, immunosuppressants, organic compounds (for drug release rate control) Substances, visualization dyes, fluorescent dyes, magnetic substance stabilizers, genes (virus vectors, DNA, RNA), carbohydrates, polysaccharides (polylactic acid, etc.), lipids, glycolipids, metals (platinum, gold, iron, Tissue such as titanium, safety, stability, magnetic control substance, etc. in the tissue may be combined.

  In addition to heparin on the fibrous structure, laminin, other heparin-binding adhesion molecules, chemokines, other heparin-binding liquid factors, and the like may be bound in addition to viruses such as AAV virus.

  This is the end of the description of the outline of the embodiment of the present invention.

[Method for producing fibrous structure]
Below, an example of the manufacturing method of a fibrous structure is demonstrated. Here, FIG. 2 is a flowchart showing an example of a method for manufacturing a fibrous structure.

  As shown in FIG. 2, synthetic fibers, natural fibers, metal fibers, and other fibers are prepared (step SA-1). The fibers may be those already formed into a thread shape, a net shape, a cloth shape, a spherical shape, or a sponge shape (for example, commercially available yarn, cloth, surgical thread, etc.). This step SA-1 or the following step SA- Molding (for example, knitting with a knitting machine) may be performed after the heparin binding treatment of No. 2.

  And heparin binding process is performed with respect to the prepared fiber (step SA-2). As the heparin binding treatment, a known heparin binding treatment method such as a chemical treatment using a commercially available reagent that binds heparin to an organic compound or the like can be used.

  Then, the AAV virus is added to the fiber to which heparin is bound (step SA-3). For example, as described in Non-Patent Document 1, a fiber-like structure to which a virus is bound via heparin can be obtained by placing a fiber to which heparin is bound into a column and adding a crude virus purified sample. . In addition, about the construction method of an AAV viral vector, and the purification method of a virus, it can carry out by the well-known method as described in a nonpatent literature 1 etc. by using a commercially available reagent. Not only AAV viruses but also AAV virus variants and other viruses can be similarly constructed and vector-purified by using known methods.

  As an optional addition, in addition to viruses, heparin-binding adhesion molecules (such as laminin) and humoral factors (such as induction / nutrition / stimulation factors such as chemokines) can be added to mix with the virus. And may be bound to heparin. By using humoral factors, etc., attracting or activating specific cells, or by attaching cells with adhesion factors such as laminin, etc. It is possible to increase the infection efficiency (gene expression efficiency) of

  Even in the case where heparin-binding proteins such as adhesion molecules and nutrient factors are not bound in Step SA-3, there are unbound heparin residues on the surface of the fibrous structure. If the fibrous structure is introduced into a tissue such as the brain, the heparin-binding protein in the tissue binds quickly, and has the effect of increasing the virus infection efficiency.

  Finally, the unbound virus is washed (step SA-4) to complete the fibrous structure. The above is an example of a manufacturing method of a fibrous structure.

[Method of injecting fibrous structure]
Next, a method for injecting the fibrous structure produced as described above into the body will be described.

(1. Injection by syringe or catheter)
For example, when used during cardiac catheterization treatment, a fibrous structure configured as a stent (AAV virus binding state via heparin) is positioned through the cardiac catheter at the vascular stenosis. A virus vector of AAV virus encodes a protein that suppresses vascular stenosis, and the protein is continuously expressed (about one month). Thereafter, it functions as a normal vasodilator stent.

  In addition, when used for the treatment of solid cancer (liver, uterus, prostate, sarcoma, kidney, ovary, brain), a linear or spherical fibrous structure to which a virus incorporating a tumor suppressor gene or a metastasis suppressor is bound. Are inserted into a plurality of locations in the cancer tissue using a syringe or a catheter.

(2. Intravascular injection)
When the desired specific site into which the virus is to be introduced is a cell or tissue having heparin binding properties, the gene can be selectively expressed by injecting it into a blood vessel using a syringe or the like. In addition, using fibrous materials and threads that dissolve in the body (for example, polyvinyl alcohol (PVA), polylactic acid fiber, polyglycolic acid fiber, collagen fiber, gelatin), etc., the working time in the body can be freely set. You may control.

(3. Movement and localization by magnetic force)
When the fibrous structure is made of a magnetic material such as a metal fiber such as a nanowire, the fibrous structure can be moved and localized from the tissue surface by a magnetic force. Here, FIG. 3 is a diagram showing an example of a fibrous structure injection control system that moves and localizes the fibrous structure by magnetic force.

  As shown in FIG. 3, the fibrous structure injection control system includes a fibrous structure injection device 100 and a magnetic field control device 200. The fibrous structure injecting apparatus 100 has a hole having an inner diameter larger than the diameter of the fibrous structure, and is a light-transmissive non-magnetic tubular fibrous structure filling cylinder, and the fibrous structure filling A photodetection device that detects light crossing the cross section near the tip of the cylinder, a shutter that controls the opening and closing of the hole of the fibrous structure filling cylinder, and a fibrous structure filling cylinder The applicator is a mechanism in which holes are formed so as to be detachable. In order to prevent the fibrous structure from adhering to the inside of the fibrous structure filling cylinder, the fibrous structure filling cylinder is suspended in a state in which the fibrous structure is suspended in artificial spinal fluid or a lubricant added thereto. May be filled.

  As shown in FIG. 3, the magnetic field control device 200 includes an induction needle 50, a control unit 60, and a magnet 70. Here, the magnet 70 is a magnetic field generator that generates a magnetic field for guiding the fibrous structure. The guide needle 50 is a needle that increases the magnetic flux density due to the magnetic field generated from the magnet 70 at the tip portion, and increases the magnetic field most at the tip of the guide needle 50. The control unit 60 is a control unit that controls the magnetic field between the magnet 70 and the guide needle 50. For example, in a state where the magnetic part of the control unit 60 is between the magnet 70 and the guide needle 50, a strong magnetic field is generated at the tip of the guide needle 50 in order to cause the magnetic field generated from the magnet 70 to reach the guide needle 50. When the magnetic part of the control unit 60 slides in the direction perpendicular to the long axis direction of the magnetic field control device 200 and moves away from between the magnet 70 and the guide needle 50, the magnetic field generated from the magnet 70 reaches the guide needle 50. Therefore, no magnetic field is generated at the tip of the guide needle 50. As a result, even when the guide needle 50 is removed or removed from the tissue, a magnetic field is not generated at the tip of the guide needle 50, so that the position of the fibrous structure that has moved in the tissue according to the magnetic field gradient in the tissue may be shifted. Disappear.

  Next, an example of processing of the fibrous structure injection control system according to the present embodiment configured as described above will be described below. First, as shown in FIG. 3, the applicator and shutter of the fibrous structure injecting apparatus 100 on the surface of a site to be injected (tissue such as brain, spinal cord, liver, heart, kidney, tumor tissue, etc.) such as an injured part in advance. The guide needle 50 is inserted or surface-contacted at a desired position in accordance with the direction in which the fibrous structure is desired to move from the site to be injected.

  Then, after the magnetic field control device 200 in which the control unit 60 is in a non-inductive state is coupled to the guide needle 50 installed at a desired position, the control unit 60 is brought into the magnetic field induction state and the fibrous structure is used as a target site. A magnetic field necessary for arranging is generated.

  Then, a fibrous structure filling cylinder is attached to the fibrous structure injecting apparatus 100 with the shutter closed, and the fibrous structure is stored under the control of a computer or the like connected to the shutter. The hole of the cylinder is opened at a desired position. As a result, the fibrous structure itself enters the tissue and is arranged at the target site along the magnetic field gradient.

  In addition, the injection | pouring condition of the fibrous structure is monitored by the optical detection apparatus of the fibrous structure injection apparatus 100, and it can confirm that injection | pouring was completed. Further, the position of the fibrous structure in the tissue may be confirmed by a surgical CT / navigator. The above is an example of a method for moving and localizing the fibrous structure by magnetic force by the fibrous structure injection control system.

(4. Introduction to brain and spinal cord tissue)
Heparin is a biomolecule (polysaccharide), particularly present in the intestinal tract wall, and has no accumulated toxicity. Heparin is also present in the brain in large amounts mainly in the form of glycans such as heparan sulfate. Heparin is widely used in daily clinical practice as a drug that suppresses blood coagulation, for example, for the dissolution treatment of intra-arterial embolism for cerebral embolism patients. In patients with cerebral embolism, since the blood-brain-barrier is broken, administered heparin penetrates into the brain tissue, but there is no brain damage to be noted.

  When this fibrous structure is introduced into the brain / spinal cord tissue, heparin binds stably to various adhesion molecules (for example, laminin) in the brain, so that heparin administered into the brain promptly becomes heparin such as laminin. It is thought to bind to binding molecules. No brain damage (nervous symptoms such as paralysis or convulsions) is observed even if a fibrous structure (wire with heparin binding) not bound to the virus is placed in the brain, and the neurons in culture It was confirmed that even when the fibrous structure was exposed for 1 month, the nerve cells were not damaged (see also the following examples).

  This is the end of the description of the present embodiment.

[Example]
Next, examples in the above-described embodiment will be described with reference to FIGS. Here, first, the background of the development of the fibrous structure of the present example will be described.

  In recent years, gene therapy is expected as a future therapeutic technique, but no method has been established for maintaining the sequence of a virus (viral vector) incorporating a gene at a specific site in a target organ accurately and over a long period of time.

  Here, to date, the following viral vector administration methods have been studied in vivo. 1) A method in which a gene-incorporated virus is injected into a blood vessel and transferred to a target organ by blood flow. 2) A method in which a virus is directly injected into a target organ with a syringe or the like.

  However, none of the above is a non-selective and inefficient virus introduction method, and it was impossible to pinpoint the virus only at a specific site. In addition, since the injected virus diffuses into the tissue in a short time, it is impossible to maintain the virus at a specific site at a high concentration for a long time, and the virus infection efficiency decreases with time. was there. Further, if the virus spreads to an unintended site, the virus integration gene is expressed in an unplanned cell or organ, and there is a risk of causing unexpected side effects.

  In particular, in the conventional gene transfer method targeting brain / spinal cord tissue, there is no method for maintaining the viral vector at a high concentration for a certain period of time (several days to several weeks) only at a specific site in the neural tissue. In addition, even if injection into the target site is accurate, the virus diffuses around the target site, and the virus concentration at the target site decreases, making it difficult to achieve the effect (so that a therapeutic effect can be obtained). It was difficult to obtain gene expression). In particular, in the cerebrospinal tissue, if the virus is expressed in a place other than the target site due to diffusion, there is a problem that there is a large risk of serious neuropathy or psychiatric symptoms. Therefore, clinical treatment using viral vectors has remained difficult in cranial nerve diseases.

  The fibrous structure of the present example has been developed as a result of intensive studies by the inventors of the present application in view of the various problems described above. In this example, an adeno-associated virus vector is used. AAV viruses have receptors for heparin (polysaccharides such as heparan sulfate) and have a high binding affinity for heparin (polysaccharides such as heparan sulfate). Therefore, if heparin is uniformly bonded to a structure (a linear structure such as a metal wire or thread (surgical thread or the like) or a spherical structure), the AAV virus is adsorbed on the surface of the structure to which heparin is bonded. Stable binding can be achieved while retaining activity. This AAV virus can bind for a long time without falling off and spreading from the heparin-binding structure. For example, even if it is left in an aqueous solution (culture solution) for 4 weeks, the AAV virus does not drop from the structure. A complex in which an AAV virus containing a vector is bound to a heparin-binding structure can be accurately and long-term localized at a specific site in the target organ.

  By using the AAV virus, 1) high affinity (infectivity) to nerve cells (neurons) (nerve selectivity), 2) little expression in glial cells, 3) nerve cells (neurons) 4) Advantages of AAV virus-infected neurons that can stably maintain a high expression level of a foreign gene incorporated into the virus vector over a long period of time. There is. In addition, it is applicable not only to a nerve cell but to all cells that express a receptor for AAV virus (eg, kidney, nerve cell, vascular endothelial cell). In addition, the type of gene to be incorporated into the viral vector is arbitrary depending on the purpose and is not limited, and any AAV virus subspecies can be used. In the following Examples, AAV2 virus incorporating GFP (Green Fluorescent Protein) was used for gene transfer confirmation (size of 2000 bps or less that can be incorporated into AAV2 vector).

  As the fibrous structure, a structure in which heparin was bonded by chemical treatment using a stainless steel wire having a diameter of 100 μm as a basic skeleton was used. In this example, the stainless steel wire was formed into a net shape by hand. The AAV2 virus containing the GFP expression vector was bound to this fibrous structure via heparin (hereinafter referred to as “GFP / AAV2-wire complex”). Here, FIG. 4 is a photograph of a GFP / AAV2-wire complex placed on 293 cells.

  Kidney-derived 293 cells are cells with extremely high AAV virus infection efficiency, and are cells that amplify and express virus vectors with high efficiency even under low concentration of AAV virus conditions. Used for amplification. In this example, 293 cells were used to verify the efficiency of gene transfer by the GFP / AAV2-wire complex. As shown in FIG. 4, 293 cells were spread over the entire surface of the culture dish in a confluent state, and a GFP / AAV2-wire complex was placed thereon and cultured for 2 weeks. Since the GFP / AAV2-wire complex has magnetism, it was fixed to the culture dish with a magnet. After culture, the green fluorescence of 293 cells expressing GFP was observed. Here, FIG. 5 is a fluorescence micrograph showing the fluorescence of GFP expressed in 293 cells by the FITC filter.

  As shown in FIG. 5, only the cells adhered to the wire network of the GFP / AAV2-wire complex showed green fluorescence. That is, in the cells that did not adhere to the wire net, the GFP gene was not expressed, indicating that the AAV2 virus was persistently bound to the wire net during the culture and did not elute into the culture medium. It proves that. This result also shows that the AAV2 virus is bound on the wire surface while retaining the biological activity (expression of the viral vector integration gene). In addition, in the lower right of the photograph in FIG. 5, the cell at a position slightly away from the wire emits green fluorescence, which is considered to have moved after contact with the wire.

  Subsequently, in order to confirm the infection ability other than the kidney-derived 293 cells, a transplanted neuron infection simulation was performed. In this simulation, a fibrous structure (GFP / AAV2-wire complex) in which a magnetic wire bound with heparin is immersed in an AAV2 virus solution containing a viral vector incorporating a GFP gene, and the AAV2 virus is bound to the wire. Was used.

  Then, rat hippocampal neurons were placed on the wire of the GFP / AAV2-wire complex, cultured for 14 days, and GFP was observed with a fluorescence microscope. Here, FIG. 6 is a fluorescence micrograph showing the fluorescence of GFP expressed by rat hippocampal neurons on the wire of the GFP / AAV2-wire complex.

  As shown in FIG. 6, the cell body and nerve fiber of the nerve cell (neuron) emits green fluorescence, and it can be seen that the GFP gene was also expressed in the nerve cell. In addition, no abnormal morphology of nerve cells was observed, and it was considered that there was no toxicity to neurons. In another experiment, it was also found that GFP fluorescence was observed in the whole nerve cell when a part of the nerve cell was brought into contact with the wire. That is, if a part of the AAV virus receptor cell (such as a nerve cell) and AAV virus is in contact with the AAV virus, the AAV virus leaves the wire and is taken into the cell while retaining its activity, and is incorporated into the virus vector. It was revealed to express the integrated gene. The above is the experimental result according to this example.

  As described above, according to the present example, an AAV virus incorporating an expression vector of a target gene is accurately placed at a specific site such as a cerebrospinal tissue, thereby expressing a foreign gene only at the specific site and exhibiting a therapeutic effect. Can be made. In addition, while maintaining the foreign gene expression activity of the virus (while maintaining the biological activity of the virus), the AAV virus can be stably bound to the magnetic wire for a long period of time, and contacted with the magnetic wire to which the AAV virus was bound. A cell (such as a nerve cell) can efficiently incorporate the AAV virus into the cell and express a foreign gene (for example, GFP) incorporated in the AAV virus vector with high efficiency. In addition, since acute toxicity to nerve cells adhered to the magnetic wire used in the examples was not observed and no virus particles were diffused in the tissues (in liquid), the AAV virus was concentrated at a high concentration at the target site. And the exposure of the virus to other than the target neurons is minimized (preventing unexpected side effects).

[Other embodiments]
Although the embodiments of the present invention have been described so far, the present invention is not limited to the above-described embodiments, but can be applied to various different embodiments within the scope of the technical idea described in the claims. It may be implemented.

  For example, among the processes described in the embodiments, all or part of the processes described as being automatically performed can be manually performed, or the processes described as being performed manually can be performed. All or a part can be automatically performed by a known method.

  In addition to the above, the processing procedures, control procedures, specific names, information including parameters such as registration data for each processing, and screen examples shown in the drawings may be changed arbitrarily unless otherwise specified. Can do. Each illustrated component is functionally conceptual and does not necessarily need to be physically configured as illustrated.

  Furthermore, the specific form of distribution / integration of the devices is not limited to that shown in the figure, and all or a part of them may be functional or physical in arbitrary units according to various additions or according to functional loads. Can be distributed and integrated. That is, the above-described embodiments may be arbitrarily combined and may be selectively implemented.

  As described above in detail, according to the fibrous structure and the fibrous structure manufacturing method of the present invention, the fibrous structure and the fibrous structure capable of selectively introducing a virus at a specific site with high efficiency. Since a structure manufacturing method can be provided, it is extremely useful in various fields such as medicine, pharmaceuticals, drug discovery, biological research, nanotechnology, and clinical examinations.

DESCRIPTION OF SYMBOLS 100 Fibrous structure injection apparatus 200 Magnetic field control apparatus 50 Guide needle 60 Control part 70 Magnet

Claims (8)

  A fibrous structure obtained by binding heparin having binding affinity to a virus to a fiber. The fibrous structure according to claim 1,
A fibrous structure characterized in that the virus is an adeno-associated virus containing a gene transfer vector. The fibrous structure according to claim 1 or 2,
The fibers are polylactic acid, polyglycolic acid, polylactone, polyvinyl alcohol, nylon, other chemical fibers, silk, cotton, collagen, gelatin, other natural fibers, magnetic nanowires, or other metal fibers A fibrous structure. The fibrous structure according to claim 1 or 2,
The fibrous structure is characterized in that the fiber has biodegradability. In the fibrous structure according to any one of claims 1 to 4,
A fibrous structure, wherein the fiber is formed into a thread shape, a net shape, a cloth shape, a spherical shape, or a sponge shape. The fibrous structure according to any one of claims 1 to 5,
A fibrous structure obtained by binding laminin, other heparin-binding adhesion molecule, chemokine, or other heparin-binding humoral factor to the above heparin. In the fibrous structure according to any one of claims 1 to 6,
A fibrous structure having a diameter of the fiber of 0.1 to 5000 μm. A first step of applying heparin binding to the fiber;
A second step of adding a virus to the fiber to which heparin is bound in the first step;
A third step of generating a fibrous structure in which the virus is bound via the heparin by washing the fiber to which the virus has been added in the second step;
The manufacturing method of the fibrous structure characterized by including.