JP2007167366A - Nerve regeneration material - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a nerve regeneration material capable of regenerating the nerve over a long distance in the living body, having elasticity and responding to a dynamic load. <P>SOLUTION: This nerve regeneration material consists of (i) a bellows-shaped tubular body composed of aliphatic polyester fiber with the mean fiber diameter of 0.05-50 μm and having axially continuing crest parts and trough parts, and (ii) marrow cells embedded inside the tubular body. <P>COPYRIGHT: (C)2007,JPO&INPIT

Description

  The present invention relates to a nerve regeneration material.

In recent years, research on regenerative medicine that uses cell differentiation and proliferation ability to reconstruct the original living tissue and organ as a treatment method for severely damaged living tissue and organ has become active. Nerve regeneration is one of them.
When the peripheral nerve is damaged, if the stumps can be pulled together at an early stage after the damage, first, the cut ends of the cut nerve are directly sutured. However, if time passes after damage, it may be impossible to pull the stumps together. In addition, if the nerve has been damaged such as traction and crushing over a long distance from the beginning, or if there is a defect between the stumps, direct stitching is impossible.
In this case, autologous nerve transplantation is performed as a treatment method, but it is necessary to collect healthy nerves in autologous nerve transplantation. Usually, autonomic nerve transplantation collects sensory nerves with less damage, but if healthy nerves are collected, there may be residual pain in the stump of the nerve, and there is a sensory disturbance in the control area of the collected nerves. There are problems, such as occurring.

Therefore, research using an artificial neural tube has been conducted. One of them is an experiment using a silicon tube by Lundborg et al. (1982), but nerve regeneration was difficult between stumps of 10 mm or more (Non-patent Document 1). This is because the silicon tube is an artificial material, has no blood flow, is hollow and is cell-free, and is impermeable so that no liquid factor is supplied from the surroundings.
More recently, research on artificial neural tubes using bioabsorbable materials such as collagen and synthetic polymers has also been conducted. In particular, in studies using a collagen gel packed in a polyglycolic acid mesh, nerve regeneration between 80 mm stumps was confirmed in dog sciatic nerves (Patent Document 1). However, the problem is that collagen is collected from other animals.

In addition, studies have been made on a tube of a bioabsorbable material in which Schwann cells, which are said to be important for the regeneration nerve regeneration, are cultured (Non-patent Document 1). When Schwann cells are used, the speed of nerve regeneration is faster than when cells are not used, but Schwann cells need to be collected from normal nerve tissue, resulting in the same problems as nerve transplantation.
WO 01/03609 Publication Arch. Otolaryngol. Head Neck Surg. 124, 1081 (1998)

  An object of the present invention is to provide a nerve regeneration material that can regenerate nerves in vivo. Another object of the present invention is to provide a nerve regeneration material in which a base material is absorbed by a living body. Furthermore, an object of the present invention is to provide a nerve regeneration material that is stretchable and can cope with a mechanical load. Another object of the present invention is to provide a method for producing the nerve regeneration material, a kit capable of producing the nerve regeneration material, and a method for regenerating damaged nerves.

  The present inventor successfully regenerates nerves by transplanting a nerve regeneration material made of an aliphatic polyester fiber having a specific average fiber diameter and having bone marrow cells embedded in a bellows-shaped cylindrical body to a nerve damage site. The present invention has been completed. Further, the present invention has been completed by finding that the nerve regeneration material has a bellows shape and can cope with a mechanical load.

  That is, the present invention comprises (i) an accordion-shaped cylindrical body made of an aliphatic polyester fiber having an average fiber diameter of 0.05 to 50 μm and having peaks and valleys continuous in the axial direction, and (ii) It is a nerve regeneration material consisting of bone marrow cells embedded inside.

  The present invention also relates to (i) a bellows-like cylindrical body comprising an aliphatic polyester fiber having an average fiber diameter of 0.05 to 50 μm and having peaks and valleys continuous in the axial direction, and (ii) a bone marrow cell solution. A kit for producing a nerve regeneration material comprising:

  Furthermore, the present invention provides (i) a step of preparing an accordion-shaped cylindrical body having a peak portion and a valley portion made of an aliphatic polyester fiber having an average fiber diameter of 0.05 to 50 μm and having axially continuous peaks and valleys; and (ii) And a method for producing a nerve regeneration material comprising the step of injecting a bone marrow cell solution into the inside of a cylindrical body.

  The present invention also includes (i) a step of preparing an accordion-shaped cylindrical body having a peak portion and a valley portion that are made of aliphatic polyester fibers having an average fiber diameter of 0.05 to 50 μm and that are continuous in the axial direction; A method for regenerating a damaged nerve comprising the steps of injecting a bone marrow cell solution into a cylindrical body to obtain a nerve regeneration material, and (iii) transplanting the obtained nerve regeneration material to a nerve damage site.

The nerve regeneration material of the present invention can regenerate nerves well in vivo. The nerve regeneration material of the present invention is effective for regeneration of a nerve having a large damaged site. In addition, the nerve regeneration material of the present invention has an advantage that the base material is composed of a bioabsorbable material and is absorbed by the living body. The nerve regeneration material of the present invention is stretchable and can cope with a mechanical load.
According to the method for producing a nerve regeneration material of the present invention, the nerve regeneration material can be easily produced. According to the kit of the present invention, a nerve regeneration material can be easily produced at the time of surgery. According to the reproduction method of the present invention, a damaged long nerve can be reproduced.

Hereinafter, the present invention will be described in detail.
<Neural regeneration material>
(Cylinder)
The cylindrical body constituting the nerve regeneration material of the present invention is made of an aliphatic polyester fiber. The cylindrical body is a three-dimensional structure formed by laminating and accumulating single or plural aliphatic polyester fibers. The average fiber diameter of the aliphatic polyester fiber is 0.05 to 50 μm, preferably 0.2 to 20 μm. An average fiber diameter of less than 0.05 μm is not preferable because the strength of the cylindrical body cannot be maintained. Moreover, when the average fiber diameter is higher than 50 μm, the specific surface area of the cylindrical body is small, and the number of engrafted cells is reduced, which is not preferable. An average fiber diameter is the average value which measured the fiber diameter in 20 places from the image by an optical microscope.
Preferred examples of the aliphatic polyester fiber include fibers made of polylactic acid, polyglycolic acid, polycaprolactone, polydioxanone, trimethylene carbonate, polybutylene succinate, polyethylene succinate, or a copolymer thereof. Of these, fibers made of polylactic acid, polyglycolic acid, polycaprolactone or copolymers thereof are preferred.

  The aliphatic polyester fiber may contain other components. The other component is preferably at least one selected from the group consisting of phospholipids, carbohydrates, glycolipids, steroids, polyamino acids, proteins, and polyoxyalkylenes. Specific components include phospholipids such as phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, phosphatidylglycerol and / or polygalacturonic acid, heparin, chondroitin sulfate, hyaluronic acid, dermatan sulfate, chondroitin, dextran sulfate, sulfated Cellulose, alginate, dextran, carboxymethylchitin, galactomannan, gum arabic, tragacanth gum, gellan gum, sulfated gellan, caraya gum, carrageenan, agar, xanthan gum, curdlan, pullulan, cellulose, starch, carboxymethylcellulose, methylcellulose, glucomannan, chitin , Carbohydrates such as chitosan, xyloglucan, lentinan, galactocerebroside, glucocerebrosi , Glycolipids such as globoside, lactosylceramide, trihexosylceramide, paragloboside, galactosyl diacylglycerol, sulfoquinovosyl diacylglycerol, phosphatidylinositol, glycosyl polyprenol phosphate, steroids such as cholesterol, cholic acid, sapogenin, digitoxin , Polyamino acids such as polyaspartic acid, polyglutamic acid, polylysine, collagen, gelatin, fibronectin, fibrin, laminin, casein, keratin, sericin, thrombin and other proteins, polyoxyethylene alkyl ether, polyoxyethylene propylene alkyl ether , Polyoxyalkylenes such as polyoxyethylene sorbitan ether, FGF (fibroblast growth factor), EG (Epidermal growth factor), PDGF (platelet-derived growth factor), TGF-β (β-type transforming growth factor), NGF (nerve growth factor), HGF (hepatocyte growth factor), BMP (bone morphogenetic factor), etc. Examples include growth factors.

An example of a cross-sectional view of the center of a plane parallel to the axial direction of the cylindrical body is shown in FIG. As shown in FIG. 4, the cylindrical body has a bellows shape, and has a crest (11) and a trough (12) that are continuous in the axial direction. FIG. 4 is a schematic diagram showing a bellows structure of a cylindrical body, and the shapes and sizes of the peaks (11) and valleys (12) are irregular.
The outer diameter (7) of the cylindrical body is preferably 0.5 to 50 mm, more preferably 1 to 20 mm. The outer diameter is measured in 10 places with a micrometer and expressed in the range of the minimum and maximum measured values. The cylindrical body preferably has a valley depth (8) of 0.1 to 10 mm. The interval between the valleys can be determined, for example, by measuring 10 points with an optical microscope and measuring the measured value.

The cylindrical body preferably has an interval (9) between peak portions of 2 mm or less. If the interval (9) between the peaks is greater than 2 mm, the stretchability may be reduced and the elastic modulus may be impaired. The interval between the peaks is measured with, for example, an optical microscope and is expressed in the range of the minimum value and the maximum value of the measurement values.
The thickness (10) of the cylindrical body is preferably 0.05 to 1 mm, more preferably 0.1 to 0.5 mm. The thickness is expressed in the range of the minimum value and the maximum value when the cylindrical body is cut open, a sample having a length of 5 cm and a width of 1 cm is prepared, and measurement is performed at 10 locations using a micrometer.
The weight of the cylindrical body is preferably 1 to 200 g / m ² , more preferably 50 to 50 g / m ² . If it is less than 1 g / m ² , a cylindrical body may not be formed. Moreover, if it exceeds 200 g / m ² , the stretchability may be impaired. The basis weight can be calculated by cutting a cylindrical body, preparing a sample having a length of 5 cm and a width of 1 cm, measuring the weight, and (weight of the sample) ÷ (area of the sample). Therefore, the cylindrical body preferably has an aliphatic polyester fiber weight per unit area of 1 to 200 g / m ² , a thickness of 0.05 to 1 mm, and an outer diameter of 0.5 to 50 mm. Moreover, it is preferable that a cylindrical body is 2 mm or less in the space | interval of a peak part, and the depth of a trough part is 0.1-10 mm. Moreover, it is preferable that a cylindrical body is a thing in which the aliphatic polyester fiber was wound spirally centering | focusing on the axis | shaft of a cylindrical body.

(Manufacture of cylindrical body)
The cylindrical body can be manufactured by an electrostatic spinning method, a spunbond method, a melt blow method, a flash spinning method, or the like. Among these, the electrospinning method is preferable. In the electrostatic spinning method, a dope in which aliphatic polyester is dissolved in a volatile solvent is discharged into an electrostatic field formed between electrodes, the dope is spun toward the electrode, and the formed fibrous material is collected. It is a method to do. The fibrous material includes not only the state in which the solvent in the dope is distilled off to form a fiber, but also the state containing a solvent.
The concentration of the aliphatic polyester in the dope is preferably 1 to 30% by weight, more preferably 2 to 20% by weight. If the concentration of the aliphatic polyester is less than 1% by weight, it may be difficult to form a cylindrical body because the concentration is too low. On the other hand, if it is larger than 30% by weight, the average fiber diameter of the obtained fiber may be too large.
The volatile solvent is preferably a substance having a boiling point of 200 ° C. or less at normal pressure and a liquid at 27 ° C. The volatile solvent is not particularly limited as long as the aliphatic polyester is dissolved. Examples of volatile solvents include methylene chloride, chloroform, acetone, methanol, ethanol, propanol, isopropanol, toluene, tetrahydrofuran, 1,1,1,3,3,3-hexafluoroisopropanol, water, 1,4-dioxane, and four. Examples thereof include carbon chloride, cyclohexane, cyclohexanone, N, N-dimethylformamide, acetonitrile and the like. Among these, methylene chloride, chloroform, and acetone are particularly preferable in view of the solubility of the aliphatic polyester. These solvents may be used alone, or a plurality of solvents may be combined. Moreover, in this invention, you may use another solvent together in the range which does not impair this objective.

The electrostatic spinning method can be performed using, for example, the apparatus shown in FIG. FIG. 1 shows an electrospinning composed of a syringe (1) having a discharge side electrode (4) and a holding tank (3), a collection side electrode (5), and a high voltage generator (6). Indicates the device. A predetermined voltage is applied between the discharge side electrode (4) and the collection side electrode (5) by the high voltage generator (6).
In the apparatus shown in FIG. 1, the dope (2) is filled in the holding tank (3), discharged into the electrostatic field through the nozzle (1), and is made into a fiber by spinning with an electric field. ) To obtain a cylindrical body.

An electrode consists of a discharge side electrode (4) and a collection side electrode (5). These electrodes are only required to be conductive with any metal, inorganic or organic material. Further, a metal, inorganic, or organic thin film exhibiting conductivity may be provided over the insulator. The electrostatic field is formed between a pair or a plurality of electrodes, and a high voltage may be applied to any of the electrodes. This includes, for example, the case where two high voltage electrodes having different voltage values (for example, 15 kV and 10 kV) and a total of three electrodes connected to the ground are used, or a case where more than three electrodes are used. Including.
While spinning the dope toward the collecting side electrode (5), the solvent evaporates depending on the conditions to form a fibrous material. At normal room temperature, the solvent completely evaporates until it is collected by the collecting side electrode (5). However, if the solvent evaporation is insufficient, the solvent may be drawn under reduced pressure.

  When a mandrel that is not mirror-finished is used as the collecting side electrode (5), a cylindrical body can be easily manufactured. That is, it is possible to easily obtain a bellows-like cylindrical body by collecting fibers until a predetermined weight is obtained on the mandrel by electrostatic spinning, and removing the cylinder from the mandrel while maintaining appropriate friction. I can do it. The surface roughness of the mandrel is preferably 0.2-S or more, more preferably 1.5 to 400-S. When removing the cylindrical body from the mandrel having an appropriate surface roughness in this way, it is preferable to apply stress to only one end of the cylindrical body. It is possible to apply stress to only one end by fixing one end of the cylindrical body and pulling out the mandrel in the direction of the fixed end.

When forming a cylindrical body on a mandrel by electrostatic spinning, it is preferable to rotate the mandrel in the circumferential direction. By rotating, a cylindrical body having a uniform thickness can be formed.
The distance between the electrodes depends on the charge amount, nozzle size, dope discharge amount, dope concentration, and the like, but a distance of 5 to 20 cm is appropriate for about 10 kV. The applied electrostatic potential is preferably 3 to 100 kV, more preferably 5 to 50 kV, and still more preferably 5 to 30 kV. When supplying the dope from the nozzle into the electrostatic field, several nozzles can be used to increase the production rate of the fibrous material. The inner diameter of the nozzle is preferably 0.1 to 5 mm, more preferably 0.1 to 2 mm.

A separate collector may be installed between the discharge side electrode (4) and the collection side electrode (5) to collect the fibrous material. The collector preferably has a surface roughness comparable to that of the mandrel used for the collection-side electrode (5) described above. The spinning temperature depends on the evaporation behavior of the solvent and the viscosity of the spinning solution, but is usually 0 to 50 ° C.
FIG. 2 shows a device in which a discharge side electrode 4 is inserted into a holding tank 3 having a nozzle 1 instead of a syringe. In this apparatus, instead of discharging the dope with a syringe, the distance between the nozzle 1 and the collecting side electrode 5 is adjusted, and the dope is scattered from the nozzle 1 to the collecting side electrode 5. FIG. 3 is a perspective view of FIG.

(Bone marrow cells)
In the nerve regeneration material of the present invention, bone marrow cells are embedded in a cylindrical body. Examples of cells that promote nerve regeneration include Schwann cells. However, Schwann cells are collected only from normal nerve tissues, and thus have a problem in that they damage normal tissues. Therefore, it is preferable to use bone marrow cells that are relatively easy to collect and have high cell differentiation ability from the viewpoint of not damaging normal nerve tissue. Furthermore, among bone marrow cells, bone marrow mononuclear cells have a high differentiation potential, so they can be used without culturing, and furthermore, since they do not require time for culturing, there is less risk of infection. preferable.
The bone marrow cells are preferably injected into the cylindrical body as a bone marrow cell solution. The bone marrow cell solution contains bone marrow cells and phosphate buffer (PBS) or saline. The number of cells in the bone marrow cell solution is preferably 5 × 10 ^{5 to} 5 × 10 ⁷ cells / ml, more preferably 1 × 10 ⁶ to 1 × 10 ⁷ cells / ml.

<Kit>
The kit of the present invention is a kit for producing a nerve regeneration material comprising (i) a cylindrical body and (ii) a bone marrow cell solution. The cylindrical body and the bone marrow cell solution are as described in the section of nerve regeneration material. At the time of use, the kit can be used as a nerve regeneration material by injecting a bone marrow cell solution into the cylinder.
<Manufacturing method>
The method for preparing a nerve regeneration material of the present invention comprises (i) a step of preparing a cylindrical body and (ii) a step of injecting a bone marrow cell solution into the inside of the cylindrical body. The cylindrical body and the bone marrow cell solution are as described in the section of nerve regeneration material.
<Playback method>
The method for regenerating damaged nerve of the present invention comprises (i) a step of preparing a cylinder, (ii) a step of injecting a bone marrow cell solution into the cylinder to obtain a nerve regeneration material, and (iii) the obtained nerve regeneration material Is transplanted to the site of nerve injury. The cylindrical body and the bone marrow cell solution are as described in the section of nerve regeneration material. After transplantation, it is preferable to maintain a rest for a predetermined period and promote nerve regeneration. The bone marrow cells are preferably bone marrow mononuclear cells. According to this regeneration method, damaged nerves of mammals such as mice, pigs, horses and humans can be regenerated.

The details of the present invention will be described more specifically by the following examples. However, the present invention is not limited to these examples.
I. The materials used in this example are as follows.
(1) Polylactic acid (LACTY9031): Shimadzu Corporation
(2) Lactic acid-glycolic acid copolymer: 50/50 Poly (DL-lactide-co-glycolide) manufactured by Absorbable Polymers International, intrinsic viscosity = 1.08 dL / g in HFIP, 30 ° C)
(3) Methylene chloride, ethanol: manufactured by Wako Pure Chemical Industries,
(4) Lewis rat: manufactured by Charles River Co., Ltd.
(5) Somnopentyl: manufactured by Kyoritsu Pharmaceutical Co., Ltd. (6) PBS: manufactured by Biomedi-cal,
(7) Histopaque-1077: manufactured by SIGMA
(8) Suture: 9-0 nylon thread manufactured by Bear Medic,
(9) Paraformaldehyde: manufactured by Merck Ltd.
(10) Sucrose: Wako Pure Chemical Industries,
(11) Anti-Neurofilament antibody: Dako Co., Ltd.
(12) Biotinylated IgG: biotinylated horse anti-mouse IgG manufactured by VECTOR
(13) Horse serum: Invitrogen (14) DAB: PIERCE (15) EUKITT: O.Kinder
(16) Tissue Tec: Made by Miles
(17) TPBS (PBS containing Tween 20): manufactured by Biomedi-cal

II. The size of each part of the cylindrical body was measured by the following method.
Average fiber diameter: 20 positions were measured with a digital microscope (KEYENCE, VHX DIGITAL MICROSCOPE), and the average value was defined as the average fiber diameter.
Outer diameter: Ten locations were measured with a micrometer (Mitsutoyo Co., Ltd.), and the range of minimum and maximum measured values was defined as the outer diameter.
Spacing between mountain parts: 10 points were measured with a digital microscope (Keyence, Inc., VHX DIGITAL MICROSCOPE), and the range between the minimum value and the maximum value of the measured values was defined as the distance range between the mountain parts.
Depth of valley: Ten points were measured with a digital microscope (KEYENCE, VHX DIGITAL MICROSCOPE, Inc.), and the range of the minimum and maximum values of the measured values was defined as the depth of the valley.
Thickness: A cylindrical body was cut open, a sample having a length of 5 cm and a width of 1 cm was prepared, and measurement was performed at 10 locations using a micrometer, and the range of the minimum value and the maximum value of the measured values was defined as the thickness.
Weight per unit area: A cylindrical body was cut open, a sample having a length of 5 cm and a width of 1 cm was prepared, weighed, and calculated from (sample weight) / (sample area).

<Example 1>
(Manufacture of cylindrical body)
The electrospinning apparatus shown in FIGS. 2 and 3 was prepared. The inner diameter of the nozzle (1) was 0.8 mm. The distance from the nozzle (1) to the collection side electrode (5) was set to 10 cm. 1 g of lactic acid-glycolic acid copolymer and 9 g of methylene chloride / ethanol = 8/1 (parts by weight / parts by weight) were mixed at room temperature (25 ° C.) to prepare a dope A having a concentration of 10% by weight. Further, 1 g of polylactic acid and 9 g of methylene chloride were mixed at room temperature (25 ° C.) to prepare a dope B having a concentration of 10% by weight.
First, the dope A was put in the holding tank (3), the voltage between the discharge side electrode 4 and the collection side electrode 5 was set to 14 kV, and the fibrous substance was discharged from the nozzle 1. During discharge, the collection side electrode 5 was rotated at a speed of 100 rpm. Discharging was performed for 1 minute. Next, in place of the dope A, the dope B was put into the holding tank 3 and discharged for 1 minute under the same conditions as the dope A. Then, dope A was put into holding tank 3 instead of dope B, and discharged for 1 minute under the same conditions. After discharging, heat treatment is performed at 70 ° C. for 10 minutes, one end of the fiber collected on the collecting side electrode 5 is held by a finger and fixed, and the collecting side electrode 5 is held by a finger and pulled to the fixed side. A bellows-like cylindrical body having peaks and valleys continuous in the direction was obtained.
The obtained cylindrical body X had an average fiber diameter of 2 to 8 μm and a cross-sectional shape generally shown in FIG. FIG. 4 is an explanatory diagram, and the actual bellows structure is not regular. The outer diameter (7) is 1.3 to 1.4 mm, the length is 100 mm, the basis weight is 20 g / m ² , the peak interval (9) is 0.3 to 0.5 mm, and the valley depth (8 ) Was 0.1 to 0.3 mm, and the thickness (10) was 0.15 to 0.2 mm. The stretchability of the cylindrical body was measured with Tensilon (EZ TEST: Shimadzu Corporation), and the yield elongation was 50%.

(Preparation of bone marrow mononuclear cell solution)
After Lewis rats were sacrificed by intraperitoneal anesthesia (Somnopentyl), both femurs were removed. The epiphysis of the femur was excised, and bone marrow cell suspension was collected by applying pressure to the bone marrow cavity with 5 ml of PBS. The bone marrow cell-PBS suspension was added with 5 ml of Histopaque-1077 and centrifuged, and the intermediate layer was collected. After the intermediate layer suspension was centrifuged again, the cell mass at the bottom of the test tube was suspended in 1 ml of PBS to prepare a bone marrow mononuclear cell solution (5 × 10 ⁶ cells / ml).
(Transplant)
Another rat was anesthetized intraperitoneally and the sciatic nerve was deployed in the thigh. A nerve defect was created by excising 10 mm of nerve at the thigh center. One end of the cylindrical body X having a length of 12 mm was sutured to the nerve stump and three needles while the inside thereof was in contact with one nerve stump. The bone marrow mononuclear cell solution prepared previously was injected from the other end of the cylindrical body X, and then the other end of the cylindrical body X and the other nerve stump were sutured in the same manner. The fascia and skin were sutured and the operation was completed.

(Evaluation)
Four weeks later, the sciatic nerve was expanded again, and the nerve was removed. Remaining cylindrical body X was confirmed visually. Frozen longitudinal sections were prepared and immunostained with anti-Neurofilament antibody. Immunostaining was performed by the following method.
Longitudinal section fixation: After fixing the rats to reflux with 4% Paraformaldehyde, the nerves are removed. The isolated nerve is fixed overnight with 4% Paraformaldehyde solution, dehydrated with 10%, 20%, 30% Sucrose solution, embedded in Tissue Tec and frozen. This was sliced and a longitudinal section having a thickness of 8 μm was prepared.
Staining: The specimen was washed with PBS, post-fixed with 4 ° C. acetone for 10 minutes, and then immersed in 0.3% H ₂ O ₂ solution for 30 minutes.
Anti-Neurofilament antibody staining: In order to suppress non-specific reaction, the specimen is immersed in Horse serum for 20 minutes and then immersed in the primary anti-Neurofilament antibody for 2 hours. After washing with TPBS, soak in the secondary antibody Biotinylated IgG for 30 minutes. After washing with TPBS, soaked in Elite ABC for 30 minutes, washed again with TPBS, stained with DAB, and then stained with Mayer's hematoxylin stain. After dehydration with ethanol, EUKITT was enclosed. As a result, the average length of the regenerated sciatic nerve was 12 mm (n = 6).

  The nerve regeneration material of the present invention is useful in the field of regenerative medicine.

It is an example of the apparatus used for an electrospinning method. It is an example of the apparatus used by the electrospinning method. FIG. 3 is a perspective view of the apparatus of FIG. 2. It is the schematic which shows the cross-sectional shape of the cylindrical body obtained in the Example.

Explanation of symbols

1. Nozzle 2. Dope Holding tank 4. 4. Discharge side electrode Collection side electrode 6. High voltage generator 7. Outer diameter 8. 8. Depth of valley Interval between peaks 10. Thickness 11. Yamabe 12. Valley

Claims (11)

(i) an accordion-shaped cylindrical body made of an aliphatic polyester fiber having an average fiber diameter of 0.05 to 50 μm and having peaks and valleys continuous in the axial direction;
(ii) bone marrow cells embedded inside the cylinder,
A nerve regeneration material consisting of The material according to claim 1, wherein the aliphatic polyester fiber is a fiber made of polylactic acid, polyglycolic acid, polycaprolactone, or a copolymer thereof. Cylindrical body, the basis weight of the aliphatic polyester fibers are 1-200 g / ^{m 2,} a thickness of 0.05 to 1 mm, and according to claim 1 or 2 having an outer diameter of 0.5~50mm Material. The material according to any one of claims 1 to 3, wherein the cylindrical body has a peak portion interval of 2 mm or less and a valley portion depth of 0.1 to 10 mm. The material according to any one of claims 1 to 4, wherein the cylindrical body is formed by winding an aliphatic polyester fiber in a spiral shape around the axis of the cylindrical body. The material according to any one of claims 1 to 5, wherein the bone marrow cell is a bone marrow mononuclear cell. The material as described in any one of Claims 1-6 containing the bone marrow cell solution of the density | concentration of 5 * 10 < ⁵ > -5 * 10 < ⁷ > piece / ml inside a cylindrical body. (i) an accordion-shaped cylindrical body made of an aliphatic polyester fiber having an average fiber diameter of 0.05 to 50 μm and having peaks and valleys continuous in the axial direction;
(ii) bone marrow cell solution,
A kit for producing a nerve regeneration material comprising: (i) a step of preparing a bellows-like cylindrical body having a peak portion and a valley portion that are made of aliphatic polyester fibers having an average fiber diameter of 0.05 to 50 μm and that are continuous in the axial direction;
(ii) a step of injecting a bone marrow cell solution into the inside of the cylindrical body,
A method for producing a nerve regeneration material comprising: (i) a step of preparing a bellows-like cylindrical body having a peak portion and a valley portion that are made of aliphatic polyester fibers having an average fiber diameter of 0.05 to 50 μm and that are continuous in the axial direction;
(ii) a step of injecting a bone marrow cell solution into a cylindrical body to obtain a nerve regeneration material; and
(iii) a step of transplanting the obtained nerve regeneration material to a nerve injury site;
A method of regenerating damaged nerves comprising: The regeneration method according to claim 10, wherein the bone marrow cells are bone marrow mononuclear cells.

Images