JP2018121751A - Collagen structure - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a collagen structure composed of a composition element of a plurality of collagen filaments arranged approximately parallel in which the adhesion strength of an adhesion part generated by bonding collagen filaments each other is strong.SOLUTION: In the collagen structure, a plurality of collagen filaments are arranged approximately parallel, the structure has an adjacent part in which collagen filaments abut each other, the adjacent part has at least one adhesion part in which the collagen filaments bond each other in a planar-wise, and whole collagen filaments is subjected to crosslinking treatment.SELECTED DRAWING: Figure 6

Description

  The present invention relates to a collagen structure formed by collagen linear bodies.

  Techniques for producing a planar shape or a three-dimensional shape using a plurality of collagen linear bodies are disclosed in Patent Documents 1 and 2, for example.

  According to claim 1 of Patent Document 1, as a method for producing a collagen fiber bundle, “(i) a step of preparing a collagen yarn produced by subjecting a water-solubilized collagen to a cross-linking treatment as a raw material (ii) the collagen yarn The step (iii) the step of immersing a plurality of wires in substantially parallel (iii) the step of immersing in water, an aqueous inorganic salt solution or a neutralized aqueous solution (iv) the step of drying is described.

  Further, Patent Document 1 describes that the crosslinking treatment may be performed again after the drying step (iv) (paragraph [0041]).

  Claim 1 of Patent Document 2 describes, as “a method for producing a collagen material having an orientation and made of string-shaped collagen gel fragments”, “a phosphate buffered saline solution containing a collagen solution through a nozzle”. While pushing out into a container containing (PBS), the nozzle is slid to give a flow in a certain direction to the collagen solution, thereby giving the orientation in the long axis direction of the string-shaped collagen gel, and the orientation Obtaining a plurality of collagen gel fragments having controlled properties, and arranging the plurality of collagen gel fragments in a desired shape and then solidifying by free drying or freeze drying to solidify the plurality of collagen gel fragments "The process of bonding each other" is described.

Japanese Patent No. 5320726 Patent No. 6031435

  One of the features of Patent Document 1 is that a collagen single yarn used as a raw material has already been cross-linked. Here, it is clear from the description of Patent Document 1 that the collagen single yarn has a circular cross-sectional shape (for example, the description on the outer diameter of the collagen yarn in paragraph [0015], paragraph [0024] ], Description of nozzle diameter in wet spinning method, description of spinning of collagen single yarn having a diameter of about 200 μm in paragraph [0047]. Accordingly, since the collagen single yarns are contacted with each other by circles and circles, they are point contacts from the viewpoint of the cross section, and are linear contacts from the viewpoint of the longitudinal direction (major axis direction). As described above, the collagen fiber bundle of Patent Document 1 has a room for improvement in improving the adhesive force because the crosslinked collagen single yarns are linearly bonded to each other.

  Patent Document 2 describes that even when a collagen material is immersed in PBS or a cell culture solution or transplanted into a living body, an effect of maintaining the shape for a necessary period can be obtained (paragraph). [0030]). However, in view of the production method of Patent Document 2, since no cross-linking treatment is performed, it is difficult to maintain a shape for a certain period when immersed in PBS or a cell culture solution or transplanted in a living body. It is done.

  The present invention provides a collagen structure having a plurality of collagen linear bodies arranged substantially in parallel as a constituent element and having a strong adhesive force at an adhesion portion where the collagen linear bodies are bonded to each other. Let it be an issue.

  As a result of intensive studies on the above problems, the present inventors have found that the above problems can be solved by making the bonding portion where the collagen linear bodies adhere to each other not a line but a plane and a crosslinking treatment. The present invention has been completed based on the above.

The present invention is as follows.
[1] A plurality of collagen linear bodies are arranged substantially in parallel, the collagen linear bodies have adjacent portions adjacent to each other, and at least one bonded portion in which the collagen linear bodies are bonded in a planar shape in the adjacent portions. A collagen structure having a portion and the entire collagen linear body being cross-linked.
[2] The collagen structure according to [1] above, wherein the adhesive portion has a deformed shape deformed by receiving an external force.
[3] The collagen structure according to the above [1] or [2], which has two or more adhesive portions.
[4] The collagen structure according to any one of [1] to [3], wherein the collagen structure has at least one loop-shaped portion formed in a loop shape by a collagenous linear body.
[5] The collagen structure according to [4], wherein the loop-shaped portion is formed at both ends of the collagen structure.
[6] The collagen structure according to any one of the above [1] to [5], wherein the collagen constituting the collagen linear body is refibrillated collagen fibril.
[7] The collagen structure according to [6] above, wherein the refibrillated collagen fibrils are oriented with substantially regularity.
[8] The collagen structure according to any one of [1] to [7], further including any one or more additional materials of resin materials, ceramic materials, and metal materials as a constituent element.
[9] The collagen structure according to the above [8], wherein the additional material has a linear structure.
[10] The collagen structure according to [8] or [9] above, wherein the additional material has biocompatibility.
[11] The method for producing a collagen structure according to any one of [1] to [10], including the following steps.
In the case where a plurality of collagen linear bodies are arranged in parallel and adjacent to each other, the entire collagen linear body is formed in a state in which a close contact portion in close contact with the surface is formed with respect to at least one of the adjacent collagen linear bodies. Step of cross-linking treatment.
[12] The method for producing a collagen structure according to the above [11], wherein the adhesion portion is formed by being deformed so as to adhere in a planar shape by an external force.
[13] The collagen according to [11] or [12], wherein the crosslinking treatment is a crosslinking treatment in which at least one of γ-ray irradiation, electron beam irradiation, UV irradiation, and plasma irradiation is performed in the presence of an aqueous solvent. Manufacturing method of structure.
[14] A medical material using the collagen structure according to any one of [1] to [10].

The schematic diagram of a collagen structure. (a) is a sheet shape in which collagen linear bodies are arranged side by side in a plane, (b) is a rod shape assembled in a parallel state, and (c) is a rod shape assembled spirally. (a) is a schematic diagram of a collagen structure having one loop-shaped portion formed by a collagen linear body. (b) is a schematic diagram of a collagen structure in which loop-shaped portions are formed at adjacent positions in a plurality of collagen linear bodies. (c) is the schematic diagram which the collagen linear body formed only by the loop-shaped part adhere | attached on the collagen linear body which comprises a collagen structure. 2 is a photograph of the collagen structure obtained in Example 1. FIG. (a) Appearance, (b) Cross section with a stereomicroscope. 2 is an appearance photograph of the collagen structure obtained in Example 2. FIG. FIG. 5 is a schematic diagram relating to the production of the collagen structure of Example 3. (a) is a figure for demonstrating winding the collagen linear body 15 around a bridge to the two acrylic sticks 12. FIG. (b) is a diagram for showing that the collagen linear body 15 is bound in a spiral shape with the nylon thread 18. The external appearance photograph of the one side terminal part vicinity of the collagen structure obtained by Example 3. FIG.

  Hereinafter, the present invention will be described in detail based on preferred embodiments. However, the present invention is not limited to the following embodiments, and various modifications can be made within the scope shown in the claims.

(Collagen structure)
The collagen structure of the present invention has a plurality of collagen linear bodies arranged in parallel, and has adjacent portions where the collagen linear bodies are adjacent to each other, and the collagen linear bodies are bonded in a planar shape at the adjacent portions. It has at least one adhesive part, and the entire collagen linear body is subjected to a crosslinking treatment.

  The collagen linear body is composed of collagen, and the shape thereof is not particularly limited as long as the shape is a linear structure, and examples thereof include a thread shape, a rod shape, and a ribbon shape. The cross-sectional shape is not limited to a circle, and may be an ellipse, and may be an oval, for example, a triangle, a rectangle (including a square, rectangle, rhombus, trapezoid, parallelogram), a pentagon, a hexagon, etc. It may be. Moreover, shapes, such as a cross shape, a star shape, C shape, H shape, L shape, O shape, T shape, U shape, V shape, Y shape, may be sufficient. In addition, one collagen linear body may have a plurality of cross-sectional shapes.

  The cross-sectional shapes of the plurality of collagen linear bodies may be the same as or different from each other. For example, some may have the same shape, and some may have different shapes. Further, the cross-sectional areas may have the same size or different sizes. For example, some may have the same shape and the same cross-sectional area, and another several may have a different shape and the same cross-sectional area. Further, for example, all have the same shape, but the cross-sectional areas may be different from each other. As described above, the shape and the cross-sectional area can be appropriately selected and set within the scope of the collagen structure of the present invention.

  There is no particular limitation on the diameter of the linear body (when the cross section is non-circular, such as a polygon or an ellipse) and the length, the length is not particularly limited, and may be appropriately set so as to obtain the effects of the present invention. That's fine. An example of the upper limit of the diameter is preferably 20 mm, more preferably 15 mm, still more preferably 10 mm, and even more preferably 5 mm. An example of the lower limit is 0.5 mm, and more preferably 1 mm. For example, the length is preferably 2 times or more of the diameter, more preferably 3 times or more, still more preferably 5 times or more, and even more preferably 10 times or more. In addition, it is desirable to determine how long the length is appropriately according to the application.

  Examples of the state in which a plurality of collagen linear bodies are arranged substantially in parallel include, for example, a state in which a plurality of prepared collagen linear bodies are arranged in substantially parallel, and one collagen linear body has a predetermined length. Examples include a state where they are reciprocated and arranged so as to be arranged substantially in parallel except at the folded portion, but are not limited thereto.

  Substantially parallel in a state where a plurality of collagen linear bodies are arranged substantially in parallel is sufficient if it is substantially parallel, and does not require geometrically parallel to each other. Further, the substantially parallel includes a spiral shape. As an example, the angle formed by each collagen linear body may be 45 degrees or less with respect to the longitudinal direction, preferably 30 degrees or less, more preferably 15 degrees or less, and most preferably 5 degrees or less. It is.

  The state in which a plurality of collagen linear bodies are arranged in parallel and have adjacent portions includes not only the state of being adjacent over the entire area of the collagen linear body but also the state having adjacent portions and non-adjacent portions. . In the latter state, a plurality of adjacent portions may be provided. Various shapes can be adopted without particular limitation as long as it is within a range arranged substantially in parallel. For example, it may be a sheet shape in which collagen linear bodies are arranged side by side in a plane (FIG. 1 (a)). However, it may be a three-dimensional shape in which collagen linear bodies are gathered. The three-dimensional shape may be, for example, a laminated sheet shape in which a planar horizontal arrangement includes a plurality of stages, or a bundle shape. Examples of bundles include rods assembled in a parallel state (FIG. 1 (b)), rods assembled in a spiral (FIG. 1 (c)), and the like. Other examples of various shapes include a dome shape and a tubular shape. The angle formed between the longitudinal direction of the tube and the longitudinal direction of the collagen linear body in the tubular shape can be in a range from 0 degree (parallel) to 90 degrees (right angle). In addition, since each figure of FIG. 1 typically shows an example of the state arrange | positioned substantially parallel, the presence or absence of an adjacent part is outside the object of illustration. Further, in each drawing of FIG. 1, the cross-sectional shape is shown as a circle, but this is merely for convenience of illustration, and it is a matter of course that the cross-sectional shape is not limited to a circle.

  The adjacent portion has at least one bonded portion where the collagen linear bodies are bonded in a planar shape. In the collagen structure of the present invention, since the bonded portion is planar, the collagen linear bodies can be bonded to each other in a larger area than the linear one. Furthermore, since the entire collagenous linear body has been subjected to a crosslinking treatment, it is a matter of course that the adhesive portion has also been crosslinked. For this reason, strong adhesion becomes possible.

  Having at least one adhesion part means that, for example, two intersecting collagen linear bodies have an adhesion part at the intersection. For example, in two collagen linear bodies adjacent in parallel, if there is an adhesive part over the entire area of the adjacent part, it has one adhesive part, and only a part of the adjacent parts You may have one adhesion part. Since the number of adhesion parts is at least one, it may be two or more. In that case, it has a non-adhesion part. The number of bonded portions is preferably 3 or more, more preferably 5 or more, still more preferably 7 or more, and even more preferably 10 or more.

  When there are two or more bonded portions, the number of each portion of the bonded portion and the non-bonded portion has one of the following relationships. <1> Number of bonded portions = Number of non-bonded portions + 1, <2> Number of bonded portions = Number of non-bonded portions, <3> Number of bonded portions = Number of non-bonded portions-1 . <1> is, for example, an arbitrary portion from each end at the both ends of the collagen linear body, and 0 or more adhesive portions at the portion excluding the adhesive portion at both ends, This applies when the part other than the part is a non-adhesive part. <2> is, for example, an adhesive part at one end of the collagen linear body from the end, and one or more adhesive parts at any part except the other end. This applies when the part other than the part is a non-adhesive part. <3> corresponds to, for example, the case where two or more adhesive portions are present in the portion excluding both ends of the collagen linear body, and the portion other than the adhesive portion is a non-adhesive portion.

  Further, for example, in FIG. 1B, all the adjacent collagenous linear bodies may adhere to each other at a specific site, or two adjacent collagenous linear bodies may adhere at an adhesive part at a certain site. Only the bodies are bonded to each other, but the bonding method may be such that the two bodies are bonded together as a whole.

  The crosslinking treatment can be used without particular limitation as long as it is suitable for collagen crosslinking. Examples of the crosslinking treatment method include a physical crosslinking method and a chemical crosslinking method. Examples of the physical crosslinking method include γ-ray irradiation, electron beam irradiation, plasma irradiation, UV irradiation, and thermal dehydration crosslinking. A typical example of the chemical crosslinking method is a method using a chemical crosslinking agent. The chemical crosslinking agent may be water-soluble or may have vaporization ability. Specific examples of chemical crosslinking agents include glutaraldehyde, polyepoxy compounds (ethylene glycol diglycidyl ether, glycerol polyglycidyl ether, etc.), carbodiimide compounds (1-ethyl-3- (3-dimethylaminopropyl) carbodiimide / hydrochloride, etc. ), Reducing sugars (such as ribose). The crosslinking treatment may be only one method or a combination of a plurality of crosslinking methods. When combining a plurality of crosslinking methods, it is desirable to appropriately design the crosslinking method and the degree of crosslinking so that the desired degree of crosslinking can be effectively obtained.

  The cross-linking treatment is performed over the entire collagen linear body, that is, the entire area of all the collagen linear bodies constituting the collagen structure (including the adhesive portion). As a result, a collagen structure having a high mechanical strength can be obtained. In addition, even when immersed in PBS, cell culture medium, or the like or transplanted into a living body, the shape can be sufficiently maintained for a necessary period. The test example for confirming this is to immerse the collagen structure in Dulbecco's phosphate buffered saline (D-PBS) at 37 ° C for 5 days. If the absolute value of the change rate of the cross-sectional area of the collagen linear body after being immersed in is within 20%, it can be evaluated that the shape retention is high. The absolute value of the change rate is more preferably within 10%. Further, in the above test example, if the dissolution rate is 10% by mass or less, it can be evaluated that it has a property that is difficult to be decomposed in a cell culture environment or an in vivo environment. The dissolution rate is the ratio (%) of the mass of the eluted component from the collagen linear body in D-PBS to the mass of the collagen linear body before immersion. The dissolution rate can be evaluated by gel permeation chromatography (GPS) by measuring the molecular weight distribution of the eluted component in D-PBS or by measuring the mass of the eluted component in D-PBS. In addition, you may use the collagen linear body cut out from the collagen structure as a sample provided to the said test example.

  Another preferred embodiment of the collagen structure of the present invention has a deformed shape in which the adhesive portion is deformed by receiving an external force. Examples of external forces that have caused deformation include tension, compression, shearing, bending, and twisting. If the cross-sectional shape of the collagen linear body is based on a circle or an ellipse, it will be accompanied by deformation in order to obtain a surface-bonded state at the bonded portion. Moreover, even if the cross-sectional shape is a collagen linear body having a planar shape such as a triangle or a quadrangle, the bonded portion may have a deformed shape. As an example of this, when one of the two collagenous linear bodies is circular and the other is a quadrangle, a rectangular flat surface portion of the two bonded portions is subjected to bending deformation. Moreover, even if it is adhesion | attachment by square corners, it can be set as a planar adhesion part by receiving bending deformation.

  Another preferred embodiment of the collagen structure of the present invention is one having at least one loop-shaped portion formed in a loop shape by a collagen linear body. Here, the loop shape means a ring shape, but is not limited to a circular shape, and may be an elliptical shape or a polygonal shape. For example, the loop-shaped portion may be formed in only one of a plurality of collagen linear bodies, and a schematic diagram thereof is shown in FIG. In FIG. 2 (a), one collagenous linear body has one loop-shaped portion, but it may have two or more.

  In addition, the loop-shaped portions are formed at adjacent positions in a plurality of collagen linear bodies (see the schematic diagram in FIG. 2 (b)), and the loop-shaped portions are connected by an adhesive portion bonded by a crosslinking process. In addition, it may be non-connected, that is, an individual loop shape portion may exist independently. In the former case, the adhesive part is preferably bonded in a planar shape, and the part of the adhesive part may be only a part of the loop shape part, or the entire loop shape part. It doesn't matter. Moreover, you may have an adhesion part in the root of a loop shape part.

  As one form of external addition of the collagen structure of the present invention, a collagen linear body formed only by a loop-shaped portion is bonded to the collagen linear body constituting the collagen structure (FIG. 2 (c ). In the said adhesion | attachment, it is preferable to adhere | attach in planar shape by the bridge | crosslinking process. Each drawing in FIG. 2 schematically shows an example of various forms having a loop shape portion. Similarly to FIG. 1, in each drawing of FIG. 2, the presence or absence of the adjacent portion is not illustrated, and the circular cross-sectional shape is merely for convenience of illustration.

  There is no restriction | limiting in particular about the formation position of a loop shape part. A preferred embodiment is one in which loop-shaped portions are formed at both ends of the collagen structure of the present invention. This is a convenient form for pulling the collagen structure itself from both. For example, a stretching force can be applied to the collagen structure by passing a wire through each of the loop-shaped portions at both ends and pulling the wire.

  The form of the collagen constituting the collagen linear body is not particularly limited, and may be in the form of collagen molecules, or refibrillation (fibrosis) having a D cycle formed by association of collagen molecules. It may be in the form of collagen fibrils (also referred to as “refibrillated collagen fibrils” in the present invention). Regardless of the form of collagen molecules and refibrillated collagen fibrils, the orientation is not particularly limited, it may be random orientation without orientation, or oriented with substantially regularity. It may be.

  Here, the collagen linear body composed of collagen molecules is “linear body A”, the collagen linear body composed of randomly oriented refibrillated collagen fibrils is “linear body B”, and is oriented with almost regularity. The collagenous linear body composed of the refibrillated collagen fibrils is referred to as “linear body C”.

  The type of collagen linear body constituting the collagen structure of the present invention may be only one of linear body A, linear body B and linear body C, or two or more types. There may be. When there are two or more kinds, for example, any kind of linear bodies are gathered as a group and the aggregates are adjacent to each other, or one kind of linear body is gathered as a group and the surroundings are separated by another kind. The form which a linear body surrounds, the form where multiple types of linear bodies are mixed, etc. are mentioned.

  From the viewpoint of biocompatibility, linear body B or linear body C having a form of refibrillated collagen fibril is preferable. Particularly preferred is a linear body C composed of refibrillated collagen fibrils oriented with substantially regularity. The linear body C is also preferable from the viewpoint of strength. From these viewpoints, the collagen structure is preferably composed only of the linear body C. Here, with respect to the linear body C, “orientated with substantially regularity” means a state in which most refibrillated collagen fibrils are arrayed substantially in parallel when the collagen linear body is observed as a whole. Of course, it will be appreciated that some refibrillated collagen fibrils are allowed to be present irregularly. Here, “substantially parallel” is intended to include not only completely parallel but also recognized as substantially parallel. Particularly preferred is a state in which the refibrillated collagen fibrils are arranged substantially parallel to the longitudinal direction of the collagen linear body. An example of a method for measuring the degree of orientation is to analyze a 10,000-fold scanning electron microscope image at an arbitrary location of the collagen linear body with Asahi Kasei Engineering Co., Ltd. image analysis software “A Image-kun (registered trademark)”. The degree is calculated by the half width method. The maximum value of the degree of orientation is 1, and a larger value indicates that the orientation is in a certain direction. The preferred range is from 0.5 to 1. If it is in the said range, it can be said that it is in the state which orientated with substantially regularity.

  According to still another preferred embodiment of the collagen structure of the present invention, the collagen structure further contains any one or more additional materials of resin materials, ceramic materials, and metal materials as components. Any additional material is not particularly limited, and a suitable material may be selected from known materials. Further, when the collagen structure of the present invention is used as, for example, a medical material, the additional material is preferably biocompatible.

  Specific examples of the additional material include a thermoplastic resin and a thermosetting resin for the resin material. Further, it may be hydrophilic or hydrophobic. For example, acrylic resin, polyethylene, polypropylene, polystyrene, ABS resin, polycarbonate, polyethylene terephthalate, polyamide, styrene resin, polyester (polylactic acid, polycaprolactone, polyglycolic acid, lactic acid-glycolic acid copolymer, lactic acid-caprolactone copolymer) Etc.), silicone resin, urethane resin, phenol resin, epoxy resin and the like. Among these, when using a biocompatible material, it is preferable to select from, for example, nylon, polyester, silicone resin and the like. Moreover, you may use what has biodegradability.

  A wide variety of materials are known as ceramic materials. However, when a material having biocompatibility is used, it is preferably selected from, for example, hydroxyapatite, tricalcium phosphate and the like.

  Examples of the metal material include titanium, titanium alloy, stainless steel, tantalum, tantalum alloy, platinum, gold, silver, cobalt chrome alloy, aluminum, iron, copper, and brass. Among these, when biocompatible ones are used, it is preferable to select from, for example, stainless steel, titanium, titanium alloy, tantalum, tantalum alloy and the like.

  The shape of the additional material is not particularly limited. Examples thereof include a linear structure and a spherical structure. Examples of the linear structure include various shapes such as a fiber shape, a rod shape, a tubular shape, and a ribbon shape. The linear structure includes those formed using a material having a linear structure such as a mesh, a woven fabric, a nonwoven fabric, and a braid. From the viewpoint of improving the strength and maintaining the shape of the collagen structure of the present invention, the additional material preferably has a linear structure. Further, the additional material having a linear structure may have a loop shape at both ends. From the viewpoint of biocompatibility, it is also preferable that the additional material is not exposed on the surface of the collagen structure by appropriately arranging the collagen linear body.

(Production method)
One embodiment of the method for producing a collagen structure of the present invention preferably includes the following steps. That is, in the case where a plurality of collagen linear bodies are arranged in parallel and adjacent to each other, at least one place between the adjacent collagen linear bodies is formed in a collagen linear state in a state in which a close contact portion in close contact with the surface is formed This is a step of crosslinking the entire body.

  As the collagen linear body, it is preferable to use an uncrosslinked one from the viewpoint of the degree of adhesion between the collagen linear bodies, but from the viewpoint of handling the collagen linear body, a cross-linked one may be used in advance. . When using a collagen linear body that has been previously cross-linked, it is preferable to use one having a low degree of cross-linking so that a cross-linking process for adhering the collagen linear bodies can be performed.

  A collagen linear body can be obtained by a known method, for example, [1] a method of electrospinning a solubilized collagen solution, [2] refibril obtained by adding an appropriate buffer to the solubilized collagen solution. And a method of spinning or injecting an aqueous solution containing a solubilized collagen fibril in the air or in an aqueous solution, and [3] a method of spinning or injecting a solubilized collagen solution into a fibrosis bath.

  Here, in the method [3] above, inorganic salt aqueous solutions, hydrophilic organic solvents such as ethanol, and the like are known as representative examples of fibrosis bath liquids. When a concentrated salt aqueous solution is used among inorganic salt aqueous solutions, or when a hydrophilic organic solvent is used, a collagen linear body composed of collagen that has not been refibrillated is obtained. On the other hand, the linear body C is obtained by using an inorganic salt aqueous solution having an ionic strength appropriately set. An example of a solution for a stabilizing bath for obtaining the linear body C is a physiological isotonic solution or a buffer solution disclosed in JP-A-2006-67983. From the viewpoint of obtaining a high-density collagen linear body, it is also a preferred embodiment to use a solubilized collagen solution having a high collagen concentration (high-concentration collagen solution). As a method for obtaining a high concentration collagen solution, a known method may be used. For example, a method of concentrating by repeating isoelectric precipitation, a method of concentrating by salting out, and the like are known.

  As described above, a plurality of collagen linear bodies are arranged substantially in parallel and adjacent to each other. In the adjacent state, as a method for forming a close contact portion in close contact with at least one place between adjacent collagenous linear bodies, for example, (i) Collagen lines that are both linear in the cross-sectional shape of the close contact portion In the case of linear bodies, a method of arranging the linear portions so that they are in close contact with each other, (ii) in the case of collagen linear bodies having one or both non-linear shapes in the cross-sectional shape of the close-contact portions, And a method of applying a strong external force to closely contact the surface. In the above (ii), it is preferable to use a collagenous linear body having elasticity.

  Here, a specific example of the above (ii) will be described with reference to FIG. By applying an external force to each collagen linear body in the direction of the upper and lower surfaces of the planar shape, the adjacent portions of the collagen linear bodies are deformed and brought into close contact with each other. Also in FIG. 1 (b), an external force is applied from an appropriate direction in order to deform adjacent portions of the collagen linear bodies and bring them into close contact with each other.

  As a method for crosslinking the entire collagen linear body, the various methods described above may be applied. The cross-linking method is a method of cross-linking the entire collagen linear body in a state in which the adhesiveness of the close contact portion in close contact with the surface is maintained, and either a physical cross-linking method or a chemical cross-linking method may be used. . In order to maintain adhesion, it is preferable to use a member. Hereinafter, such a member is referred to as a “contact member”. It is preferable to select and design the shape of the contact member as appropriate so that a desired contact force can be obtained. The material of the adhesion member may be appropriately selected in consideration of compatibility with collagen and a crosslinking method. For example, it is also one of preferred embodiments to select a material that is difficult to adhere to collagen or a material that has high durability against irradiation crosslinking. Specific examples of the material include a resin material and an inorganic material. Examples of the resin material include acrylic resin, polyethylene, polypropylene, ABS resin, polycarbonate, polyethylene terephthalate, polyamide, styrene resin, silicone resin, urethane resin, phenol resin, and epoxy resin. Moreover, as an inorganic material, a metal, glass, etc. are mentioned, for example. Among these, a resin material is preferable, and a preferable example is a urethane resin, a silicone resin, or the like, and a urethane resin is particularly preferable. It is also a preferred embodiment to use a close contact member having water permeability or air permeability.

  A particularly preferred embodiment of the crosslinking treatment method is one in which at least one irradiation crosslinking among γ-ray irradiation, electron beam irradiation, UV irradiation and plasma irradiation is performed in the presence of an aqueous solvent. Hereinafter, this aspect will be described.

  When a collagen linear body produced in an aqueous solvent is used as the collagen linear body, the wet state of the aqueous linear solvent or the collagen linear body is maintained as it is in the presence of water without performing a drying treatment. Under the condition, a plurality of collagen linear bodies are arranged in parallel and adjacent to each other, and after forming a close contact portion in close contact with at least one place between the adjacent collagen linear bodies, It is preferable to carry out radiation crosslinking in the presence of. The aqueous solvent used in the irradiation crosslinking is preferably the same as the aqueous solvent used in the production of the collagen linear body.

  The aqueous solvent applied to the irradiation crosslinking is not limited as long as it contains water. For example, water, physiological saline, buffer solution or physiological saline thereof, acidic salt aqueous solution, neutral salt aqueous solution, alkaline Examples thereof include an aqueous salt solution. Furthermore, an aqueous solvent to which an organic solvent is added can also be used. Aqueous solvents suitable for maintaining the morphology of refibrillated collagen fibrils during irradiation cross-linking are those having moderate ionic strength and moderate pH. About pH, it is preferable to set suitably according to the kind of collagen within the range of 3-10, for example. Examples of the suitable aqueous solvent include a buffer solution or physiological saline thereof, and specific examples include a phosphate buffer solution, a Tris buffer solution, a HEPES buffer solution, an acetate buffer solution, a carbonate buffer solution, a citrate buffer solution, and a phosphate solution. Buffered saline (PBS), Dulbecco's phosphate buffered saline (D-PBS), Tris buffered saline, HEPES buffered saline, and the like. In addition, even if it is an aqueous solvent in which refibrillated collagen fibrils are relatively easy to dissolve, it can be used when the immersion and crosslinking treatment in this aqueous solvent is performed in a short time.

  Irradiation crosslinking may be carried out by only one of γ-ray irradiation, electron beam irradiation, UV irradiation and plasma irradiation, or may be performed by combining two or more. Further, one type of irradiation crosslinking may be performed twice or more. For example, when irradiation crosslinking is performed twice, it is preferable to set so that a low crosslinking degree is obtained at the first time and a high crosslinking degree is obtained at the second time. In addition, when performing a combination of two or more, it is basically preferable to carry out an irradiation method with a high degree of crosslinking after an irradiation method with a low degree of crosslinking, for example, a combination in which γ-ray irradiation is performed after UV irradiation. . Preferably, it is a method of carrying out irradiation crosslinking once by γ-ray irradiation which has high permeability and can be uniformly crosslinked. In particular, in the crosslinking treatment by γ-ray irradiation, a high-strength collagen linear body can be obtained by appropriately setting the irradiation dose. In γ-ray irradiation, a predetermined irradiation dose can be easily obtained by using a radiation source with a fixed dose rate and appropriately setting conditions such as irradiation time. For example, when a cobalt 60 radiation source is used, the crosslinking treatment can be performed with an irradiation dose of 5 to 75 kGy. The irradiation dose is preferably 5 to 50 kGy, more preferably 10 to 50 kGy, and further preferably 15 to 30 kGy. Furthermore, if irradiation conditions are set appropriately, sterilization can be performed simultaneously with the crosslinking treatment. Therefore, by maintaining the sealed state during the crosslinking treatment and after the crosslinking treatment, it can be distributed as a sterilized product to the market as it is.

  Although the mechanism of action when irradiation crosslinking is performed in the presence of an aqueous solvent is not clear, the radicals of water generated by irradiation (γ rays, etc.) act on the uncrosslinked portion of collagen, thereby causing a crosslinking reaction. Presumed to start or progress. Therefore, it is preferable that the fluidity of the aqueous solvent is secured at least at the molecular level at the site intended for crosslinking, and it is desirable to select an adhesion member having water permeability suitable for it. The amount of the aqueous solvent is not particularly limited, and may be adjusted according to the outer shape and size of the collagen structure. For example, at least the entire surface of the collagen structure is covered with an aqueous solvent, and preferably the collagen structure is immersed in the aqueous solvent. Further, even when the collagen structure is not completely immersed in the aqueous solvent, for example, even when a part of the collagen structure is not immersed in the aqueous solvent, the infiltrating property in the part can be secured. It can be said that the collagen structure is immersed in an aqueous solvent. In this specification, the state of the aqueous solvent for the collagen structure as exemplified above is referred to as “in the presence of an aqueous solvent”. The amount of the aqueous solvent is, for example, preferably in the range of 2 to 100 times, more preferably in the range of 5 to 100 times, and still more preferably in the range of 10 to 50 times the volume of the collagen structure.

  By the way, one characteristic of a collagen structure composed of a collagen linear body cross-linked by irradiation in the presence of an aqueous solvent is, for example, as described in Japanese Patent No. 5633880, as a cell culture environment or an in vivo environment. It is difficult to disassemble. The collagen structure produced by suitable irradiation cross-linking conditions has the dissolution rate when the collagen structure is immersed in Dulbecco's phosphate buffered saline (D-PBS) at 37 ° C. for 5 days. Is 10% by mass or less.

  The collagen structure of the present invention can be widely used as a medical material. Among them, the entire collagen linear body is constituted by the linear body C, and has a loop-shaped portion formed in a loop shape by the collagen linear body at both ends of the collagen structure, and other than the root portion of the loop-shaped portion. In addition to the high biocompatibility due to the properties of re-fibrillated collagen fibrils oriented with a regularity, the collagen structure having a plurality of bonded portions (and thus also a plurality of non-adhered portions) is high by cross-linking treatment. It has strength and also has elasticity because the collagenous linear body can bend at the non-adhered portion. This collagen structure is a material that can be applied as a transplanted ligament in ligament reconstruction since a wire can be passed through each of the loop-shaped portions at both ends and the wire can be pulled. Moreover, it is good also as a collagen structure provided with the additional material for the purpose of the further strength improvement.

  EXAMPLES The present invention will be described in more detail with reference to the following examples, but the present invention is not limited thereto. In Examples, “%” means “% by mass” unless otherwise specified.

[Production of collagen linear body]
As a solubilized collagen solution, “Cell Campus FD-08G” (freeze-dried product) manufactured by Taki Chemical Co., Ltd. manufactured from tilapia scales was dissolved in a pH 3 HCl solution, and the collagen concentration was adjusted to 15%. A clear solution was used. PBS was used as the fibrosis bath. The solubilized collagen solution was filled into a syringe equipped with an 18G non-bevel needle, and then injected into a fibrosis bath to obtain a collagen linear body having a circular cross section and a length of about 200 cm. This collagenous linear body was used in the following examples while maintaining a wet state.

[Example 1]
Two pieces of collagen linear bodies, which are cut to a length of about 7 cm, are arranged in parallel in a plane and the collagen linear bodies are adjacent to each other, and the upper and lower surfaces are pre-immersed in PBS. By pressing with a polyurethane sponge (adhering member), the sheet was brought into close contact with the entire area of the adjacent portion. This was immersed in PBS and irradiated with 25 kGy of γ rays. The obtained collagen structure was obtained by bonding the bonded portions of the collagen linear bodies in a planar shape, and the entire collagen linear body including the bonded portions was subjected to a crosslinking treatment. FIG. 3 (a) is a photograph of the appearance of the entire collagen structure, and FIG. 3 (b) is a photograph of a cross section taken with a stereomicroscope. Note that FIG. 3B is a diagram in which the cross-sectional portions are separated from each other so that the cross-sectional shapes of the individual collagenous linear bodies can be easily confirmed.

[Example 2]
In order to produce a bundle-like (rod-like) collagen structure, a single collagenous linear body was reciprocated at a length of 8 cm, and was arranged so as to be arranged substantially in parallel at the portion excluding the folding. Nylon yarn was bound to this in a spiral shape to form a bundle, and a close contact portion and a non-contact portion that were in close contact with each other were formed. This was immersed in PBS and irradiated with 25 kGy of γ rays.
The obtained collagen structure (FIG. 4) was obtained by bonding the bonded portions of the collagen linear bodies in a planar shape, and the entire collagen linear bodies including the bonded portions were subjected to a crosslinking treatment. This collagen structure had stretchability in the longitudinal direction due to the presence of the non-adhesive portion.

Example 3
As shown in the schematic diagram of FIG. 5 (a), one collagen linear body 15 was wound around a bridge around two acrylic rods 12 installed at a distance of about 8 cm. Nylon thread 18 was bound in a spiral shape (see the schematic diagram in FIG. 5 (b)) to form a close contact portion and a non-contact portion that were in close contact with each other. This was irradiated with 25 kGy of γ-rays while immersed in PBS with an acrylic rod attached.
The obtained collagen structure had loop-shaped portions at both ends by removing the acrylic rod. Moreover, the adhesion part of collagen linear bodies adhere | attached planarly, and the whole collagen linear body including the adhesion part was bridge | crosslinked. This collagen structure had stretchability in the longitudinal direction due to the presence of the non-adhesive portion. FIG. 6 is a photograph of the vicinity of one end of the collagen structure, and a spiral pattern formed by binding the loop shape portion and the nylon thread can be confirmed.

  12 acrylic rods, 15 collagen filaments, 18 nylon threads.

Claims (14)

A plurality of collagen linear bodies are arranged substantially in parallel, the collagen linear bodies have adjacent portions adjacent to each other,
In the adjacent portion, the collagen linear bodies have at least one adhesive portion bonded in a planar shape,
The entire collagenous linear body is cross-linked.
Collagen structure. The collagen structure according to claim 1, wherein the adhesive portion has a deformed shape deformed by receiving an external force. The collagen structure according to claim 1 or 2, wherein the collagen structure has two or more adhesion portions. The collagen structure according to any one of claims 1 to 3, wherein the collagen structure has at least one loop-shaped portion formed in a loop shape by a collagen linear body. The collagen structure according to claim 4, wherein the loop-shaped portion is formed at both ends of the collagen structure. The collagen structure according to any one of claims 1 to 5, wherein the collagen constituting the collagen linear body is a refibrillated collagen fibril. The collagen structure according to claim 6, wherein the refibrillated collagen fibrils are oriented with substantially regularity. Furthermore, the collagen structure of any one of Claims 1-7 containing any 1 or more types of additional material among a resin material, a ceramic material, and a metal material as a component. The collagen structure according to claim 8, wherein the additional material has a linear structure. The collagen structure according to claim 8 or 9, wherein the additional material has biocompatibility. The manufacturing method of the collagen structure of any one of Claims 1-10 including the following processes.
In the case where a plurality of collagen linear bodies are arranged in parallel and adjacent to each other, the entire collagen linear body is formed in a state in which a close contact portion in close contact with the surface is formed with respect to at least one of the adjacent collagen linear bodies. Step of cross-linking treatment. The method for producing a collagen structure according to claim 11, wherein the close contact portion is formed by being deformed so as to be in close contact with the surface by an external force. The method for producing a collagen structure according to claim 11 or 12, wherein the crosslinking treatment is a crosslinking treatment in which at least one of γ-ray irradiation, electron beam irradiation, UV irradiation, and plasma irradiation is performed in the presence of an aqueous solvent. The medical material using the collagen structure of any one of Claims 1-10.