Gel material for treating tendons or ligaments

Abstract

To provide a gel material that suppresses intrusion of a scar structure in treatment of a tendon or a ligament, promotes healing due to inter-tendon cells, and makes an intrinsic function of the tendon efficiently recover, and a treatment method.SOLUTION: A gel material containing a polymer gel for suppressing expression of type III collagen in treatment of injury or rupture of tendons or ligaments, in which the polymer gel is a hydro gel having a structure in which a three-dimensional network structure is formed by crosslinking a hydrophilic polymer, has a polymer content in a range of 1 to 5 wt.%, and an elastic modulus in a range of 100 to 10000 Pa.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present invention relates to a gel material suitable for treating tendons or ligaments, and a method of treating tendons or ligaments using said gel material. [Background technology]

[0002] Generally, there is no established effective treatment for tendon and ligament injuries. On the other hand, surgical treatments such as suture surgery are selected for tendon ruptures and ligament reconstruction surgery for ligament ruptures, while conservative treatments are selected for partial injuries.

[0003] However, in both treatments, healing by scar tissue consisting of extratendinous cells (fibroblasts and inflammatory cells) is mixed in the healing process of the tendon and ligament, and the healing of the tendon and ligament itself is inhibited (for example, Non-Patent Document 1). As a result, the healed tendon and ligament loses strength equivalent to that of scar tissue and is unable to exert the original function of the tendon. This causes re-rupture after surgery and hinders return to sports, lowering the quality of life (QOL) of many patients. [Prior art documents] [Non-patent literature]

[0004] [Non-Patent Document 1] Best et.al,The FASEB Journal,vol.33,July 2019 Summary of the Invention [Problem to be solved by the invention]

[0005] Therefore, an objective of the present invention is to provide a method for inhibiting the invasion of scar tissue in the treatment of tendons or ligaments, promoting healing by cells within the tendon, and efficiently restoring the tendon's original function. [Means for solving the problem]

[0006] As a result of intensive research aimed at solving the above-mentioned problems, the inventors discovered that by using a hydrogel having a structure in which hydrophilic polymers form a three-dimensional mesh structure by cross-linking between molecules, and covering the area around a damaged or ruptured tendon ligament with said hydrogel, it is possible to inhibit the invasion of scar tissue, which is primarily composed of type III collagen, and to efficiently bring about endogenous healing by cells within the tendon, thereby completing the present invention.

[0007] That is, in one aspect, the present invention relates to a gel material suitable for treating injuries or ruptures of tendons or ligaments, more specifically, <1> A gel material containing a polymer gel for suppressing expression of type III collagen in the treatment of injury or rupture of a tendon or ligament, the gel material being characterized in that the polymer gel is a hydrogel having a three-dimensional network structure formed by crosslinking a hydrophilic polymer, the gel material having a polymer content in the range of 2 to 5% by weight and an elastic modulus in the range of 100 to 10,000 Pa; <2> The hydrophilic polymer has a polyether skeleton or a polyvinyl skeleton. <1> 2. A gel material according to claim 1 ; <3> The hydrophilic polymer is a di-, tri- or tetra-branched polyethylene glycol. <1> or <2> 2. A gel material according to claim 1 ; <4> The hydrophilic polymer is composed of a first polymer unit having one or more nucleophilic functional groups at a side chain or at a terminal, and a second polymer unit having one or more electrophilic functional groups at a side chain or at a terminal. <1> ~ <3> 13. A gel material according to any one of claims 1 to 12. <5> The nucleophilic functional group is selected from the group consisting of a thiol group and an amino group, and the electrophilic functional group is selected from the group consisting of a maleimidyl group, an N-hydroxy-succinimidyl (NHS) group, a sulfosuccinimidyl group, a phthalimidyl group, an imidazoyl group, an acryloyl group, a nitrophenyl group, and -CO2PhNO2. <4> 2. A gel material according to claim 1 ; <6> The above-mentioned method is used to cover a tendon or ligament after a tendon or ligament suturing operation. <1> ~ <5> 13. A gel material according to any one of claims 1 to 12. <7> By covering the tendon or ligament, the expression of type III collagen can be suppressed and the expression of type I collagen can be maintained. <6> A gel material according to claim 1, <8> the above <1> ~ <7> A kit for producing the gel material according to any one of the above, comprising two or more solutions containing a hydrophilic polymer. This provides:

[0008] In another aspect, the present invention relates to the treatment of tendons or ligaments using the gel material described above, more specifically, <9> the above <1> ~ <7> A method for treating an injury or rupture of a tendon or ligament, comprising covering the periphery of the tendon or ligament after a suturing operation with the gel material according to any one of the above items; <10> the above <1> ~ <7> 2. A method for suppressing the expression of type III collagen in a tendon or ligament, comprising covering the tendon or ligament after suturing surgery with the gel material according to any one of the above items 1 to 10. <11> A method for treating injury or rupture of a tendon or ligament, comprising the steps of: preparing a polymer solution A containing a first hydrophilic polymer having one or more nucleophilic functional groups at a side chain or end, and a polymer solution B containing a second hydrophilic polymer having one or more electrophilic functional groups at a side chain or end; applying polymer solutions A and B to the periphery of a tendon or ligament after suturing surgery to crosslink the hydrophilic polymers to form a polymer gel having a three-dimensional network structure, which coats the periphery of the tendon or ligament; and suppressing the expression of type III collagen in the tendon or ligament by the coating; <12> The polymer gel has a polymer content in the range of 1 to 5% by weight and an elastic modulus in the range of 100 to 10,000 Pa. <11> The method of treatment according to claim 1, <13> The first and second hydrophilic polymers have a polyether backbone or a polyvinyl backbone. <11> or <12> The method of treatment according to claim 1, <14> The first and second hydrophilic polymers are di-, tri- or tetra-branched polyethylene glycols. <11> ~ <13> any one of the methods of treatment according to any one of the preceding claims; and <15> the nucleophilic functional group is selected from the group consisting of a thiol group and an amino group, and the electrophilic functional group is selected from the group consisting of a maleimidyl group, an N-hydroxy-succinimidyl (NHS) group, a sulfosuccinimidyl group, a phthalimidyl group, an imidazoyl group, an acryloyl group, a nitrophenyl group, an isothiocyanate group, an aldehyde group, an iodoacetamide group, a vinylsulfone group, and -CO2PhNO2; <11> ~ <14> The method for treatment according to any one of the preceding claims. This provides:

[0009] In a further aspect, the present invention also relates to a screening method using the above-mentioned gel material, more specifically, <16> A method for screening for a drug effective for treating injury or rupture of a tendon or ligament, comprising: <1> ~ <7> The screening method includes a step of searching for and identifying a compound having type I collagen proliferation activity in a tendon or ligament covered with the gel material according to any one of the above. This provides: Effect of the Invention

[0010] The gel material and treatment method of the present invention can inhibit the infiltration of inflammatory cells and fibroblasts from around the affected area, and inhibit or eliminate the infiltration of scar tissue composed mainly of type III collagen, which has been considered unavoidable in the healing process of tendons and ligaments. This promotes endogenous healing based on the expression of type I collagen by cells in the tendon, and efficiently restores the original function of the tendon, an effect not achieved by conventional methods. [Brief description of the drawings]

[0011] [Figure 1] FIG. 1 shows images of flexor tendons coated with a gel of the present invention (gel group) and flexor tendons not coated with a gel (control group) 21 days after surgery. [Diagram 2] FIG. 2 is an image showing tissue cross-sections of flexor tendons coated with a gel of the present invention (gel group) and flexor tendons not coated with a gel (control group). DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0012] Hereinafter, embodiments of the present invention will be described. The scope of the present invention is not limited to these descriptions, and other than the following examples, the present invention can be appropriately modified and implemented without departing from the spirit of the present invention.

[0013] 1. Gel material of the present invention The gel material of the present invention is a gel material containing a polymer gel for suppressing the expression of type III collagen in the treatment of injury or rupture of tendons or ligaments. The polymer gel is a hydrogel having a three-dimensional network structure formed by crosslinking of hydrophilic polymers, and is characterized by having a polymer content in the range of 1 to 5% by weight and an elastic modulus in the range of 100 to 10,000 Pa. Here, "elastic modulus" refers to the storage elastic modulus G'.

[0014] (1-1) Hydrophilic polymer The hydrophilic polymer constituting the polymer gel is capable of forming a hydrogel by crosslinking, more specifically, is capable of forming a network structure, particularly a three-dimensional network structure, in the final gel by crosslinking the polymers with each other or via any low molecular weight compound. As the hydrophilic polymer, any known hydrophilic polymer in the art can be used as long as it is capable of forming a hydrogel by a gelation reaction (crosslinking reaction, etc.) in an aqueous solution, but preferably is a hydrophilic polymer having a polyether skeleton or a polyvinyl skeleton.

[0015] Representative examples of polymers having a polyether skeleton include polymers having a polyalkylene glycol skeleton. Preferably, polymers having a plurality of polyethylene glycol skeleton branches are used, and in particular, di-, tri- or tetra-branched polyethylene glycols are preferred. Gels consisting of a tetrabranched polyethylene glycol skeleton are generally known as Tetra-PEG gels, and a meshwork network is constructed by an AB-type cross-end coupling reaction between two kinds of tetrabranched polymers each having an electrophilic functional group such as an active ester structure and a nucleophilic functional group such as an amino group at the end (Matsunaga et al., Macromolecules, Vol. 42, No. 4, pp. 1344-1351, 2009). Tetra-PEG gels can be easily prepared on the spot by simple two-liquid mixing of each polymer solution, and the gelation time can be controlled by adjusting the pH and ionic strength during gel preparation. Since this gel is mainly composed of PEG, it is also excellent in biocompatibility.

[0016] Examples of hydrophilic polymers having a polyvinyl skeleton include polyalkyl methacrylates such as polymethyl methacrylate, polyacrylates, polyvinyl alcohols, poly N-alkylacrylamides, and polyacrylamides.

[0017] The hydrophilic polymer is 1x10 3 ~1x10 5 range, preferably 5x10 3 ~5x10 4 in the range of 1x10 4 ~4x10 4 The weight average molecular weight (Mw) ranges from 0.1 to 1.0.

[0018] In a preferred embodiment, the hydrophilic polymer is a combination of a first polymer unit having one or more nucleophilic functional groups at the side chain or end and a second polymer unit having one or more electrophilic functional groups at the side chain or end. For example, it is preferred that the first polymer unit has one or more nucleophilic functional groups at the side chain or end, and the second polymer unit has one or more electrophilic functional groups at the side chain or end. A gel having a three-dimensional network structure is formed by crosslinking such nucleophilic functional groups and electrophilic functional groups. Here, the total number of nucleophilic functional groups and electrophilic functional groups is preferably 5 or more. It is more preferred that these functional groups are present at the end.

[0019] Nucleophilic functional groups present in the first and second polymer units include thiol groups (-SH), amino groups, etc., and those skilled in the art can use known nucleophilic functional groups as appropriate. Preferably, the nucleophilic functional group is a -SH group. The nucleophilic functional groups may be the same or different, but are preferably the same. By using the same functional groups, the reactivity with the electrophilic functional groups that form crosslinks becomes uniform, making it easier to obtain a gel with a uniform three-dimensional structure.

[0020] As the electrophilic functional group present in the first and second polymer units, an active ester group can be used. Examples of such active ester groups include a maleimidyl group, an N-hydroxy-succinimidyl (NHS) group, a sulfosuccinimidyl group, a phthalimidyl group, an imidazoyl group, an acryloyl group, a nitrophenyl group, an isothiocyanate group, an aldehyde group, an iodoacetamide group, a vinylsulfone group, and -CO2PhNO2 (Ph represents an o-, m-, or p-phenylene group), and other known active ester groups can be appropriately used by those skilled in the art. Preferably, the electrophilic functional group is a maleimidyl group. The electrophilic functional groups may be the same or different, but are preferably the same. By using the same functional group, the reactivity with the nucleophilic functional group that forms the crosslinked bond becomes uniform, and it becomes easier to obtain a gel having a uniform three-dimensional structure.

[0021] A non-limiting specific example of a preferred polymer unit having a nucleophilic functional group at its terminal is a compound represented by the following formula (I) having four branches of a polyethylene glycol skeleton and a thiol group at its terminal. [ka]

[0022] In formula (I), R 11 ~R 14 are the same or different, and each represents a C1-C7 alkylene group, a C2-C7 alkenylene group, -NH-R 15 -, -CO-R 15 -, -R 16 -OR 17 -, -R 16 -NH-R 17 -, -R 16 -CO2-R 17 -, -R 16 -CO2-NH-R 17 -, -R 16 -CO-R 17 - or -R 16 -CO-NH-R 17 - where R 15 represents a C1-C7 alkylene group, R 16 represents a C1-C3 alkylene group, R 17 represents a C1-C5 alkylene group.

[0023] n 11 ~n 14 may be the same or different. 11 ~n 14 The closer the values ​​of n are, the more uniform the three-dimensional structure can be and the higher the strength will be. Therefore, in order to obtain a gel with high strength, it is preferable that they are the same. 11 ~n 14 If the value of is too high, the gel strength will be weak and 11 ~n 14 If the value of n is too low, the gel is difficult to form due to the steric hindrance of the compound. 11 ~n 14is an integer value of 25 to 250, preferably 35 to 180, more preferably 50 to 115, and particularly preferably 50 to 60. The molecular weight is 1x10 3 ~1x10 5 range, preferably 5x10 3 ~5x10 4 in the range of 1x10 4 ~4x10 4 A weight average molecular weight (Mw) in the range is preferred.

[0024] In the above formula (I), R 11 ~R 14 R is a linker moiety that connects the functional group to the core moiety. 11 ~R 14 may be the same or different, but are preferably the same in order to produce a high-strength gel having a uniform three-dimensional structure. 11 ~R 14 represents a C1-C7 alkylene group, a C2-C7 alkenylene group, -NH-R 15 -, -CO-R 15 -, -R 16 -OR 17 -, -R 16 -NH-R 17 -, -R 16 -CO2-R 17 -, -R 16 -CO2-NH-R 17 -, -R 16 -CO-R 17 - or -R 16 -CO-NH-R 17 - where R 15 R represents a C1-C7 alkylene group. 16 R represents a C1-C3 alkylene group. 17 represents a C1-C5 alkylene group.

[0025] Here, the term "C1-C7 alkylene group" refers to an alkylene group having 1 to 7 carbon atoms which may have branches, and refers to a linear C1-C7 alkylene group or a C2-C7 alkylene group having one or more branches (having 2 to 7 carbon atoms including the branches). Examples of the C1-C7 alkylene group include a methylene group, an ethylene group, a propylene group, and a butylene group. Examples of C1-C7 alkylene groups include -CH2-, -(CH2)2-, -(CH2)3-, -CH(CH3)-, -(CH2)3-, -(CH(CH3))2-, -(CH2)2-CH(CH3)-, -(CH2)3-CH(CH3)-, -(CH2)2-CH(C2H5)-, -(CH2)6-, -(CH2)2-C(C2H5)2-, and -(CH2)3C(CH3)2CH2-.

[0026] The term "C2-C7 alkenylene group" refers to a linear or branched alkenylene group having 2 to 7 carbon atoms and having one or more double bonds in the chain, and examples thereof include divalent groups having double bonds formed by removing 2 to 5 hydrogen atoms from adjacent carbon atoms of the alkylene group.

[0027] On the other hand, non-limiting specific examples of preferred polymer units having an electrophilic functional group at their terminals include, for example, a compound represented by the following formula (II) having four branches of a polyethylene glycol skeleton and an N-hydroxy-succinimidyl (NHS) group at its terminal. [ka]

[0028] In the above formula (II), n 21 ~n 24 may be the same or different. 21 ~n 24 The closer the values ​​of n are, the more uniform the gel can be in a three-dimensional structure and the higher the strength, which is preferable, and it is more preferable that they are the same. 21 ~n 24 If the value of is too high, the gel strength will be weak and 21 ~n 24If the value of n is too low, the gel is difficult to form due to the steric hindrance of the compound. 21 ~n 24 is an integer value of 5 to 300, preferably 20 to 250, more preferably 30 to 180, further preferably 45 to 115, and further preferably 45 to 55. The molecular weight of the second four-branching compound of the present invention is 1x10 3 ~1x10 5 range, preferably 5x10 3 ~5x10 4 in the range of 1x10 4 ~4x10 4 A weight average molecular weight (Mw) in the range is preferred.

[0029] In the above formula (II), R 21 ~R 24 R is a linker moiety that connects the functional group to the core moiety. 21 ~R 24 may be the same or different, but are preferably the same in order to produce a high-strength gel having a uniform three-dimensional structure. 21 ~R 24 are the same or different, and each represents a C1-C7 alkylene group, a C2-C7 alkenylene group, -NH-R 25 -, -CO-R 25 -, -R 26 -OR 27 -, -R 26 -NH-R 27 -, -R 26 -CO2-R 27 -, -R 26 -CO2-NH-R 17 -, -R 26 -CO-R 27 - or -R 26 -CO-NH-R 27 - where R 25 R represents a C1-C7 alkylene group. 26 R represents a C1-C3 alkylene group. 27 represents a C1-C5 alkylene group.

[0030] In this specification, the alkylene group and the alkenylene group may have one or more optional substituents. Examples of the substituents include, but are not limited to, an alkoxy group, a halogen atom (which may be a fluorine atom, a chlorine atom, a bromine atom, or an iodine atom), an amino group, a mono- or di-substituted amino group, a substituted silyl group, an acyl group, or an aryl group. When an alkyl group has two or more substituents, they may be the same or different. The same applies to the alkyl portion of other substituents containing an alkyl portion (e.g., an alkyloxy group, an aralkyl group, etc.).

[0031] In addition, in the present specification, when a functional group is defined as "optionally having a substituent", the type of the substituent, the substitution position, and the number of the substituent are not particularly limited, and when it has two or more substituents, they may be the same or different. Examples of the substituent include, but are not limited to, an alkyl group, an alkoxy group, a hydroxyl group, a carboxyl group, a halogen atom, a sulfo group, an amino group, an alkoxycarbonyl group, and an oxo group. These substituents may further have a substituent.

[0032] In another embodiment, a low molecular weight compound can be additionally used to crosslink the hydrophilic polymer. More specifically, one of the first polymer unit or the second polymer unit can be replaced with a low molecular weight compound. In this case, the low molecular weight compound has one or more nucleophilic functional groups or electrophilic functional groups in the molecule. This allows, for example, a low molecular weight compound having a nucleophilic functional group in the molecule to be used instead of the first polymer, and reacted with a second polymer having one or more electrophilic functional groups in the side chain or end, thereby crosslinking between the second polymers and gelling them. Examples of such "low molecular weight compounds having a nucleophilic functional group in the molecule" include compounds having a thiol group in the molecule, for example, dithiothreitol can be used.

[0033] (1-2) Hydrogel As described above, the gel material of the present invention includes a hydrogel (polymer gel) having a structure in which the hydrophilic polymer is crosslinked to form a three-dimensional network structure. In this specification, the term "gel" generally refers to a dispersion system of polymers that has high viscosity and has lost fluidity, and has a state in which the storage modulus G' and loss modulus G" satisfy the relationship G'≧G". In addition, the term "hydrogel" refers to a gel that contains water.

[0034] The hydrogel contained in the gel material of the present invention has a polymer content in the range of 1 to 5% by weight, preferably in the range of 2 to 4% by weight. The hydrogel also has an elastic modulus in the range of 100 to 10,000 Pa, preferably in the range of 500 to 5,000 Pa. By setting the polymer content and elastic modulus within an appropriate range, undesirable effects such as excessive gel expansion when the hydrogel is used to cover the periphery of tendons or ligaments can be suppressed, and the hydrogel can have an appropriate strength to remain in the affected area for a certain period of time.

[0035] Moreover, the hydrogel contained in the gel material of the present invention preferably has an osmotic pressure of 500 to 10,000 Pa. In general, the osmotic pressure (Π os ) can be calculated by subjecting a sample prepared in a dialysis device to reverse osmosis using polyvinylpyrrolidone (PVP) solutions of different concentrations and using the Flory-Rehner equilibrium equation shown below.

number

[0036] 2. Treatment Method of the Present Invention In another aspect, the present invention relates to a method for treating an injury or rupture of a tendon or ligament using the gel material. More specifically, the gel material is used to cover the affected area of ​​a tendon or ligament that needs treatment due to injury or rupture, thereby promoting healing. Covering such an affected area is typically performed after a tendon or ligament suturing operation.

[0037] By covering the tendon or ligament after suturing surgery with the above-mentioned gel material, it is possible to inhibit the infiltration of inflammatory cells and fibroblasts from around the affected area and inhibit or eliminate the infiltration of scar tissue. Surprisingly, the present invention has revealed that tendon or ligament healing proceeds efficiently even in a state where scar tissue, which has been thought to be unavoidable in the healing process of tendons and ligaments, is inhibited or eliminated.

[0038] In the present specification, the tendon or ligament to be treated is a tendon or ligament of a mammal selected from the group consisting of humans, horses, pigs, dogs, cats, guinea pigs, rabbits, rats and mice. Typically, it is a tendon or ligament in the human body.

[0039] As used herein, "injury or tear" of a tendon or ligament can include traumatic injury or tear (e.g., due to a sports injury, overuse, or medical or surgical intervention), injury of genetic origin or disease, and disease. "Tear" also includes partial tears.

[0040] As used herein, "tendon" includes the Achilles tendon, rotator cuff, hand and wrist, patellar tendon, flexor tendon, and extensor tendon. "Injury" to a tendon can include tendinosis, tendinitis, synovitis, tenosynovitis, and other injuries and tendon-related disorders such as avulsion.

[0041] In addition, in this specification, the term "ligament" includes ligaments in the knee joint, shoulder, elbow joint, ankle joint, and spine. For example, "damage" to the knee ligament includes lateral collateral ligament injury, medial collateral ligament injury, anterior cruciate ligament injury, and posterior cruciate ligament injury.

[0042] Without necessarily being bound by theory, as shown in the examples below, when the periphery of a tendon or ligament is covered with the above-mentioned gel material, the expression of type III collagen characteristic of scar tissue is significantly suppressed, while the expression of type I collagen by cells in the tendon is maintained, and it is believed that the repair of the tendon or ligament occurs by endogenous healing due to the expression of type I collagen. This healing process had not been elucidated before, and is a mechanism that has been revealed for the first time by the present invention. About 95% of the collagen that constitutes the tendon is composed of type I collagen, and this healing process is rational and ideal, and it can be said that this is the effect that has been achieved for the first time in the present invention in that healing occurs while maintaining the original function of the tendon or ligament.

[0043] Therefore, the method of the present invention can be said to be a method for suppressing the expression of type III collagen in tendons or ligaments, which includes covering the periphery of the tendons or ligaments after suturing surgery with the above-mentioned gel material. Alternatively, it can be expressed as being able to suppress the expression of type III collagen and maintain the expression of type I collagen by covering the periphery of the tendons or ligaments after suturing surgery with the above-mentioned gel material.

[0044] In an exemplary embodiment, the method of the present invention for treating tendon or ligament injuries or ruptures comprises the steps of: a) preparing a polymer solution A containing a first hydrophilic polymer having one or more nucleophilic functional groups at a side chain or at an end, and a polymer solution B containing a second hydrophilic polymer having one or more electrophilic functional groups at a side chain or at an end; b) applying the polymer solutions A and B to the periphery of the tendon or ligament after suturing surgery, thereby forming a polymer gel having a three-dimensional network structure formed by crosslinking the hydrophilic polymer, and coating the periphery of the tendon or ligament; and c) inhibiting expression of type III collagen in the tendon or ligament by means of said coating.

[0045] The types of hydrophilic polymers used in steps a) and b) are as already described for the gel material of the present invention.

[0046] The solvent in the polymer solutions A and B is water, but in some cases, it may be a mixed solvent containing alcohols such as ethanol or other organic solvents. Preferably, the polymer solutions A and B are aqueous solutions containing water as the sole solvent. The volumes of the polymer solutions A and B can be appropriately adjusted depending on the area of ​​the affected area to which they are applied, the complexity of the structure, etc., but are typically in the range of 0.1 to 20 ml, preferably 1 to 10 ml, respectively.

[0047] The pH of the polymer solutions A and B is typically in the range of 4 to 8, and preferably in the range of 5 to 7. The pH of the polymer solutions A and B can be adjusted using a pH buffer known in the technical field. For example, the pH can be adjusted to the above range by using a citrate-phosphate buffer (CPB) and changing the mixing ratio of citric acid and disodium hydrogen phosphate.

[0048] In step b), polymer solutions A and B may be applied independently to the periphery of the tendon or ligament, or a solution may be prepared by mixing polymer solutions A and B immediately beforehand, and the mixed solution may be applied to the periphery of the tendon or ligament, thereby allowing a polymer gel (hydrogel) to be formed in situ.

[0049] The gelation time in this case is preferably in the range of 10 to 300 seconds, and more preferably in the range of 30 to 100 seconds. To repeat, the gelation time can be adjusted mainly by appropriately setting the polymer concentration, pH, and ionic strength of the polymer solution. Here, the "gelation time" refers to the time required for the storage modulus G' and the loss modulus G" to reach G'=G".

[0050] Furthermore, the polymer gel formed around the tendon or ligament in step b) preferably remains around the affected area for the period required for the tendon or ligament to be repaired or regenerated. For example, the polymer gel preferably has a decomposition rate in vivo of 10 to 90 days, more preferably 20 to 50 days.

[0051] The polymer solutions A and B can be mixed using, for example, a two-liquid mixing syringe as disclosed in International Publication WO2007 / 083522. The temperature of the two liquids during mixing is not particularly limited, and may be a temperature at which the precursor units are dissolved and each liquid has fluidity. For example, the temperatures of the two liquids may be different, but it is preferable that the temperatures are the same because the two liquids are more easily mixed.

[0052] Step c) is a step of suppressing the expression of type III collagen in tendons or ligaments by covering the tendons or ligaments with a polymer gel. This is mainly based on the fact that by covering the tendons or ligaments with a polymer gel, the infiltration of inflammatory cells and fibroblasts from the surrounding area is suppressed, and the infiltration of scar tissue in which type III collagen is abundant is suppressed or eliminated. Without necessarily being bound by theory, it is believed that the treatment method of the present invention can maintain the expression of type I collagen by cells in tendons while suppressing the exogenous expression of type III collagen due to the infiltration of scar tissue, thereby resulting in endogenous healing.

[0053] In another aspect, the present invention also relates to a kit for preparing the above-mentioned gel material (polymer gel), which contains two or more solutions containing a hydrophilic polymer. Such a kit is suitable for use in the above-mentioned treatment method. The type of hydrophilic polymer contained in the solution is as described above, and any solution that can form a hydrogel by crosslinking can be used. Preferably, the solution can be a polymer solution A containing a first hydrophilic polymer having one or more nucleophilic functional groups at the side chain or end, and a polymer solution B containing a second hydrophilic polymer having one or more electrophilic functional groups at the side chain or end. Typically, the first and second hydrophilic polymers are polymers having a polyether skeleton or a polyvinyl skeleton, and are preferably dibranched, tribranched, or tetrabranched polyethylene glycol.

[0054] Such a kit is suitable for use in the above-mentioned treatment method. In this case, the kit may further include a medical device for covering the periphery of the tendon or ligament covered with the gel material. Such a medical device is suitable for stably fixing the tendon or ligament after suturing surgery, and is, for example, a device that can form a hollow cylindrical shape. However, a device of an appropriate shape can be used depending on the location of the affected area, the postoperative condition, etc. EXAMPLES

[0055] The present invention will be described in more detail below with reference to examples, but the present invention is not limited to these examples.

[0056] 1. Preparation of Polymer Solutions The raw polymers used were Tetra-PEG-SH (tetrathiol-polyethylene glycol) having -SH groups at the ends and Tetra-PEG-MA (tetramaleimidyl-polyethylene glycol) having maleimidyl groups at the ends. These raw polymers were commercially available from NOF Corporation. The molecular weight (molar mass) of both was 20,000. Sodium citrate buffer (pH 5.10; Fujifilm Wako Pure Chemical Industries, Ltd.) was used as a buffer for the polymer solution.

[0057] The solution conditions used were as follows: [Polymer solution A] Concentration: 2.5wt% Tetra-PEG-SH pH: 5.10 [Polymer solution B] Concentration: 2.5wt% Tetra-PEG-MA pH:5,10

[0058] 2. Application to a rat flexor tendon rupture model A rat flexor tendon rupture model was created, and 25 μl of polymer solution A and 25 μl of polymer solution B were injected around the tendon after tendon suture surgery, and gelation was confirmed after 90 seconds. It was confirmed that covering the sutured tendon with the gel successfully blocked the infiltration of inflammatory cells and fibroblasts from the surrounding area. On the other hand, when the tendon was not covered with the gel, the tendon was strongly covered with scar tissue. Images of the repaired tendon covered with the gel of the present invention (gel group) and the repaired tendon not covered with the gel (control group) are shown in Figure 1.

[0059] We therefore histologically evaluated repaired tendons covered with the gel of the present invention (gel group) and repaired tendons not covered with the gel (control group). On the 21st day after surgery, in the gel group, strong cell proliferation along the long axis of the tendon was observed in tissues within the tendon, such as the epitenon and paratenon, whereas in the control group, disorderly abnormal proliferation of fibroblasts and strong neovascularization were observed around the tendon (Figure 2). This suggests that in the gel group, progenitor cells present within the tendon are primarily involved in healing.

[0060] Furthermore, a comparative analysis of gene expression was performed on the healing process in both groups using organ-cultured tendon tissue. In the control group, Col3a1 (Type III Collagen), a characteristic of scar tissue, was strongly expressed from the early stages after surgery, whereas in the gel group, Col3a1 was expressed in small amounts, and strong expression of Col1a1 (Type 1 Collagen) was observed from 2 to 3 weeks after surgery. From the above, it can be seen that in the control group, healing of scar tissue, mainly composed of Type III Collagen, occurs early, whereas in the gel group, healing is initiated by cells within the tendon. Type I It was found that healing was occurring due to collagen.

Claims

1. A gel material comprising a polymer gel for inhibiting expression of type III collagen in the treatment of tendon or ligament injury or rupture, comprising: The polymer gel is a hydrogel having a structure in which a hydrophilic polymer is crosslinked to form a three-dimensional network structure, and has a polymer content in the range of 1 to 5 wt % and an elastic modulus in the range of 100 to 10,000 Pa; The hydrophilic polymer is a di-, tri- or tetra-branched polyethylene glycol. The gel material is characterized by:

2. 2. The gel material of claim 1, wherein the hydrophilic polymer comprises a first polymer unit having one or more nucleophilic functional groups at a side chain or terminal thereof and a second polymer unit having one or more electrophilic functional groups at a side chain or terminal thereof.

3. The nucleophilic functional group is selected from the group consisting of a thiol group and an amino group, and the electrophilic functional group is selected from the group consisting of a maleimidyl group, an N-hydroxy-succinimidyl (NHS) group, a sulfosuccinimidyl group, a phthalimidyl group, an imidazoyl group, an acryloyl group, a nitrophenyl group, an isothiocyanate group, an aldehyde group, an iodoacetamide group, a vinylsulfone group, and a -CO ２ PhNO ２ 3. The gel material of claim 2, selected from the group consisting of:

4. The gel material according to any one of claims 1 to 3, which is used to cover a tendon or ligament after a tendon or ligament suturing surgery.

5. The gel material described in claim 4, which is capable of suppressing expression of type III collagen and maintaining expression of type I collagen by covering the tendon or ligament.

6. A kit for preparing a gel material described in any one of claims 1 to 5, which is used to inhibit expression of type III collagen in the treatment of tendon or ligament damage or rupture, the kit containing two or more solutions containing a hydrophilic polymer.

7. A method for screening for a drug effective in treating injury or rupture of a tendon or ligament, comprising a step of searching for and identifying a compound having type I collagen proliferation activity in a tendon or ligament covered with the gel material according to any one of claims 1 to 5.

Images