JP2003245341A - Anti-synechia membrane - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a modified anti-synechia membrane containing a collagen which makes it usable as filling or prosthetic membranes for defective parts or cut surfaces of endogenous membrane-like tissues such as pleura, pericardium, endocranium and serosa, and of various organs, or the like, yields sufficient anti-synechia effects for a longer period of time with little side effects, and moreover, achieves higher biocompatibility and better degradation and absorption in vivo while satisfying mechanical strengths enough to enable suture fixation. <P>SOLUTION: In this anti-synechia membrane, a cover layer containing a mixture of the collagen and hyaluronic acid is arranged on the surface of a laminate membrane-like matter having a nonwoven fabric layer comprising collagen fibers and a sponge layer comprising the collagen to maintain anti- synechia effects for a longer period of time. <P>COPYRIGHT: (C)2003,JPO

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a collagen-containing adhesion prevention film having a tissue regeneration-inducing and promoting action.
The adhesion-preventing membrane of the present invention has good biocompatibility, exhibits a stable adhesion-preventing effect in the living body for a long period of time, and can be sutured. It is used as a prosthetic film for filling in incomplete tissues or cut surfaces of various organs or in cut surfaces.

[0002]

2. Description of the Related Art In various surgical operations, the affected part is often resected and the damaged part is repaired, especially when targeting various organs such as the lung, heart, liver, brain, digestive organs and gallbladder. In many cases, the fundamental function of the organ is often impaired unless the cut surface or defective portion is supplemented or prosthetic with the membranous material covering the tissue of the organ. If these treatments are performed imperfectly, it is often seen that the prognosis tends to be extremely poor even if the organ dies due to dysfunction of the organ or escapes a life threat. In addition, if the prosthesis or suture fixation at the prosthetic site is poor, even if the function of the treated organ itself can barely be maintained, body fluid, digestive fluid, contents, etc. exuded or leaked from these organs, etc. May cause infection, attack other organs, or cause erosion, resulting in a life threat.

Further, adhesion between the prosthesis or the supplemented membranous material and the organ may occur frequently, and as a result, the dysfunction of the organ may be induced over time. For the purpose of solving such various problems, a film-like material or an adhesion preventing film covering an organ or a tissue of the organ has been developed with various materials.

The simplest and most effective method for preventing adhesion is to prevent the tissue damaged by damage or defect from contacting another tissue physically accessible to the tissue with a septum. That is. However, when this is performed with synthetic fibers or the like, various inconveniences such as excessive calcification, foreign body reaction, and inflammatory reaction occur due to lack of biocompatibility. Further, even when a biocompatible material is used as the partition wall, the material itself must not mediate adhesion between a damaged or defective tissue and another corresponding tissue. Examples of those which satisfy these conditions include hyaluronic acid and gelatin. Both can be handled as a viscous, water-soluble liquid, and can be used as a gel by various processing methods.

Since these materials are mainly extracted and purified from living bodies such as animals, they have good biocompatibility and have already been put to practical use in various medical fields including pharmaceuticals. For example, an anti-adhesion membrane using hyaluronic acid, medical materials, etc. are disclosed in Patent Document 1, Patent Document 2, Patent Document 3, Patent Document 4, Patent Document 5, Patent Document 6, Patent Document 7, Patent Document 8, Patent Document 9 , Patent Document 10 and the like.

[0006]

[Prior Art Document]

[Patent Document 1] JP-A-6-73103

[Patent Document 2] Japanese Patent Publication No. 7-30124

[Patent Document 3] Registered Patent No. 2670996

[Patent Document 4] Japanese Patent Laid-Open No. 8-333402

[Patent Document 5] Japanese Patent Application Laid-Open No. 61-234864

[Patent Document 6] Registered Patent No. 2648308

[Patent Document 7] Japanese Patent Laid-Open No. 8-157378

[Patent Document 8] Japanese Patent Laid-Open No. 9-296005

[Patent Document 9] JP-A-7-102002

[Patent Document 10] Japanese Patent Publication No. 7-509386

[0007] Patent Document 11 describes a medical collagen film in which at least one surface of a non-woven fabric layer made of cross-linked collagen fibers is covered with a collagen film as an improved medical film using only animal-derived collagen. Has been done. Patent Document 12 describes a technique in which gelatin or hyaluronic acid alone is used as an anti-adhesion film on a nonwoven fabric of collagen fibers. Patent Document 13 describes an adhesion preventing film having a coating layer of gelatin or hyaluronic acid on a nonwoven fabric layer of collagen fibers and a sponge layer of collagen.

[0008]

[Patent Document 11] Japanese Patent Laid-Open No. 2000-93497

[Patent Document 12] Japanese Unexamined Patent Publication No. 2000-271207

[Patent Document 13] Japanese Patent Laid-Open No. 2000-210376

[0009]

However, the above-mentioned techniques are sufficiently satisfactory in terms of biocompatibility, biodegradability / absorption, physical / mechanical strength, ease of handling, and long-term adhesion prevention effect. Not a thing. It exerts a sufficient anti-adhesion effect for a long period of time, has few side effects, and has good biocompatibility, biodegradability and absorbability,
There is a need for improved medical materials that simultaneously satisfy the mechanical strength required for suture fixation.

[0010]

[Means for Solving the Problems] In order to solve the above problems, as a result of various studies, the present inventors have found that a laminated film-like product having a nonwoven fabric layer made of collagen fibers and a sponge layer made of collagen. By providing a coating layer containing a mixture of collagen and hyaluronic acid on the surface, it is possible to construct an anti-adhesion layer with significantly improved degradation resistance, and an anti-adhesion membrane that maintains the anti-adhesion effect for a long period of time. They have found that they can be obtained and have reached the present invention.

That is, according to the present invention, (1) a coating layer containing a mixture of collagen and hyaluronic acid is provided on the surface of a laminated membranous material having a nonwoven fabric layer made of collagen fibers and a sponge layer made of collagen. Anti-adhesion membrane,
(2) The anti-adhesion membrane as described in 1 above, wherein the coating layer containing a mixture of collagen and hyaluronic acid is sponge-like or film-like. (3) The coating layer containing a mixture of collagen and hyaluronic acid contains collagen and hyaluronic acid. 2. The adhesion preventive film according to 1 above, which is a layer formed by subjecting the mixture to a cross-linking reaction, and (4) the adhesion according to 1 above, wherein the coating layer containing the mixture of collagen and hyaluronic acid has a thickness of about 50 μm to 20 mm. An anti-adhesion film, (5) The adhesion-prevention film according to the above 1, wherein the collagen constituting the anti-adhesion film is enzyme-solubilized collagen, acid-solubilized collagen, alkali-solubilized collagen or neutral solubilized collagen, (6) anti-adhesion film (1) The adhesion preventive membrane according to the above (1), wherein a part or all of the collagen constituting the above is crosslinked, (7) a nonwoven fabric layer comprising collagen fibers (1) Lagen or (2) the anti-adhesion membrane according to the above 1, which is a nonwoven fabric layer in which collagen fibers are bonded to each other with a mixture of collagen and hyaluronic acid, (8) the thickness of the nonwoven fabric layer made of collagen fibers is about 50 μm to 10 mm, The sponge layer made of collagen has a thickness of 50 μm to 20 mm, and the coating layer containing a mixture of collagen and hyaluronic acid has a thickness of about 50 μm.
(10) The anti-adhesion membrane as described in 1 above, which is 20 mm, (9) The anti-adhesion membrane as described in 1 above, wherein the nonwoven fabric layer made of collagen fibers is a laminate of 1 to 6 nonwoven fabrics made of collagen fibers. Collagen fiber diameter is about 1
0 to 1,000 μm, the adhesion-preventing membrane according to 1 above, wherein the nonwoven fabric layer made of collagen fibers has a bulk density of about 5 × 10 ^{−4 to} 50 g / cm ³ , and (11) the overall thickness of the membrane is about 150 μm to The adhesion-preventing membrane according to 1 above, which has a thickness of 50 mm.

[0012]

BEST MODE FOR CARRYING OUT THE INVENTION In the present invention, a non-woven fabric layer made of collagen fibers is a structure having a secondary or tertiary structure made of collagen fibers, and some or all of the collagen fiber voids are gas, liquid, It may be filled with a solid or the like, or a part or all of the outer peripheral surface of the collagen fiber may be covered with a solid or a liquid. The collagen fibers may be joined together with a binder such as collagen or a mixture of collagen and hyaluronic acid. The collagen constituting the collagen fiber may be partially or entirely crosslinked. In addition, collagen in the collagen constituting the collagen fiber or collagen in the mixture of collagen and hyaluronic acid is, when acid-solubilized collagen is used, alkali metal (eg, sodium, potassium, etc.), alkaline earth metal (eg, magnesium, calcium). Etc.) and the like, and when alkali-solubilized collagen is used, an inorganic acid (eg, hydrochloric acid,
It may be used as a salt with sulfuric acid, nitric acid, etc.) and an organic acid (eg, acetic acid, citric acid, etc.). Hyaluronic acid may be used as a salt of an alkali metal (eg, sodium, potassium, etc.), an alkaline earth metal (eg, magnesium, calcium, etc.).

The diameter of collagen fibers is usually about 10
The thickness is 1,000 μm, preferably about 20 to 250 μm. The non-woven fabric layer composed of collagen fibers is usually 1 to 6
One, preferably 2 to 4 collagen nonwoven fabrics are laminated. The bulk density of the non-woven fabric layer made of collagen fibers is usually about 5 × 10 ^{−4 to} 50 g / cm ³ , preferably about 0.05 to 50 g / cm ³ . The thickness is usually
About 50 μm to 10 mm, preferably about 0.2 to 2 mm
Is.

The sponge layer made of collagen is a layered product containing collagen and usually having a sponge-like form. When acid-solubilized collagen is used, the collagen may be a salt of an alkali metal (eg, sodium, potassium, etc.) or an alkaline earth metal (eg, magnesium, calcium, etc.). When alkali-solubilized collagen is used, it may be a salt with an inorganic acid (eg, hydrochloric acid, sulfuric acid, nitric acid, etc.) or an organic acid (eg, acetic acid, citric acid, etc.). Further, the collagen may be subjected to a crosslinking reaction treatment. In order to create a laminated film having a non-woven fabric layer made of collagen fibers and a sponge layer made of collagen, an aqueous solution containing collagen, hyaluronic acid, a mixture of collagen and hyaluronic acid, etc. is made from a sponge layer made of collagen and collagen fibers. The non-woven fabric layer may be applied to the surface to be adhered, and the non-woven fabric layer made of collagen fibers and the sponge layer made of collagen may be adhered. The thickness of the collagen sponge layer is usually about 50 μm to 20 mm.
And preferably about 0.1 to 1 mm. The collagen sponge layer may be laminated on one side or both sides of the non-woven fabric layer formed of collagen fibers.If the non-woven fabric layer formed of collagen fibers is composed of a plurality of non-woven fabrics, it is laminated between the non-woven fabrics. May be.

The coating layer containing a mixture of collagen and hyaluronic acid is a layered product containing collagen and hyaluronic acid and usually having a sponge-like or film-like form. When acid-solubilized collagen is used, the mixture of collagen and hyaluronic acid may be a salt of an alkali metal (eg, sodium, potassium, etc.) or an alkaline earth metal (eg, magnesium, calcium, etc.). When alkali-solubilized collagen is used, it may be a salt with an inorganic acid (eg, hydrochloric acid, sulfuric acid, nitric acid, etc.) or an organic acid (eg, acetic acid, citric acid, etc.). The mixing ratio (weight ratio) of collagen and hyaluronic acid is about 1: 100 to about 100 :.
A range of 1 is used, preferably from about 3: 7 to about 7: 3.
And more preferably about 1: 1. The mixture of collagen and hyaluronic acid may be subjected to a crosslinking reaction treatment. In order to bond a laminated membrane having a coating layer containing a mixture of collagen and hyaluronic acid, a nonwoven fabric layer made of collagen fibers and a sponge layer made of collagen, collagen, hyaluronic acid, and a mixture of collagen and hyaluronic acid are used. An aqueous solution containing the same may be applied to the surfaces of the coating layer and the laminated film-like material to be bonded. The thickness of the coating layer containing a mixture of collagen and hyaluronic acid is usually about 50 μm to 20 mm, preferably about 0.
It is 1 to 1 mm. The coating layer may be laminated on one side or both sides of the laminated film material.

The adhesion-preventing membrane of the present invention has a laminated structure in which a nonwoven fabric layer made of collagen fibers, a sponge layer made of collagen, and a coating layer containing a mixture of collagen and hyaluronic acid are laminated. The total thickness of the anti-adhesion membrane of the present invention is usually about 150 μm to 50 mm,
It is preferably about 0.5 to 9 mm. The adhesion-preventing membrane of the present invention has a non-woven fabric layer mainly composed of collagen fibers in charge of sufficiently sutureable membrane strength, and a sponge layer mainly composed of collagen is in charge of induction and promotion of tissue regeneration. A coating layer containing a mixture of collagen and hyaluronic acid exerts an effect of preventing adhesion with a tissue.

The adhesion-preventing membrane of the present invention, in which collagen and hyaluronic acid are bio-derived materials and has not only excellent biocompatibility, but also gradually decomposed and absorbed in the transplanted living body, and finally Will be all decomposed and absorbed. In particular, the non-woven fabric layer consisting of collagen fibers exists as a scaffold for filling, prosthesis, and sealing until the tissue regeneration of the in vivo defect site is completed, and after maintaining the membrane strength for a certain period of suture fixation, all It is decomposed and absorbed. Further, the outermost layer, the coating layer containing a mixture of collagen and hyaluronic acid, prevents adhesion between the tissue at the damaged or defective site and the surrounding tissue due to its viscosity and sustained release action. The period of preventing the adhesion lasts until the tissue at the damaged or defective site is regenerated and healed to the extent that adhesion with surrounding tissues does not occur in the natural state. While exhibiting these adhesion prevention effects, they are gradually decomposed and absorbed in the body,
Finally, the coating layer containing the mixture of collagen and hyaluronic acid also disappears.

The method for producing the anti-adhesion membrane of the present invention is described below. Examples of typical collagen raw materials used in the present invention include enzyme-solubilized collagen, acid-solubilized collagen, alkali-solubilized collagen, neutral solubilized collagen and the like. These solubilized collagens are collagens that have been treated so that they can be dissolved in a solvent. Examples include solubilized collagen such as acid solubilized collagen, alkali solubilized collagen, enzyme solubilized collagen, and neutral solubilized collagen. In particular, atelocollagen, which is subjected to a solubilization treatment and a treatment to remove telopeptide, which is an antigenic determinant of collagen, is preferable. Regarding the solubilization method of these collagens, Japanese Patent Publication No. 46-15003 and Japanese Patent Publication No. 43-25
9839, Japanese Patent Publication No. 43-27513, etc. The origin of collagen is extracted from the skin, tendons, bones, cartilage, organs and the like of animal species such as cow, pig, bird, fish, rabbit, sheep, rat, and human. Collagen type I, III
The type I is not limited to any type that can be classified, but the type I is particularly preferable from the viewpoint of handling.

When acid-solubilized collagen is used, the collagen is coagulated and then, in water, alkali metal carbonate (eg, sodium carbonate, potassium carbonate, etc.), alkali metal hydrogen carbonate (eg, sodium hydrogen carbonate, etc.), alkali metal hydroxide ( (Eg, sodium hydroxide, etc.) and the like. Also, when alkali-solubilized collagen is used, the collagen is coagulated and then, in water, an inorganic acid (eg, hydrochloric acid, sulfuric acid, nitric acid, etc.), an organic acid (eg, acetic acid, citric acid, etc.)
It may be used as a salt neutralized by the like (the same applies in the following cases).

Hyaluronic acid may be derived from animals or microorganisms, but medical grade products are particularly suitable. Hyaluronic acid may be used as a salt of an alkali metal (eg, sodium, potassium, etc.). The solvent for solubilizing collagen is not particularly limited as long as it is a solvent capable of solubilizing collagen, but preferably water, an aqueous solution of a neutral salt (eg, sodium chloride, sodium hydrogen phosphate, etc.) or a dilute solution. It may contain a mineral acid (eg, dilute hydrochloric acid, dilute nitric acid, etc.) and a hydrophilic organic solvent (eg, monohydric or polyhydric alcohols such as ethyl alcohol and ethylene glycol). Water is suitable for handling.

A non-woven fabric composed of collagen fibers is preferably produced by continuously spinning an aqueous solution of collagen according to a conventional method (for example, the method described in Japanese Patent Laid-Open No. 2000-93497) to obtain a collagen yarn and processing it into a non-woven fabric. . The concentration of the collagen aqueous solution used in the present invention is arbitrary depending on the type of collagen used and may be any concentration as long as it can be spun, but it is usually about 0.1 to 20% by weight, About 1% by weight in wet spinning
About 10% by weight is particularly suitable. The collagen discharge speed during spinning is arbitrary as long as it is within the range in which spinning is possible.

Any device such as a general-purpose gear pump, a dispenser, and various extrusion devices may be used for discharging the collagen solution during spinning, but in order to perform uniform spinning, the pulsation is small and the collagen is stable. A device that can dispense a fixed amount of solution is preferable. Further, the pore size of the spinneret during spinning is not particularly limited as long as it can be spun,
If the pore size is too large, it becomes difficult to change the fibrous material into a film by the binder treatment, and if the diameter is extremely small, it is difficult to improve the film strength.
m to 1000 μm, preferably about 50 μm to 700 μm
The range of is used. Furthermore, the number of holes in the die may be single or plural. The shape of the die is not particularly limited, and if it is spinnable, for example, a slit shape or various shapes may be used. Further, the hole length of the die is not particularly limited as long as it can be spun, but it is preferably as long as possible for the purpose of orienting as many collagen molecules in the solubilized collagen molecules as possible.

The coagulation bath used in the wet spinning is not particularly limited, and may be a solvent, suspension, emulsion or solution capable of coagulating collagen in general, but it is not particularly limited, but may be an inorganic salt aqueous solution or an inorganic salt-containing organic solvent. , Alcohols, ketones or any combination thereof. The inorganic salt aqueous solution is preferably an aqueous solution of sodium sulfate, sodium chloride, ammonium sulfate, calcium chloride, magnesium chloride or the like, particularly an aqueous solution of sodium chloride, sodium sulfate, ammonium sulfate or the like. An inorganic salt-containing organic solvent in which these inorganic salts are dissolved / dispersed in alcohol or acetone may be used. In this case, an ethanol-dissolved / dispersed solution of sodium chloride is particularly preferable. As alcohols, methanol, ethanol,
Isopropanol, amyl alcohol, pentanol,
Examples thereof include alcohols having 1 to 6 carbon atoms such as hexanol, glycols such as ethylene glycol, and preferably ethanol. Examples of the ketones include acetone and methyl ethyl ketone.

The above coagulation bath is not only for coagulation of collagen, but also in combination with various crosslinking agents described below,
A processing method capable of simultaneously performing coagulation and cross-linking of collagen is also effective. For example, when a solution obtained by mixing ethanol and glutaraldehyde is used as a coagulation bath having both coagulation treatment and crosslinking treatment, both steps can be performed at one time, and the spun collagen yarn is directly subjected to crosslinking treatment. These simultaneous treatments are very effective not only for streamlining the process but also for spinning a dilute collagen solution or spinning a thin yarn.

In the above method for forming the non-woven fabric layer, a particularly preferred specific example will be described below. A collagen discharge die having a hole diameter of about 200 μm and a hole length of about 15 to 20 mm can be used, and the collagen solution can be discharged with a dispenser or the like without pulsation, and wet spinning can be performed in a coagulation bath of about 99.5% by volume ethanol. desirable. When collagen is extruded into a coagulation bath of ethanol of about 99.5% by volume, the ejection nozzle is moved at any time to continuously extrude the spun yarn so that the spun yarn can intersect in any direction, and the yarn is in a multi-layered state. By removing the coagulating liquid after washing, washing again with ethanol, and drying under reduced pressure, a very good cotton-like fibrous material can be easily obtained. This method is particularly effective in terms of process simplification, shortening, and economic efficiency, since spinning and nonwoven fabric formation can be performed at the same time.

This example is a typical one, and is not limited to this as long as a fibrous material can be obtained.
For example, the above-mentioned collagen fiber staple may be used, the type of coagulation bath, the use of a coagulation bath and a cross-linking agent mixed bath,
Further, the drying method or the like may be changed, and the combination thereof may be changed.

On the other hand, short staple fibers are produced in both continuous and discontinuous spinning by cutting the discontinuous spinning from the aqueous collagen solution by intermittent discharge or the thread obtained after performing ordinary continuous spinning. A product is obtained. It is also possible to produce a nonwoven fabric (fibrous material) which is dried by a method such as reduced pressure drying or natural drying in a state where these are uniformly dispersed in a container of an appropriate size.

The non-woven fabric obtained by the above method may be further cross-linked if necessary in order to obtain sufficient suture strength. By this cross-linking treatment, the physical strength especially when wet is improved, and the strength required for suturing can be sufficiently secured. It is also to dramatically delay the time of decomposition / absorption when transplanted into a living body, as compared with the case of uncrosslinked. By this cross-linking treatment, the defective part of the living body is filled or prosthetic, the dysfunction of organs / tissues due to the defect is prevented, and the membrane strength required in the body until the repair of the wound surface and the regeneration of the tissue is completed. It is possible to remain while maintaining.

This cross-linking method is roughly classified into a physical cross-linking method and a chemical cross-linking method. Examples of the physical crosslinking treatment method include γ-ray irradiation, ultraviolet ray irradiation, electron beam irradiation, plasma irradiation, and thermal dehydration crosslinking treatment. Of these, thermal dehydration crosslinking treatment is preferable. In the heat dehydration cross-linking treatment, the collagen single yarn is physically wound by being heat-treated under reduced pressure in a wound state. In this cross-linking treatment, it is possible to control biocompatibility and degradability by the cross-linking temperature and cross-linking time. Physical cross-linking and chemical cross-linking may be carried out individually or in combination, and when they are combined, the order is not limited. As the cross-linking agent used in the chemical cross-linking reaction, any cross-linking agent can be used as long as it can cross-link with collagen, and examples thereof include aldehydes, epoxies, carbodiimides, and isocyanates. Aldehydes include formaldehyde, glutaraldehyde, glyoxal,
Examples include dialdehyde starch, epoxies such as glycerol diglycidyl ether, carbodiimides such as water-soluble carbodiimide, and isocyanates such as hexamethylene diisocyanate. Glutaraldehyde is preferred. Crosslinking of the collagen single yarn is usually performed by immersing the collagen single yarn in a solution of a crosslinking agent. The solvent of the cross-linking agent solution is not particularly limited, but water, ethanol and the like are suitable, and ethanol is particularly suitable. Degradation and absorption and biocompatibility can be controlled by the concentration of the crosslinking agent solution and the immersion time. When the cross-linking agent is glutaraldehyde,
The concentration of the solution is usually about 0.001 to 25% by volume,
Preferably, it is about 0.01% to 1.0% by volume.

The non-woven fabric layer comprising collagen fibers obtained by the above method may be subjected to a binder treatment for the purpose of improving its physical strength. The binder treatment means that the non-woven fabric layer is impregnated with (1) collagen or (2) an aqueous solution of a mixture of collagen and hyaluronic acid, and is then dried by air, dried under blast, dried under reduced pressure, or dried under low temperature. This is a treatment for drying fibers by a drying method to bond the fibers of the nonwoven fabric layer together. The non-woven fabric layer obtained by this binder treatment has a much higher physical strength than the untreated non-woven fabric layer, and therefore the suture strength is also significantly improved.
This impregnation / drying step may be repeated once to several tens of times or more depending on the required physical strength.

However, when the binder treatment is carried out, if the nonwoven fabric layer made of only collagen fibers is not subjected to the crosslinking treatment, it is treated with (1) collagen or (2) an aqueous solution of a mixture of collagen and hyaluronic acid. The non-woven fabric layer itself may be dissolved when impregnated. Therefore, it is desirable to carry out the crosslinking treatment in advance by the above-mentioned method or the like. The binder treatment method is (1)
In addition to the method of impregnating the nonwoven fabric layer with an aqueous collagen solution or (2) an aqueous solution of a mixture of collagen and hyaluronic acid, collagen or a mixture of collagen and hyaluronic acid is cast or filled in a suitable container or mold together with the nonwoven fabric layer. Or a method of directly applying a collagen solution or a solution of a mixture of collagen and hyaluronic acid to the non-woven fabric layer.

In the present invention, a binder treatment method using a sponge layer may be used. For example, a membranous material obtained by sandwiching a non-woven fabric layer between collagen layers that have been dried in a sponge shape, or a membranous product obtained by simultaneously compressing the sponge layer and the non-woven fabric layer and embedding the non-woven fabric layer in the sponge layer. The substance is placed under normal pressure or reduced pressure in the presence of dilute collagen or a solution of the above-mentioned base salt or acid addition salt or water, to dissolve the collagen in the sponge layer and sufficiently dissolve the solution in the non-woven fabric layer. A method in which the particles are permeated and then dried by various drying methods is used. In the binder treatment method using this sponge layer, since the fibers of the non-woven fabric layer are bonded to each other, a very small amount of solvent component such as water is required with respect to the amount of collagen actually used. Therefore, there is a great merit that the drying is completed in a short time in a later step and the shrinkage and deformation during the drying are extremely small. Further, in the usual impregnation step, the amount of substantial collagen in the impregnating collagen solution is limited to about several% due to the practical solution viscosity, and the remaining 90% or more is a solvent component such as water. Therefore, the operation of impregnation / drying takes time and it is necessary to repeat the operation itself, but there is an advantage that the operation is very simple. Of course, these methods are typical examples, and any method may be used as long as it is a method of joining the fibers of the non-woven fabric layer or the compressed non-woven fabric layer using collagen, and the above representative examples are not particularly limited. Not done. Also, for binder processing,
In addition to the above collagen aqueous solution, a hyaluronic acid solution, an aqueous solution of a mixture of collagen and hyaluronic acid, or the like can be used.

Next, a sponge layer made of collagen and a coating layer containing a mixture of collagen and hyaluronic acid may be formed on the nonwoven fabric layer made of collagen fibers obtained by the above method. The forming method can be easily performed by a conventional method such as freeze-drying. There is no particular limitation on the order, method, etc. in the manufacturing process for forming. As a specific manufacturing method, for example, a nonwoven fabric layer made of collagen fibers, a sponge layer made of collagen, and a sponge-like or film-like coating layer containing a mixture of collagen and hyaluronic acid are laminated, or a sponge layer made of collagen. And a sponge-like coating layer containing a mixture of collagen and hyaluronic acid is independently prepared, and then the coating layer, the sponge layer, and the non-woven fabric layer composed of collagen fibers are treated with collagen, hyaluronic acid, or collagen and hyaluronic acid. There is a method of bonding using an aqueous solution of a mixture with an acid. In addition, the nonwoven fabric layer made of collagen fibers is immersed in an aqueous solution of collagen and then frozen once, and then again immersed in an aqueous solution containing a mixture of collagen and hyaluronic acid, and these are frozen and integrated and then frozen. There is a method of obtaining an adhesion-preventing membrane having three layers of a nonwoven fabric layer made of collagen fibers, a sponge layer made of collagen, and a coating layer containing a mixture of collagen and hyaluronic acid by drying at the same time.

Further, after immersing the collagen nonwoven fabric in a collagen solution filled in a container and a solution consisting of a mixture of collagen and hyaluronic acid, it is frozen in a freezer and further freeze-dried to form a collagen sponge layer and collagen. It is also possible to form the collagen nonwoven fabric in a sponge layer made of a mixture of hyaluronic acid. However, these are merely examples of the processing method of the present invention, and the purpose of these processing is to form a sponge layer made of collagen and a sponge-like or film-like coating layer made of a mixture of collagen and hyaluronic acid and collagen fibers. The purpose is to integrate the resulting non-woven fabric layer without being easily peeled and separated during transplantation in vivo, and any processing sequence and method may be used as long as this is achieved.

For producing a sponge layer made of collagen and a sponge-like coating layer containing a mixture of collagen and hyaluronic acid, a collagen solution or an aqueous solution containing a mixture of collagen and hyaluronic acid is cast into a container, or a desired solution is cast. A uniform sponge layer can be obtained by filling the product to a thickness and freezing it sufficiently with a general-purpose freezer and then drying it with a freeze dryer. At this time, the pore size of the fine pores formed in the sponge layer made of collagen or the sponge layer made of a mixture of collagen and hyaluronic acid depends on the concentration of the collagen solution or the solution made of a mixture of collagen and hyaluronic acid and its solvent, when frozen. It changes depending on the temperature and freezing time.

With respect to the total amount of various raw materials and the thickness of the sponge-like coating layer in the sponge-like coating layer containing collagen and a mixture of collagen and hyaluronic acid, the anti-adhesion effect and the damage of the target site can be determined. -It is desirable that the sponge layer remains in the body for about 1 to 4 weeks so as not to interfere with repair of the cut site and induction of tissue regeneration. The thickness of the sponge-like coating layer composed of this mixture of collagen and hyaluronic acid, and the total amount of raw materials used for forming the sponge layer are
It can be arbitrarily controlled in consideration of the degradation / absorption time when transplanted in the living body and the influence on the induction of tissue regeneration. The thickness of each sponge layer is specifically about 50 μm to 20 mm, preferably 100 to 1000 μm.
In particular, the thickness at the end of drying is 1 to 10 mm,
When these are compressed and used, 100 μm to
A thickness of 1 mm is preferred. Considering these situations, the concentration range of the aqueous collagen solution and the solution containing the mixture of collagen and hyaluronic acid is 0.1 to 60.
It is desirable that the content is wt%, preferably 0.5 to 10 wt%. The freezing temperature is -200 to -10 ° C, preferably -80 to -10 ° C, which can be set by a general-purpose freezer or deep freezer. The freeze dryer is not particularly limited as long as it can stably dry.

Further, the filling amount of the aqueous collagen solution and the aqueous solution containing the mixture of collagen and hyaluronic acid into the container is such that the thickness of the finished sponge is about 50 μm to 20 μm.
mm, preferably about 10 to 1000 μm. These values can be changed at any time depending on the purpose of use and are not limited to these examples.

The coating layer containing a mixture of collagen and hyaluronic acid that exerts an adhesion preventing effect is not limited to the shape of a sponge, and is processed into a shape such as a film obtained by an ordinary casting method or the like. It may be done. In addition, when forming a coating layer containing a mixture of collagen and hyaluronic acid to impart an adhesion preventing effect, various forms such as one side or both sides of the membrane or partial or whole surface coating can be selected according to the purpose. The method of forming the coating layer containing a mixture of collagen and hyaluronic acid or the site thereof is not particularly limited, and any combination is possible.

As the objects to be cross-linked by the above-mentioned various cross-linking methods, a non-woven fabric layer constituting a membrane, a sponge layer made of collagen, a coating layer containing a mixture of collagen and hyaluronic acid, and these are integrated. Part or all of the laminated and laminated anti-adhesion film is a target. Further, the order of crosslinking, the combination of the crosslinking method is arbitrary,
There is no particular limitation. However, most preferably, the non-woven fabric layer made of collagen fibers is subjected to cross-linking using aldehydes such as glutaraldehyde, and then a sponge layer made of collagen and a coating layer containing a mixture of collagen and hyaluronic acid are formed. Finally, heat dehydration crosslinking is applied. These methods include a method of mixing a coagulant such as ethanol and a cross-linking agent represented by glutaraldehyde in the steps of spinning collagen and forming a non-woven fabric, and performing the steps of spinning and cross-linking at the same time. . If the acid-solubilized collagen is used as it is in the living body as a material for the adhesion-preventing membrane, it may acidify surrounding body fluids. To avoid this, use alkali metal carbonate (eg, sodium carbonate, potassium carbonate, etc.), alkali metal hydrogen carbonate (eg, sodium hydrogen carbonate, etc.), alkali metal hydroxide (eg, sodium hydroxide, etc.) in an aqueous solution. It may be used as a salt. The alkali-solubilized collagen may be used as a salt neutralized with an inorganic acid (eg, dilute hydrochloric acid, dilute nitric acid, etc.), an organic acid (eg, acetic acid, citric acid, etc.) (the same applies in the following cases). You may neutralize with an acid (dilute hydrochloric acid, dilute nitric acid, acetic acid, etc.) before use. The neutral solubilized collagen may be used as it is.

The adhesion-preventing membrane of the present invention is an excellent adhesion-preventing membrane in terms of suture strength, biocompatibility, biodegradability / absorption, and promotion / induction of tissue regeneration. When the binder treatment is performed on the non-woven fabric layer made of collagen fibers, it is preferable that the layer formed thereby is also thermally dehydrated and crosslinked. However, this is merely an example, and for example, even if all layers are treated by thermal dehydration crosslinking, there is no problem, and both sterilization and crosslinking may be performed and, for example, γ-ray irradiation may be performed.

The anti-adhesion membrane having the non-woven fabric layer obtained by the above method, the sponge layer made of collagen, and the coating layer containing the mixture of collagen and hyaluronic acid can be further compression-molded. The sponge layer made of collagen and the coating layer containing a mixture of collagen and hyaluronic acid may be compressed alone and then combined with the non-compressed non-woven layer to be integrated. Particularly preferably, in the final step of film formation, a nonwoven fabric layer, a sponge layer made of collagen, and a coating layer containing a mixture of collagen and hyaluronic acid, which are integrated, are simultaneously compressed. This is because the film thickness is reduced by compression and thinned, so when actually using the anti-adhesion film at the surgical site, handling such as penetrating the suture needle in suturing and cutting into an arbitrary shape Is particularly improved, and transplant surgery can be performed more smoothly. The method of compression can be performed with a general-purpose press, but since it is intended for medical use, it is aseptically prepared with a sufficiently durable sterilized packaging material, such as an aluminum pack or a high-strength resin packaging material. It is desirable to be compressed in a packaged state. The pressure for compressing the anti-adhesion membrane is not particularly limited as long as it does not destroy the membrane body, but is usually 10 to 1000 k.
It is preferably gf / cm 2.

The adhesion-preventing membrane of the present invention has good biocompatibility, has almost no side effects, exhibits a stable adhesion-preventing effect in vivo for a long time, and can be sutured. It is used as a prosthetic membrane for filling incomplete portions or cut surfaces of in vivo membranous tissues such as brain dura and serosa and various organs. The anti-adhesion membrane of the present invention can be safely used for humans and animals by a method known per se.

[0043]

EXAMPLES The present invention will be described in detail below with reference to examples and experimental examples. Hereinafter, the present invention will be described in detail with reference to examples. The collagen used in the examples of the present invention is pig-derived type I mixed collagen powder (SOFD type, Lot No. 0 manufactured by Nippon Ham Co., Ltd.) which is atherotized acid-solubilized collagen.
102226) and the hyaluronic acid used was a Hilnate injection kit (Hishiyama Pharmaceutical Co., Ltd.).

(Example 1) Preparation of anti-adhesion membrane (three layers) (1) Preparation of non-woven fabric layer consisting of collagen fibers 99 mL of 7% by weight aqueous solution of acid-solubilized collagen was added to 99.
5% by volume ethanol (manufactured by Wako Pure Chemical Industries, special grade) was extruded into a 3 L coagulation bath, dehydrated and coagulated, and the obtained collagen yarn was laminated according to the method described in JP-A-2000-93497 to obtain a collagen nonwoven fabric. Next, after air-drying the obtained collagen non-woven fabric in a clean bench, as it is in a vacuum dry oven (EYELA: VOS-300VD type) under high vacuum (1 torr or less), 120 ° C, 24 hours condition Then, a thermal dehydration crosslinking reaction was performed. After the cross-linking reaction was completed, as a binder treatment, a collagen 1 wt% aqueous solution was applied to the collagen non-woven fabric to fill the gaps between the cross-linked collagen non-woven fabric yarns, and then dried. By repeating the coating operation and the drying operation three times, a nonwoven fabric layer made of collagen fibers was obtained. Then, it was heated at 120 ° C. for 12 hours in a vacuum dry oven (manufactured by EYELA: VOS-300VD type) under high vacuum (1 torr or less) to subject the applied collagen to a thermal dehydration crosslinking reaction. After the crosslinking reaction, the collagen film was immersed in a 0.1N sodium hydroxide aqueous solution for 30 minutes for neutralization treatment, and then taken out from the sodium hydroxide aqueous solution. Sodium hydroxide remaining on the surface of the nonwoven fabric layer made of collagen fibers was washed with distilled water and air-dried in a clean bench.

(2) Preparation of a sponge-like coating layer containing a mixture of collagen and hyaluronic acid, and a sponge layer made of collagen A 1% by weight aqueous solution of hyaluronic acid and an equal amount of 1% by weight aqueous collagen solution were mixed to form collagen and hyaluronic acid. 250 mL of an equal mixture with acid was obtained. After neutralizing the mixture in an acidic state with a 0.1N aqueous sodium hydroxide solution, a metal container having a rectangular parallelepiped space (length 11 cm, width 11 c)
m) and frozen overnight at -20 ° C. The frozen product was decompressed under a freeze dryer (EYELA: FDU-830 type) (1
Lyophilized for about 24 hours under a torr) to obtain a sponge-like coating layer containing a mixture of collagen and hyaluronic acid. A sponge layer made of collagen was also produced from 250 mL of a 1% by weight collagen aqueous solution using the same method.

(3) The sponge layer made of collagen obtained in the above (2) was compressed at a pressure of 100 kgf / cm ² with a compressor (15t press, manufactured by Inei Seieido Co., Ltd.), and this and the above ( The non-woven fabric layer composed of collagen fibers obtained in 1) was adhered with a 1% by weight collagen aqueous solution to obtain a two-layer structure. The sponge-like coating layer containing the mixture of collagen and hyaluronic acid obtained in the above (2) is compressed in the same manner as the compression operation of the sponge layer made of collagen, and then it is made of the collagen fiber of the two-layer structure. It adhered on the nonwoven fabric layer with the solution containing the mixture of collagen and hyaluronic acid. Thus, a laminated structure composed of three types of layers, that is, a sponge layer made of collagen on one surface of the non-woven fabric layer made of collagen fibers and a sponge-like coating layer containing a mixture of collagen and hyaluronic acid on the opposite surface. An anti-adhesion film having Then, the collagen and hyaluronic acid of the anti-adhesion membrane were treated with a vacuum dry oven (EYELA: VOS-300
24 V at 110 ° C under high vacuum (1 torr or less) in VD type)
After heating for a time, a thermal dehydration crosslinking reaction was performed. In this way, the spongy coating layer containing a mixture of collagen and hyaluronic acid as the adhesion prevention layer is composed of the scaffold for cell adhesion / growth and the collagen which promotes the induction of tissue regeneration in the other outer layer. An anti-adhesion membrane having a thickness of about 3 mm was obtained, which had three types of layers, a sponge layer and a non-woven fabric layer made of collagen fibers for maintaining the strength capable of withstanding suture fixation.

Example 2 Preparation of Adhesion Prevention Membrane (5 Layers) Using the same method as in Example 1, a nonwoven fabric layer made of collagen fibers and a sponge layer 2 made of compressed collagen were prepared.
Two sponge-like coating layers each containing a mixture of compressed collagen and hyaluronic acid were prepared. A sponge layer made of collagen was adhered to both surfaces of the non-woven fabric layer made of collagen fibers in the same manner as in Example 1 to obtain a laminated structure. Further, a sponge-like coating layer containing a mixture of collagen and hyaluronic acid was adhered to both surfaces of the laminated structure in the same manner as in Example 1, and heat dehydration crosslinking treatment was performed by a vacuum dry oven (EYELA: VOS-300VD type). ) Under high vacuum (1
(torr or less), 110 ° C., and 24 hours. In this way, a non-woven fabric layer made of collagen fibers that bears the strength to withstand suturing, a sponge layer made of collagen that promotes tissue regeneration and induction on both surfaces, and a collagen that exerts an adhesion preventive function on the outer layer. An anti-adhesion membrane having a total thickness of about 5 mm and having a five-layer structure having a sponge-like coating layer containing a mixture of hyaluronic acid was obtained.

(Example 3) Production of adhesion preventing film (4 layers) Using the same method as in Example 1, a nonwoven fabric layer made of collagen fibers and a sponge layer 2 made of compressed collagen were prepared.
One sheet and one sponge-like coating layer containing a mixture of compressed collagen and hyaluronic acid were prepared. A sponge layer made of collagen was adhered to both surfaces of the nonwoven fabric layer made of collagen fibers in the same manner as in Example 1 to obtain a three-layer laminated structure. Further, a sponge-like coating layer containing a mixture of collagen and hyaluronic acid was adhered to one surface of the laminated structure in the same manner as in Example 1, and the heat dehydration crosslinking treatment was carried out by a vacuum dry oven (EYELA: VOS-300VD. Mold) under high vacuum (1 torr or less) at 110 ° C. for 24 hours. In this way, a nonwoven fabric layer consisting of collagen fibers that bears the strength to withstand suturing in the center, a sponge layer consisting of collagen that promotes tissue regeneration and induction on both surfaces,
An adhesion-preventing film having a total thickness of about 4 mm and having a sponge-like coating layer containing a mixture of collagen and hyaluronic acid exhibiting an adhesion-preventing ability was obtained on one surface of the laminated structure. .

Example 4 Preparation of Sponge and Application to Nonwoven Fabric After a nonwoven fabric layer made of collagen fibers was prepared in the same manner as in Example 1, 250 mL of a 1% by weight collagen solution (solution A) and a 1% by weight collagen solution were used. Hyaluronic acid 1
250 mL of an aqueous solution consisting of an equal amount mixture with a wt% aqueous solution
(Solution B) was prepared and neutralized with a 0.1N aqueous sodium hydroxide solution. Next, as shown in FIG. 1, a non-woven fabric layer 2 made of collagen fibers is placed on a flat plate 1 made of metal, and a polyfluorinated ethylene fiber rubber packing 3 for preventing liquid leakage is placed thereon and opened at the top. A metallic filling container 4 having a shape having a portion 41 was covered and firmly fixed (FIG. 1).
Subsequently, the solution A is filled into the filling container 4 through the uppermost opening 41 of the filling container 4 (length 15 cm, width 15 cm), and −2
Frozen for 12 hours at 0 ° C. The frozen product is freeze-dried (EYEL
Approx. 24 under reduced pressure (1 torr or less) with A company: FDU-830 type)
After freeze-drying for an hour, a sponge layer made of collagen was produced and adhered on a nonwoven fabric layer made of collagen fiber to obtain a two-layer structure. The two-layer structure was compressed at a pressure of 100 kgf / cm ² with a compressor (15 t press, manufactured by Iuchi Seieidou Co., Ltd.). Next, the nonwoven fabric layer on the side opposite to the side filled with the solution A was filled with the solution B in the same manner, and a sponge layer was prepared and adhered in the same manner as the solution A, and compressed. In this way, the sponge layer was prepared and adhered to the non-woven fabric layer at the same time, and the sponge layer made of collagen and the sponge-like coating layer containing the mixture of collagen and hyaluronic acid were provided on the non-woven fabric layer made of collagen fibers. A three layer structure was obtained.

(Example 5) Preparation of anti-adhesion membrane (5 layers) (1) After cross-linking reaction, a non-woven fabric layer composed of collagen fibers was formed.
After neutralizing by soaking in 7.5% sodium hydrogen carbonate aqueous solution for 30 minutes, it is taken out from the sodium hydrogen carbonate aqueous solution, the sodium hydrogen carbonate remaining on the surface of the non-woven fabric is washed with distilled water, and air-dried in a clean bench. A non-woven fabric layer made of collagen fibers was obtained in the same manner as in Example 1 except for the operation. (2) Separately, in the same manner as in Example 1, two sponge-like coating layers containing a mixture of collagen and hyaluronic acid and two sponge layers made of collagen were prepared. (3) Next, the sponge-like coating layers are adhered to both sides of the membranous material made of collagen nonwoven fabric in the same manner as in Example 1 and subjected to heat dehydration cross-linking, so that the center is strong enough to withstand suture. A non-woven fabric layer consisting of collagen fibers, a sponge layer consisting of collagen that promotes tissue regeneration and induction on both surfaces, and a sponge-like coating layer that contains a mixture of collagen and hyaluronic acid that exerts an adhesion prevention function on the outer layer. An adhesion preventive film having a total thickness of 5 layers and having a thickness of about 5 mm was obtained.

Example 6 Preparation of Adhesion Prevention Membrane (5 Layers) (1) In the same manner as in Example 5, a nonwoven fabric layer made of collagen fibers was prepared. (2) Separately, two sponge-like coating layers containing a mixture of collagen and hyaluronic acid and two sponge layers made of collagen were prepared by the following procedure. 125mL collagen 1% by weight aqueous solution and hyaluronic acid 1
125% by weight aqueous solution was prepared, and after mixing, 1N
The pH was adjusted to 1.5 with HNO ₃ . Mixture-2
By freezing at 0 ° C. for 24 hours and thawing at room temperature, a gel-like substance composed of a mixture of collagen and hyaluronic acid was obtained. The obtained gel-like material was immersed in a phosphate buffer for 30 minutes to be neutralized, and then washed in sterile distilled water for 30 minutes, and the washing was performed 3 times in total. The neutralized gel-like material was freeze-dried in the same procedure as in Example 1 to obtain a sponge-like coating layer containing a mixture of collagen and hyaluronic acid.
The sponge layer made of collagen was prepared from 250 mL of a 1% by weight collagen aqueous solution by the same method as in Example 1.

(3) Other operations are carried out in the same manner as in the embodiment, and a non-woven fabric layer made of collagen fiber having a strength capable of withstanding suturing is provided at the center, and a sponge made of collagen having a function of promoting regeneration and induction of tissue on both surfaces thereof. An anti-adhesion film having a thickness of about 5 mm was obtained, which was composed of a total of five layers having a layer and a sponge-like coating layer containing a mixture of collagen and hyaluronic acid exhibiting anti-adhesion ability in the outer layer.

Example 7 Preparation of Adhesion Prevention Membrane (5 Layers) (1) In the same manner as in Example 5, a nonwoven fabric layer made of collagen fibers was prepared.

(2) Separately, two sponge layers made of collagen were prepared in the same manner as in Example 1. In addition, a film-like coating layer containing a mixture of collagen and hyaluronic acid was prepared by the following procedure. Hyaluronic acid 1
An aqueous mixture of 1% by weight of collagen and an equal amount of an aqueous solution of 1% by weight of collagen is mixed to obtain an equal mixture of hyaluronic acid and collagen of about 125
mL was obtained. The mixture in an acidic state is neutralized with a 0.1N aqueous sodium hydroxide solution, and then the mixture is filled in a filling container having an opening at the top and air-dried in a clean bench to contain a mixture of collagen and hyaluronic acid. One sheet of film-shaped coating layer was prepared. Similarly, another film-like coating layer containing another mixture of collagen and hyaluronic acid was prepared.

(3) 100 k of a sponge layer made of collagen was compressed by a compressor (15-ton press manufactured by Inei Seieidou Co., Ltd.)
After compressing with a pressure of gf / cm ² ,
The non-woven fabric layer composed of collagen fibers obtained in the above step was adhered to both surfaces with a 1% by weight collagen aqueous solution to obtain a three-layer structure.
The film-like coating layer containing the mixture of collagen and hyaluronic acid obtained in (2) above is provided on the outer layer of the three-layer structure,
Bonding was done with a solution containing a mixture of collagen and hyaluronic acid. Then, each layer and the collagen used for adhesion and the mixture of collagen and hyaluronic acid were subjected to high vacuum (1 torr or less) in a vacuum dry oven (EYELA: VOS-300VD type) in the same manner as the above crosslinking reaction treatment. 1
The mixture was heated at 10 ° C. for 24 hours to carry out a thermal dehydration crosslinking reaction. In this way, a film-shaped coating layer containing a mixture of collagen and hyaluronic acid as an adhesion preventing layer, a scaffold for cell adhesion and growth in its inner layer, and a sponge layer made of collagen that promotes the induction of tissue regeneration. An anti-adhesion membrane having a 5-layer laminated structure with a thickness of about 5 mm having three layers of a compressed layer and a non-woven fabric layer made of collagen fiber for maintaining the strength to withstand suture fixation in the central layer was obtained. .

(Experimental Example 1) As anti-degradation test samples 1 to 3, adhesion preventing films having sponge-like coating layers in which the mixing ratio of collagen and hyaluronic acid was changed (3: 7, 5: 5, 7: 3) Was manufactured in the same manner as in Example 1. The sample was cut into 1 cm × 1 cm square and used for the test. As a comparative sample, an adhesion-preventing film having a sponge-like coating layer made of hyaluronic acid was prepared and cut in the same manner as in Samples 1 to 3 to be used for the test. Each sample was placed in a vacuum dry oven (EYELA: VOS-300VD type) under high vacuum (1 to
rr or less), 110 ° C., and 24 hours for thermal dehydration crosslinking. Store each sample in 10 mL of distilled water for injection at 37 ° C in a constant temperature bath, and change with time for 30 minutes, 24 hours, 3 hours.
The area of the coating layer was measured at three points after the day, the degree of decomposition was observed from the reduction rate, and the decomposition resistance was evaluated according to the evaluation criteria shown in Table 1. The evaluation results are shown in Table 2.

[0057]

[Table 1] Decomposition resistance scoring

[0058]

[Table 2] Decomposition resistance test

As is clear from Table 2, the comparative sample (hyaluronic acid alone) was completely dissolved after 1 day. Actually, it was completely dissolved within 30 minutes to 1 hour. Also,
Sample 3 with a high ratio of hyaluronic acid was completely decomposed after 3 days. It was observed that Samples 1 and 2 remained to retain their shape even after 3 days. Further, when the sample 2 after 3 days was examined manually, it was confirmed that the viscosity peculiar to hyaluronic acid was retained. From this, it was confirmed that the decomposition resistance was dramatically improved by mixing hyaluronic acid and collagen.

(Experimental Example 2) Implantation test The adhesion-preventing membrane obtained in Example 1 was implanted in the back muscle of a rabbit (n = 8), and the tissue reaction was observed with the naked eye and an optical microscope. , Biocompatibility was evaluated. The implant sample is
The anti-adhesion membrane obtained in Example 1 was treated with 1.5 mm × 10 mm
It was cut to the size of and used. Further, as a comparative sample, a high-density polyethylene plate was cut into the same size as the sample and used. The comparative sample was sterilized by EOG before use. The sample film was irradiated with 25 kGy of γ-rays and sterilized, and then used for an implantation test. Implantation was carried out by first performing normal inhalation anesthesia on a rabbit (body weight: about 2.5 kg to 3.0 kg), and then aseptically sandwiching the rabbit dorsal spinal cord, and a sample for comparison was placed on the left, and an interval was placed on the right. Each type was implanted. Implantation was carried out by inserting a sterilized 15-gauge injection needle from the skin surface at an angle of about 30 °, and extruding the sample film and comparative sample filled in the injection needle. Implanted in muscle. After this, 4 weeks after implantation 1 week, 4 more
Four weeks later, 4 birds were used as observation targets. At each observation time, the sample implantation site was dissected under anesthesia in two of the four rabbits, and the implantation site and surrounding tissues were visually observed centering on the inflammatory reaction.

From these observation results, at any observation time and in all rabbits, the sample membrane did not show a marked inflammatory reaction or the like to the comparative sample, and the biocompatibility of the anti-adhesion membrane obtained by the present invention was confirmed. It was found that the property was good. If 4 weeks have passed since the planting,
It was found that part of the sample film was decomposed and absorbed.

(Experimental Example 3) Adhesion-preventing action test An anti-adhesion membrane (hereinafter referred to as Sample 1) having a sponge-like coating layer composed of a mixture of collagen and hyaluronic acid, obtained in Example 1, was used for rabbits (n = 8) The abdominal wall defect part (1 cm square) and the part where the intestinal serosa was peeled off with a scalpel were embedded and suture-fixed. One week and six weeks after the implantation, the abdomen was opened, and the antiadhesion effect was observed from the residual amount of the antiadhesive membrane and the degree of adhesion. At this time, comparative evaluation was carried out by using, as a comparative sample, a portion having only a defective portion (hereinafter, Comparative 1) and an adhesion preventive film having a coating layer made of hyaluronic acid sponge (hereinafter, Comparative 2).

As a result of comparing the sample 1 and the comparison 2, regarding the residual amount after one week, the coating layer remained in the sample 1, but in the comparison 2, all the coating layers made of hyaluronic acid were decomposed and absorbed. I was sick. After 6 weeks, not only the coating layer but also the anti-adhesion film itself decomposed in Sample 1
It was absorbed. From this, it was found that the decomposition resistance was improved by mixing collagen and hyaluronic acid. That is, it is clear that the durability of the anti-adhesion effect is improved.

The anti-adhesion effect was evaluated based on the criteria shown in Table 3. The evaluation results are shown in Table 4.

[0065]

[Table 3] Criteria for determining the degree of adhesion

[0066]

[Table 4] Evaluation results of adhesion prevention effect

As is clear from Table 4, there was no case in which adhesion of Grade 2 or higher was observed in Sample 1 after 6 weeks, whereas Comparative 1 was Grade 4 or higher, and Comparative 2 was Grade 3
The above adhesion was recognized. From the above, it was revealed that the anti-adhesion film of the present invention has an effective anti-adhesion effect and high resistance to decomposition. From the above, it is clear that the anti-adhesion membrane of the present invention is excellent in anti-adhesion effect and decomposition resistance by having the coating layer containing the mixture of collagen and hyaluronic acid on the surface thereof.

[0068]

EFFECTS OF THE INVENTION The anti-adhesion membrane of the present invention has suture strength, biocompatibility, and anti-adhesion performance. In particular, the anti-adhesion layer containing a mixture of collagen and hyaluronic acid has a high resistance to decomposition and dramatically extends the duration of the anti-adhesion effect. Therefore, it is very useful as a sutureable artificial biological membrane capable of preventing the injured site or the bleeding site in the living body in surgery or the like, or the adhesion between the site and the normal site.

[Brief description of drawings]

FIG. 1 is an explanatory diagram of a fourth embodiment of the present invention.

[Explanation of symbols]

1. Flat plate 2. Nonwoven layer made of collagen fiber 3. Packing 4. Filling container 41. Aperture

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Claims (11)

[Claims] 1. An adhesion-preventing film having a coating layer containing a mixture of collagen and hyaluronic acid on the surface of a laminated film material having a non-woven fabric layer made of collagen fibers and a sponge layer made of collagen. 2. The adhesion preventing film according to claim 1, wherein the coating layer containing a mixture of collagen and hyaluronic acid is sponge-like or film-like. 3. The adhesion preventive film according to claim 1, wherein the coating layer containing a mixture of collagen and hyaluronic acid is a layer obtained by subjecting a mixture of collagen and hyaluronic acid to a crosslinking reaction. 4. The anti-adhesion membrane according to claim 1, wherein the coating layer containing a mixture of collagen and hyaluronic acid has a thickness of about 50 μm to 20 mm. 5. The adhesion-preventing membrane according to claim 1, wherein the collagen constituting the adhesion-preventing membrane is enzyme-solubilized collagen, acid-solubilized collagen, alkali-solubilized collagen or neutral solubilized collagen. 6. The anti-adhesion film according to claim 1, wherein a part or all of collagen constituting the anti-adhesion film is crosslinked. 7. The anti-adhesion membrane according to claim 1, wherein the non-woven fabric layer made of collagen fibers is a non-woven fabric layer in which collagen fibers are bonded to each other with (1) collagen or (2) a mixture of collagen and hyaluronic acid. 8. A non-woven fabric layer made of collagen fibers has a thickness of 50 μm to 10 mm, a sponge layer made of collagen has a thickness of 50 μm to 20 mm, and a coating layer containing a mixture of collagen and hyaluronic acid has a thickness of 50 μm to 20 mm. 50 μm
The anti-adhesion membrane according to claim 1, which has a thickness of -20 mm. 9. The adhesion-preventing film according to claim 1, wherein the non-woven fabric layer made of collagen fibers is formed by laminating 1 to 6 non-woven fabrics made of collagen fibers. 10. A collagen fiber having a diameter of about 10-1,
The adhesion-preventing membrane according to claim 1, wherein the non-woven fabric layer made of collagen fibers has a bulk density of about 5 × 10 ^{−4 to} 50 g / cm ³ . 11. The total thickness of the film is about 150 μm to 50 m.
The anti-adhesion membrane according to claim 1, which is m.