JP3463596B2 - Suture-resistant adhesion prevention membrane - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION [0001] TECHNICAL FIELD The present invention is capable of suturing
The present invention relates to an adhesion preventing film. Specifically, healing in vivo
Biological materials such as pleura, pericardium, dura mater, and serosa as anti-adhesion films
Defects or cut surfaces such as intimal tissue and various organs
It is used for filling, prosthesis, etc., and is particularly sutureable.
First, an anti-adhesion membrane with good biocompatibility and bioabsorbability
It is about. [0002] 2. Description of the Related Art In various surgical operations, an affected part is resected.
And often repairs damaged areas, especially in the lungs and heart.
Targets various organs such as the organs, liver, brain, digestive organs, and gallbladder
In the case of a surgical operation that does,
Do not supplement or prosthetic the membrane covering the tissue of the organ.
If so, it often impairs the underlying function of that organ.
If these procedures are performed incompletely, organ dysfunction may occur.
If you die or escape from a life threat, your prognosis will be great
The tendency to get worse is often seen. In addition, these supplements
If the suturing and fixing of the film at the site of
The function of the treated organ itself was barely maintained
Even if body fluids exude or leak from these organs,
Infection or attack on other organs due to
It can cause fire and erosion, resulting in a life-threatening crisis. In addition, these prostheses or supplemented sites
In some cases, adhesion of membranes occurs frequently,
Inducing organ dysfunction over time as a result of
There is also. To solve these various problems,
Film or anti-adhesion membrane covering organ or tissue of said organ
Has been developed with various materials. Conventionally, a nonwoven fabric made of collagen fibers
The layer is water-resistant with aldehydes, and the fibers are
Wound dressing material in which fibers are bonded to each other (Japanese Patent Laid-Open No. 50-141190)
Is known. However, the coating is
Is a layer using a cross-linking agent.
There is a problem in inducing weaving promotion. Also like this
The dressing only covers the wound and is sutureable.
It has a function to prevent film adhesion and adhesion.
Or not. [0005] SUMMARY OF THE INVENTION In the above prior art,
Is sufficient adhesion prevention effect, suture strength,
Lack of compatibility, biodegradation and absorption
Have the problem that they cannot be satisfied at the same time.
Was. That is, the present invention has the disadvantages of the conventional anti-adhesion film.
Sufficient suture strength and biocompatibility,
Insufficient solution / absorptivity and insufficient adhesion prevention
It is intended to solve these problems at the same time. [0006] That is, the present invention provides a
Gelatin or non-woven fabric layer
Sewing having a coating layer containing hyaluronic acid
It is an adhesion preventing film that can be combined. By the way, as a mechanism for preventing adhesion, the most
Simple and effective methods are also based on damage or loss
Physical contact is possible with the damaged tissue.
Another tissue is not brought into contact with the partition wall. I
However, if this is done with synthetic fibers, etc., lack of biocompatibility
From various calcifications, foreign body reactions, inflammatory reactions, etc.
Inconvenience arises. Also, the material used for the partition itself
Is the damaged or missing tissue and the corresponding other tissue
Do not mediate adhesions with. These conditions
Hyaluronic acid or gelatin
And the like. Both are viscous water-soluble liquids
Can be handled by various processing methods.
It can be used as a gel. These materials are mainly extracted from living bodies such as animals.
・ Because it is purified, its biocompatibility is good.
Practical application in various medical fields, including pharmaceuticals
I have. Also, when implanted in a living body, it is decomposed and absorbed
During the process, gelatin or hyaluronic acid molecules are released
Continue to exhibit viscosity while maintaining water-based properties,
Contact with tissue at the damaged or defective site
Combined with the characteristic that adhesion and adhesion hardly occur
One. Applying these features, hyaluronic acid
As a technology for manufacturing anti-adhesion films and medical materials
Is disclosed in JP-A-6-73103, JP-B-7-3012.
No. 4, Japanese Patent No. 2670996, Japanese Unexamined Patent Application Publication No.
-333402, JP-A-61-234864
, Registered Patent No. 2648308, JP-A-8-15
No. 7378, JP-A-9-296005, JP-A-9-296005
JP-A-7-102002, JP-T-7-509386
Gazettes and the like. But including these technologies
Mechanical strength for sufficient adhesion prevention and suturing
There is no technology that combines [0010] Similar to the above-mentioned hyaluronic acid, it cures gelatin.
Japanese Patent Laid-Open Publication No.
JP-103479, JP-A-8-52204
Those described are mentioned. However, these technologies
In all cases, as with hyaluronic acid, adhesion prevention
The two points of mechanical strength that can be stopped and suture fixed
It has yet to provide a satisfactory medical material. [0011] BEST MODE FOR CARRYING OUT THE INVENTION The adhesion preventing film of the present invention is a synthetic fiber.
Collar is a bio-based material that does not use any fibers or synthetics
Gen and gelatin or hyaluronic acid as main ingredients
These are processed into a non-woven or sponge shape
2 to 8 layers, preferably 3 to 5 layers as a whole
And a laminated structure composed of The anti-adhesion film is
The collagen non-woven fabric layer is responsible for the strength of the membrane that can be sutured in minutes.
The biocompatibility and the effect of preventing adhesion to surrounding tissue
The layer formed by tin or hyaluronic acid exerts
It is considered something. [0012] The anti-adhesion film of the present invention is a
To collagen and gelatin or hyaluronic acid
Is very biocompatible
It is not degraded and absorbed gradually in the transplanted living body.
In the end, all will be decomposed and absorbed. In particular
The collagen non-woven fabric layer can be used to regenerate tissue
Until completion, as a scaffold to refill, prosthetic and seal
Present and maintained its membrane strength for a period of time after suture fixation
Later, they are all decomposed and absorbed. Also, the outermost layer of gelatin
Or the hyaluronic acid layer, due to its viscosity and sustained release action,
Prevents adhesion of damaged or defective tissue to surrounding tissue
You. Note that the period during which this adhesion is prevented is
The tissue at the site does not naturally adhere to surrounding tissue
It lasts until it is regenerated and healed to a certain extent. these
Decomposes and absorbs gradually in the body while exhibiting the adhesion prevention effect of
Finally, these gelatin and hyaluronic acid layers
Lost. The typical collagen used in the present invention is
In addition, enzyme-solubilized collagen, acid-solubilized collagen,
Alkaline or neutral solubilized collagen
And soluble collagen. , These solubilized
Collagen is a particular type of proteolytic enzyme (eg, pep
Or solubilization with trypsin, etc.)
It has been solubilized by Lucari,
At the same time, the determinant of telopeptide,
Removed, usually called atelocollagen
Those suitable for medical use are particularly preferred. Solubilization of these
About the collagen which was done, from the following animal species,
Known technologies (JP-B-46-15033, JP-B-43-259839)
Gazettes and Japanese Patent Publication No. 43-27513).
It is. [0014] The reason for the collagen used in the present invention is as follows.
There are no particular restrictions on the coming, but in general, cattle,
Pigs, birds, fish, rabbits, sheep, rats, humans, etc.
You. Collagen can also be found in these skins, tendons, bones, cartilage,
Obtained by using various known extraction methods from organs, etc.
It is something that can be done. Also, about the type of collagen
Is any of the types that can be classified, such as type I and type III
Crab is not particularly limited, but handling
To type I collagen are particularly preferred. Also Collage
For solvents that solubilize water, water is preferred for handling.
Suitable. Gelatin is normal Japanese Pharmacopoeia compliant gelatin
Using hyaluronic acid from animals or microorganisms
However, medical grade products are particularly suitable. Nonwoven layer having sufficient film strength to be sutured
Spin the collagen solution into a coagulation bath to obtain
And crossed in multiple layers at the bottom of the coagulation bath to reduce the fiber diameter.
10 to 1000 μm, preferably 20 to 300 μm, bulk
Density (fiber density) is 5 × 10 ^-Four~ 5g / cm^Three, Preferred
1.0x10^-3~ 2.0g / cm^ThreeCollection of fibers
Get united. That is, the solution of the solubilized collagen
Continuous wet spinning, and put this long yarn in a suitable container
Move and arrange in a crossed state so that it is not arranged only in a certain direction
And place it. Next, vacuum drying, natural drying, low temperature drying,
Non-woven fabric (fiber
State). Collagen in any drying method
Below the denaturation temperature of the collagen used to prevent denaturation
It is important to perform drying at a temperature
Depending on the type of lagen, the temperature is about 35-45 ° C.
Below the temperature range is desirable. In particular, low-temperature drying or vacuum drying is preferable.
Good. On the other hand, discontinuous spinning by intermittent ejection or normal spinning
After continuous spinning, the obtained yarn is cut.
In both continuous and discontinuous spinning,
A short staple-like fibrous material is obtained. These are appropriate
In a state of being uniformly dispersed in a container of
Drying by non-woven fabric (fiber
State) can also be obtained. The concentration of the solubilized collagen solution used for spinning
The degree is optional depending on the type of collagen used.
Any concentration is possible as long as it is possible, but usually
0.1-20 wt%, of which 1-10 in wet spinning
Particularly preferred is about wt%. In addition, the solubilizing core
The discharge speed of the lagen and the winding speed of the obtained yarn
Any range is possible as long as the yarn can be formed. For discharging a solubilized collagen solution during spinning
The equipment used is a general-purpose gear pump, dispenser,
Any type of equipment such as a seed extrusion device may be used, but uniform spinning is performed.
For stable quantification of collagen solution with low pulsation
A device that can discharge is good. The size of the hole diameter of the spinneret when spinning is
There is no particular limitation as long as spinning is possible, but too much
For large pores when converting from fibrous to membrane-like material in a later process
This is difficult to achieve, and extremely small
Usually, 10 to 1000 μm is preferable because it is difficult to improve
More preferably, it is in the range of 50 to 700 μm.
The number of holes in the base may be singular or plural. Also the base
The shape is not particularly limited as long as spinning is possible.
Lit-shaped or various shapes may be used. More mouth
Regarding the hole length of gold, it is particularly limited as long as it can be spun
But not collagen molecules in the solubilized collagen molecules
As long as possible for the purpose of aligning
Is preferred. The coagulation bath of the wet spinning method is generally
If it is possible to coagulate the lagen
Is not, but an inorganic salt aqueous solution, an inorganic salt dissolving organic solvent,
Selected from alcohols, ketones or any of them
Combinations can be employed. With these coagulation baths
For example, as an aqueous solution of inorganic salts, sodium sulfate
, Sodium chloride, ammonium sulfate, calcium chloride
And magnesium chloride.
Thorium, sodium sulfate and ammonium sulfate for spinning
Is preferred. In addition, these inorganic salts are
Use of organic solvents such as inorganic salts dissolved / dispersed in
It can be used especially for dissolving sodium chloride in ethanol /
Dispersed solutions are preferred. In addition, as alcohols,
Tanol, ethanol, isopropanol, amylal
Call, etc.
Are particularly preferred. As ketones, aceto
And methyl ethyl ketone. In addition, not only as a coagulation bath,
Collagen with various cross-linking agents described below
A processing method having both coagulation and cross-linking treatment is also effective.
For example, a mixture of ethanol and glutaraldehyde
The liquid is used as a coagulation bath that has both coagulation and cross-linking.
If used, both processes can be performed at once and spinning
Cross-linked by immersing collagen yarn as it is
Processing is also possible. These simultaneous processes streamline the process
Not only spinning and small diameter with dilute collagen solution
It is very effective when spinning the yarn of In the above-mentioned method for forming various nonwoven fabric layers,
Particularly preferred examples are described below. Collar ejection mouth
The diameter is about 200μm and the hole length is about 15-20mm.
Dispense solubilized collagen solution without pulsation
In a 99.5 vol% ethanol coagulation bath.
It is desirable to perform wet spinning. 99.5vol% ethanol
When extruding solubilized collagen into the coagulation bath,
Move the gold at any time, and the spun yarn
Continuously extrude in a state where it can be
You. After this, remove the coagulating liquid and wash again with ethanol.
By cleaning and drying under reduced pressure, a very good floc
We can get fibers. This method simplifies the process and shortens the process.
This is particularly effective in terms of reduction and economic efficiency. Note that this example is a typical one,
It is not limited to this as long as it is obtained.
For example, the above-mentioned staple-like short fibers may be used.
Type of coagulation bath, use of coagulation bath and cross-linking agent mixed bath,
Drying methods, etc., and their combinations.
May be changed. The nonwoven fabric layer obtained by the above method is sufficiently sutured.
To be further cross-linked to obtain strength
Is desirable. This is due to the crosslinking treatment, especially when wet.
Physical strength and ensure that the strength required for suturing is sufficient.
Because it can be maintained. Also, when implanted in vivo,
Dramatically shortens the time required for dissolution and absorption compared to uncrosslinked
It is also to delay. By this cross-linking treatment,
To replace or replace the missing part of the organ
To prevent dysfunction and to repair wound surfaces and tissue
Maintains necessary membrane strength in the body until regeneration is completed
It is possible to remain as it is. This crosslinking method is roughly divided into physical crosslinking methods.
There are methods and chemical cross-linking methods. Examples of physical crosslinking methods and
Γ-ray, ultraviolet ray, electron beam, plasma, thermal dehydration crosslinking
On the other hand, as a typical example of the chemical crosslinking method,
Is an aldehyde such as dialdehyde or polyaldehyde
, Epoxy, carbodiimide, isocyanate
Tannin treatment, chromium treatment and the like. this
Of the aldehydes, formaldehyde, glue
Talaldehyde, acid aldehyde, glyoxal, malo
Acid dialdehyde, succinic acid dialdehyde, phthalic acid
Aldehyde, dialdehyde starch, polyacrolein,
Limethacrolein, etc.
Any available aldehydes
I do not care. In addition, as epoxies, glycerol di
Glycidyl ether, sorbitol diglycidyl ether
, Ethylene glycol diglycidyl ether, polyether
Tylene glycol diglycidyl ether, polyglycero
Polyglycidyl ether, etc.
Epoxy compounds that can crosslink with
Is available. Further, as an example of carbodiimides
Is particularly preferably a water-soluble carbodiimide,
Anything is possible as long as the reaction mechanism is possible. Also, isocyanates
Hexamethylene diisocyanate is a typical example of nates.
And tolylene diisocyanate
Has at least two isocyanate groups involved in the crosslinking reaction.
Can be cross-linked with collagen
Yes, and not particularly limited to these. By the above-mentioned method, the kola obtained by spinning or the like is used.
-Gen nonwoven fabric (fibrous material) obtains higher suture strength
For this purpose, it can be further compressed. Compress
This increases the fiber density of the non-woven fabric layer,
It is possible to obtain a highly preferable film-like material. Compressed
The non-woven fabric has a bulk density of particularly 0.05 to 5 g / cm.^ThreeIs
Is preferred. The compression can be performed by a general-purpose press.
Although it is intended for medical use, it is
Sterile packaging materials, such as aluminum packs and high-strength resin packages
Compressed aseptically packaged with equipment, etc.
Is desirable. At this time, regarding the pressure for compressing the nonwoven fabric layer,
Is not particularly limited as long as it does not destroy the non-woven fabric layer body.
Not usually, but usually 10-1000kgf / cm^TwoIs
It is desirable. Non-woven fabric layer has fiber density for the purpose of improving strength
With a laminated structure that is the final form
Handling of medical treatment when using an adhesion prevention film
From the point of view, it is compressed and thinned as much as possible
It is desirable. The nonwoven fabric layer thus thinned is
By laminating this to form a multilayer structure,
It becomes possible to obtain an adhesion preventing film having an increased joint strength. The nonwoven fabric layer or pressure obtained by the above method
The purpose of further improving the physical strength of the shrinked nonwoven fabric layer
Thus, it is possible to perform a binder treatment. Binder
-As for collagen sponge layer and / or soluble
Collagenized collagen solution is preferred. This is a non-woven layer or pressure
The collagen sponge layer was dissolved in the shrinked nonwoven fabric layer
After impregnated with the solubilized collagen solution, air dry and blow
Under appropriate drying methods such as drying under reduced pressure, drying under reduced pressure, and drying under low temperature
After drying, the fibers of the non-woven fabric layer are bonded together to form a film.
It is a technique that The film obtained by this operation is non-woven
The physical strength is much higher than that of the fabric layer alone,
The suture strength is also significantly improved. Note that the required physical
Depending on the degree of strength, this impregnation and drying step is performed once to several times.
It can be repeated zero or more times. However, when performing the binder treatment,
Cross-linking is applied to the woven or compressed non-woven layer
If not, it will not work when impregnated with the solubilized collagen solution.
The woven fabric layer itself may be dissolved. Therefore,
Cross-linking treatment can be performed in advance by the method described above.
desirable. In addition, other than the treatment method of impregnating the collagen solution,
In a suitable container or mold, solubilize
The method of casting or filling the lage or directly to the non-woven fabric layer
There is also a method of applying a solubilized collagen solution. The film-like material of the present invention comprises collagen fibers.
1 to 6 layers, preferably 1 to 3 layers
Having. The nonwoven fabric has a thickness of 100 μm to 50 mm, preferably
Preferably, the nonwoven fabric layer has a thickness of 200 μm to 4.0 mm.
It consists of 1 to 6 layers, preferably 1 to 3 layers. Film form of the present invention
To get the product, for example, dry it in advance into a sponge
A membrane-like material obtained by sandwiching the nonwoven fabric layer with the collagen layer
Alternatively, the sponge layer and the non-woven
The film material in which the cloth layer is embedded in the sponge layer is
Pressure or reduced pressure in the presence of activated collagen solution or water
By placing it under pressure, the solubilized collagen of the sponge layer
Dissolve and let it mix well with the non-woven fabric layer
There is a method of drying by a drying method. Use this sponge layer
The binder treatment method used is to bind the fibers of the nonwoven fabric layer to each other.
To the amount of collagen actually used
And a very small amount of solvent components such as water
In addition to short time when drying in the process,
The great merit of very little contracture and deformation
is there. If it is a normal impregnation step, solubilized collagen for impregnation
The actual amount of collagen in the solution is related to the practical solution viscosity.
Therefore, the limit is about several percent, and the remaining 90% or more is moisture
And so on. Therefore, for impregnation and drying operations
Not only is it time-consuming, but the operation itself needs to be repeated
Is a simple method, but lacks rationality.
You. Of course, these methods are representative examples,
Solubilizing fiber between layers or compressed non-woven layers
If it is a method of binding using a gene to form a film,
Such a method may be used, and the above representative example is not particularly limited.
No. In addition, for the binder treatment, the collagen solution
In addition, gelatin solution, hyaluronic acid solution, etc. can be used
is there. In the present invention, the cola obtained by the above method is used.
A non-woven fabric layer consisting of
It is necessary to form a hyaluronic acid layer. The formation
The method is easy to carry out by freeze drying etc.
It is possible. , The order and method in the manufacturing process to form
The method is not particularly limited. Examples of specific manufacturing methods
For example, collagen non-woven fabric layer and gelatin or hyaluron
Laminate an acid sponge or film layer, or
Independently manufactures a gelatin or hyaluronic acid sponge layer
Then, after that, the sponge layer and the collagen nonwoven fabric layer
Solubilized collagen, gelatin solution, hyaluronic acid solution
There is a method of bonding using any method. Also, collagen non-woven
Once the fabric layer is immersed in the solubilized collagen solution,
Frozen and dissolve gelatin or hyaluronic acid again
Immersion in a liquid, freeze them and integrate them, then freeze-dry
By drying, the collagen layer and gelatin or
Is a method of obtaining a laminated sponge of hyaluronic acid. The gelatin solution filled in the container or
Dipped collagen nonwoven fabric in hyaluronic acid solution
Later, freeze in a freezer, freeze-dry and
Collagen non-woven fabric in the sponge or hyaluronic acid sponge layer
It is also possible to mold to a state in which is contained. I
However, these are merely examples of the processing method of the present invention.
The purpose of these processing is gelatin or hyaluronic acid
Sponge layer or film layer and collagen nonwoven layer
Can be integrated without being easily separated or separated when implanted in vivo
The goal is to be
Any processing order and method may be used. Gelatin or hyaluronic acid sponge layer
For preparation, gelatin solution or hyaluronic acid solution
Is cast into a container or filled to a desired thickness, and
After freezing sufficiently in a freezer, etc., dry with a freeze dryer.
By drying, various uniform sponge layers can be obtained.
At this time, formed on gelatin or hyaluronic acid sponge layer
The pore size of the fine porosity is determined by the gelatin solution, hyaluronic
Acid solution concentration and solvent, freezing temperature and freezing time, etc.
It changes with. Further, a gelatin sponge layer, hyaluronic acid
Total amount of various raw materials in sponge layer and various sponges
About the thickness of the layer, the adhesion prevention effect of the target site,
Support for repairing damaged / cut sites and inducing tissue regeneration
Each sponge layer for about 1 to 4 weeks so as not to interfere
Is desirably in the body. This gelatin
Or the thickness of the hyaluronic acid sponge layer and the sponge layer
The total amount of raw materials used for the formation of
Considering the effect on the decomposition / absorption time and induction of tissue regeneration
And can be controlled arbitrarily. Spong
The thickness of the bilayer is specifically about 50 μm to 20 mm, preferably
100 to 1000 μm, especially at the end of drying.
Thickness is 1-10mm, these are compressed and used
In some cases, a sponge having a thickness of 100 μm to 1 mm
Di is preferred. Considering these situations, the gelatin solution
Is 0.5 to 60% by weight, preferably 5 to 4% by weight.
0 wt%. Hyaluronic acid solution concentration range
Is 0.1 to 50 wt%, preferably 0.5 to 5 wt%
It is desirable that The freezing temperature is -196 to -1
0 ° C, preferably general-purpose freezer or deep free
-80 to -10 ° C, which can be set by the user
Good. Also, freeze dryers can be dried stably.
It is not particularly limited. Further, a gelatin solution or hyaluronic acid
The filling amount of the solution into the styrene
The thickness of the flange is about 50 μm to 20 mm, preferably 10 μm.
What is necessary is just to fill so that it may be set to about 0-1000 micrometers. This
These values can be changed at any time according to the purpose of use.
However, the present invention is not limited to these examples. A hyaluron exhibiting an adhesion preventing effect
Acid or gelatin layer is limited to sponge shape only
However, for example, a file obtained by a normal casting method or the like is used.
It may be processed into a film shape or the like. Also prevents adhesion
Gelatin or hyaluronic acid layer to give effect
When forming, depending on the purpose, one side or both sides of the membrane
Various forms such as surface, partial or full coverage
The method of forming the gelatin or hyaluronic acid layer is optional.
The law or its site is particularly limited
, And any combination is possible. The cross-linking treatment is performed by the various cross-linking methods described above.
The target is that the anti-adhesion film of the present invention
And gelatin or hyaluronic acid
The non-woven fabric layer, gelatin layer or
Hyaluronic acid layers, and these are integrated and laminated
Part or all of the anti-adhesion film is targeted. Also, the frame
The order of the bridges and the combination of the crosslinking methods are arbitrary, and
Not determined. However, most preferably, collagen non-woven
Glutaraldehyde and other aldehydes in the fabric layer
Cross-linking, followed by a gelatin layer or hyal
Form a layer of lonic acid, integrate with it, and finally heat dehydration rack
Apply a bridge. In these methods, collagen spinning
-In the process of nonwoven fabric, a coagulant such as ethanol,
Mix crosslinking agents such as glutaraldehyde and spin
It also includes a method of performing the yarn and cross-linking steps at once. The healing finally obtained by the method of the present invention
Anti-adhesion membrane has suture strength, biocompatibility, biodegradation and absorption
It is an anti-adhesion film excellent in terms of properties. In addition,
When performing binder treatment on the non-woven fabric layer
Can be thermally dehydrated and crosslinked also in the layer formed by this.
good. However, this is only an example, for example,
No problem when all layers are treated by thermal dehydration crosslinking
In addition, for both sterilization and cross-linking, for example,
Is also good. The nonwoven fabric layer obtained by the above method and the gelatin
An anti-adhesion film having a hyaluronic acid or hyaluronic acid layer,
It is possible to compress. Gelatin or hyaluronic
After compressing the acid sponge layer alone or the nonwoven fabric layer alone,
Uncompressed nonwoven layer or gelatin or hyal
You may integrate with combining with a lonic acid sponge layer. Special
Preferably, the nonwoven fabric layer and
Tin or hyaluronic acid sponge layer is integrated
To compress things at the same time. This is to compress
The thickness was reduced by the
When actually using the anti-adhesion membrane at the site,
Of penetrating suture needles, cutting into arbitrary shapes, etc.
Handling is particularly improved, and transplant surgery and the like can be performed more smoothly. Pressure
The method of shrinking is exactly the same as when compressing the collagen nonwoven layer
Can be performed with a general-purpose press,
Sterile packaging that is strong enough for the intended purpose
Materials such as aluminum packs and high-strength resin packaging materials
Desirably, it is compressed in a bacterially packaged state. Ma
As for the pressure to compress the adhesion prevention membrane,
There is no particular limitation as long as it does not break, but usually 1
0-1000kgf / cm^TwoIt is desirable that [0040] Next, the present invention will be described with reference to examples.Example 1 First, in an Erlenmeyer flask (Corning)
Magnetically transforms chicken-derived atelocollagen
While gently stirring with a stirrer, add distilled water for injection,
Collagen concentration of 3 wt% or 5 wt%
Prepare the collagen solution aseptically in a clean bench.
Was. Next, dispens 40 ml of 5 wt% collagen solution.
Sensor (San-A-Tech: EFD900)
Needle tip of 27 gauge size (200 μm hole diameter)
From the end, 99.5 vol under a constant pressure condition of 4.0 bar
% Ethanol solution (Wako Pure Chemical Industries, special grade) in a coagulation bath
Continuous extrusion spinning was performed. During continuous extrusion spinning,
The needle tip randomly on an ethanol coagulation bath.
While moving, the precipitated and coagulated collagen yarn is
Spun so as to cross multiple layers at the bottom of
(Aggregate of fibrous collagen) was obtained. Then this non-woven
Leave the cloth (aggregate of fibrous collagen) for 1 hour
After coagulation, the coagulation liquid is exchanged twice with the same ethanol.
And washed. The above nonwoven fabric (aggregate of fibrous collagen)
As it is in a vacuum dry oven (EYELA
Oil rotary vacuum pump (ULVA) in VOS-300VD
C Company: GCD135-XA) at room temperature under reduced pressure (less than 1 Torr)
Full) dried for 4 hours, non-woven fabric (fibrous collagen non-woven)
Cloth). Fiber diameter is 60μm, bulk density of non-woven fabric is
4.0 × 10^-2g / cm^ThreeIt is. Next, this non-woven fabric
Put in sterilized aluminum packaging, high pressure jack
100 kgf with a (15-ton press machine manufactured by Iuchi Seieido)
/ Cm^TwoCompressed at a pressure of approximately 8cm x 5cm
Cloth (fiber diameter is 60μm, bulk density of non-woven fabric is 0.9g /
cm^Three) Got. Further, the compressed non-woven fabric was used in 5% glutar.
Aldehyde solution (Wako Pure Chemical, first grade glutaraldehyde
25% solution diluted with distilled water for injection) for 4 hours,
Crosslinking treatment was performed. After the reaction is complete, use distilled water for injection.
After washing, immerse for 1 hour in distilled water bath for injection
Change the water three times to remove excess glutaraldehyde.
I left. Reduce the pressure of the non-woven fabric after cross-linking treatment again
Dry and compress, 0.6mm thick, about 8cm in size
5 cm compressed nonwoven fabric (fiber diameter is 60 μm, bulk of nonwoven fabric
The density is 0.91 g / cm^Three) Got. Separately, in order to perform binder processing,
A dried collagen sponge was prepared. This is 3w
Put the t% solubilized collagen solution into a styrene square container
After filling to a thickness of about 17 mm, the freezer (SA
NYO: MEDICALFREEZER) at -20 ° C
It was frozen for about an hour. Next, the frozen solubilized collagen
Lyophilizer (EYELA, Inc.)
Manufactured by FDU-830) and an oil rotary vacuum pump (ULV
AC company: GCD200-XA) under reduced pressure (0.05 Torr or less)
Full) freeze-dried for about 24 hours and remove collagen sponge
Obtained. The film thickness at the end of freeze-drying was about 15 mm,
Rate was 80%. One sheet of the obtained crosslinked nonwoven fabric (thickness:
0.6 mm), separately prepared collagen sponge
100kgf / cm again with one sheet (15mm thick)
^TwoCompressed under the pressure of
3 mm thick, buried in a ponge layer, approximately 7 cm in size x 4.
A 5 cm-thick collagen film having a two-layer structure was obtained. Next, the above film was solubilized in a 3 wt%
Vacuum while immersed in about 15 ml
In a dry oven (manufactured by EYELA: VOS-300VD)
To decompress the air in the film,
Solution is forcibly impregnated into the sponge layer of the film.
Was. The collagen in the sponge layer is water in the collagen film
And a one-layered film was obtained. The film obtained
The product is dried at low temperature (4 ° C.) for 24 hours,
Treated collagen membrane (0.18 mm thick, large
(Approximately 7 cm × 4.5 cm). Next, the obtained Photoshop
Transfer the genogen film to a styrene square container and place it on the container.
Pour a 30 wt% gelatin solution from
Adjust the position with sterile tweezers etc. so that it comes to the middle layer
After that, freezer (Sanyo: MEDICALFREEZER)
At -20 ° C for about 12 hours. In this way
Freezing with integrated gelatin layer and compressed collagen nonwoven layer
A three-layer laminated structure film in a bonded state was obtained. Next, the three-layer structure film was placed in a container.
Similarly, freeze-drying was performed at −20 ° C. for about 24 hours.
The obtained compressed collagen nonwoven layer and gelatin sponge layer
Was again integrated into a 400 kgf / c
m^TwoCompressed with a pressure of about 1.6mm thick, about 7c in size
An mx 5 cm three-layer laminated film was obtained. Next, the obtained film is vacuum-dried.
Oven and oil rotary vacuum pump (ULVAC GCD1)
35-XA) at 135 ° C under reduced pressure (less than 1 Torr)
The thermal dehydration crosslinking treatment was performed for 12 hours. In this way,
Only the fibers of the collagen nonwoven fabric layer are glutaraldehyde frames
Bridge treated, one side of the surface is about 0.4mm thick
Anti-adhesion film having gelatin sponge layer (thickness of about 1.6
mm, size about 7 cm × 5 cm). The adhesion obtained
Suture strength, biocompatibility, inducibility of tissue regeneration, healing
The anti-adhesion properties are shown in Examples 3 and 4 below. [0048]Example 2 As in Example 1, chicken atelocollagen was injected.
Dissolve in distilled water for injection, and collagen concentration is 5 wt%
Solution was aseptically prepared in a clean bench. next,
40 ml of a 5 wt% collagen solution is dispensed
20 Technics: EFD900)
From the tip of the needle of the needle size (pore diameter about 600μm)
Under constant pressure conditions of 2.0 bar, 99.5 vol% ethanol
1000ml / 25% glue solution (made by Wako Pure Chemical Industries, special grade)
Continuous extrusion spinning into a mixing bath of 42 ml of taraldehyde solution
Yarn went. Fiber diameter 200μm, non-woven fabric bulk density
0.01 g / cm^ThreeWas obtained. Examples below
Operate in the same way as in Step 1 and contain about 1% glutaraldehyde
The above non-woven fabric at 4:00 in a mixed ethanol coagulation and crosslinking bath
Reaction. After the completion of the reaction,
Remove the mixed solution of rualdehyde / ethanol and add 99.5
Wash the nonwoven fabric three times with a 3% ethanol solution and remove excess gluta.
The rualdehyde was removed. Next, the pressure was reduced in the same manner as in Example 1.
After drying, compress this collagen non-woven fabric
0.7mm thick and about 7cm x 5cm
Woven cloth (fiber diameter 200 μm, non-woven fabric bulk density 0.8
g / cm^Three) Got. The compressed nonwoven thus obtained
A laminated structure was formed using two cloths. Next, these two nonwoven fabric layers were
Put into a square container, and further add 20wt% hyal from the top of the container
After filling with the acid solution, a freezer (manufactured by Sanyo:
MEDICALFREEZER), freeze at -20 ° C for 12 hours,
After that, freeze-drying is performed for about 24 hours,
Membrane of four-layer structure containing collagen nonwoven layer in dilayer
I got something. 500 kgf / cm^Twoof
Compressed by pressure, thickness about 1.9mm, size about 7cm × 5
cm of adhesion preventing film was obtained. Vacuum the resulting membrane
Dry oven and oil rotary vacuum pump (ULVAC)
(GCD135-XA type) at 135 ° C under reduced pressure (1 Torr)
) For 12 hours. in this way
And the non-woven fabric layer has a two-layer structure,
Cross-linking, and the thickness of one side is about 0.25m
m hyaluronic acid sponge layer is integrated with the nonwoven fabric layer,
Thermally crosslinked four-layer adhesion prevention film (about 1.9m thick)
m, size about 7 cm × 5 cm). Anti-adhesion obtained
Suture strength of the stop membrane, biocompatibility, inducibility of tissue regeneration, adhesion
The preventive properties are shown in Examples 3 and 4 below. [0050]Example 3 Measurement of suture strength Of the anti-adhesion film prepared in Example 1 and Example 2.
The suture strength was measured. As a control sample
Is Gore-Tex pericardium (Gore-Tex Corp .: Gore-Tex)
S EPTFE patch II (pericardial sheet)), pig extirpation
Pericardium and porcine isolated brain dura were used. The porcine isolated pericardium
And excised cerebral dura, excised about 20 kg of pig under anesthesia
After surgery, each membrane is removed and immersed in saline.
And immediately measured fresh. The measurement method is as follows.
I went there. First, each sample membrane and control membrane
Were cut into 1 cm x 2.5 cm plate sections
At the distance of 5 mm from one end in the long side direction, and the center of the film
In the part, the suture (4-0 pro-lean thread, ETHICON,
INC). Next, knot the suture
The sliced section was immersed in physiological saline at 37 ° C. for 30 minutes.
After that, take it out immediately and measure the tensile strength (manufactured by Shimadzu Corporation).
The tensile strength was measured by the autograph S-500D).
Was. The measurement conditions are as follows: the end opposite to the side where the suture passed
Chuck to a distance of about 10mm from
Hook a looped suture on the measuring hook, 10mm / min
At a constant speed. At this time, the membrane
Twist or when the suture separates from the membrane section to be measured
The change in stress up to the point was measured. Within the recorded stress,
The suture strength of the membrane to be measured (the
, N). Table 1 shows the results. [0051] [Table 1] Unit: (N) As is clear from Table 1, the adhesion prevention of the present invention
The stop membrane is suture-resistant enough to withstand normal suturing
Strength. [0053]Example 4 Implanting test The anti-adhesion membrane obtained in Example 1 was used for rabbits (n = 8).
Implanted in the back muscles, the tissue response is visually and optically examined.
Observation with a microscope was performed to evaluate biocompatibility. Implanted sump
The adhesion preventing film obtained in Example 1 was 1.5 mm
It was cut into a size of × 10 mm and used. In addition,
As a roll, a high-density polyethylene plate is sampled.
Cut into the same size as The control
Was used after sterilizing ethylene oxide gas. Sa
The sample membrane is sterilized by irradiation with 25 kGy γ-rays,
Used for implantation tests. Implantation should be done with rabbits (weight approx.
2.5 kg-3.0 kg), perform normal inhalation anesthesia,
Aseptically sandwich the spinal cord at the back of the rat and control to the left
The sample was implanted on the right. Implantation method is sterilized 1
Using a 5 gauge needle, diagonally to the skin surface
Inserted at an angle of 30 degrees and filled inside the injection needle
Extrude sample membranes and controls into rabbit intramuscular
Buried. One week after implantation, four birds, and another four weeks
Four hours later, four birds were used as observation targets. At each observation time
And two of the four rabbits were under anesthesia.
Make an incision in the implanted site, and observe the implanted area and surrounding tissue.
Observations were made visually with a focus on inflammatory reactions and the like. Ma
The other two birds were killed by excessive anesthesia.
And remove the surrounding tissue, including the embedded plant, and
Perform phosphorus fixation, prepare sections, and observe with a microscope.
Was. From these observation results, any observation time
, And in all rabbits, the sample membrane
No significant inflammatory response, etc. to the control
Good biocompatibility of anti-adhesion membrane obtained by light
I understood that. If 4 weeks have passed since implantation,
That part of the sample membrane has been decomposed and absorbed
Was found. [0055]Example 5 Examination of adhesion prevention effect 10 rats (body weight 250 g-300 g), 5
Into two groups, one group serving as a control and the other group serving as a control.
And a sample group. The sample group was manufactured in Example 2.
An anti-adhesion film was used. Control group, sample
In any of the groups, rats were anesthetized by intramuscular injection.
After the irradiation, the state of anesthesia was maintained by inhalation anesthesia. Conte
In the roll group, an abdominal incision was made under anesthesia to expose the cecum and
After that, the serosa of about 5 mm square was peeled off. Also, peel off
Separate the separated cecal serosa and the corresponding abdominal wall in the same manner.
Adhesion model where the cecal injury surface and the abdominal wall injury surface
Was prepared. After this, in the control group, nothing
The abdomen was closed without any treatment. On the other hand, in the sample group,
After creating an adhesion model as in the roll group,
The surface is coated with the anti-adhesion film prepared in Example 2 and fixed.
Specified. The size of the fixed anti-adhesion membrane is about 10 mm x
It is about 10mm and the four corners are sutures (5-0
The thread was lightly hung over the intestinal tract with suture and fixed. Sample group
Rats were re-operated after 2 weeks in both control and control groups
The abdomen was opened, and the state of adhesion was visually observed. Eye
The degree of visual adhesion is determined based on the criteria in Table 2 below.
Perform scoring to compare the sample and control groups.
Comparative evaluation. If it is judged that adhesion has been recognized, grade 3 or higher
Adopted above. [0056] [Table 2][0057] [Table 3] As is clear from Table 3, the sample group and the
As a result of comparative study of the control group, adhesion was found in the sample group.
No cases were observed, whereas the control group
All cases showed adhesion of grade 3 or higher. Also sample
In almost all cases in the group, the anti-adhesion membrane was
It remains and can move damaged surfaces without moving to other places.
It served as a quarantine. Furthermore, residual adhesion prevention
Carefully peel off the stop membrane and visually observe the damaged surface of the cecum,
It was apparent that the damaged surface had begun to regenerate. [0059] The adhesion preventing film of the present invention can be sutured.
Therefore, it can be fixed directly to the defect or injury site in the living body
This allows the body to be seen in the conventional anti-adhesion membrane.
Healing without moving, dropping, or deviating from the target site in
Good adhesion by reliably isolating the site of attachment from defects and injuries
It is possible to exhibit the effect of prevention, further preventing adhesion
After the necessary period of time has passed, all will be decomposed and absorbed in the body
It has the feature of.

Claims (1)

(57) [Claims 1] A solution of solubilized collagen is continuously wet-spun, and the long fibers are arranged in a crossed state so as not to be arranged only in a certain direction, and dried. A non-woven fabric layer comprising collagen fibers obtained by the above method, having a coating layer containing gelatin or hyaluronic acid on the surface thereof.