WO2015076252A1

WO2015076252A1 - Tissue adhesive porous film, method for producing same, and tissue adhesive porous film tape

Info

Publication number: WO2015076252A1
Application number: PCT/JP2014/080477
Authority: WO
Inventors: 田口　哲志
Original assignee: 独立行政法人物質・材料研究機構
Priority date: 2013-11-22
Filing date: 2014-11-18
Publication date: 2015-05-28
Also published as: JP6120392B2; JPWO2015076252A1

Abstract

This tissue adhesive porous film (1) is a film having a porous structure in which hydrophobized gelatin (10) is crosslinked, wherein the hydrophobized gelatin (10) is gelatin (13) in which a part of an amino group (12) of Lys is substituted with a hydrophobic functional group (11). By using this tissue adhesive porous film (1), a tissue adhesive film which has high adhesion strength to wet tissue and adequate strength, and a method for producing the same can be provided.

Description

Tissue-adhesive porous membrane, method for producing the same, and tissue-adhesive porous membrane tape

The present invention relates to a tissue adhesive porous membrane, a method for producing the same, and a tissue adhesive porous membrane tape.

The tissue adhesive film is a polymer film capable of adhering to a living tissue (hereinafter also referred to as “tissue”) such as intestine, blood vessel, and skin during operations such as digestive surgery and cardiovascular surgery. By applying this to an anastomosis such as an intestine or a blood vessel, leakage of contents can be prevented, and the safety of surgery can be improved.

There are roughly three types of biomaterials for adhering tissues: cyanoacrylate-based tissue adhesives, tissue adhesives composed of biopolymers and aldehyde-based crosslinking agents, fibrin-based tissue adhesives, and tissue adhesive films.
None of these tissue adhesives and tissue adhesive films have sufficient properties of either adhesive strength or biocompatibility.

Since fibrin-based tissue adhesives and tissue adhesive films are blood products that use human blood as a raw material, they are classified as pharmaceuticals and require a great deal of effort in terms of approval. Moreover, when it became a pharmaceutical classification | category, after authorization, the usage history must be continued for 20 years, and there existed a subject that a great effort was required.

On the other hand, a tissue adhesive film prepared from a medical material (Patent Document 1) prepared by cross-linking gelatin, which is a non-blood preparation, with succinimidated poly-L-glutamic acid (Patent Document 1) and gelatin or Collagen (Patent Document 2) ), A tissue adhesive composition (Patent Document 3) in which a polymerizable and / or crosslinkable material in the form of particles and a particulate material are mixed, and gelatin having an alkyl group introduced in the side chain have been reported. However, all of these have a problem that the adhesive force to the wet tissue is not sufficient.

Under such circumstances, there is a demand for a tissue adhesive film having high adhesive strength and appropriate strength with respect to wet tissue. Until now, it has been clarified that medical films using hydrophobized gelatin show adhesiveness to wet tissues, but water cannot be sufficiently absorbed from the wet tissues, and the medical film itself swells. There has been a concern that peeling may occur in a short time (Non-Patent Document 2).

JP-A-9-103479 JP 2008-284256 A Special table 2006-523113 gazette

An object of the present invention is to provide a tissue adhesive film having a high adhesive strength to a wet tissue and an appropriate strength, and a method for producing the same.

The inventors of the present invention have prepared a porous tissue-adhesive porous membrane containing a hydrophobicized gelatin in which a hydrophobic group is introduced into the amino group of gelatin, thereby sufficiently absorbing moisture from the wet tissue, On the other hand, it was discovered that a tissue adhesive film having high adhesive strength and appropriate strength and a method for producing the same can be provided, and the present invention has been completed.
The present invention has the following configuration.

(1) A membrane in which hydrophobic gelatin is crosslinked and has a porous structure, wherein the hydrophobic gelatin is gelatin in which a part of the amino group of Lys is substituted with a hydrophobic functional group Tissue-adhesive porous membrane.

(2) The tissue-adhesive porous membrane according to (1), wherein the porous structure is composed of pores between a plurality of hydrophobic gelatins provided in the crosslinked structure of the hydrophobic gelatin.
(3) The porous structure includes voids between a plurality of hydrophobic gelatins provided in the crosslinked structure of the hydrophobic gelatin, and porogen holes provided in the crosslinked structure using a porogen as a template. The tissue-adhesive porous membrane according to (1).
(4) The tissue-adhesive porous membrane according to (3), wherein the porogen pores are provided using a porogen having a particle size of 50 to 100 μm as a template.

(5) The tissue adhesive porous membrane according to any one of (1) to (3), wherein the hydrophobic gelatin is crosslinked with a crosslinking agent or a condensing agent.
(6) The cross-linking agent or condensing agent is trisuccinimidyl citrate, disuccinimidyl tartrate, disuccinimidyl malate, disuccinimidyl succinate, glutaraldehyde, formaldehyde, polyethylene glycol diglycidyl ether, genipin The tissue-adhesive porous membrane according to (5), which is one or a combination of two or more water-soluble carbodiimides.
(7) The tissue adhesive porous membrane according to any one of (1) to (3), wherein the hydrophobized gelatin is crosslinked by thermal crosslinking.

(8) The hydrophobic functional group is a saturated fatty acid ethyl group (carbon number 2), propyl group (carbon number 3), butyl group (carbon number 4), pentyl group (carbon number 5), hexanoyl group (carbon 6), heptanoyl group (carbon number 7), octanoyl group (carbon number 8), nonanoyl group (carbon number 9), decanoyl group (carbon number 10), undecanoyl group (carbon number 11), dodecanoyl group (carbon number 12) ), Tridecanoyl group (carbon number 13), tetradecanoyl group (carbon number 14), pentadecanoyl group (carbon number 15), hexadecanoyl group (carbon number 16), heptadecanoyl group (carbon number 17), stearoyl group (18 carbon atoms), branched saturated fatty acid isopropyl group (3 carbon atoms), isobutyl group (4 carbon atoms), isopentyl group (5 carbon atoms), isohexanoyl group (6 carbon atoms) Isoheptanoyl group (7 carbon atoms), isooctanoyl group (8 carbon atoms), isononanoyl group (9 carbon atoms), isodecanoyl group (10 carbon atoms), isoundecanoyl group (11 carbon atoms), isododecanoyl group (12 carbon atoms) , Isotridecanoyl group (carbon number 13), isotetradecanoyl group (carbon number 14), isopentadecanoyl group (carbon number 15), isohexadecanoyl group (carbon number 16), isopalmityl group (carbon number) 16), isoheptadecanoyl group (carbon number 17), isostearoyl group (carbon number 18), unsaturated fatty acid oleyl group (carbon number 18, unsaturated carbon 1), linolenyl group (carbon number 18, non-carbon) Saturated carbon 2), α-linolenyl group (18 carbon atoms, 3 unsaturated carbons), cholesteryl group which is a cell membrane component, or a combination of two or more The tissue-adhesive porous membrane according to (1), which is characterized in that it exists.

(9) The tissue adhesive porous material according to (1), wherein the gelatin is one or a combination of two or more of gelatin derived from human, pig, cow, tilapia, cod or genetically modified gelatin The membrane.
(10) The tissue adhesive porous membrane according to any one of (1) to (9), wherein the tissue adhesive porous membrane contains a drug.
(11) The agent is a cell differentiation inducer, anticancer agent, immunosuppressant, cell growth factor, cytokine, thrombin inhibitor, antithrombotic agent, thrombolytic agent, fibrinolytic agent, vasospasm inhibitor, calcium channel Blocking agents, vasodilators, antihypertensive agents, antibacterial agents, antibiotics, surface glycoprotein receptor inhibitors, antiplatelet agents, cytostatics, microtubule inhibitors, antisecretory agents, actin inhibitors, remodeling inhibitors Drugs, antisense nucleotides, antimetabolites, antiproliferative substances, anticancer chemotherapeutic drugs, anti-inflammatory steroids or non-steroidal anti-inflammatory drugs, immunosuppressants, growth hormone antagonists, dopamine agonists, radiotherapy drugs, Peptides, proteins, enzymes, extracellular matrix components, inhibitors, free radical scavengers, chelators, antioxidants, antipolymers , Antiviral agents, photodynamic therapy agents, and one or a combination of two or more thereof, characterized in (10) tissue adhesive porous membrane according to selected from gene therapy.

(12) A step of dissolving hydrophobized gelatin in a water-soluble organic solvent to prepare a hydrophobized gelatin-containing solution, and adding a cross-linking agent or a condensing agent to the hydrophobized gelatin-containing solution to cross-link the hydrophobized gelatin to make water Forming a water-soluble organic solvent-containing hydrophobized gelatin cross-linked body, placing the water-soluble organic solvent-containing hydrophobized gelatin cross-linked body in water, substituting the water-soluble organic solvent with water, and water-containing hydrophobized gelatin cross-linked body And a step of lyophilizing the water-containing hydrophobized gelatin cross-linked body to form a tissue-adhesive porous membrane. A method for producing a tissue-adhesive porous membrane comprising the steps of:

(13) A step of dissolving hydrophobized gelatin in a water-soluble organic solvent to prepare a hydrophobized gelatin-containing solution, and dispersing a porogen in the hydrophobized gelatin-containing solution to prepare a porogen-dispersed hydrophobized gelatin-containing solution A step of adding a cross-linking agent or a condensing agent to the porogen-dispersed hydrophobized gelatin-containing solution to cross-link the hydrophobized gelatin to form a hydrophobized gelatin cross-linked body containing a water-soluble organic solvent and a porogen; Water-containing hydrophobized gelatin in which porogen pores are formed by placing a hydrophobized gelatin cross-linker containing a water-soluble organic solvent and a porogen into water, replacing the water-soluble organic solvent with water, and dissolving and removing the porogen A step of forming a crosslinked body, and a step of lyophilizing the water-containing hydrophobized gelatin crosslinked body in which the porogen pores are formed to form a tissue-adhesive porous membrane. Method for producing a tissue adhesive porous membrane characterized by and.

(14) The method for producing a tissue-adhesive porous membrane according to (13), wherein the porogen is particles made of any material of NaCl, KCl, MgCl, ice, and polyethylene glycol.
(15) The method for producing a tissue-adhesive porous membrane according to (12) or (13), wherein the water-soluble organic solvent is dimethyl sulfoxide.
(16) A tissue-adhesive porous membrane tape, characterized in that the tissue-adhesive porous membrane according to any one of (1) to (11) is formed into a strip shape.

The tissue-adhesive porous membrane of the present invention is a membrane in which hydrophobic gelatin is crosslinked and has a porous structure, wherein the hydrophobic gelatin has a portion of the amino group of Lys substituted with a hydrophobic functional group. Since the structure is gelatin, the hydrophobic functional group introduced into the gelatin simultaneously with the absorption of moisture contained in the tissue can be driven into the tissue and cell membrane (anchoring) to form a physically strong bond, The adhesive strength can be increased even in a humid environment. Moreover, since it has a physical porous structure provided using porogen as a template, it can increase the adhesive strength by more effectively absorbing the moisture contained in the tissue. Furthermore, after application to a living tissue, the porous structure is a scaffold for tissue regeneration. In addition, since the gelatin is easily degraded by an enzyme (collagenase) in the wound healing process, the biocompatibility of the porous membrane can be increased.

The method for producing a tissue-adhesive porous membrane of the present invention comprises a step of preparing a hydrophobized gelatin-containing solution by dissolving hydrophobized gelatin in a water-soluble organic solvent, and a crosslinking agent or a condensing agent in the hydrophobized gelatin-containing solution. And forming a water-soluble organic solvent-containing hydrophobized gelatin cross-linked body by cross-linking the hydrophobized gelatin, and placing the water-soluble organic solvent-containing hydrophobized gelatin cross-linked body in water, And a step of forming a water-containing hydrophobized gelatin cross-linked body and a step of lyophilizing the water-containing hydrophobized gelatin cross-linked body to form a tissue-adhesive porous membrane. However, it is possible to easily produce a tissue-adhesive porous membrane having high adhesive strength and appropriate strength.

The method for producing a tissue-adhesive porous membrane of the present invention comprises a step of dissolving a hydrophobic gelatin in a water-soluble organic solvent to prepare a hydrophobic gelatin-containing solution, and dispersing a porogen in the hydrophobic gelatin-containing solution. A step of preparing a porogen-dispersed hydrophobized gelatin-containing solution, and a hydrophobizing agent containing a water-soluble organic solvent and a porogen by cross-linking the hydrophobized gelatin by adding a cross-linking agent or a condensing agent to the porogen-dispersed hydrophobized gelatin-containing solution. A step of forming a gelatin cross-linked product, and the hydrophobized gelatin cross-linked product containing the water-soluble organic solvent and porogen is placed in water, the water-soluble organic solvent is replaced with water, and the porogen is dissolved and removed. A step of forming a water-containing hydrophobized gelatin cross-linked body having porogen pores formed thereon, and a tissue adhesive property by freeze-drying the water-containing hydrophobized gelatin cross-linked body having porogen pores formed therein Since the structure has a step of forming a porous membrane, it has a physical porous structure provided with porogen as a template, so it can increase the adhesive strength by absorbing moisture contained in the tissue, and is wet A tissue-adhesive porous membrane having high adhesive strength and moderate strength can be easily produced even under an environment.

Since the tissue-adhesive porous membrane tape of the present invention has a structure in which the tissue-adhesive porous membrane described above is formed in a strip shape, the cut portion of the intestine can be easily wound, and the tissue adhesive can be obtained by leaving it as it is. Can be made.

It is the schematic which shows the structure | tissue adhesive porous membrane which is the 1st Embodiment of this invention. It is the schematic which shows adhesion | attachment of the tissue adhesive porous membrane at the time of applying the tissue adhesive porous membrane which is the 1st Embodiment of this invention to a biological tissue. It is process drawing explaining an example of the manufacturing method of the structure | tissue adhesive porous membrane which is the 1st Embodiment of this invention. It is the schematic which shows the structure | tissue adhesive porous membrane which is the 2nd Embodiment of this invention. It is the schematic which shows adhesion | attachment of the tissue adhesive porous membrane at the time of applying the tissue adhesive porous membrane which is the 2nd Embodiment of this invention to a biological tissue. It is process drawing explaining an example of the manufacturing method of the structure | tissue adhesive porous membrane which is the 2nd Embodiment of this invention. It is the schematic of the adhesion test of a tissue adhesive porous membrane and porcine large intestine epidermis. 2 is an electron micrograph of a cross section of the tissue adhesive porous membrane of Example 1. FIG. 4 is an electron micrograph of the surface of the tissue adhesive porous membrane of Example 2. FIG. 4 is an electron micrograph of the surface of the tissue adhesive porous membrane of Example 3. FIG. 4 is an electron micrograph of the surface of the tissue-adhesive porous membrane of Example 4. 6 is an electron micrograph of the surface of a tissue adhesive porous membrane of Example 5. FIG. 4 is an electron micrograph of the surface of the tissue adhesive porous membrane of Example 6. FIG. 4 is an electron micrograph of a cross section of a porous film of Comparative Example 1. 4 is an electron micrograph of the surface of a porous film of Comparative Example 2. 4 is an electron micrograph of the surface of a porous film in Comparative Example 3. 4 is an electron micrograph of the surface of a porous film in Comparative Example 4. 6 is an electron micrograph of the surface of a porous membrane of Comparative Example 5. 6 is an electron micrograph of the surface of a porous membrane in Comparative Example 6. 2 is an electron micrograph of a cross section of a tissue adhesive porous membrane of Examples 13 to 17. 2 is an electron micrograph of the surface of a tissue-adhesive porous membrane of Examples 13 to 17. 6 is a graph showing the results of adhesion tests between the tissue adhesive porous membranes of Examples 7 to 12 and the porous membrane of Comparative Example 11 and the porcine large intestine epidermis with respect to the chain length of the hydrophobic functional group in the hydrophobized gelatin. FIG. 7 is a graph showing the results of an adhesion test between the tissue adhesive porous membranes of Examples 13 to 17 and the porous membranes of Comparative Examples 7 to 11 and the porcine large intestine epidermis with respect to the particle size of NaCl, which is a porogen. 6 is a graph showing the results of water absorption rate tests of the tissue adhesive porous membranes of Examples 13 to 17 and the porous membranes of Comparative Examples 7 to 11. 6 is a graph showing the results of L929 cell culture tests of the tissue adhesive porous membranes of Examples 1 to 6 and the porous membranes of Comparative Examples 1 to 6. 2 is a photomicrograph showing the extension of L929 cells on the tissue adhesive porous membranes of Examples 1 to 6 and the porous membranes of Comparative Examples 1 to 6. It is a graph which shows the result of angiogenesis evaluation using the tissue adhesive porous membrane of Examples 2 and 5 and the porous membrane of Comparative Examples 2 and 5.

(Embodiment of the present invention)
Hereinafter, a tissue-adhesive porous membrane, a production method thereof, and a tissue-adhesive porous membrane tape that are embodiments of the present invention will be described with reference to the accompanying drawings.

(First embodiment of the present invention)
<Tissue-adhesive porous membrane>
First, the tissue adhesive porous membrane which is the 1st Embodiment of this invention is demonstrated.
FIG. 1 is a schematic view showing a tissue-adhesive porous membrane according to the first embodiment of the present invention.
As shown in FIG. 1, the tissue adhesive porous membrane 1 is a membrane having a porous structure in which a hydrophobic gelatin 10 is cross-linked and is composed of a plurality of holes 10c1. The tissue adhesive porous membrane 1 is formed by integrating only the hydrophobic gelatin 10 into a film shape.
The shape of the tissue adhesive porous membrane 1 is, for example, a substantially rectangular shape in plan view. One side and the other side of the film are each flat. The size of the tissue adhesive porous membrane 1 is not particularly limited. For example, the tissue adhesive porous membrane 1 may be a membrane having a side length of 8 mm and a thickness of 100 μm.
However, the shape of the tissue adhesive porous membrane 1 is not limited to this, and may be a substantially circular shape in plan view or a substantially elliptical shape in plan view.

The hydrophobized gelatin 10 has a main chain 13 made of gelatin, an amino group 12 and a hydrophobic functional group 11 on its side chain. By using the gelatin skeleton, it can be easily decomposed by an enzyme in the wound healing process, and the biocompatibility of the porous membrane can be increased.
The hydrophobized gelatin 10 is gelatin in which a part of the amino group 12 of Lys contained as an amino acid is substituted with a hydrophobic functional group 11.

Lys is one of the α-amino acids constituting the protein and is an essential amino acid. Lys is an amino acid having an ε-amino group in the side chain.
A part of the amino group of Lys can be easily substituted with the hydrophobic functional group 11 by a known method. In this embodiment, a part of the Lys amino group 12 of the hydrophobized gelatin 10 is substituted with the hydrophobic functional group 11 using a generally known method.

Examples of gelatin to be the main chain 13 include human, porcine, bovine, tilapia, cod-derived gelatin and genetically modified gelatin. One or a combination of two or more of these gelatins may be used.

The molecular weight of the hydrophobized gelatin 10 is preferably more than 50,000 and less than 100,000, more preferably more than 60,000 and less than 100,000, and still more preferably more than 70,000 and less than 100,000. By making the molecular weight of the hydrophobized gelatin 10 within this range, the strength of the porous membrane can be maintained to a level that can be practically used, and the degree of freedom of movement of the main chain made of gelatin is increased, and the hydrophobicity described later The degree of freedom of anchoring of the functional group 11 to the tissue can be increased, and the adhesion strength of the porous membrane to the tissue can be improved.
When the molecular weight of the hydrophobized gelatin 10 is 50000 or less, the strength as a porous film is low and it is difficult to maintain the shape.

The tissue adhesive porous membrane 1 has a porous structure composed of a plurality of holes 10c1. The shape of the hole 10c1 is not particularly limited, and includes holes having various shapes. The pore diameter is preferably 30 μm or more and 500 μm or less, and more preferably 50 μm or more and 100 μm or less in order to efficiently absorb moisture contained in the tissue. The porous structure is composed of gap holes 10 c 1 between the plurality of hydrophobic gelatins 10 provided in the crosslinked structure of the hydrophobic gelatin 10.

In this embodiment, it is preferable that the hydrophobized gelatin 10 is crosslinked by a crosslinking agent or a condensing agent.
Cross-linking agents or condensing agents include trisuccinimidyl citrate, disuccinimidyl tartrate, disuccinimidyl malate, disuccinimidyl succinate, glutaraldehyde, formaldehyde, polyethylene glycol diglycidyl ether, genipin, water-soluble One type or a combination of two or more types of carbodiimides can be mentioned.

As the hydrophobic functional group 11, an ethyl group (2 carbon atoms), a propyl group (3 carbon atoms), a butyl group (4 carbon atoms), a pentyl group (5 carbon atoms), a hexanoyl group (6 carbon atoms), which are saturated fatty acids. ), Heptanoyl group (carbon number 7), octanoyl group (carbon number 8), nonanoyl group (carbon number 9), decanoyl group (carbon number 10), undecanoyl group (carbon number 11), dodecanoyl group (carbon number 12), Tridecanoyl group (carbon number 13), tetradecanoyl group (carbon number 14), pentadecanoyl group (carbon number 15), hexadecanoyl group (carbon number 16), heptadecanoyl group (carbon number 17), stearoyl group (carbon) 18), isopropyl group (carbon number 3), isobutyl group (carbon number 4), isopentyl group (carbon number 5), isohexanoyl group (carbon number 6) which are branched saturated fatty acids , Isoheptanoyl group (carbon number 7), isooctanoyl group (carbon number 8), isononanoyl group (carbon number 9), isodecanoyl group (carbon number 10), isoundecanoyl group (carbon number 11), isododecanoyl group (carbon number 12) ), Isotridecanoyl group (carbon number 13), isotetradecanoyl group (carbon number 14), isopentadecanoyl group (carbon number 15), isohexadecanoyl group (carbon number 16), isopalmityl group (carbon) 16), isoheptadecanoyl group (17 carbon atoms), isostearoyl group (18 carbon atoms), oleyl group (18 carbon atoms, 1 unsaturated carbon) which is an unsaturated fatty acid, linolenyl group (18 carbon atoms, 2 or more unsaturated carbon atoms), α-linolenyl group (18 carbon atoms, 3 unsaturated carbon atoms), one or a combination of two or more cholesteryl groups that are cell membrane components Can be mentioned.

In the present embodiment, the hydrophobized gelatin 10 is preferably cross-linked by thermal cross-linking 14. When the hydrophobized gelatin 10 is thermally cross-linked, peptide bonds that link the linear polymers are formed in the hydrophobized gelatin 10. Thereby, it can be insolubilized in water, the strength of the membrane is increased, and a tissue-adhesive porous membrane having high adhesive strength and appropriate strength even in a wet environment can be obtained.

When the tissue-adhesive porous membrane 1 is affixed to a living tissue, the hydrophobic functional group 11 introduced into the hydrophobic gelatin 10 anchors to the hydrophobic domain of the extracellular matrix and the lipid bilayer of the cell contained in the tissue. By doing so, the tissue adhesive porous membrane 1 is firmly fixed to the tissue.
Moreover, since the tissue adhesive porous membrane 1 has a porous structure, it can absorb moisture in the living tissue. Thereby, the water | moisture content is removed in the adhesive surface of the tissue adhesive porous membrane 1 and a biological tissue, and the adhesion | attachment to the structure | tissue of the tissue adhesive porous membrane 1 becomes stronger.
When the chain length of the hydrophobic functional group 11 is short (1 or less carbon atoms), the hydrophobicity of the hydrophobic gelatin 10 is low, so that the anchoring effect of the hydrophobic functional group 11 cannot be sufficiently obtained, and the tissue adhesive porous The membrane 1 cannot be firmly fixed to the tissue.

<Adhesion to tissue using tissue-adhesive porous membrane of this embodiment>
Next, the adhesiveness to the tissue using the tissue adhesive porous membrane of the first embodiment of the present invention will be described.
FIG. 2 is a schematic view showing adhesion of the tissue adhesive porous membrane when the tissue adhesive porous membrane according to the first embodiment of the present invention is applied to a living tissue.
As shown in FIG. 2, the hydrophobic functional group 11 having a certain molecular weight and size pierces the tissue 21 due to hydrophobic interaction (this is called anchoring). Thereby, the tissue adhesive porous membrane 1 is firmly fixed to the tissue 21.
Moreover, the water | moisture content of a biological tissue is absorbed in the hole 10c of the tissue adhesive porous membrane 1 which has a porous structure, and the water | moisture content of the adhesion | attachment site | part of the tissue adhesive porous membrane 1 and the structure | tissue 21 is removed. . Thereby, the tissue adhesive porous membrane 1 can be physically and firmly adhered to the tissue 21.
There are also hydrophobic functional groups 11 that are not anchored.

When two tissues are bonded using both surfaces of the tissue adhesive porous membrane 1, the bonding operation is performed by attaching one surface of the tissue adhesive porous membrane 1 to one surface of the tissue 21 and then bonding the tissue. It is only necessary to attach another structure to the other surface of the porous porous membrane 1 and leave it at room temperature.
The standing time is a time sufficient for the tissue-adhesive porous membrane 1 to absorb moisture in the tissue 21, and the kind of gelatin that becomes the main chain 13 of the hydrophobic gelatin 10 in the tissue-adhesive porous membrane 1 Depending on the type of the hydrophobic functional group 11, etc. For example, it can be about 3 minutes.

<The manufacturing method of the structure | tissue adhesive porous membrane which is the 1st Embodiment of this invention>
Next, the manufacturing method of the tissue adhesive porous membrane which is the 1st Embodiment of this invention is demonstrated.
FIG. 3 is a process diagram for explaining an example of a method for producing a tissue-adhesive porous membrane according to the first embodiment of the present invention.
The method for producing a tissue adhesive porous membrane according to the first embodiment of the present invention includes a hydrophobized gelatin-containing solution preparation step S1, a water-soluble organic solvent-containing hydrophobized gelatin crosslinked body forming step S2, and a water-containing hydrophobizing step. It has gelatin crosslinked body formation process S3 and structure | tissue adhesive porous film formation process S4. The hydrophobized gelatin is preferably prepared by the following hydrophobized gelatin synthesis step S21.

(Hydrophobic gelatin synthesis step S21)
In the hydrophobized gelatin synthesis step S21, an organic molecule having a hydrophobic functional group is added to a solution in which gelatin is dissolved in the presence of triethylamine, and a part of the amino group of the side chain of the gelatin is substituted with the hydrophobic functional group. This is the step of synthesizing hydrophobic gelatin.
As the gelatin, a gelatin having a molecular weight of hydrophobized gelatin of more than 50,000 and not more than 100,000 is selected.

First, an organic molecule having a hydrophobic functional group having reactivity with an amino group is mixed with gelatin dissolved in an organic solvent in the presence of triethylamine to prepare a mixed solution in a container.
For example, dimethyl sulfoxide (DMSO) is used as the organic solvent.
Examples of the organic molecule having a hydrophobic functional group having reactivity with an amino group include hexanoyl chloride represented by the following chemical formula (1).

Next, the mixed solution is heated and stirred in an inert gas atmosphere. For example, under a nitrogen atmosphere, the heating temperature is 80 ° C., and the stirring time is day and night.
Next, this mixed solution is dropped into an ice-cooled ethanol solvent. Next, this solution is filtered with a glass filter or the like.
Further, the filtrate is washed with an organic solvent. Thereby, impurities in the filtrate can be removed, and the purity of the hydrophobized gelatin can be improved. As the organic solvent for washing, for example, ethanol is used.
Through the above steps, hydrophobic gelatin in which a part of the amino group of the side chain of gelatin is substituted with a hydrophobic functional group can be synthesized.

(Hydrophobized gelatin-containing solution preparation step S1)
The hydrophobized gelatin is dissolved in a water-soluble organic solvent to prepare a hydrophobized gelatin-containing solution 51 in a container 50 as shown in FIG. Examples of the water-soluble organic solvent include dimethyl sulfoxide (DMSO).

(Water-soluble organic solvent-containing hydrophobized gelatin crosslinked product forming step S2)
The hydrophobized gelatin-containing solution 51 is cross-linked by adding a cross-linking agent or a condensing agent. Thereby, as shown in FIG. 3B, the hydrophobized gelatin 10 is cross-linked to form a water-soluble organic solvent-containing hydrophobized gelatin cross-linked body 53. The water-soluble organic solvent-containing hydrophobized gelatin cross-linked body 53 is obtained by forming the hydrophobized gelatin 10 into a network and contains a water-soluble organic solvent 55. The hydrophobized gelatin 10 is a hydrophobized gelatin molecular chain, and has a main chain made of gelatin and an amino group and a hydrophobic functional group in its side chain.

(Water-containing hydrophobized gelatin crosslinked product forming step S3)
The water-soluble organic solvent-containing hydrophobized gelatin cross-linked body 53 is immersed in a large excess of water. For example, put it in water. As a result, as shown in FIG. 3C, the water-soluble organic solvent 55 of the water-soluble organic solvent-containing hydrophobized gelatin cross-linked body 53 is replaced with water 20 to form a water-containing hydrophobized gelatin cross-linked body 58. At this time, in addition to the water-soluble organic solvent 55, an unreacted crosslinking agent and a crosslinked byproduct are removed.

(Tissue-adhesive porous film forming step S4)
The water-containing hydrophobized gelatin crosslinked body 58 is lyophilized. As a result, as shown in FIG. 3 (d), water molecules are removed, and a porous structure is formed which includes pores 10c1 between the plurality of hydrophobic gelatins 10 provided in the crosslinked structure of the hydrophobic gelatin 10. A tissue adhesive porous membrane 1 is formed.

(Second embodiment of the present invention)
<Tissue-adhesive porous membrane according to the second embodiment of the present invention>
First, the tissue adhesive porous membrane which is the 2nd Embodiment of this invention is demonstrated.
FIG. 4 is a schematic view showing a tissue adhesive porous membrane according to a second embodiment of the present invention.
The tissue adhesive porous membrane 2 according to the second embodiment of the present invention is the tissue adhesive property according to the first embodiment of the present invention except that the porogen hole 10c2 is provided in addition to the void hole 10c1. The configuration is the same as that of the porous membrane 1.

A porogen is a particle that serves as a template for a pore, and examples of the porogen material include NaCl, KCl, MgCl, ice, and polyethylene glycol.
A porogen hole is a hole provided in a crosslinked structure using porogen as a template.
Since the porogen materials exemplified above are water-soluble, for example, when these porogen materials are used, in the production method described later, the water-soluble organic solvent of the hydrophobized gelatin crosslinked product containing the water-soluble organic solvent and the porogen is used. When replacing with water, the porogen dissolves in water, so that the porogen is easily removed from the hydrophobized gelatin crosslinked product. Thereby, the porogen hole 10c2 is formed in the tissue adhesive porous membrane 2.

Since the porogen hole 10c2 is a hole provided using porogen as a template, the porogen hole 10c2 is defined to have substantially the same shape and size as the porogen used. Further, by making the shape and size of each porogen the same, the shape and size of the porogen hole can be made the same. FIG. 4 illustrates a spherical porogen hole. However, the shape of the porogen hole is not limited to this.
The size of the porogen can be appropriately set according to the shape and size of the tissue-adhesive porous membrane to be produced, the type of gelatin serving as the main chain of the hydrophobized gelatin, the type of hydrophobic functional group, and the like. For example, a porogen having a particle size of 50 to 100 μm can be used.

<Adhesion to tissue using tissue-adhesive porous membrane of this embodiment>
Next, the adhesiveness to the tissue using the tissue adhesive porous membrane of the second embodiment of the present invention will be described.
FIG. 5 is a schematic view showing adhesion of the tissue adhesive porous membrane when the tissue adhesive porous membrane according to the second embodiment of the present invention is applied to a living tissue.
The tissue adhesive porous membrane 2 has the same configuration as that of the tissue adhesive porous membrane 1 according to the first embodiment of the present invention except that the porogen hole 10c2 is provided.
The tissue-adhesive porous membrane 2 absorbs moisture in the living tissue more effectively than the tissue-adhesive porous membrane 1 by providing the porogen hole 10c2, and the tissue-adhesive porous membrane 2 The moisture at the bonding site between the membrane 2 and the tissue 21 can be removed more effectively, and the tissue-adhesive porous membrane 2 can be physically and firmly bonded to the tissue 21.

<The manufacturing method of the tissue adhesive porous membrane which is the 2nd Embodiment of this invention>
Next, the manufacturing method of the tissue adhesive porous membrane which is the 2nd Embodiment of this invention is demonstrated.
FIG. 6 is a process diagram for explaining an example of a method for producing a tissue-adhesive porous membrane according to the second embodiment of the present invention.
The method for producing a tissue adhesive porous membrane according to the second embodiment of the present invention includes a hydrophobic gelatin-containing solution preparation step S11, a porogen-dispersed hydrophobic gelatin-containing solution preparation step S12, a water-soluble organic solvent, and a porogen. Hydrophobized gelatin crosslinked body forming step S13, water-containing hydrophobized gelatin crosslinked body forming step S14 in which porogen pores are formed, and tissue-adhesive porous membrane forming step S15.

(Hydrophobized gelatin-containing solution preparation step S11)
Hydrophobized gelatin is dissolved in a water-soluble organic solvent to prepare a hydrophobized gelatin-containing solution 51 in a container 50 as shown in FIG. Examples of the water-soluble organic solvent include dimethyl sulfoxide (DMSO).

(Porogen-dispersed hydrophobized gelatin-containing solution preparation step S12)
Porogen 62 is dispersed in the hydrophobized gelatin-containing solution 51 to prepare a porogen-dispersed hydrophobized gelatin-containing solution 60 in the container 50 as shown in FIG.

(Hydrophobized gelatin cross-linking body forming step S13 containing a water-soluble organic solvent and a porogen)
The porogen dispersed hydrophobized gelatin-containing solution 60 is cross-linked by adding a cross-linking agent or a condensing agent. Thereby, as shown in FIG.6 (c), the hydrophobized gelatin crosslinked body 63 containing a water-soluble organic solvent and a porogen is formed. The hydrophobized gelatin cross-linked body 63 containing a water-soluble organic solvent and a porogen is formed by networking the hydrophobized gelatin 10 in the form of a network, and contains the water-soluble organic solvent 55 and the porogen 62. The hydrophobized gelatin 10 is a hydrophobized gelatin molecular chain, and has a main chain made of gelatin and an amino group and a hydrophobic functional group in its side chain.

(Water-containing hydrophobized gelatin crosslinked product forming step S14 in which porogen pores are formed)
A hydrophobized gelatin cross-linked body 63 containing a water-soluble organic solvent and a porogen is immersed in a large excess of water. For example, put it in water. Thereby, as shown in FIG.6 (d), the water-soluble organic solvent 55 can be substituted by the water 20, and the unreacted crosslinking agent, the bridge | crosslinking by-product, and the porogen 62 can be melt | dissolved and removed. The porogen hole 10c2 is physically provided by dissolving and removing the porogen 62. Thereby, the water-containing hydrophobized gelatin crosslinked body 68 in which the porogen hole 10c2 is formed is formed. The water-containing hydrophobized gelatin cross-linked body 68 in which the porogen hole 10c2 is formed contains water 20 between the hydrophobized gelatin molecular chains of the hydrophobized gelatin 10, and also contains water 20 in the porogen hole 10c2.

(Tissue-adhesive porous film forming step S15)
The water-containing hydrophobized gelatin crosslinked product 68 is lyophilized. Thereby, water is removed from the pores of the crosslinked structure, and the tissue adhesiveness having a porous structure composed of the void holes 10c1 and the porogen holes 10c2 between the plurality of hydrophobic gelatins 10 provided in the crosslinked structure of the hydrophobic gelatin 10 The porous membrane 2 can be obtained.

The tissue adhesive

porous membranes

1 and 2 that are embodiments of the present invention may further contain a drug. Thereby, when used for tissue adhesion, the porous membrane not only firmly adheres to the tissue, but also can be treated with a drug.

Examples of the drug include cell differentiation inducer, anticancer agent, immunosuppressive agent, cell growth factor, cytokine, thrombin inhibitor, antithrombotic agent, thrombolytic agent, fibrinolytic agent, vasospasm inhibitor, calcium channel blocker , Vasodilator, antihypertensive, antibacterial, antibiotic, surface glycoprotein receptor inhibitor, antiplatelet agent, cytostatic, microtubule inhibitor, antisecretory, actin inhibitor, remodeling inhibitor, Antisense nucleotide, antimetabolite, antiproliferative substance, anticancer chemotherapeutic agent, anti-inflammatory steroid or non-steroidal anti-inflammatory agent, immunosuppressant, growth hormone antagonist, dopamine agonist, radiotherapy agent, peptide, Protein, enzyme, extracellular matrix component, inhibitor, free radical scavenger, chelating agent, antioxidant, antipolymer , Antiviral agents, mention may be made of photodynamic therapy, and one or more combinations selected from the gene therapy. By using these drugs, it is possible to appropriately treat the tissue.

<Tissue-adhesive porous membrane tape>
Next, the tissue adhesive porous membrane tape according to an embodiment of the present invention will be described.
The tissue-adhesive porous membrane tape is formed by forming the tissue-adhesive

porous membranes

1 and 2 into a strip shape. Thereby, according to the magnitude | size and shape of the cutting | disconnection part of a structure | tissue, a tape piece can be created and tissue adhesion | attachment can be carried out easily. For example, in the case of an intestine, it can be wound and joined to the cut surface.

Tissue-adhesive

porous membranes

1 and 2 according to an embodiment of the present invention are membranes in which hydrophobic gelatin 10 is crosslinked and has a porous structure, and hydrophobic gelatin 10 is a part of amino group 12 of Lys. Is the gelatin 13 substituted with the hydrophobic functional group 11, so that the hydrophobic functional group 11 introduced into the gelatin simultaneously with the absorption of moisture contained in the tissue is driven (anchored) into the physical structure Strong bond can be formed, and the adhesive strength can be increased even in a wet environment. Moreover, since the tissue adhesive porous membrane 2 has a physical porous structure provided using porogen as a template, the adhesive strength can be increased by more effectively absorbing moisture contained in the tissue. Furthermore, the tissue-adhesive

porous membranes

1 and 2 are characterized in that the porous structure becomes a scaffold for tissue regeneration after application to a living tissue. Moreover, since gelatin 13 is easily decomposed by an enzyme (collagenase) in the wound healing process, the biocompatibility of the tissue adhesive

porous membranes

1 and 2 can be increased.

The tissue-adhesive porous membrane 1 according to an embodiment of the present invention has a structure in which the porous structure is composed of gap holes 10c1 between a plurality of hydrophobic gelatins 10 provided in a crosslinked structure of the hydrophobic gelatin 10. The holes 10c1 can absorb water contained in the tissue, have high adhesive strength even in a moist environment and have an appropriate strength, and can easily enter the tissue and can be used as a scaffold excellent in tissue regeneration.

In the tissue adhesive porous membrane 2 according to an embodiment of the present invention, the porous structure is composed of a void hole 10c1 and a porogen hole 10c2 provided with a porogen as a template in the cross-linked structure. As well as the porogen hole 10c2, the water contained in the tissue can be absorbed more effectively, and can be used as a film having a high adhesive strength and an appropriate strength even in a wet environment, making the invasion of the tissue easier, It can be used as an excellent scaffold for tissue regeneration.

The tissue-adhesive porous membrane 2 according to the embodiment of the present invention has a configuration in which the porogen hole 10c2 is provided using a porogen having a particle size of 50 to 100 μm as a template, so that the moisture absorption rate can be appropriately controlled, and the tissue Can be controlled appropriately, and can be used as a film with high adhesive strength and moderate strength even in a wet environment, and as a scaffold with high adhesive strength and moderate strength and excellent tissue regeneration. it can.

Since the tissue-adhesive

porous membranes

1 and 2 according to the embodiment of the present invention have a configuration in which the hydrophobic gelatin 10 is crosslinked by a crosslinking agent or a condensing agent, it can be insolubilized in water and the strength of the membrane can be increased. A tissue-adhesive porous membrane having high adhesive strength and moderate strength even in a moist environment can be obtained.

In the tissue adhesive

porous membranes

1 and 2 according to the embodiment of the present invention, the crosslinking agent or condensing agent is triscinimimidyl citrate, disuccinimidyl tartrate, disuccinimidyl malate, disuccinimidyls. Since it is composed of one or more of succinate, glutaraldehyde, formaldehyde, polyethylene glycol diglycidyl ether, genipin, and water-soluble carbodiimide, it can be insolubilized in water, increasing the strength of the film, and in a humid environment However, a tissue adhesive porous membrane having high adhesive strength and appropriate strength can be obtained.

The tissue-adhesive

porous membranes

1 and 2 according to an embodiment of the present invention have a structure in which the hydrophobic gelatin 10 is crosslinked by thermal crosslinking 14, so that it can be insolubilized in water, the strength of the membrane is increased, and the wetness is increased. A tissue-adhesive porous membrane having high adhesive strength and moderate strength even in an environment can be obtained.

In the tissue adhesive

porous membranes

1 and 2 according to an embodiment of the present invention, the hydrophobic functional group 11 is an ethyl group (carbon number 2), propyl group (carbon number 3), butyl group (carbon number) which is a saturated fatty acid. 4), pentyl group (5 carbon atoms), hexanoyl group (6 carbon atoms), heptanoyl group (7 carbon atoms), octanoyl group (8 carbon atoms), nonanoyl group (9 carbon atoms), decanoyl group (10 carbon atoms) , Undecanoyl group (carbon number 11), dodecanoyl group (carbon number 12), tridecanoyl group (carbon number 13), tetradecanoyl group (carbon number 14), pentadecanoyl group (carbon number 15), hexadecanoyl group ( Carbon number 16), heptadecanoyl group (carbon number 17), stearoyl group (carbon number 18), branched saturated fatty acid isopropyl group (carbon number 3), isobutyl group (carbon number 4), isopentyl (Carbon number 5), isohexanoyl group (carbon number 6), isoheptanoyl group (carbon number 7), isooctanoyl group (carbon number 8), isononanoyl group (carbon number 9), isodecanoyl group (carbon number 10), isoun Decanoyl group (carbon number 11), isododecanoyl group (carbon number 12), isotridecanoyl group (carbon number 13), isotetradecanoyl group (carbon number 14), isopentadecanoyl group (carbon number 15), Isohexadecanoyl group (16 carbon atoms), isopalmityl group (16 carbon atoms), isoheptadecanoyl group (17 carbon atoms), isostearoyl group (18 carbon atoms), unsaturated fatty acid oleyl group (18 carbon atoms) , Unsaturated carbon 1), linolenyl group (18 carbon atoms, 2 unsaturated carbons), α-linolenyl group (18 carbon atoms, 3 unsaturated carbons), cell membrane component co Since the structure is a combination of one type or two or more types of lesteryl groups, the hydrophobic functional group 11 can be driven into the tissue (anchored) to form a physically strong bond to increase the adhesive strength. it can.

In the tissue adhesive

porous membranes

1 and 2 according to an embodiment of the present invention, the gelatin 13 is one or a combination of two or more of gelatin derived from human, pig, cow, tilapia and cod or genetically modified gelatin. Because of the structure, the strength of the film can be increased.

Since the tissue-adhesive

porous membranes

1 and 2 according to the embodiment of the present invention include a drug, the therapeutic effect on the tissue can be enhanced.

In the tissue adhesive

porous membranes

1 and 2 according to the embodiment of the present invention, the drug is a cell differentiation inducer, anticancer agent, immunosuppressant, cell growth factor, cytokine, thrombin inhibitor, antithrombotic agent, thrombus Solubilizer, fibrinolytic agent, vasospasm inhibitor, calcium channel blocker, vasodilator, antihypertensive agent, antibacterial agent, antibiotic, surface glycoprotein receptor inhibitor, antiplatelet agent, cytostatic agent, micro Vascular inhibitor, antisecretory drug, actin inhibitor, remodeling inhibitor, antisense nucleotide, antimetabolite, antiproliferative substance, anticancer chemotherapeutic agent, anti-inflammatory steroid or non-steroidal anti-inflammatory drug, immunosuppression Drugs, growth hormone antagonists, dopamine agonists, radiation therapy drugs, peptides, proteins, enzymes, extracellular matrix components, inhibitors, free radical scavenges Since the composition is one or a combination of two or more selected from a jar, a chelating agent, an antioxidant, an anti-polymerase, an antiviral agent, a photodynamic therapeutic agent, and a gene therapy agent, the specific therapeutic effect of the tissue on the above-mentioned agent Can be raised appropriately.

The method for producing a tissue-adhesive porous membrane 1 according to an embodiment of the present invention includes a step of preparing hydrophobized gelatin-containing solution 51 by dissolving hydrophobized gelatin in a water-soluble organic solvent, and hydrophobized gelatin-containing solution 51. Adding a crosslinking agent or a condensing agent to form a water-soluble organic solvent-containing hydrophobized gelatin cross-linked body 53 by cross-linking the hydrophobized gelatin; and putting the water-soluble organic solvent-containing hydrophobized gelatin cross-linked body 53 in water; Replacing the water-soluble organic solvent with water to form a water-containing hydrophobized gelatin cross-linked body 58; and lyophilizing the water-containing hydrophobized gelatin cross-linked body 58 to form the tissue adhesive porous membrane 1; Therefore, a porous structure can be easily formed by forming a plurality of pores 10c1 between a plurality of hydrophobized gelatins provided in the cross-linked structure of the hydrophobized gelatin 10, and an adhesive strength can be obtained even in a wet environment. Tissue adhesive porous film having a high appropriate strength can be manufactured easily.

The method for producing a tissue adhesive porous membrane 2 according to an embodiment of the present invention includes a step of preparing a hydrophobized gelatin-containing solution 51 by dissolving hydrophobized gelatin in a water-soluble organic solvent, and a hydrophobized gelatin-containing solution 51. A step of preparing a porogen-dispersed hydrophobized gelatin-containing solution 60 by dispersing porogen 62 in the solution; and a cross-linking agent or a condensing agent in the porogen-dispersed hydrophobized gelatin-containing solution 60 to cross-link the hydrophobized gelatin to form a water-soluble organic solvent A step of forming a hydrophobized gelatin cross-linked body 63 containing water and a porogen, and the hydrophobized gelatin cross-linked body 63 containing a water-soluble organic solvent and a porogen is placed in water, and the water-soluble organic solvent is replaced with water. And a step of dissolving and removing the porogen to form a water-containing hydrophobized gelatin cross-linked body 68 in which porogen pores are formed, and a water-containing hydrophobizing gelatin in which porogen pores are formed. And the step of forming the tissue-adhesive porous membrane 2 by freeze-drying the crosslinked body 68, so that not only the pores 10c1 but also the porogen pores 10c2 can contain more water contained in the tissue. A tissue-adhesive porous membrane that absorbs effectively and has high adhesive strength and moderate strength even in a moist environment can be easily produced.

In the method for producing a tissue adhesive porous membrane 2 according to an embodiment of the present invention, the porogen is a particle made of any material of NaCl, KCl, MgCl, ice, and polyethylene glycol. A porogen hole provided as a mold can be easily formed.

The method for producing tissue-adhesive

porous membranes

1 and 2 according to an embodiment of the present invention has a structure in which the water-soluble organic solvent is dimethyl sulfoxide, so that a stable crosslinked structure can be formed and water can be easily replaced. .

Since the tissue adhesive tape according to the embodiment of the present invention has a band-shaped configuration of the tissue adhesive

porous membranes

1 and 2 described above, the cut portion of the intestine can be easily wound and left as it is. Thus, tissue adhesion can be achieved.

The tissue-adhesive porous membrane, the method for producing the tissue-adhesive porous membrane tape and the tissue-adhesive porous membrane tape that are embodiments of the present invention are not limited to the above-described embodiments, It can be changed and implemented. Specific examples of this embodiment are shown in the following examples. However, the present invention is not limited to these examples.

Example 1
(Production of tissue adhesive porous membrane)
<Synthesis of hydrophobized AlGltn and confirmation of introduction rate>
First, alkali-treated gelatin (hereinafter also referred to as AlGltn) was prepared. The alkali-treated gelatin is gelatin obtained by converting asparagine and glutamine present in gelatin into aspartic acid and glutamic acid by deamidation.

Next, according to the method of Matsuda et al. (Non-patent Document 1), hydrophobized AlGltn (Hx: C ₆ ) was introduced by hydrophobizing alkali-treated gelatin (AlGltn) using Hexanoyl chloride. HxAlGltn) was prepared.

Absorbance measurement using TNBS method (DMSO solvent) was performed. This confirmed that the introduction ratio of hexanoyl groups in HxAlGltn was 27%. Hereinafter, this is expressed as 27HxAlGltn.

<Film formation>
In order to enhance the fluidity of the HxAlGltn-containing solution, the 27HxAlGltn was mixed with AlGltn to prepare a relatively 25HxAlGltn (hereinafter referred to as 25 ′ (27) HxAlGltn). When 25 ′ (27) HxAlGltn was dissolved in a 10% lactic acid-DMSO mixed solvent at a content ratio of 25 w / v%, it had fluidity suitable for film formation.
Therefore, a porous membrane was prepared using 25 w / v% 25 ′ (27) HxAlGltn / 10% lactic acid-DMSO.

As a cross-linking agent, trisuccinimidyl citrate (TSC) is used, and 25 w / v% 25 ′ is used so that the molar ratio of the amino group of HxAlGltn to the succinimidyl group contained in TSC is 1: 1. (27) HxAlGltn / 10% lactic acid-DMSO 8 mL and TSC / 10% lactic acid-DMSO 2 mL were sufficiently mixed and stirred with a hand stirrer to obtain a 20 w / w% HxAlGltn solution.
Next, NaCl (particle size 250 to 500 μm: average particle size 375 μm) was used as the porogen, and the HxAlGltn solution and NaCl were mixed so that the weight ratio was 1: 0.
Next, the mixture obtained in Unipack was packed, clipped with a 1 mm silicone sheet on a glass plate, and allowed to stand overnight to crosslink the amino group of HxAlGltn with TSC.

After cross-linking, the obtained gel-like DMSO / NaCl-containing HxAlGltn cross-linked product was released into ultrapure water cooled to 4 ° C. and immersed for 3 days while exchanging the ultrapure water in a large excess amount. At the same time, NaCl, lactic acid, DMSO, and N-hydroxysuccinimide, which is a cross-linking byproduct, were removed.
The obtained water-containing HxAlGltn crosslinked product was frozen at −80 ° C. and then dried under reduced pressure to obtain a tissue-adhesive porous membrane.

(Example 2)
A tissue-adhesive porous membrane was prepared in the same manner as in Example 1 except that the weight ratio of the HxAlGltn solution and NaCl (particle size 250 to 500 μm: average particle size 375 μm) was 1: 1.

Example 3
A tissue-adhesive porous membrane was prepared in the same manner as in Example 1 except that the weight ratio of the HxAlGltn solution and NaCl (particle size 250 to 500 μm: average particle size 375 μm) was 1: 2.

Example 4
A tissue-adhesive porous membrane was prepared in the same manner as in Example 1 except that the weight ratio of the HxAlGltn solution and NaCl (particle size 250 to 500 μm: average particle size 375 μm) was 1: 3.

(Example 5)
A tissue-adhesive porous membrane was prepared in the same manner as in Example 1 except that the weight ratio of the HxAlGltn solution and NaCl (particle size 250 to 500 μm: average particle size 375 μm) was 1: 4.

(Example 6)
A tissue-adhesive porous membrane was prepared in the same manner as in Example 1 except that the weight ratio of the HxAlGltn solution and NaCl (particle size 250 to 500 μm: average particle size 375 μm) was 1: 5.

8 to 13 are electron micrographs of the tissue adhesive porous membranes of Examples 1 to 6. FIG.
From these electron micrographs, it can be seen that a porous structure is formed and that more pores are present as the amount of NaCl mixed at the time of production increases.

(Comparative Example 1)
Using non-hydrophobized AlGltn, 8 mL of 25 w / v% AlGltn / 10% lactic acid-DMSO and 2 mL of TSC / 10% lactic acid-DMSO were thoroughly mixed and stirred with a hand stirrer to prepare a 20 w / w% AlGltn solution. Obtained.
Next, NaCl (particle size: 250 to 500 μm: average particle size: 375 μm) was used as a porogen and mixed so that the weight ratio of the AlGltn solution to NaCl was 1: 0.
Next, the mixture obtained in Unipack was packed, clipped with a 1 mm silicone sheet on a glass plate, and allowed to stand overnight to crosslink the amino group of AlGltn with TSC.

After cross-linking, the obtained gel-like DMSO / NaCl-containing AlGltn cross-linked product is released in ultrapure water cooled to 4 ° C. and immersed for 3 days while exchanging the ultrapure water in a large excess amount. At the same time, NaCl, lactic acid, DMSO, and N-hydroxysuccinimide, which is a cross-linking byproduct, were removed.
The obtained water-containing AlGltn crosslinked product was frozen at −80 ° C. and dried under reduced pressure to obtain a porous membrane made of non-hydrophobized alkali-treated gelatin.

(Comparative Examples 2 to 6)
The weight ratio of the AlGltn solution to NaCl (particle size 250 to 500 μm: average particle size 375 μm) is 1: 1 (Comparative Example 2), 1: 2 (Comparative Example 3), 1: 3 (Comparative Example 4), 1: A porous membrane made of non-hydrophobized alkali-treated gelatin was prepared in the same manner as in Comparative Example 1 except that the ratio was 4 (Comparative Example 5) and 1: 5 (Comparative Example 6).

14 to 19 are electron micrographs of the porous membranes of Comparative Examples 1 to 6.
From these electron micrographs, it can be seen that a porous structure is formed and that more pores are present as the amount of NaCl mixed at the time of production increases.

The properties (porosity and water content, membrane strength, membrane-adhesive porous membrane (Examples 1 to 6) and non-hydrophobized alkali-treated gelatin porous membrane (Comparative Examples 1 to 6) were as follows. Adhesion strength) was measured.

<Porosity and moisture content measurement>
The porosity and moisture content were determined by punching a porous membrane swollen by immersing in water for a predetermined time to φ1 cm, W ₂ : weight of moisture in the pores (the porous membrane after swelling), W ₁ : in the pores Weight when not containing water (porous film before swelling), W ₀ : Measured as weight when dried (porous film dried under vacuum after swelling), and calculated using the following equations (1) and (2) did.

<Membrane strength measurement>
First, a tissue-adhesive porous membrane was molded into 5 mm × 10 mm, and 2.5 mm × 5 mm at both ends were bonded to a 5 mm × 10 mm plastic piece with a cyanoacrylate adhesive.
Next, the plastic piece was sandwiched between texture analyzer jigs and the breaking strength was compared under the condition of 10 mm / min. The average value of three measurements was taken as the measurement value.

<Measurement of adhesive strength between tissue-adhesive porous membrane and porcine large intestine epidermis>
FIG. 7 is a schematic diagram of an adhesion test between a tissue-adhesive porous membrane and porcine large intestine epidermis.
As shown in FIG. 7, first, the tissue-adhesive porous membranes of Examples 1 to 6 punched to a diameter of 7 mm or the porous membranes of Comparative Examples 1 to 6 were placed on a plate set at 37 ° C. It was fixed with cello tape (registered trademark) punched to 4 mm.
Next, the porcine large intestine epidermis, which is a wet biological tissue, was punched into a diameter of 4 mm, and fixed to a jig (probe) with a cyanoacrylate adhesive.
Next, the surface of the porcine large intestine epidermis fixed to the probe and the tissue adhesive porous membrane fixed on the plate were brought into contact with each other, pressurized at 40 g / mm ² for 3 minutes, and then subjected to a tensile test at 10 mm / sec. The experiment was performed three times, and the average value was taken as the measured value.

Table 1 shows the preparation conditions and property evaluation results of a tissue adhesive porous membrane made of hexanoylated alkali-treated gelatin (hereinafter also referred to as “(P) 25 ′ (27) HxAlGltn”).

Table 2 shows the preparation conditions and property evaluation results of a non-hydrophobized alkali-treated gelatin (AlGltn) porous film (hereinafter also referred to as “(P) AlGltn”).

(Characteristic measurement results of tissue adhesive porous membrane)
<Porosity and moisture content measurement results>
From Tables 1 and 2, it was confirmed that the porosity increased with increasing NaCl mixing ratio and increased to about 64%, and the porosity was controlled by the NaCl mixing amount. In addition, it was confirmed that the water content tends to increase slightly as the NaCl mixing ratio or porosity increases.

<About film strength measurement results>
From Tables 1 and 2, the tissue adhesive porous membrane using hexanoylated alkali-treated gelatin tends to have lower membrane strength than the porous membrane using non-hydrophobized alkali-treated gelatin at the same NaCl mixing ratio. It was seen. Further, in the porous membrane made of non-hydrophobized alkali-treated gelatin, the membrane strength decreased as the NaCl mixing amount increased. In addition, if it has film | membrane intensity | strength of 0.1 Mpa or more as a structure | tissue adhesive film, it is thought that it can be used without a problem practically.

<About adhesive strength measurement results>
As can be seen from Tables 1 and 2, the tissue-adhesive porous membrane prepared using hexanoylated alkali-treated gelatin was about 10 times that of the porous membrane prepared using non-hydrophobized alkali-treated gelatin. High adhesive strength.
Moreover, as shown in Table 1, the tissue adhesive porous membrane prepared using hexanoylated alkali-treated gelatin had a maximum adhesion strength to the porcine large intestine with a porosity of about 60%.

(Production of tissue-adhesive porous membranes with various hydrophobic functional groups)
(Example 7)
<Synthesis of hydrophobized AlGltn and confirmation of introduction rate>
Hydrophobized AlGltn (DecAlGltn) into which a decanoyl group (Dec: C ₁₀ ) was introduced was prepared by subjecting AlGltn to a hydrophobic treatment using Decanoyl chloride in the same manner as in Example 1.
Absorbance measurement using the TNBS method (DMSO solvent) confirmed that the decanoyl group introduction rate in DecAlGltn was 25%. Hereinafter, this is expressed as 25DecAlGltn.

<Film formation>
A porous membrane was prepared using 25 w / v% 25DecAlGltn / 10% lactic acid-DMSO.
As a cross-linking agent, triscinnimidyl citrate (TSC) is used, and 25 w / v% 25 DecAlGltn / so that the molar ratio of the amino group of DecAlGltn to the succinimidyl group contained in TSC is 1: 1. 8 mL of 10% lactic acid-DMSO and 2 mL of TSC / 10% lactic acid-DMSO were sufficiently mixed and stirred with a hand stirrer to obtain a 20 w / w% DecAlGltn solution.
Next, NaCl (particle size: 250 to 500 μm: average particle size: 375 μm) was used as a porogen and mixed so that the weight ratio of the DecAlGltn solution to NaCl was 1: 4.
Next, the mixture obtained in Unipack was packed, clipped with a 1 mm silicone sheet on a glass plate, and allowed to stand overnight to crosslink the amino group of DecAlGltn with TSC.

After cross-linking, the obtained gel-like DMSO / NaCl-containing DecAlGltn cross-linked product was released into ultrapure water cooled to 4 ° C. and immersed for 3 days while exchanging the ultrapure water in a large excess amount. At the same time, NaCl, lactic acid, DMSO, and N-hydroxysuccinimide, which is a cross-linking byproduct, were removed.
The obtained water-containing DecAlGltn crosslinked product was frozen at −80 ° C. and then dried under reduced pressure to obtain a tissue-adhesive porous membrane.

(Example 8)
Using Stearoyl chloride, the AlGltn was hydrophobized in the same manner as in Example 7 to prepare a hydrophobized AlGltn (SteAlGltn) into which a stearoyl group (Ste: C ₁₈ ) was introduced.
The introduction rate of stearoyl groups in SteAlGltn was 22%. Hereinafter, this is expressed as 22 SteAlGltn.
A tissue-adhesive porous membrane was prepared in the same manner as in Example 7 using 25 w / v% 22 SteAlGltn / 10% lactic acid-DMSO.

Example 9
Hydrogelation of AlGltn was carried out in the same manner as in Example 7 using Oleyl chloride to prepare hydrophobic AlGltn (OleAlGltn) into which an oleyl group (Ole: C ₁₈ , one unsaturated carbon) was introduced.
The introduction rate of oleyl group in OleAlGltn was 34%. Hereinafter, this is expressed as 34OleAlGltn.
In film formation, in order to enhance the fluidity of the OleAlGltn-containing solution, 34OleAlGltn was mixed with AlGltn to prepare a relatively 25OleAlGltn. Hereinafter, this is expressed as 25 ′ (34) OleAlGltn.
A tissue-adhesive porous membrane was prepared in the same manner as Example 7 using 25 w / v% 25 ′ (34) OleAlGltn / 10% lactic acid-DMSO.

(Example 10)
Hydrochlorination of AlGltn was performed using Cholesteryl chloroform as in Example 7 to prepare hydrophobic AlGltn (CholAlGltn) into which a cholesteryl group (Chol: cell membrane component) was introduced.
The introduction rate of cholesteryl group in CholAlGltn was 27%. Hereinafter, this is expressed as 27CholAlGltn.
In film formation, in order to improve the fluidity of the CholAlGltn-containing solution, 27CholAlGltn was mixed with AlGltn to prepare a relatively 25CholAlGltn. Hereinafter, this is expressed as 25 ′ (27) CholAlGltn.
A tissue-adhesive porous membrane was prepared in the same manner as Example 7 using 25 w / v% 25 ′ (27) CholAlGltn / 10% lactic acid-DMSO.

(Example 11)
Hydrophobized AlGltn (ProAlGltn) into which a propyl group (Pro: C ₃ ) was introduced was prepared by hydrophobizing AlGltn in the same manner as in Example 7 using Propyl Chloride.
The introduction rate of propyl group in ProAlGltn was 30%. Hereinafter, this is expressed as 30ProAlGltn.
In film formation, in order to improve the fluidity of the ProAlGltn-containing solution, 30 ProAlGltn was mixed with AlGltn to prepare a relatively 25ProAlGltn. Hereinafter, this is referred to as 25 ′ (30) ProAlGltn.
A tissue-adhesive porous membrane was prepared in the same manner as in Example 7, using 25 w / v% 25 ′ (30) ProAlGltn / 10% lactic acid-DMSO.

Example 12
Hydroxylized AlGltn (HxAlGltn) into which a hexanoyl group (Hx: C ₆ ) was introduced was prepared by subjecting AlGltn to a hydrophobic treatment using Hexanoyl chloride in the same manner as in Example 7.
The introduction ratio of hexanoyl groups in HxAlGltn was 28%. Hereinafter, this is expressed as 28HxAlGltn.
In film formation, in order to improve the fluidity of the HxAlGltn-containing solution, 28HxAlGltn was mixed with AlGltn to prepare a relatively 25HxAlGltn. Hereinafter, this is expressed as 25 ′ (28) HxAlGltn.
A tissue-adhesive porous membrane was prepared in the same manner as in Example 7, using 25 w / v% of 25 ′ (28) HxAlGltn / 10% lactic acid-DMSO.

(Production of tissue adhesive porous membranes using porogens with various particle sizes)
(Example 13)
<Synthesis of hydrophobized AlGltn and confirmation of introduction rate>
Hydroxylized AlGltn (HxAlGltn) into which a hexanoyl group (Hx: C ₆ ) was introduced was prepared by subjecting AlGltn to a hydrophobic treatment using Hexanoyl chloride in the same manner as in Example 1.
The introduction ratio of hexanoyl groups in HxAlGltn was 34%. Hereinafter, this is expressed as 34HxAlGltn.

<Film formation>
In order to increase the fluidity of the HxAlGltn-containing solution, 34HxAlGltn was mixed with AlGltn to prepare a relatively 25HxAlGltn. Hereinafter, this is expressed as 25 ′ (34) HxAlGltn.
As a cross-linking agent, trisuccinimidyl citrate (TSC) is used, and 25 w / v% 25 ′ is used so that the molar ratio of the amino group of HxAlGltn to the succinimidyl group contained in TSC is 1: 1. (34) HxAlGltn / 10% lactic acid-DMSO (4.5 mL) and TSC / 10% lactic acid-DMSO (0.5 mL) were sufficiently mixed and stirred with a hand stirrer to obtain a 20 w / w% HxAlGltn solution.
Next, NaCl (particle size: 32-53 μm: average particle size: 43 μm) was used as the porogen, and the HxAlGltn solution and NaCl were mixed so that the weight ratio was 1: 4.
Next, the mixture was packed in a unipack, clipped with a 1 mm silicone sheet on a glass plate, and allowed to stand overnight to crosslink the amino group of HxAlGltn with TSC.

(Example 14)
A tissue-adhesive porous membrane was prepared in the same manner as in Example 13 except that NaCl (particle size 53 to 100 μm: average particle size 77 μm) was used as the porogen.

(Example 15)
A tissue-adhesive porous membrane was prepared in the same manner as in Example 13 except that NaCl (particle size: 100 to 212 μm: average particle size: 156 μm) was used as the porogen.

(Example 16)
A tissue-adhesive porous membrane was prepared in the same manner as in Example 13 except that NaCl (particle size 212 to 250 μm: average particle size 231 μm) was used as the porogen.

(Example 17)
A tissue-adhesive porous membrane was prepared in the same manner as in Example 13 except that NaCl (particle size 250 to 500 μm: average particle size 375 μm) was used as the porogen.

FIG. 20 is an electron micrograph of a cross section of the tissue-adhesive porous membrane of Examples 13 to 17. FIG. 21 is an electron micrograph of the surface of the tissue-adhesive porous membrane of Examples 13 to 17.
From these electron micrographs, it can be seen that a porous structure is formed and that the tissue-adhesive porous film has porogen pores corresponding to the particle diameter of NaCl at the time of production.

(Comparative Example 7)
Using non-hydrophobized AlGltn, 4.5 w / v% AlGltn / 10% lactic acid-DMSO (4.5 mL) and TSC / 10% lactic acid-DMSO (0.5 mL) were thoroughly mixed and stirred with a hand stirrer to give 20 w / w% Of AlGltn was obtained.
Next, NaCl (particle size: 32 to 53 μm: average particle size: 43 μm) was used as a porogen and mixed so that the weight ratio of the AlGltn solution to NaCl was 1: 4.
Next, the mixture obtained in Unipack was packed, clipped with a 1 mm silicone sheet on a glass plate, and allowed to stand overnight to crosslink the amino group of AlGltn with TSC.

After cross-linking, the obtained gel-like DMSO / NaCl-containing AlGltn cross-linked product is released in ultrapure water cooled to 4 ° C. and immersed for 3 days while exchanging the ultrapure water in a large excess amount. At the same time, NaCl, lactic acid, DMSO, and N-hydroxysuccinimide, which is a cross-linking byproduct, were removed.
The obtained water-containing hydrophobized AlGltn crosslinked product was frozen at −80 ° C. and then dried under reduced pressure to obtain a porous membrane made of non-hydrophobized alkali-treated gelatin.

(Comparative Example 8)
A porous membrane made of non-hydrophobized alkali-treated gelatin was prepared in the same manner as in Comparative Example 7, except that NaCl (particle size 53 to 100 μm: average particle size 77 μm) was used as the porogen.

(Comparative Example 9)
A porous membrane made of non-hydrophobized alkali-treated gelatin was prepared in the same manner as in Comparative Example 7 except that NaCl (particle size: 100 to 212 μm: average particle size: 156 μm) was used as the porogen.

(Comparative Example 10)
A porous membrane made of non-hydrophobized alkali-treated gelatin was prepared in the same manner as in Comparative Example 7 except that NaCl (particle size 212 to 250 μm: average particle size 231 μm) was used as the porogen.

(Comparative Example 11)
A porous membrane made of non-hydrophobized alkali-treated gelatin was prepared in the same manner as in Comparative Example 7 except that NaCl (particle size 250 to 500 μm: average particle size 375 μm) was used as the porogen.

<Measurement of adhesive strength between tissue-adhesive porous membrane with various hydrophobic functional groups and porcine colonic epidermis>
First, the tissue adhesive porous membrane of Example 7 to Example 12 punched to a diameter of 7 mm or the porous membrane of Comparative Example 11 was placed on a plate set at 37 ° C., and the cello tape punched to a diameter of 4 mm (registered) Trademark).
Next, the porcine large intestine epidermis was punched out to a diameter of 4 mm and fixed to a jig (probe) with a cyanoacrylate adhesive.
Next, the surface of the porcine large intestine epidermis fixed to the probe is brought into contact with the tissue adhesive porous membrane (or porous membrane) fixed on the plate, pressurized at 500 g for 3 minutes, and then subjected to a tensile test at 10 mm / min. went. The experiment was performed three times, and the average value was taken as the measured value.
The results are shown in FIG.

FIG. 22 shows that the tissue adhesive porous membranes (Examples 7 to 12) made of hydrophobized alkali-treated gelatin into which hydrophobic functional groups have been introduced are treated with non-hydrophobized alkali treated in any of the hydrophobic functional groups. Compared to the porous membrane made of gelatin (Comparative Example 11), the adhesion to the pig large intestine epidermis was at least about 6 times higher.

<Measurement of adhesive strength between tissue-adhesive porous membrane having various porogen pores and porcine large intestine epidermis>
First, the tissue adhesive porous membranes of Examples 13 to 17 punched to a diameter of 7 mm or the porous membranes of Comparative Examples 7 to 11 were placed on a plate set at 37 ° C., and the cellophane tape punched to a diameter of 4 mm (registered) Trademark).
Next, the porcine large intestine epidermis was punched out to a diameter of 4 mm and fixed to a jig (probe) with a cyanoacrylate adhesive.
Next, the surface of the porcine large intestine epidermis fixed to the probe is brought into contact with the tissue adhesive porous membrane (or porous membrane) fixed on the plate, pressurized at 40 g / mm ² for 3 minutes, and then at 10 mm / min. A tensile test was performed. The experiment was performed three times, and the average value was taken as the measured value.
The results are shown in FIG.

As shown in FIG. 23, in the porous membranes made of non-hydrophobized alkali-treated gelatin of Comparative Examples 7 to 11, the adhesive strength to the porcine large intestine epidermis surface decreased as the average particle size of the porogen NaCl increased. On the other hand, in the tissue adhesive porous membranes of Examples 13 to 17, when the NaCl particle size was 50 to 100 μm (average particle size 77 μm), the adhesion strength to the surface of the porcine large intestine epidermis showed a maximum value.
Therefore, in addition to the introduction of hydrophobic functional groups into the amino groups of gelatin, by controlling the pore size in the porous structure, a tissue-adhesive porous membrane having higher adhesion strength to the surface of porcine large intestine epidermis is obtained. It became clear that it was possible.

(Water absorption rate test of tissue adhesive porous membrane with various porogen pores)
Based on the water absorption test method (JIS L 1907: 2010) by the dropping method, the system was reduced as follows, and the tissue adhesive porous membranes of Examples 13 to 17 and the porous membranes of Comparative Examples 7 to 11 were used. The water absorption rate test was conducted.
Specifically, 2 μL of ultrapure water is dropped on each sample, and the side of the sample is photographed with a contact angle measuring device every 100 ms, and the time until the convex surface of the water droplet is no longer observed from the surface of the film is measured. did. The experiment was performed three times, and the average value was taken as the measured value.
The results are shown in FIG.

24, the tissue-adhesive porous membranes of Examples 13 to 17 all had a higher water absorption rate than the non-hydrophobized alkali-treated gelatin porous membranes of Comparative Examples 7 to 11. This is presumed to be due to the change in the polarity of gelatin molecules due to the presence of hexanoyl groups in the tissue adhesive porous membranes of Examples 13 to 17.
In the porous films of Comparative Examples 7 to 11, the water absorption rate showed a simple drop as the average particle diameter of the porogen NaCl increased, whereas the tissue adhesive porous films of Examples 13 to 17 In the membrane, when the particle diameter of NaCl was 50 to 100 μm (average particle diameter 77 μm), the water absorption rate showed a maximum value.
In general, as the pore size decreases, the capillary phenomenon tends to occur and the water absorption rate increases. However, in the tissue adhesive porous membrane of the present invention, the pore size in the porous structure and the hydrophobic functional group in the porous membrane ( For example, a synergistic effect with the distribution of hexanoyl groups) causes a significant increase in the water absorption rate of the tissue-adhesive porous membrane when porogen holes are provided using a porogen having a specific particle size as a template. It is thought that it was done.

(Cell proliferation test of tissue-adhesive porous membrane using L929 cells)
On the tissue adhesive porous membranes of Examples 1-6 and Comparative Examples 1-6, L929 cells were cultured for 24 hours under serum-free conditions.
Thereafter, 10 μL of WST-8 was added on each porous membrane, the absorbance (450 nm) was measured, and the number of cells was counted.
The results are shown in FIGS. 25 and 26.

As shown in FIG. 25, the L929 cells cultured on the tissue-adhesive porous membranes of Examples 1 to 6 had a cell number at any porosity as compared with the case of culturing on the porous membranes of Comparative Examples 1 to 6. Is about 4 times.
This is because the tissue adhesive porous membrane of the present invention has a high affinity with L929 cells due to the hexanoyl group (hydrophobic functional group), and also has a porous structure. This is considered to be due to the improved passability.

In addition, from FIG. 26, the extension was observed in the L929 cells cultured on the tissue adhesive porous membranes of Examples 1 to 6, while the extension was observed in the L929 cells cultured on the porous membranes of Comparative Examples 1 to 6. It can be seen that cannot be seen.
From this result, it was suggested that the interaction between the tissue adhesive porous membrane made of hydrophobized alkali-treated gelatin and the surface of the L929 cell may have occurred.

(Angiogenesis evaluation of tissue adhesive porous membrane by immunostaining of CD34, α-SMA)
After the tissue adhesive porous membranes of Examples 2 and 5 and the porous membranes of Comparative Examples 2 and 5 were implanted subcutaneously in rats for 7 days, the tissue sections containing each porous membrane were subjected to CD34 and α-SMA. Immunostaining was performed using (α smooth muscle actin), and evaluation was performed using Image J. In the evaluation, the total area of the portion immunostained in brown was defined as “Vessel-Area” and the field of view was defined as “Area”.
The results are shown in FIG.

FIG. 27 shows that the tissue-adhesive porous film made of hexanoylated alkali-treated gelatin (25 ′ (27) HxAlGltn) compared with the non-hydrophobized alkali-treated gelatin (AlGltn) porous film ((P) AlGltn). It can be seen that by embedding ((P) HxAlGltn) subcutaneously in rats, angiogenesis is promoted significantly. Furthermore, it is clear that (P) HxAlGltn having a higher porosity further improves the effect of promoting angiogenesis.

Tissue-adhesive porous membrane of the present invention, method for producing the same, and tissue-adhesive porous membrane tape are tissue-adhesive porous membranes having high adhesive strength (adhesive strength) and appropriate strength even in a wet environment, and methods for producing the same In addition, the present invention relates to a tissue adhesive porous membrane tape, and may be used in the manufacturing industry of tissue adhesives, tissue sealants, hemostatic agents, and the like.

DESCRIPTION OF

SYMBOLS

1, 2 ... Tissue-adhesive porous membrane, 10 ... Hydrophobized gelatin (hydrophobized gelatin molecular chain), 10c1 ... Pore pore (pore), 10c2 ... Porogen pore (pore), 11 ... Hydrophobic functional group, 12 ... Amino Group (-NH ₂ ), 13 ... (consisting of gelatin), 14 ... thermal cross-linking (peptide bond), 20 ... water, 21 ... biological tissue (tissue), 50 ... container, 51 ... hydrophobized gelatin-containing solution, 53 ... water-soluble organic solvent-containing hydrophobized gelatin cross-linked product, 55 ... water-soluble organic solvent, 58 ... water-containing hydrophobized gelatin cross-linked product, 60 ... porogen-dispersed hydrophobized gelatin-containing solution, 62 ... porogen, 63 ... water-soluble organic solvent And a hydrophobized gelatin cross-linked product containing porogen, 68... Water-containing hydrophobized gelatin cross-linked product in which porogen pores are formed.

Claims

A membrane in which hydrophobic gelatin is crosslinked and has a porous structure,
The tissue-adhesive porous membrane, wherein the hydrophobized gelatin is gelatin in which a part of the amino group of Lys is substituted with a hydrophobic functional group.
2. The tissue-adhesive porous membrane according to claim 1, wherein the porous structure comprises voids between a plurality of hydrophobic gelatins provided in the crosslinked structure of the hydrophobic gelatin.
The porous structure is composed of pores between a plurality of hydrophobic gelatins provided in the crosslinked structure of the hydrophobic gelatin, and porogen holes provided in the crosslinked structure using a porogen as a template. Item 2. The tissue-adhesive porous membrane according to Item 1.
4. The tissue-adhesive porous membrane according to claim 3, wherein the porogen pores are provided using a porogen having a particle size of 50 to 100 μm as a template.
The tissue-adhesive porous membrane according to any one of claims 1 to 3, wherein the hydrophobized gelatin is crosslinked with a crosslinking agent or a condensing agent.
The cross-linking agent or condensing agent is trisuccinimidyl citrate, disuccinimidyl tartrate, disuccinimidyl malate, disuccinimidyl succinate, glutaraldehyde, formaldehyde, polyethylene glycol diglycidyl ether, genipin, water-soluble The tissue-adhesive porous membrane according to claim 5, which is one type or a combination of two or more types of carbodiimides.
The tissue-adhesive porous membrane according to any one of claims 1 to 3, wherein the hydrophobized gelatin is crosslinked by thermal crosslinking.
The hydrophobic functional group is a saturated fatty acid ethyl group (2 carbon atoms), propyl group (3 carbon atoms), butyl group (4 carbon atoms), pentyl group (5 carbon atoms), hexanoyl group (6 carbon atoms). , Heptanoyl group (carbon number 7), octanoyl group (carbon number 8), nonanoyl group (carbon number 9), decanoyl group (carbon number 10), undecanoyl group (carbon number 11), dodecanoyl group (carbon number 12), tridecanoyl Group (carbon number 13), tetradecanoyl group (carbon number 14), pentadecanoyl group (carbon number 15), hexadecanoyl group (carbon number 16), heptadecanoyl group (carbon number 17), stearoyl group (carbon number) 18) isopropyl group (carbon number 3), isobutyl group (carbon number 4), isopentyl group (carbon number 5), isohexanoyl group (carbon number 6), i.e., branched saturated fatty acid Heptanoyl group (carbon number 7), isooctanoyl group (carbon number 8), isononanoyl group (carbon number 9), isodecanoyl group (carbon number 10), isoundecanoyl group (carbon number 11), isododecanoyl group (carbon number 12) , Isotridecanoyl group (carbon number 13), isotetradecanoyl group (carbon number 14), isopentadecanoyl group (carbon number 15), isohexadecanoyl group (carbon number 16), isopalmityl group (carbon number) 16), isoheptadecanoyl group (carbon number 17), isostearoyl group (carbon number 18), unsaturated fatty acid oleyl group (carbon number 18, unsaturated carbon 1), linolenyl group (carbon number 18, non-carbon) Saturated carbon 2), α-linolenyl group (18 carbon atoms, 3 unsaturated carbons), cholesteryl group which is a cell membrane component, or a combination of two or more. The tissue-adhesive porous membrane according to claim 1, wherein:
The tissue-adhesive porous membrane according to claim 1, wherein the gelatin is one or a combination of two or more of gelatin derived from human, pig, cow, tilapia, cod or genetically modified gelatin.
The tissue-adhesive porous membrane according to any one of claims 1 to 9, further comprising a drug.
The agent is a cell differentiation inducer, anticancer agent, immunosuppressant, cell growth factor, cytokine, thrombin inhibitor, antithrombotic agent, thrombolytic agent, fibrinolytic agent, vasospasm inhibitor, calcium channel blocker, Vasodilator, Antihypertensive, Antibacterial, Antibiotic, Surface glycoprotein receptor inhibitor, Antiplatelet agent, Cytostatic, Microtubule inhibitor, Antisecretory, Actin inhibitor, Remodeling inhibitor, Anti Sense nucleotides, antimetabolites, antiproliferative substances, anticancer chemotherapeutic drugs, anti-inflammatory steroids or nonsteroidal anti-inflammatory drugs, immunosuppressants, growth hormone antagonists, dopamine agonists, radiotherapeutic drugs, peptides, proteins , Enzyme, extracellular matrix component, inhibitor, free radical scavenger, chelating agent, antioxidant, anti-polymerase, anti-polymer -Virus agents, photodynamic therapy agents, and tissue adhesive porous membrane according to claim 10, characterized in that the one or more combinations selected from the gene therapy.
A step of preparing a hydrophobic gelatin-containing solution by dissolving hydrophobic gelatin in a water-soluble organic solvent, and adding a crosslinking agent or a condensing agent to the hydrophobic gelatin-containing solution to cross-link the hydrophobic gelatin to form a water-soluble organic solvent Forming a water-containing hydrophobized gelatin cross-linked body, and forming the water-containing hydrophobized gelatin cross-linked body by placing the water-soluble organic solvent-containing hydrophobized gelatin cross-linked body in water and substituting the water-soluble organic solvent with water. And a step of lyophilizing the water-containing hydrophobized gelatin cross-linked body to form a tissue-adhesive porous membrane.
Dissolving a hydrophobized gelatin in a water-soluble organic solvent to prepare a hydrophobized gelatin-containing solution, dispersing a porogen in the hydrophobized gelatin-containing solution, and preparing a porogen-dispersed hydrophobized gelatin-containing solution; Adding a cross-linking agent or a condensing agent to the porogen-dispersed hydrophobized gelatin-containing solution to cross-link the hydrophobized gelatin to form a hydrophobized gelatin cross-linked body containing a water-soluble organic solvent and a porogen; and the water-soluble organic solvent Water-containing hydrophobized gelatin cross-linked product in which porogen pores are formed by placing a hydrophobized gelatin cross-linked product containing selenium and porogen in water, replacing the water-soluble organic solvent with water, and dissolving and removing the porogen. And a step of lyophilizing the water-containing hydrophobized gelatin crosslinked body in which the porogen pores are formed to form a tissue adhesive porous membrane. Method for producing a tissue adhesive porous film whose symptoms.
14. The method for producing a tissue-adhesive porous membrane according to claim 13, wherein the porogen is particles made of any material of NaCl, KCl, MgCl, ice, and polyethylene glycol.
14. The method for producing a tissue-adhesive porous membrane according to claim 12 or 13, wherein the water-soluble organic solvent is dimethyl sulfoxide.
A tissue-adhesive porous membrane tape comprising the tissue-adhesive porous membrane according to any one of claims 1 to 11 in a strip shape.