JP2006150072A - Wound dressing - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a wound dressing having small variation (high reproducibility) of wound healing effect. <P>SOLUTION: This wound dressing comprises a single layer film (S) with surface texture parameters of arithmetic mean roughness (Ra) 1-1,000 μm regulated by JIS B0601:2001 "4.2.1 Arithmetic mean deviation of the assessed profile" and mean length (RSm) 10 μm to 5 mm regulated by JIS B0601:2001 "4.3.1 Mean length of the profile elements". The wound dressing preferably has a compound (AM) containing at least one functional group (A) selected from a group composed of secondary amino group (A1), tertiary amino group (A2), ammonio group (A3), phosphatidyl group (A4), and lysophosphatidyl group (A5) and/or polypeptide (XM) with at least one cell-adhering minimum amino acid sequence (X) in one molecule. <P>COPYRIGHT: (C)2006,JPO&NCIPI

Description

  The present invention relates to a wound dressing. More particularly, the present invention relates to a wound dressing that promotes the regeneration of the epidermis of the skin and is particularly effective for healing of skin wounds.

As wound dressings that promote skin regeneration in wound healing, wound dressings comprising a cell-adhesive polypeptide (Patent Document 1) and the like are known.
JP 2004-049921 A

  However, although the above wound dressing may show a high wound healing effect, there is a problem that the effect varies. That is, an object of the present invention is to provide a wound dressing material having a small variation in wound healing effect (high reproducibility).

  The wound dressing of the present invention is characterized by an arithmetic average roughness (Ra) defined in JIS B0601: 2001 “4.2.1 Arithmetic mean height of contour curve” of 1 μm to 1000 μm, and JIS B0601: 2001 “ The point is that it is composed of a monolayer film (S) having a surface property parameter in which the average length (RSm) of the contour curve element defined in “4.3.1 Average length of contour curve element” is 10 μm to 5 mm. To do.

  The wound dressing of the present invention can exhibit an excellent wound healing effect with high reproducibility.

In the present invention, the surface property parameter means an arithmetic average roughness (Ra) and an average length (RSm) of contour curve elements.
The arithmetic average roughness (Ra) is preferably 1 to 1000 μm, more preferably 10 to 500 μm, and particularly preferably 20 to 300 μm, from the viewpoint of variation (reproducibility) of the wound healing effect. Within this range, the reproducibility of the wound healing effect is further improved.
Further, the average length (RSm) of the contour curve element is preferably 10 μm to 5 mm, more preferably 20 μm to 1 mm, and particularly preferably 50 μm to 600 μm from the viewpoint of variation (reproducibility) of the wound healing effect. Within this range, the reproducibility of the wound healing effect is further improved.

  As long as the wound dressing of the present invention is within the range of the above surface property parameters, the surface shape of the single layer film (S) is not limited. For example, on a flat surface, a bulge of a uniform or non-uniform size {hemisphere, flat (elliptical) shape, irregular shape, etc.} and / or a depression {hemispherical, flat (elliptical) shape, irregular shape, etc.} The shape may be scattered uniformly or non-uniformly, or may have a shape or irregular shape of ridges and / or grooves on a flat surface, and there is no flat portion, and the entire surface is uneven {hemispherical, flat (elliptical) ) Shape, irregular shape, etc.}.

基準長さ｛Ｕ｝及び輪郭曲線｛Ｚ（ｘ）｝については単層フィルム（Ｓ）の断面を模式的に表した図１を参照できる。 The arithmetic average roughness (Ra) is defined in “4.2.1 Arithmetic average height of contour curve” of JIS B0601: 2001, and the ordinate {Z ( x)} (the height of the contour curve at an arbitrary position) means the average of absolute values (represented by the following equation).

For the reference length {U} and the contour curve {Z (x)}, reference can be made to FIG. 1 schematically showing the cross section of the single-layer film (S).

基準長さ（Ｕ）及び輪郭曲線要素｛ＸＳｉ｝については、単層フィルム（Ｓ）の断面を模式的に表した図２を参照できる。 The average length (RSm) of the contour curve element is defined in “4.3.1 Average length of the contour curve element” of JIS B0601: 2001, and the contour curve element at the reference length {U} It means the average of the length {XSi} (represented by the following formula).

For the reference length (U) and the contour curve element {XSi}, FIG. 2 schematically showing the cross section of the single layer film (S) can be referred to.

Arithmetic average roughness (Ra) and average length of contour curve element (RSm) can be measured by a known method, for example, a surface shape microscope {surface shape measuring microscope VF-7500 manufactured by Keyence Corporation} and the like An atomic force microscope {atomic force microscope SFA-300 manufactured by Seiko Electronics Co., Ltd.} is used.
The measurement is performed by leaving the measurement sample in an atmosphere of 25 ° C. and a relative humidity of 65% for at least 12 hours and at the same temperature and humidity.

In the present invention, the single layer film (S) means not a film in which two or more films are combined, but a film composed of a single layer.
In addition, you may have a fine hole in the whole surface or a part in a single layer film (S). In the case of having fine pores, the size of the pores is preferably a size that allows air and water vapor to pass therethrough but does not allow liquid or bacteria to pass through. As the size of the hole, the area of the opening is preferably 1 μm ² or less. In addition, the shape of the hole includes a circle, an ellipse, a polygon, an indeterminate shape, and a line shape (slit), and any shape can be used as long as the amount of exudate stored in the wound can be appropriately maintained. May be.

The shape and size of the single layer film (S) are not particularly limited as long as the wound can be covered. For example, a polygon, an ellipse, etc. may be sufficient, and the part can also be cut out and used. In addition, the wound dressing of the present invention may be fixed using a cover with an adhesive or a pressure-sensitive adhesive, a bandage, a stretchable mesh, an eye patch, or the like, and may be sutured or bonded.
The thickness of the monolayer film (S) is not particularly limited as long as the wound dressing material can cover the wound without being displaced or twisted on the wound surface, but preferably 1 μm to 1 cm, more preferably 5 to 3000 μm, Especially preferably, it is 15-500 micrometers.

The material of the single-layer film (S) is not limited as long as it does not adversely affect cells and living bodies, and is a material that is easily dispersed, dissolved or absorbed in body fluids when applied to wounds, etc. Any of the degradable material (S1)}, a material that is difficult to be dispersed, dissolved, or absorbed in a body fluid or the like (hereinafter referred to as a hardly biodegradable material (S2)) when applied to a wound or the like can be used. Among these, the hardly biodegradable biomaterial (S2) is preferable from the viewpoint of ease of peeling of the wound dressing from the wound surface. Moreover, you may combine an easily biodegradable material (S1) and a hardly biodegradable material (S2).
When combining the easily biodegradable material (S1) and the hardly biodegradable material (S2), the combination form and the combination method are not particularly limited, but easily biodegradable on the film surface made of the hardly biodegradable material (S2). Examples include a form in which fine particles made of the functional material (S1) are scattered.

As the readily biodegradable material (S1), a natural polymer (S1A), a synthetic polymer (S1B), an inorganic substance (S1C), and the like can be used.
Natural polymers (S1A) include collagen, gelatin, glycosaminoglycan, hyaluronic acid, chondroitin sulfate, keratan sulfate, dermatan sulfate, heparin, elastin, chitin, chitosan, fibrin, alginic acid, starch, dextran, albumin, polyhydroxy Examples include butyric acid, pectin, pectic acid, galactan, pullulan, agarose, cellulose, gluten, and fibroin.

  As the synthetic polymer (S1B), a monomer selected from the group consisting of lactic acid, leucine, glycolic acid, ε-caprolactone, dioxanone, malic acid, lactide and glycolide is an essential monomer (co) polymer (Polyglycolic acid), and synthetic polypeptides other than the polypeptide (XM) described below.

  Examples of inorganic substances (S1C) include calcium carbonate (light calcium carbonate, heavy calcium carbonate, etc.), calcium phosphate {hydroxyapatite, tricalcium phosphate, and a mixture of these with other calcium phosphates (monocalcium hydrogen phosphate, etc.)} Etc. are included.

  Of these, natural polymer (S1A) and synthetic polymer (S1B) are preferable, more preferably natural polymer (S1A), particularly preferably collagen, gelatin, hyaluronic acid, chitin, chitosan, fibrin, alginic acid, starch, Dextran, agarose, cellulose and fibroin.

As the hardly biodegradable material (S2), a natural polymer (S2A), a synthetic polymer (S2B), an inorganic substance (S2C), and the like can be used.
Examples of the natural polymer (S2A) include natural fibers (cotton, hair, hemp, silk, etc.).

  Synthetic polymer (S2B) includes polyolefin (polyethylene, polypropylene, modified products thereof), olefin copolymer {ethylene-vinyl acetate copolymer, ethylene-ethyl (meth) acrylate copolymer, ethylene-methyl ( Meth) acrylate copolymer, ethylene- (meth) acrylic acid copolymer, etc.}, polyurethane, polyester, polyacrylic acid, polyamide (nylon, etc.), polyvinyl chloride, polyvinylidene chloride, polystyrene, fluororesin, silicone resin, Cellulose, viscose rayon, cupra rayon, polyacetate, polyacrylonitrile, vinylon, vinylidene and the like are used.

  As the inorganic substance (S2C), metal (gold, silver, platinum, titanium, nickel, etc.), ceramics (alumina, zirconia, aluminum nitride, etc.), etc. are used.

  Of these, the synthetic polymer (S2B) and the inorganic substance (S2C) are preferable, the synthetic polymer (S2B) is more preferable, and the polyolefin, polyurethane, polyester, polyacrylic acid, polyamide (nylon), polystyrene, and silicone resin are particularly preferable. Most preferably, it is polyurethane.

The single layer film (S) may contain fine particles.
The fine particles are not limited as long as they do not adversely affect the cells and the living body, and may be inorganic fine particles or organic fine particles. Moreover, the microparticles | fine-particles which consist of an easily biodegradable material (S1) or a hardly biodegradable material (S2) may be sufficient.
As inorganic fine particles, fine particles made of an easily decomposable material (S1) (calcium carbonate, etc.) and a hardly decomposable material (S2) (calcium silicate, magnesium silicate, silica, barium sulfate, zinc oxide, titanium oxide, clay and alumina) Etc.).
Examples of the organic fine particles include fine particles made of an easily decomposable material (S1) (polyglycolic acid, glucosaminoglycan, etc.), and a hardly decomposable material (S2) (polyurethane resin, polyester resin, polyacrylic acid resin, polyamide ( Nylon) resin, polystyrene resin, silicone resin, etc.).
Of these, organic fine particles are preferable, and polystyrene resin is more preferable.

  Although there is no restriction | limiting in the shape of microparticles | fine-particles, For example, spherical shape, needle shape, flat (ellipse) shape, flake shape, amorphous crushing shape, fiber shape, etc. are mentioned. Of these, a spherical shape, a flat (elliptical) shape and a flake shape are preferable, a spherical shape and a flat (elliptical) shape are more preferable, and a spherical shape is particularly preferable.

  Although there is no restriction | limiting in the magnitude | size of microparticles | fine-particles, as for the volume average particle diameter (micrometer) of microparticles | fine-particles, 2-2000 are preferable, More preferably, it is 80-1200, Most preferably, it is 40-600. The volume average particle diameter is measured with a laser type particle size distribution measuring apparatus, for example, LA-920 (dispersion medium: methanol, measurement temperature: 25 ° C.) manufactured by HORIBA, Ltd. in accordance with JIS Z8825-1: 2001. The

  When the fine particles are contained, the content (% by weight) is preferably 30 to 200, more preferably 40 to 150, particularly preferably based on the weight of the single layer film (S) (including the weight of the fine particles). 50-110.

  The wound dressing of the present invention is further selected from the group consisting of secondary amino group (A1), tertiary amino group (A2), ammonio group (A3), phosphatidyl group (A4), and lysophosphatidyl group (A5). It may have a compound (AM) containing at least one functional group (A).

As the secondary amino group (A1), an aliphatic amino group, an aromatic amino group, a group represented by —NHCH ₂ — and a group represented by —C (═NH) — can be used. 1-16 are preferable, as for these carbon numbers, More preferably, it is 1-12, Most preferably, it is 1-6.

  Examples of the aliphatic amino group include methylamino, ethylamino, propylamino, butylamino, cyclohexylamino, 2-ethylhexylamino, dodecylamino, hexadecylamino, benzylamino and 2-phenylethylamino.

Examples of the aromatic amino group include phenylamino (anilino), 4-methylphenylamino (4-methylanilino), naphthylamino and the like.
Among these, an aliphatic amino group, a group represented by —NHCH ₂ — and a group represented by —C (═NH) — are preferred, and more preferably a methylamino group, an ethylamino group, a propylamino group, — A group represented by NHCH ₂ — and a group represented by —C (═NH) —, particularly preferably a group represented by —NHCH ₂ — and a group represented by —C (═NH) —.

Examples of the tertiary amino group (A2) include an aliphatic amino group, an aromatic amino group, a group represented by —N (CH ₂ —OH) CH ₂ — or —N (CH ₃ ) CH ₂ —. It is done. These carbon numbers are preferably 2 to 32, more preferably 2 to 24, and particularly preferably 2 to 12.

  Examples of the aliphatic amino group include dimethylamino, diethylamino, methylpropylamino, diisopropylamino, dibutylamino, dicyclohexylamino, didodecylamino, dihexadecylamino and dibenzylamino.

Examples of the aromatic amino group include diphenylamino and dinaphthylamino.
Among these, an aliphatic amino group, a group represented by —N (CH ₂ —OH) CH ₂ — or —N (CH ₃ ) CH ₂ — is preferable, and dimethylamino, diethylamino, dipropylamino and A group represented by —N (NH ₂ —OH) CH ₂ — or —N (CH ₃ ) CH ₂ —, particularly preferably dimethylamino, diethylamino and —N (CH ₂ —OH) CH ₂ — or —N ( CH ₃ ) is a group represented by CH ₂ —.

The ammonio group (A3) includes an aliphatic ammonio group, an aromatic ammonio group, a group represented by> C = N ⁺ = C <, and a group represented by —CH ₂ N ⁺ (CH ₃ ) ₂ CH ₂ —. Etc. As for these carbon number, 1-48 are preferable, More preferably, it is 2-36, Most preferably, it is 2-18.

  Aliphatic ammonio groups include methylammonio, ethylammonio, propylammonio, butylammonio, cyclohexylammonio, 2-ethylhexylammonio, dodecylammonio, hexadecylammonio, benzylammonio, 2-phenylethyl Ammonio, Dimethylammonio, Diethylammonio, Methylpropylammonio, Diisopropylammonio, Dibutylammonio, Dicyclohexylammonio, Didodecylammonio, Dihexadecylammonio, Dibenzylammonio, Trimethylammonio, Triethylammonio , Methyldiethylammonio, methyldiisopropylammonio, methyldibutylammonio, methyldicyclohexylammonio, methyldidodecylammonio and trihexade Ruanmonio, and the like.

  Examples of the aromatic ammonio group include methylphenylammonio, methylnaphthylammonio, methyldiphenylamino, methyldinaphthylamino, and tribenzylammonio.

Among these, an aliphatic ammonio group, a group represented by> C═N ⁺ = C <, and a group represented by —CH ₂ N ⁺ (CH ₃ ) ₂ CH ₂ — are preferable, and methylammonio is more preferable. , ethyl ammonio, butyl ammonio, trimethylammonio, triethyl ammonio, methyl diethyl ammonio, methyl diisopropyl ammonio, methyl dibutyl Ruan ^{ammonio,> C = N + = C} < group represented by the and -CH ₂ N A group represented by ⁺ (CH ₃ ) ₂ CH ₂ —, particularly preferably represented by methylammonio, ethylammonio, trimethylammonio, triethylammonio, methyldiethylammonio,> C═N ⁺ = C < And a group represented by —CH ₂ N ⁺ (CH ₃ ) ₂ CH ₂ —.

As the phosphatidyl group (A4), a group represented by HOCH ₂ CH (OH) CH ₂ OP (═O) (O ⁻ ) — (unsubstituted phosphatidyl group) or R ¹ OCH ₂ CH (OR ² ) CH ₂ OP A group represented by (═O) (O ⁻ ) — (substituted phosphatidyl group, R ¹ and R ² are the same or different organic groups (acyl group, alkyl group or alkenyl group)) and the like can be used. The organic group (acyl group, alkyl group or alkenyl group) preferably has 1 to 22 carbon atoms, more preferably 2 to 18 carbon atoms, and particularly preferably 12 to 18 carbon atoms.

  Examples of acyl groups include saturated acyl groups (acetyl, butanoyl, hexanoyl, decanoyl, lauroyl, myristoyl, palmitoyl, stearoyl, arachidonoyl, etc.), and unsaturated acyl groups (linoleoyl, octadecenoyl, oleoyl, etc.). .

  Examples of the alkyl group include methyl, hexadecyl, butyl, hexyl, decyl, dodecyl, tetradecyl, hexadecyl, octadecyl and icosanoyl.

Examples of the alkenyl group include linoleyl, octadecadienyl, octadecatrienyl and octadecenyl.
Of these, an unsubstituted phosphatidyl group and a substituted phosphatidyl group (R ¹ and R ² are acyl groups) are preferable, and an unsubstituted phosphatidyl group and a substituted phosphatidyl group (R ¹ and R ² are unsaturated acyl groups) are more preferable. Particularly preferably a substituted phosphatidyl group (R ¹ and R ² are the same unsaturated acyl group).

As the lysophosphatidyl group (A5), a group represented by R ³ OCH ₂ CH (OH) CH ₂ OP (═O) (O ⁻ ) — (R ³ is a saturated or unsaturated acyl group) or the like is used. it can. As for carbon number of an acyl group, 1-22 are preferable, More preferably, it is 2-18, Most preferably, it is 12-18.

As an acyl group, the same thing as the case of a phosphatidyl group (A4) can be used.
Of these, a substituted lysophosphatidyl group in which R ³ is an unsaturated acyl group is preferred.

The weight average molecular weight (MW) of the compound (AM) is preferably 1,000 to 100,000, more preferably 1,200 to 50,000, particularly preferably low in toxicity to cells and high in cell attractiveness. Preferably, it is 1,800-30,000.
The weight average molecular weight (MW) of the compound (AM) can be measured by gel permeation chromatography (GPC) (the reference material is a polystyrene standard having a molecular weight of 420 to 20,600,000 (for example, manufactured by Tosoh Corporation). ).)
The compound (AM) includes at least one primary amine group or secondary amine group polyamine (2 to 56 carbon atoms) described in Patent Document 1 {aliphatic polyamine, alicyclic polyamine, heterocyclic Polyamine, aromatic polyamine} and the like can be used as they are.

  Among the functional groups (A), from the viewpoint of promoting wound healing, the secondary amino group (A1) and the tertiary amino group (A2) are preferable, and a secondary amino group is more preferable. That is, among the compounds (AM), the compound (AM1) containing the secondary amino group (A1) and the compound (AM2) containing the tertiary amino group (A2) are preferable from the viewpoint of high wound healing effect, More preferably (AM1), particularly preferably an aliphatic polyamine (PA) and an aromatic polyamine (PAr). {These compounds are described in detail in Patent Document 1. }.

The aliphatic polyamine (PA) includes a compound having a plurality of alkylene groups and at least one plurality selected from the group consisting of primary to tertiary amino groups and ammonio groups.
The aromatic amine (PAr) includes a compound having a plurality of arylene groups and at least one plurality selected from the group consisting of primary to tertiary amino groups and ammonio groups.
In the aliphatic polyamine (PA), a part of the alkylene group may be replaced with an arylene group. In the aromatic polyamine (PAr), part of the arylene group may be replaced by an alkylene group.

The aliphatic polyamine (PA) and the aromatic polyamine (PAr) may be either linear or branched.
As the linear polyamine, the following chemical formula (R represents an alkylene group or an arylene group which may be the same or different, R ′ represents a hydrogen atom or an alkyl group having 1 to 4 carbon atoms, and n represents a polymer. In which the weight average molecular weight is 1,000 to 100,000.).

  The alkylene group of the aliphatic polyamine (PA) and the aromatic polyamine (PAr) includes an alkylene group having 2 to 6 carbon atoms (ethylene, propylene, isopropylene, butylene, hexylene, etc.) and the like. The arylene group includes an arylene group having 4 to 8 carbon atoms (flangedyl, phenylene, toluenediyl, xylenediyl, etc.).

  Aliphatic polyamines (PA) include alkylene diamines (ethylene diamine, propylene diamine, trimethylene diamine, tetramethylene diamine, hexamethylene diamine, etc.), polyalkylene polyamines having 2 to 6 carbon atoms in the alkylene group (diethylene triamine, iminobis). Propylamine, triethylenetetramine, tetraethylenepentamine, pentaethylenehexamine, etc.), alkyl (carbon number 1-18) substitution products thereof (dimethylaminopropylamine, diethylaminopropylamine, dipropylaminopropylamine, methylethylaminopropyl) Amines, trimethylhexamethylenediamine, N, N-dioctadecylethylenediamine, trioctadecylethylenediamine and methyliminobispropylamine ), Polyethyleneimine, poly (ethyleneimine / N-methylethyleneamine), poly (N-methylethyleneamine), poly (ethyleneimine / N-ethylethyleneamine), polypropyleneimine, polybutyleneimine, poly (ethyleneimine Propyleneimine), poly (ethyleneimine / hexyleneimine) and the like.

As aromatic amine (PAr), phenylenediamine, N, N′-dimethylphenylenediamine, N, N, N′-trimethylphenylenediamine, diphenylmethanediamine, 2,6-diaminopyridine, tolylenediamine, diethyltolylenediamine 4,4′-bis (methylamino) diphenylmethane, 1-methyl-2-methylamino-4-aminobenzene, polyphenyleneimine, poly (phenyleneimine / ethyleneimine), polyfuranylimine, poly (flangylimine Ethyleneimine) and the like.
A compound (AM) can use 1 type (s) or 2 or more types.
Of these aliphatic polyamines (PA) and aromatic polyamines (PAr), (PA) is preferred, and polyethyleneimine is more preferred.

The aliphatic polyamine (PA) and the aromatic polyamine (PAr) can be produced by known methods as shown in the following (1) to (4).
(1) Open an alkyleneimine (ethyleneimine, propyleneimine, etc.) in the presence of a catalyst (carbon dioxide, hydrochloric acid, hydrobromic acid, p-toluenesulfonic acid, methanesulfonic acid, aluminum chloride, boron trifluoride, etc.). A method of ring polymerization.
(2) A method in which a halogenated alkylene and / or a halogenated arylene (for example, ethylene chloride, propylene bromide, etc.) and an alkylene diamine and / or an arylene diamine (for example, ethylene diamine, propylene diamine, etc.) are subjected to a polycondensation reaction.
(3) A method of heating axazolidone-2 or the like.
(4) A method of extracting from a living body.

The wound dressing of the present invention may further have a polypeptide (XM) having at least one cell adhesion minimal amino acid sequence (X) in one molecule.
When the polypeptide (XM) contains the minimal amino acid sequence (X) that adheres to cells, the cell adhesion activity is increased, the adhesion of cells (especially skin cells) is promoted, the adhered cells grow, and wound healing can be promoted. It becomes.

  The content of the minimum amino acid sequence (X) is preferably 1 to 50, more preferably 3 to 30, particularly preferably 4 to 1 molecule of polypeptide (XM) from the viewpoint of cell adhesion / proliferation. 20. Within this range, the cell adhesion activity is further increased, and the proliferation of cells (particularly skin cells) can be further facilitated.

  Any cell adhesion minimal amino acid sequence (X) may be used as long as it functions as an adhesion signal. For example, “Pathophysiology” published by Nagai Publishing Co., Ltd., Vol. 9, no. 7, 1990, page 527, etc. can be used.

  Of these, Arg Gly Asp sequence, Leu Asp Val sequence, Leu Arg Glu sequence, His Ala Val sequence, Arg Glu Asp Val sequence (1), Tyr Ile Gly Ser Arg sequence (2), Pro Asp Ser Gly Arg sequence (3), Arg Tyr Val Val Leu Pro Arg sequence (4), Leu Gly Thr Ile Pro Gly sequence (5), Arg Asn Ile Ala Glu Ile Ile Lys Asp Ile sequence (6), Ile Lys Val Ala Val sequence (7) and Asp Gly Glu Ala sequence (8) are preferred, Arg Gly Asp sequence, His Ala Val sequence and Ile Lys Val Ala Val sequence (7) are particularly preferred. Is Arg Gly Asp sequence.

  The polypeptide (XM) preferably has an auxiliary amino acid sequence (Y) in addition to the cell adhesion minimal amino acid sequence (X) from the viewpoint of improving the thermal stability of the polypeptide (XM).

As the auxiliary amino acid sequence (Y), an amino acid sequence other than the minimum amino acid sequence (X) can be used. From the viewpoint of improving the heat resistance of the polypeptide (XM), a sequence having Gly and / or Ala is preferable.
The auxiliary amino acid sequence (Y) includes (Gly Ala) a sequence, (Gly Ala Gly Ala Gly Ser) b sequence, (Gly Ala Gly Ala Gly Tyr) c sequence, (Gly Ala Gly Val Gly Tyr) d sequence, ( Gly Ala Gly Tyr Gly Val) e sequence, {Asp Gly Gly (Ala) f Gly Gly Ala} g sequence, (Gly Val Pro Gly Val) h sequence, (Gly) i sequence, (Ala) j sequence, (Gly Gly Ala) k sequence, (Gly Val Gly Val Pro) m sequence, (Gly Pro Pro) n sequence, (Gly Ala Gln Gly Pro Ala Gly Pro Gly) o sequence, (Gly Ala Pro Gly Ala Pro Gly Ser Gln Gly Ala Pro Gly Leu Gln) p sequence and / or (Gly Ala Pro Gly Thr Pro Gly Pro Gln Gly Leu Pro Gly Ser Pro) sequence having q sequence and the like are included. Among these, (Gly Ala) a sequence, (Gly Ala Gly Ala Gly Ser) b sequence, (Gly Ala Gly Ala Gly Tyr) c sequence, (Gly Ala Gly Val Gly Tyr) d sequence, (Gly Ala Gly Tyr Gly Val) e, {Asp Gly Gly (Ala) f Gly Gly Ala} g sequence, (Gly Val Pro Gly Val) h sequence, (Gly Val Gly Val Pro) m sequence and / or (Gly Pro Pro) n sequence Those having a (Gly Ala Gly Ala Gly Ser) b sequence, (Gly Val Pro Gly Val) h sequence, (Gly Val Gly Val Pro) m sequence and / or (Gly Pro Pro) n sequence are preferred. Particularly preferred are those having the (Gly Ala Gly Ala Gly Ser) b sequence.
A is an integer of 5 to 100, b, c, d, and e are integers of 2 to 33, f is an integer of 1 to 194, and g is rounded down to the nearest decimal point of {1} to {200 / (6 + f)}. , H is an integer of 2 to 40, i and j are integers of 10 to 200, k is an integer of 3 to 66, m is an integer of 2 to 40, n is an integer of 3 to 66, o is 1 to 22 , P and q are integers of 1-13.

The auxiliary amino acid sequence (Y) preferably contains glycine (Gly) and / or alanine (Ala). When glycine (Gly) and alanine (Ala) are included, the total content (%) thereof is preferably 10 to 100, more preferably 20 to 95, based on the total number of amino acids in the auxiliary amino acid sequence (Y). Especially preferably, it is 30-90, Most preferably, it is 40-85. Within this range, the heat resistance is further improved.
When both glycine (Gly) and alanine (Ala) are included, the content ratio (Gly / Ala) is preferably 0.03 to 40, more preferably 0.08 to 13, and particularly preferably 0.2. ~ 5. Within this range, the heat resistance is further improved.

  When the auxiliary amino acid sequence (Y) is included, the content of (Y) is preferably 2 to 51, more preferably 3 to 35, particularly 1 to 35 molecules in the polypeptide (XM) molecule from the viewpoint of improving heat resistance. Preferably it is 4-20. In addition, the polypeptide (XM) may contain two or more auxiliary amino acid sequences (Y).

Examples of the auxiliary amino acid sequence having the (Gly Ala) a sequence include amino acid sequences represented by SEQ ID NOs: (9) to (11).
Examples of the auxiliary amino acid sequence having (Gly Ala Gly Ala Gly Ser) b sequence include amino acid sequences represented by SEQ ID NOs: (12) to (14).
Examples of the auxiliary amino acid sequence having (Gly Ala Gly Ala Gly Tyr) c sequence include amino acid sequences represented by SEQ ID NOs: (15) to (17).
Examples of the auxiliary amino acid sequence having the (Gly Ala Gly Val Gly Tyr) d sequence include amino acid sequences represented by SEQ ID NOs: (18) to (20).
Examples of the auxiliary amino acid sequence having the (Gly Ala Gly Tyr Gly Val) e sequence include amino acid sequences represented by SEQ ID NOs: (21) to (23).
Examples of the auxiliary amino acid sequence having the {Asp Gly Gly (Ala) f Gly Gly Ala} g sequence include amino acid sequences represented by SEQ ID NOs: (24) to (26).
Examples of the auxiliary amino acid sequence having (Gly Val Pro Gly Val) h sequence include amino acid sequences represented by SEQ ID NOs: (27) to (29) and (54).
Examples of the auxiliary amino acid sequence having the (Gly) i sequence include amino acid sequences represented by SEQ ID NOs: (30) to (32).
Examples of the auxiliary amino acid sequence having the (Ala) j sequence include the amino acid sequences represented by SEQ ID NOs: (33) to (35).
Examples of the auxiliary amino acid sequence having the (Gly Gly Ala) k sequence include amino acid sequences represented by SEQ ID NOs: (36) to (38).
Examples of the auxiliary amino acid sequence having the (Gly Val Gly Val Pro) m sequence include amino acid sequences represented by SEQ ID NOs: (39) to (41).
Examples of the auxiliary amino acid sequence having (Gly Pro Pro) n sequence include amino acid sequences represented by SEQ ID NOs: (42) to (44).
Examples of the auxiliary amino acid sequence having (Gly Ala Gln Gly Pro Ala Gly Pro Gly) o sequence include amino acid sequences represented by SEQ ID NOs: (45) to (47).
(Gly Ala Pro Gly Ala Pro Gly Ser Gln Gly Ala Pro Gly Leu Gln) Examples of the auxiliary amino acid sequence having the p sequence include amino acid sequences represented by SEQ ID NOs: (48) to (50).
(Gly Ala Pro Gly Thr Pro Gly Pro Gln Gly Leu Pro Gly Ser Pro) Examples of the auxiliary amino acid sequence having the q sequence include amino acid sequences represented by SEQ ID NOs: (51) to (53).

  Among these SEQ ID NOs: (9), (10), (12), (13), (14), (15), (16), (18), (19), (21), (22) , (24), (25), (26), (27), (28), (30), (31), (33), (34), (36), (37), (39), ( 40), (42), (43), (45), (46), (48), (49), (51), (52) and (54) are preferred, and (10), (12) are more preferred. ), (13), (14), (16), (19), (22), (26), (27), (28), (29), (31), (34), (37), (40), (43), (46), (49), (52) and (54), particularly preferably (12), (13) and (54).

  Polypeptide (XM) may contain a branched chain, may be partially crosslinked, or may contain a cyclic structure. However, the polypeptide (XM) is preferably not cross-linked, more preferably a non-cross-linked linear structure (which may contain a cyclic structure), and particularly preferably a non-cross-linked structure without a cyclic structure. It has a linear structure. The linear structure also includes a β structure (secondary structure in which linear peptides are bent and the portions are arranged in parallel and a hydrogen bond is formed therebetween).

The polypeptide (XM) preferably has a structure in which the minimum amino acid sequence (X) and the auxiliary amino acid sequence (Y) are alternately chemically bonded from the viewpoint of cell adhesion and heat resistance. In this case, the number (units) of repeating units (XY) of the minimum amino acid sequence (X) and the auxiliary amino acid sequence (Y) is preferably 2 to 50, more preferably 3 from the viewpoint of cell adhesion. -40, particularly preferably 4-30, most preferably 5-20.
Further, the number of the minimum amino acid sequence (X) and the auxiliary amino acid sequence (Y) may be the same or different. When they are different from each other, it is preferable that one of the contained numbers is smaller than the other contained number {in this case, the auxiliary amino acid sequence (Y) is preferably small}. The content ratio (X / Y) between the minimum amino acid sequence (X) and the auxiliary amino acid sequence (Y) in the polypeptide (XM) is preferably 0.5 to 2, more preferably 0.9 to 1.4. Especially preferably, it is 1-1.3.
Further, other amino acids may be contained in the terminal portion of the polypeptide (XM) (from the minimum amino acid sequence (X) or auxiliary amino acid sequence (Y) to the end of the peptide). When other amino acids are included, the content is preferably 1 to 1000, more preferably 3 to 300, and particularly preferably 10 to 100 per polypeptide (XM).

The number average molecular weight (Mn) of the polypeptide (XM) is preferably 1,000 to 1,000,000, more preferably 2,000 to 700,000, particularly preferably 3,000 to 400,000, and most preferably. Is 4,000 to 200,000.
The number average molecular weight (Mn) of the polypeptide (XM) is determined by separating the measurement sample {polypeptide etc.} by SDS-PAGE (SDS polyacrylamide gel electrophoresis) and comparing the migration distance with a standard substance. And the like (hereinafter the same).

A part of preferable polypeptide (XM) is illustrated below.
(1) When the minimum amino acid sequence (X) is Arg Gly Asp sequence (x1), it has 13 (x1) and (Gly Ala Gly Ala Gly Ser) ₉ sequence (13) (13) (y1) A polypeptide having an Mn of about 110,000 having a structure in which these are alternately chemically bonded ("pronectin F", pronectin: a registered trademark of Sanyo Chemical Industries, Ltd. (Japan and the United States), manufactured by Sanyo Chemical Industries Co., Ltd. < The same applies below));
A polypeptide having an Mn of about 20,000 (5) of (x1) and 5 of (GlyAlaGlyAlaGlySer) ₃ sequence (12) (y2) and having a structure in which these are alternately chemically bonded ( “Pronectin F2”);
(X1) and 3 (Gly Val Pro Gly Val) ₂ Gly Gly (Gly Ala Gly Ala Gly Ser) ₃ Sequence (54) (y3) 3 A polypeptide having a Mn of about 10,000 (“pronectin F3”) and the like.

(2) When the minimum amino acid sequence (X) is the Ile Lys Val Ala Val sequence (x2) The Arg Gly Asp sequence (x1) of pronectin F, pronectin F2 or pronectin F3 is converted into the Ile Lys Val Ala Val sequence (7) (x2) “Pronectin L”, “pronectin L2”, “pronectin L3”, etc.

(3) When the minimum amino acid sequence (X) is the Tyr Ile Gly Ser Arg sequence (x3) The Arg Gly Asp sequence (x1) of pronectin F, pronectin F2 or pronectin F3 was changed to the Tyr Ile Gly Ser Arg sequence (x3) “Pronectin Y”, “Pronectin Y2”, “Pronectin Y3” and the like.

In addition to the polypeptides (1) to (3), RetroNectin (recombinant human fibronectin CH-296) manufactured by Takara Shuzo Co., Ltd. {Arg Gly Asp sequence (x1) and Leu Asp Val sequence as the minimum amino acid sequence (X) Containing Mn of about 60,000 polypeptide} and RGDS-Protein A {polypeptide having Mn of about 30,000 containing Arg Gly Asp sequence (x1) as the minimum amino acid sequence (X)} can be preferably used. This polypeptide is derived from nature and does not contain the auxiliary amino acid sequence (Y). Therefore, heat resistance etc. are inferior to said (1)-(3). The amino acid sequences of these polypeptides are disclosed in JP-A-2-31498. }.

  The production method of the polypeptide (XM) is not particularly limited, and can be produced in the same manner as a conventionally known method for synthesizing a peptide. For example, an organic synthesis method (solid phase synthesis method, liquid phase synthesis method, etc.) and It can be synthesized by a biochemical synthesis method [genetically modified microorganism (yeast, bacteria, Escherichia coli, etc.)] or the like. Regarding the organic synthesis method, for example, in the Biochemistry Society of Japan, “Sequel Biochemistry Experiment Course 2, Protein Chemistry (below)” pages 641-694 (May 20, 1987; published by Tokyo Chemical Co., Ltd.) The described methods are used. As for the biochemical synthesis method, for example, the method described in JP-T-3-502935 is used. A biochemical synthesis method using a genetically modified microorganism is preferable in that a high molecular weight polypeptide (XM) can be easily synthesized, and a method using a genetically modified Escherichia coli is particularly preferable.

  The compound (AM) and / or polypeptide (XM) is preferably all or part of the compound (AM) and / or polypeptide (XM) on the surface of the monolayer film (S), more preferably (AM) and / or (XM) are all present on the surface of the monolayer film (S).

When the wound dressing of the present invention has the compound (AM) and / or polypeptide (XM) containing the functional group (A), the monolayer film (S) is produced and then the functional group (A) is contained. Method (1) for binding compound (AM) and / or polypeptide (XM) to the surface of (S), or compound (AM) and / or polypeptide (XM) containing functional group (A), monolayer After mixing with the raw material of film (S), it can manufacture by the method (2) etc. which are made into a single layer film.
In the method (1), the monolayer film (S) can be produced by using a known method. For example, (a) the raw material of the monolayer film (S) is melted or made into a solution, and this is coated on a release paper. A film obtained by processing and drying, or a film obtained by injection molding, using an embossing device {for example, an embosser (manufactured by Cyto Engineering Co., Ltd.)} or the like to form irregularities on the film surface. (B) The raw material of the single layer film (S) is melted or made into a solution, and this is mixed with fine particles, and this mixture is coated on release paper using a coating machine. Or a method of injection molding this mixture to obtain a monolayer film (S) having irregularities on the surface, (c) making the raw material of the monolayer film (S) into a dispersion or solution, On the surface by drying The method or the like to obtain a single layer film (S) having a convex can be applied.

When the raw material is made into a solution, the solvent is not particularly limited as long as the raw material can be made into a uniform solution or an emulsified state. For example, in the case of a urethane resin, an organic solvent such as isopropyl alcohol, methyl ethyl ketone and toluene can be used, and an emulsifying urethane. In the case of a resin, an emulsified liquid can be prepared using water or the like as a solvent. When the solvent is water, emulsifiers and other additives may be used as necessary. When using an organic solvent, two or more organic solvents may be used.
When the raw material is used as a dispersion, the dispersion medium is not particularly limited as long as the raw material can be uniformly dispersed, but the above organic solvents and water can be used.
Coating equipment includes roll coaters (size press, gate roll coater, etc.), bar coaters (K paint applicator; manufactured by Matsuo Sangyo Co., Ltd.), gravure coaters, air knife coaters, blade coaters, etc. Can be used.
The release paper is not particularly limited as long as it can be easily peeled off from the produced film, and examples thereof include polypropylene, polyethylene, or paper coated with a release agent (EV130TPD; manufactured by Lintec Corporation, etc.). .
Coating amount is preferably 50~450g / m ² as the weight after drying, more preferably from 100 to 300 g / m ^2.
Drying is performed by room temperature drying or hot air drying. The drying temperature is preferably 20 to 200 ° C, more preferably 70 to 150 ° C.

As an apparatus for forming irregularities on the film surface, in addition to using an embossing apparatus, sandblasting (a method for forming irregularities by colliding hard fine particles against the film surface), a chemical method for etching the surface with acid or alkali Etc. are also applicable.
A method similar to a paper making method such as Japanese paper or Western paper can be applied as a method for squeezing.

  When the wound dressing of the present invention comprises a polypeptide (XM), the polypeptide (XM) and the monolayer film (S) have chemical bonds (ionic bonds, hydrogen bonds and / or covalent bonds, etc.) and They can be bonded by physical adsorption (adsorption by van der Waals force or the like).

As a method for covalently bonding the polypeptide (XM) and the monolayer film (S), the following (1) to (3) can be applied.
(1) Polypeptide (XM) having a primary amino group or a secondary amino group {Arginine, histidine, isoleucine, leucine, methionine, phenylalanine, threonine, tryptophan, tyrosine, valine, alanine, asparagine, aspartic acid, Glutamine, glutamic acid, proline, cysteine, lysine, serine, glycine, ornithine, histidine, 3-aminopropionic acid, 8-aminooctanoic acid and / or 20-aminoeicosanoic acid as a structural unit} Layer film (S) having a carboxyl group (polyglycolic acid, synthetic polypeptide not containing cell adhesion minimal amino acid sequence (X), modified maleic acid of polyethylene or polypropylene, ethylene- (meth) acrylic acid copolymer) Heavy Body, polyacrylic acid, collagen, gelatin, glycosaminoglycan, hyaluronic acid, chondroitin sulfate, dermatan sulfate, heparin, elastin, fibrin, alginic acid, albumin, pectin, pectic acid, gluten and / or fibroin) The method of reacting (amide bond method).

(2) Polypeptide (XM) having a primary amino group or secondary amino group and monolayer film (S) having a hydroxyl group (those containing a hydroxyl group are not particularly limited) For example, polyglycolic acid, ethylene-vinyl acetate copolymer, polyurethane, polyester, cellulose, viscose rayon, cupra rayon, polyacetate, vinylon, collagen, gelatin, glycosaminoglycan, hyaluronic acid, chondroitin sulfate, keratan sulfate , Dermatan sulfate, heparin, elastin, chitin, chitosan, fibrin, alginic acid, starch, dextran, albumin, polyhydroxybutyric acid, pectin, pectic acid, galactan, pullulan, agarose, gluten and / or fibroin A method of reacting Irumu etc.) (N-C bond is how to).

(3) Polypeptide (XM) having a hydroxyl group (polypeptide containing aspartic acid, glutamic acid, serine, threonine, tyrosine, thyronine and / or hydroxypurine as a structural unit) and monolayer film (S) Among them, a method of reacting with one having a carboxyl group (method of ester bonding).

These reactions can be carried out by known methods (such as “the basis and experiment of peptide synthesis, published on Mar. 5, 1997, published by Maruzen Co., Ltd.”). Specifically, it is as the following (1) to (3).
(1) When reacting a polypeptide (XM) having a primary amino group or a secondary amino group with a monolayer film (S) having a carboxyl group, the carboxyl of the monolayer film (S) After reacting the group in advance with a carbodiimide compound to obtain acylisourea {R′—N═C (OCOR) —NH—R ′ (part where —OCOR is derived from the monolayer film (S))}, polypeptide ( The monolayer film (S) and the polypeptide (XM) can be amide-bonded by adding one having a primary amino group or a secondary amino group among XM) to the acylisourea.
Examples of the carbodiimide compound include N, N′-dicyclohexylcarbodiimide, 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride, and the like.

(2) When reacting a polypeptide (XM) having a primary amino group or a secondary amino group with a monolayer film (S) having a hydroxyl group, the hydroxyl of the monolayer film (S) After reacting a group with a carbonyldiimidazole compound in advance to obtain an imidazole derivative {R-Im, Im is an imidazoline ring, and R is derived from a monolayer film (S)}, a primary amino group of the polypeptide (XM) Alternatively, by adding a compound having a secondary amino group to the imidazole derivative, the monolayer film (S) and the polypeptide (XM) can be N—C bonded.
Examples of the carbonyldiimidazole compound include N, N′-carbonyldiimidazole.

  (3) When reacting a polypeptide (XM) having a hydroxyl group with a monolayer film (S) having a carboxyl group, the carboxyl group of the monolayer film (S) is reacted with a carbodiimide compound in advance. After the acylisourea is obtained, the monolayer film (S) and the polypeptide (XM) can be ester-bonded by adding the polypeptide (XM) having a hydroxyl group to the acylisourea.

  As a method of physically adsorbing, ion-bonding and / or hydrogen-bonding the polypeptide (XM) to the monolayer film (S), the polypeptide (XM) and (S) are put into a solvent or the like and mixed. And the like. Although there is no restriction | limiting in particular as a solvent, The aqueous solution etc. which contain 0.001-50 weight% (preferably 0.01-10 weight%) of inorganic salt, organic acid salt, an acid, and / or a base can be used.

Examples of inorganic salts include metal halide salts, sulfate metal salts, phosphate metal salts, hydrogen phosphate metal salts, nitrate metal salts, carbonate metal salts, metal perhalogenates, and the like. Sodium, sodium sulfate, sodium phosphate, calcium chloride, iron nitrate, potassium chloride, magnesium sulfate, sodium carbonate, sodium hydrogen phosphate, potassium phosphate, potassium hydrogen phosphate, copper sulfate, iron sulfate, lithium chloride, sodium bromide Lithium bromide, sodium perchlorate, lithium perchlorate and the like.
Examples of the organic acid salt include sodium formate, sodium acetate, lithium acetate, and sodium tartrate.
Examples of the acid include inorganic acids, organic acids having 1 to 6 carbon atoms, and specific examples include hydrochloric acid, phosphoric acid, acetic acid, formic acid, phenol, and sulfuric acid.
Examples of the base include inorganic bases, organic bases having 2 to 6 carbon atoms, and specific examples include sodium hydroxide, potassium hydroxide, ammonia, and triethylamine.
Examples of water include distilled water, ion exchange water, tap water, and ion exchange distilled water.
Among these solvents, an aqueous solution containing an inorganic salt, an acid and / or a base, and water are preferred, and an aqueous solution containing an inorganic salt, an acid and / or a base is more preferred.

  A chemical bond is preferable and a covalent bond is more preferable in that the polypeptide (XM) and the monolayer film (S) are firmly bonded.

When the polypeptide (XM) is contained in the wound dressing, the content is 0.1 ng / cm ^{2 to} 100 mg / cm per 1 cm × 1 cm square monolayer film (S) from the viewpoint of promoting wound healing. ² , more preferably 1 ng / cm ^{2 to} 10 mg / cm ² , particularly preferably 10 ng / cm ^{2 to 1} mg / cm ² , most preferably 100 ng / cm ^{2 to} 100 μg / cm ² .

  Although the measuring method of content of polypeptide (XM) per square-shaped monolayer film (S) of 1 cm x 1 cm is not particularly limited, for example, an immunological measuring method can be used. In the present invention, the content of the polypeptide (XM) per 1 cm × 1 cm square-shaped monolayer film (S) was cut off a part of the surface of the wound dressing (for example, 1 cm × 1 cm square shape). It is indicated by a value obtained by reacting a test piece with an antibody labeled with an antibody that binds to the polypeptide (XM) and measuring the amount of the enzyme labeled antibody reacted.

  The enzyme-labeled antibody is usually an antibody obtained by chemically bonding an enzyme and a specific antibody (can be chemically bonded by a known method). A method of chemically binding an enzyme (peroxidase, β-D-galactosidase, alkaline phosphatase, glucose-6-phosphate dehydrogenase, etc.) and a specific antibody by the glutaraldehyde method, periodate method, maleimide method, pyridyl disulfide method, etc. Etc. (Ultra-sensitive enzyme immunoassay, written by Yuji Ishikawa, Society Publishing Center, 1993; Enzyme immunoassay, translated by Yuji Ishikawa, Tokyo Kagaku Dojin, 1989; and enzyme antibody method, edited by Keiichi Watanabe et al., Interdisciplinary) Planning Co., Ltd., 1992) is applicable.

  The specific antibody is an antibody that specifically binds to the polypeptide (XM) {known method; polyclonal antibody production method and monoclonal antibody production method (enzyme immunoassay, translated by Junji Ishikawa, Tokyo Kagaku Dojin, 1989; and enzymes) Antibody method, edited by Keiichi Watanabe et al., Interdisciplinary Planning Co., Ltd., 1992) can be applied. The smaller the affinity constant of the specific antibody for the cross-reactive antigen, the better. For example, when the affinity constant of the specific antibody to the polypeptide (XM) is 1, the affinity constant for the cross-reactive antigen is preferably 1 or less, more preferably 0.1 or less, particularly preferably 0.01 or less. is there. This affinity constant can be obtained by the method described in Enzyme Immunoassay (translated by Yuji Ishikawa, Tokyo Kagaku Dojin, 1989).

  When using a compound (AM), it is preferable that the compound (AM) is couple | bonded with the monolayer film (S) and / or polypeptide (XM).

  Compound (AM) can be bonded to monolayer film (S) and / or polypeptide (XM) by chemical reaction, physical adsorption {polypeptide (XM) and monolayer film (S) described above] A method similar to the method of combining the two and the like can be applied.

When the compound (AM) is contained in the wound dressing, this content (pieces / cm ² ) is expressed as the average number of amino groups per 1 cm × 1 cm square monolayer film (S) from the viewpoint of promoting wound healing. 10 ⁸ to 10 ²² is preferable, and 10 ¹⁰ to 10 ²⁰ is more preferable.

  The average number of amino groups is determined by a known method, for example, trinitrobenzene sulfonic acid (TNBS) method {Protein Chemistry IV (published by Tokyo Chemical Industry Co., Ltd., 1981)} or hydrochloric acid / indicator (bromphenol blue, etc.) titration method. It can be measured by a total amine number measuring method (JIS K7237-1986, ASTM D2074-66, etc.) and the like.

In the present invention, specifically, the average number (number / cm ² ) of amino groups per 1 cm × 1 cm square single layer film (S) is measured as follows. A calibration curve (a graph of the number of amino groups and absorbance) of a solution of a compound (AM) or (AM) having a known number of amino groups is prepared by the TNBS method.

  The absorbance of a test piece obtained by cutting a part of the surface of the wound dressing (for example, a 1 cm × 1 cm square shape) is measured by the TNBS method and converted to the number of amino groups using a calibration curve.

  The wound dressing of the present invention may be sterilized as necessary. As a sterilization method, a sterilization method using radiation, ethylene oxide gas, plasma, γ rays, alcohol, autoclave, dry heat, or the like can be applied. These may be performed only by one type of method, or two or more types may be combined.

The wound dressing of the present invention can be applied to the wound surface without limitation as long as it can cover the wound surface, for example, a cover with an adhesive or an adhesive, a bandage, a stretchable mesh, or an eye patch and the like. It may also be sutured or glued. In addition, when the wound is a chronic wound such as an ulcer, it is preferably applied to a wound in the yellow phase to the red phase, and when the wound is an acute wound such as a burn or traumatic skin defect, a wound from the inflammation phase to the granulation phase Application to is preferable.
The wound dressing material of the present invention can be applied to human body surface sites such as human wound sites, burn sites, pressure ulcer sites, ulcer sites, skin defect sites such as skin removal sites, as well as suture sites and puncture sites such as catheters. .

  As cells applicable to the wound dressing of the present invention, human-derived cells are suitable. For example, cells involved in the skin (epithelial cells, fibroblasts, vascular endothelial cells, smooth muscle cells, etc.) and blood vessels are involved. Cells (such as vascular endothelial cells, smooth muscle cells and fibroblasts), cells involved in muscle (such as muscle cells), cells involved in fat (such as fat cells), cells involved in nerves (such as nerve cells), Cells involved in liver (liver parenchymal cells, etc.), cells involved in pancreas (pancreatic islet cells, etc.), cells involved in kidney (kidney epithelial cells, proximal tubular epithelial cells, mesangial cells, etc.), lung / bronchi Cells (epithelial cells, fibroblasts, vascular endothelial cells, smooth muscle cells, etc.), cells involved in the eyes (visual cells, corneal epithelial cells, corneal endothelial cells, etc.), cells involved in the prostate (epithelial cells, Stromal cells and smooth muscle cells) Cells involved in bone (such as osteoblasts, bone cells and osteoclasts), cells involved in cartilage (such as chondroblasts and chondrocytes), cells involved in teeth (such as periodontal ligament cells and osteoblasts), Blood-related cells (white blood cells and red blood cells) and stem cells (eg, bone marrow undifferentiated mesenchymal stem cells, skeletal muscle stem cells, hematopoietic stem cells, neural stem cells, hepatic stem cells (oval cells, small hepatocytes, etc.), adipose tissue stem cells Embryonic stem (ES) cells, epidermal stem cells, intestinal stem cells, sperm stem cells, embryonic germ stem (EG) cells, pancreatic stem cells (pancreatic duct epithelial stem cells, etc.), leukocyte stem cells, lymphocyte stem cells, corneal stem cells, progenitor cells (Adipose precursor cells, vascular endothelial precursor cells, cartilage precursor cells, lymphocyte precursor cells, NK precursor cells, etc.)} and the like}. These cells can also be suitably applied when using the wound dressing of the present invention as a substrate for cell culture in vitro.

Hereinafter, the present invention will be described in more detail with reference to examples. However, the present invention is not limited to these examples. Unless otherwise specified, “%” means “% by weight”.
<Example 1>
<Preparation of single layer film (S1)>
6.67 g of urethane emulsion (trade name: Permarin UA200, manufactured by Sanyo Chemical Industries, Ltd.) and 3.33 g of ion-exchanged water were mixed to prepare a urethane emulsion. 6.67 g of this urethane emulsion and 12 nylon beads (trade name: Microcomposite Sphere, manufactured by TRI Advance Laboratory, diameter 150 μm to 250 μm) are mixed, and a polypropylene sheet (stock) 20 cm long × 20 cm wide × 1 mm thick The product was uniformly spread over a medical agent (made by a company Medical Agent) and cast using a glass rod wrapped with vinyl tape at a height of 300 μm at both ends so that the film thickness was 300 μm, and left at room temperature (about 25 ° C.). 24 hours after standing at room temperature, it was dried in a circulating air dryer at 120 ° C. for 1 hour. After drying, the urethane film formed on the polypropylene sheet was peeled from the polypropylene sheet to obtain a single layer film (S1).
When the film thickness after drying was measured with a digital film thickness meter, the average film thickness of the film was 58 μm. The arithmetic average roughness (Ra) was 40 μm, and the average length (RSm) of the contour curve elements was 300 μm.

<Preparation of cell adhesion polypeptide (P2-1)>
According to the method described in the examples in JP-T-3-502935, it has about 13 Arg Gly Asp sequences (1) and (Gly Ala Gly Ala Gly Ser) 9 sequences (47). A peptide “SLPF” having an average molecular weight of about 100,000 was produced by genetically modified E. coli and purified by column chromatography to obtain a cell adhesion polypeptide (P2-0). Furthermore, this (P2-0) was autoclaved (120 ° C., 20 minutes) to obtain a cell adhesion polypeptide (P2-1).

<Preparation of wound dressing (H1)>
0.479 g of 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (manufactured by Sigma) was dissolved in 50 mL of ion exchange water to prepare a carbodiimide aqueous solution. 50 mL of this carbodiimide aqueous solution and 10 cm × 10 cm of the single layer film (S1) were put into a glass petri dish and allowed to stand at 25 ° C. for 1 hour. Thereafter, it was washed 5 times with 100 mL of deionized water. Next, 50 mL of an aqueous solution of the cell adhesion polypeptide (P2-1) {(P2-1) concentration: 100 ng / mL} was added, and allowed to stand at 25 ° C. for 1 hour, and (P1− 1) was bound. Further, 50 mL of a polyethyleneimine-containing aqueous solution {concentration of polyethyleneimine (MW = 10000): 50 ng / mL} was added and allowed to stand at 25 ° C. for 1 hour, and polyethyleneimine (MW = 10000) was bound to (S1). It was. Then, it wash | cleaned 5 times with 100 mL ion-exchange water, it was made to dry for 12 hours in a 37 degreeC circulating air dryer, and the wound dressing (H1) was prepared. (H1) The adhesion amount (content) of (P2-1) per 1 cm × 1 cm was 1 μg.

The adhesion amount (content) of the cell adhesive polypeptide (P2-1) of the wound dressing (H1) was measured by the following procedure.
(1) A standard wound dressing (N1) with a known adhesion amount (content) of the cell adhesion polypeptide (P2-1) and a wound dressing (H1) with an unknown adhesion amount are each 1 cm × 1 cm square shape One was placed in 3 mL of PBS containing 1% by weight of bovine serum albumin and immersed for 2 hours at room temperature (25 ° C.).
The standard wound dressing is adjusted in the same manner as in the preparation of the wound dressing (H1). The amount of adhesion is obtained by concentrating and lyophilizing the P2-1 aqueous solution to which the cell adhesive polypeptide has adhered. The weight of unattached cell adhesive polypeptide was determined, and the weight of unattached cell adhesive polypeptide was subtracted from the weight of cell adhesive polypeptide before attachment.

(2) Each wound dressing is taken out, and each wound dressing 1 in 2 ml of PBS containing 10 μg / mL of peroxidase-labeled anti-P2-1 antibody, 1% by weight of bovine serum albumin and 0.2% by weight of Tween 20 The sheet was put in and reacted at 37 ° C. for 2 hours. After the reaction, each wound dressing was washed three times with 5 mL of PBS containing Tween 20 at 0.2 wt%.
The peroxidase-labeled anti-P2-1 antibody was prepared by immunizing a rabbit with a cell-adhesive polypeptide (P2-1) according to a polyclonal antibody production method (enzyme immunoassay, translated by Junji Ishikawa, Tokyo Chemical Co., Ltd., 1989). Thus, the anti-P2-1 antibody is obtained, and the anti-P2-1 antibody and peroxidase (manufactured by Toyobo Co., Ltd.) are bound by the maleimide method (enzyme immunoassay, translated by Junji Ishikawa, Tokyo Kagaku Dojin, 1989). As a result, a peroxidase-labeled anti-P2-1 antibody was obtained.

(3) Take out each wound dressing, and put one wound dressing in a mixed solution of 0.2 mL of OLYDAS exclusive reagent coloring solution set (manufactured by Sanyo Chemical Industries, Ltd.) and 1.8 mL of ion-exchanged water. The reaction was carried out at 37 ° C. for 1 hour. After the reaction, the absorbance was measured at a wavelength of 380 nm.
(4) A calibration curve was created using the absorbance of the standard wound dressing (N1), and the amount of adhesion of the wound dressing (H1) was obtained from the calibration curve. Hereinafter, the amount of polypeptide attached was measured in the same manner.

<Example 2>
<Preparation of wound dressing (H2)>
0.479 g of 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (manufactured by Sigma) was dissolved in 50 mL of ion exchange water to prepare a carbodiimide aqueous solution. 50 mL of this carbodiimide aqueous solution and 10 cm × 10 cm of the single layer film (S1) were put into a glass petri dish and allowed to stand at 25 ° C. for 1 hour. Thereafter, it was washed 5 times with 100 mL of deionized water. Next, 50 mL of an aqueous solution of cell adhesion polypeptide (P2-1) {concentration of (P2-1): 100 ng / mL} was added, and allowed to stand at 25 ° C. for 1 hour to form a single layer film (S1). (P2-1) was bound. Then, it wash | cleaned 5 times with 100 mL ion-exchange water, it was made to dry for 12 hours in a 37 degreeC circulating air dryer, and the wound dressing (H2) was prepared. (H2) The adhesion amount (content) of (P2-1) per 1 cm × 1 cm was 1 μg.

<Example 3>
<Preparation of wound dressing (H3)>
0.479 g of 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (manufactured by Sigma) was dissolved in 50 mL of ion exchange water to prepare a carbodiimide aqueous solution. 50 mL of this carbodiimide aqueous solution and 10 cm × 10 cm of the single layer film (S1) were put into a glass petri dish and allowed to stand at 25 ° C. for 1 hour. Thereafter, it was washed 5 times with 100 mL of deionized water. Next, 50 mL of a polyethyleneimine-containing aqueous solution {polyethyleneimine (MW = 10000) concentration: 50 ng / mL} is added and allowed to stand at 25 ° C. for 1 hour, and polyethyleneimine (MW = 10000) is bound to (S1). I let you. Then, it wash | cleaned 5 times with 100 mL ion-exchange water, it was made to dry for 12 hours in a 37 degreeC circulating air dryer, and the wound dressing material (H3) was prepared.

<Example 4>
<Preparation of single layer film (S2)>
A monolayer film (S2) was obtained in the same manner as in Example 1 except that 12 nylon beads (diameter 150 to 250 μm) were changed to 12 nylon beads (diameter 400 to 500 μm). When the film thickness after drying was measured with a digital film thickness meter, the average film pressure was 138 μm. The arithmetic average roughness (Ra) was 280 μm, and the average length (RSm) of the contour curve elements was 190 μm.
<Preparation of wound dressing (H4)>
A wound dressing (H4) was prepared in the same manner as in Example 1 except that the single layer film (S1) was changed to the single layer film (S2). (H4) The adhesion amount (content) of (P2-1) per 1 cm × 1 cm was 0.9 μg.

<Example 5>
<Preparation of single layer film (S3)>
A single layer film (S3) was obtained in the same manner as in Example 1 except that 6.67 g of 12 nylon beads were changed to 2.75 g. When the film thickness after drying was measured with a digital film thickness meter, the average film pressure was 47 μm. The arithmetic average roughness (Ra) was 32 μm, and the average length (RSm) of the contour curve elements was 588 μm.
<Preparation of wound dressing (H5)>
A wound dressing (H5) was produced in the same manner as in Example 1 except that the single layer film (S1) was changed to the single layer film (S3). (H5) The adhesion amount (content) of (P2-1) per 1 cm × 1 cm was 0.9 μg.

<Example 6>
<Preparation of cell adhesion polypeptide (P3-1)>
Example 1 except that the peptide sequence was changed to a peptide having about 5 Arg Gly Asp sequences (1) and (Gly Ala Gly Ala Gly Ser) 9 sequences (47), each having a number average molecular weight of about 20,000. In the same manner as above, a cell adhesion polypeptide (P3-1) was obtained.
<Preparation of wound dressing (H6)>
A wound dressing (H6) was prepared in the same manner as in Example 1 except that the cell adhesive polypeptide (P2-1) was changed to the cell adhesive polypeptide (P3-1). (H6) The adhesion amount of (P3-1) of 1 cm × 1 cm was 0.8 μg.

<Example 7>
<Preparation of cell adhesion polypeptide (P4-1)>
Example 1 except that the peptide sequence was changed to a peptide having about 3 Arg Gly Asp sequences (1) and (Gly Ala Gly Ala Gly Ser) 9 sequences (47) each and a number average molecular weight of about 10,000. In the same manner as above, a cell adhesive polypeptide (P4-1) was obtained.
<Preparation of wound dressing (H7)>
A wound dressing (H7) was prepared in the same manner as in Example 1 except that the cell adhesive polypeptide (P2-1) was changed to the cell adhesive polypeptide (P4-1). (H7) The adhesion amount of (P4-1) of 1 cm × 1 cm was 1.0 μg.

<Example 8>
<Preparation of single layer film (S4)>
A single layer film (S4) was obtained in the same manner as in Example 1 except that 6.67 g of 12 nylon beads were changed to 8.25 g. When the film thickness after drying was measured with a digital film thickness meter, the average film pressure was 61 μm. The arithmetic average roughness (Ra) was 20 μm, and the average length (RSm) of the contour curve elements was 52 μm.
<Preparation of wound dressing (H8)>
A wound dressing (H8) was produced in the same manner as in Example 1 except that the single layer film (S1) was changed to the single layer film (S4). (H8) The adhesion amount (content) of (P2-1) per 1 cm × 1 cm was 0.7 μg.

<Example 9>
<Preparation of single layer film (S5)>
A monolayer film (S5) was obtained in the same manner as in Example 1 except that 12 nylon beads (diameter 150 to 250 μm) were changed to 12 nylon beads (diameter 50 to 200 μm). When the film thickness after drying was measured with a digital film thickness meter, the average film pressure was 50 μm. The arithmetic average roughness (Ra) was 11 μm, and the average length (RSm) of the contour curve elements was 920 μm.
<Preparation of wound dressing (H9)>
A wound dressing (H9) was produced in the same manner as in Example 1 except that the single layer film (S1) was changed to the single layer film (S5). (H9) The adhesion amount (content) of (P2-1) per 1 cm × 1 cm was 0.7 μg.

<Example 10>
<Preparation of single layer film (S6)>
A monolayer film (S6) was obtained in the same manner as in Example 1 except that 12 nylon beads (diameter 150 to 250 μm) were changed to 12 nylon beads (diameter 750 to 1500 μm). When the film thickness after drying was measured with a digital film thickness meter, the average film pressure was 80 μm. The arithmetic average roughness (Ra) was 510 μm, and the average length (RSm) of the contour curve elements was 80 μm.
<Preparation of wound dressing (H10)>
A wound dressing (H10) was prepared in the same manner as in Example 1 except that the single layer film (S1) was changed to the single layer film (S6). (H10) The adhesion amount (content) of (P2-1) per 1 cm × 1 cm was 0.8 μg.

<Example 11>
<Preparation of single layer film (S7)>
A monolayer film (S7) was obtained in the same manner as in Example 1 except that 6.67 g of 12 nylon beads (diameter 150 to 250 μm) was changed to 12.0 g of 12 nylon beads (diameter 50 to 100 μm). When the film thickness after drying was measured with a digital film thickness meter, the average film pressure was 94 μm. The arithmetic average roughness (Ra) was 14 μm, and the average length (RSm) of the contour curve elements was 21 μm.
<Preparation of wound dressing (H11)>
A wound dressing (H11) was produced in the same manner as in Example 1 except that the single layer film (S1) was changed to the single layer film (S7). (H11) The adhesion amount (content) of (P2-1) per 1 cm × 1 cm was 0.9 μg.

<Example 12>
<Preparation of wound dressing (H12)>
0.479 g of 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (manufactured by Sigma) was dissolved in 50 mL of ion exchange water to prepare a carbodiimide aqueous solution. 50 mL of this carbodiimide aqueous solution and 10 cm × 10 cm of the single layer film (S1) were put into a glass petri dish and allowed to stand at 25 ° C. for 1 hour. Thereafter, it was washed 5 times with 100 mL of deionized water. Next, 50 mL of an aqueous solution of the cell adhesion polypeptide (P2-1) {(P2-1) concentration: 100 ng / mL} was added, and allowed to stand at 25 ° C. for 1 hour, and (P1− 1) was bound. Further, 50 mL of a polyethyleneimine-containing aqueous solution {concentration of polyethyleneimine (MW = 1800): 50 ng / mL} was added and allowed to stand at 25 ° C. for 1 hour to bind polyethyleneimine (MW = 1800) to (S1). It was. Then, it wash | cleaned 5 times with 100 mL ion-exchange water, it was made to dry for 12 hours in a 37 degreeC circulating air dryer, and the wound dressing material (H12) was prepared. (H12) The adhesion amount (content) of (P2-1) per 1 cm × 1 cm was 1 μg.

<Example 13>
<Preparation of wound dressing (H13)>
0.479 g of 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (manufactured by Sigma) was dissolved in 50 mL of ion exchange water to prepare a carbodiimide aqueous solution. 50 mL of this carbodiimide aqueous solution and 10 cm × 10 cm of the single layer film (S1) were put into a glass petri dish and allowed to stand at 25 ° C. for 1 hour. Thereafter, it was washed 5 times with 100 mL of deionized water. Next, 50 mL of polyethyleneimine-containing aqueous solution {concentration of polyethyleneimine (MW = 1800): 50 ng / mL} is added and allowed to stand at 25 ° C. for 1 hour, and polyethyleneimine (MW = 1800) is bound to (S1). I let you. Then, it wash | cleaned 5 times with 100 mL ion-exchange water, it was made to dry for 12 hours in a 37 degreeC circulating air dryer, and the wound dressing material (H13) was prepared.

<Example 14>
<Preparation of wound dressing (H14)>
0.479 g of 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (manufactured by Sigma) was dissolved in 50 mL of ion exchange water to prepare a carbodiimide aqueous solution. 50 mL of this carbodiimide aqueous solution and 10 cm × 10 cm of the single layer film (S1) were put into a glass petri dish and allowed to stand at 25 ° C. for 1 hour. Thereafter, it was washed 5 times with 100 mL of deionized water. Next, 50 mL of an aqueous solution of the cell adhesion polypeptide (P2-1) {(P2-1) concentration: 100 ng / mL} was added, and allowed to stand at 25 ° C. for 1 hour, and (P1− 1) was bound. Further, 50 mL of a poly-L-ornithine-containing aqueous solution {Poly-L-ornithine (MW = 11900) concentration: 50 ng / mL} was added and allowed to stand at 25 ° C. for 1 hour, and poly-L- was added to (S1). Ornithine (MW = 11900) was conjugated. Then, it wash | cleaned 5 times with 100 mL ion-exchange water, it was made to dry for 12 hours in a 37 degreeC circulating air dryer, and the wound dressing material (H14) was prepared. (H14) The adhesion amount (content) of (P2-1) per 1 cm × 1 cm was 1.1 μg.

<Comparative Example 1>
<Preparation of single layer film (S8)>
A single layer film (S8) was obtained in the same manner as in Example 1 except that 12 nylon beads were not mixed. When the film thickness after drying was measured with a digital film thickness meter, the average film thickness of the film was 60 μm.
<Preparation of wound dressing (H15)>
A wound dressing (H15) was produced in the same manner as in Example 1 except that the single layer film (S1) was changed to the single layer film (S8). (H15) The adhesion amount (content) of (P2-1) per 1 cm × 1 cm was 0.7 μg.

<Comparative example 2>
<Preparation of single layer film (S9)>
Single nylon film (S9) in the same manner as in Example 1 except that 12 nylon beads (diameter 150 μm to 250 μm) were changed to glass beads having a diameter of 1500 to 2500 μm (trade name: Glass beads # 2, manufactured by Tokyo Glass Instrument Co., Ltd.). )
When the film thickness after drying was measured with a digital film thickness meter, the average film thickness of the film was 80 μm. The arithmetic average roughness (Ra) was 1020 μm, and the average length (RSm) of the contour curve elements was 510 μm.
<Preparation of wound dressing (H16)>
A wound dressing (H16) was produced in the same manner as in Example 1 except that the single layer film (S1) was changed to the single layer film (S9). (H16) The adhesion amount (content) of (P2-1) per 1 cm × 1 cm was 0.7 μg.

<Evaluation 1 (cell culture: in vitro)>
The wound dressings (H1) to (H16) of Examples 1 to 14 and Comparative Examples 1 and 2 were each cut to a size of 1.2 cm in diameter, and UV irradiation was performed for 1 hour each on the front and back sides in a clean bench. Sterilized. In addition, 6 sheets were prepared for each type of wound dressing and evaluated 6 times.
In addition, 15,000 normal human dermal fibroblasts (manufactured by Kurashiki Spinning Co., Ltd.) pre-cultured in 10 mL of a low serum medium for proliferation of normal human skin fibroblasts (manufactured by Kurashiki Spinning Co., Ltd.) for 5 days. Was seeded on a collagen type I gel (trade name: Cell Matrix, Nitta Gelatin Co., Ltd.) placed in each hole of a 24-hole polystyrene plate (Becton Dickinson), and the carbon dioxide gas concentration was 5% by volume at 37 ° C. The cells were cultured in an incubator for 7 days to prepare a cultured dermis model.

Seven days later, the cultured dermis model was transferred to another 24-well polystyrene plate containing 150 μL of serum-free epidermal keratinocyte growth medium (Kurashiki Boseki Co., Ltd.) in each well. 3 μL of human epidermal keratinocytes (manufactured by Kurashiki Boseki Co., Ltd.) were seeded, and wound dressings (H1) to (H16) previously sterilized were pasted on the seeded normal human epidermal keratinocytes. Pasting is performed without using an adhesive or the like, and cultured dermis seeded with normal human epidermal keratinocytes so that the cultured dermis model seeded with normal human epidermis keratinocytes and the wound dressings (H1) to (H16) are in contact with each other. The wound dressings (H1) to (H16) were placed on the top using tweezers.
The cultured dermis model to which the wound dressings (H1) to (H16) were attached was returned again to an incubator at 37 ° C. and a carbon dioxide gas concentration of 5% by volume, and further cultured for 7 days.

  After 7 days of culturing, the wound dressings (H1) to (H16) were peeled off from the upper surface of the cultured dermis, and wound dressings (H1) to (H16) were added to a new 24-hole polystyrene plate at 1 sheet / hole. 125 μL of a phosphate buffer (0.02 M, pH 7.2) containing 0.85% by weight of sodium chloride and 25 μL of Tetracolor One (Seikagaku Corporation) are added one by one at 37 ° C. And left in an incubator with a carbon dioxide gas concentration of 5% by volume for 4 hours.

  After 4 hours, 60 μL of the liquid was extracted from each well of the 24-well plate that had been incubated, and the amount of formazan produced in the liquid was measured using a spectrophotometer at an absorbance of 492 nm (control wavelength: 630 nm). The cell activity of the cells adhering to the wound dressing surface was defined. This measurement was performed twice for each hole, and as a result, 12 pieces of measurement data were obtained for (H1) to (H16). These measurement data, arithmetic mean, standard deviation, and coefficient of variation (%) are shown in Table 1 and It was shown in 2. In addition, since cell activity is proportional to the height of the light absorbency, it means that cell activity is so high that the arithmetic mean of light absorbency is large. Further, tetracolorium tetrazolium salt is reduced by intracellular mitochondrial dehydrogenase to produce formalin, thereby producing color.

  From the above results, the coefficient of variation of the wound dressing of the comparative example is as high as 31 or 41%, whereas the coefficient of variation of the wound dressing of the present invention {Examples 1 to 14} is 7 to 20%. Very low. This indicates that the cells grow and adhere to the surface of the wound dressing of the present invention with good reproducibility, that is, the wound dressing of the present invention has little variation in wound healing effect (reproducible). High).

<Evaluation 2 (Animal Experiment)>
Inhalation anesthesia with diethyl ether was performed on 12 DM mice (C57BLK Jcl db / db, manufactured by Clea Japan Co., Ltd.), and the entire back portion was shaved with a feather razor. 0.0 cm) full thickness skin defect wound was prepared. The DM mouse used was a diabetes confirmation strip (UroPiece, manufactured by Fujisawa Pharmaceutical Co., Ltd.) that has been confirmed to develop diabetes.
What was obtained by cutting the wound dressing material (H1) of Example 1 and the wound dressing material (H15) of Comparative Example 1 into a size of 1.5 cm × 1.5 cm, respectively, was used as an adhesive film (trade name: Tegaderm, 3M Healthcare). The wound dressing was attached so that the wound dressing side was in contact with the wound surface. Further, the whole circumference of the trunk was wound and fixed with an adhesive bandage (trade name: Silky Tex, manufactured by Alcare Corporation). The number of observed individuals was 6 for each of the (H1) application group and the (H15) application group.
The growth environment was a room temperature of 24 ° C., and both feed and water were freely ingested. The observation point was the seventh day, and the wound healing state was confirmed by macroscopic findings. Evaluation was made into a two-step evaluation of ○ and × in accordance with the following criteria. These results are shown in Table 3.
<Criteria>
○: The epidermis is smoothly regenerated from the wound edge toward the center of the wound, and the unregenerated part is less than 64 mm ² .
X: An unregenerated part of the wound is present at 64 mm ² or more, and the regeneration of the epidermis is insufficient.

  From the above results, in the wound dressing of the present invention (Example 1), all of the six animals became ◯, whereas in the comparative wound dressing (Comparative Example 1), 3 animals were ◯ and 3 animals were x. there were. This indicates that Example 1 has high reproducibility of the wound healing effect.

  The wound dressing of the present invention can be used not only as a wound dressing for humans but also as a wound dressing for non-human animals. Furthermore, it can be used as a base material for cell culture in vitro.

It is sectional drawing of the single layer film (S) which represented typically the relationship between reference | standard length {U} and the contour curve {Z (x)}. It is sectional drawing of the single layer film (S) which represented typically the relationship between reference | standard length (U) and an outline curve element {XSi}. It is the photograph which observed the surface of the wound dressing material [H1] of the present invention obtained in Example 1 with a digital microscope (HD microscope “VH-8000” manufactured by Keyence Corporation) at a magnification of 50 times.

Claims (5)

The arithmetic average roughness (Ra) specified in JIS B0601: 2001 “4.2.1 Arithmetic mean height of contour curve” is 1 μm to 1000 μm, and JIS B0601: 2001 “4.3.1 Mean of contour curve element” A wound dressing comprising a single layer film (S) having a surface property parameter having an average length (RSm) of a contour curve element defined in "Length" of 10 m to 5 mm. At least one functional group (A) selected from the group consisting of a secondary amino group (A1), a tertiary amino group (A2), an ammonio group (A3), a phosphatidyl group (A4), and a lysophosphatidyl group (A5); The wound dressing according to claim 1, comprising a compound (AM) containing and / or a polypeptide (XM) having at least one cell adhesion minimal amino acid sequence (X) per molecule. A polypeptide (XM) having a cell adhesion minimal amino acid sequence (X) of Arg Gly Asp sequence, Leu Asp Val sequence, Leu Arg Glu sequence, His Ala Val sequence and SEQ ID NOs: (1) to (8) The wound dressing according to claim 2, which is at least one selected from the group consisting of the amino acid sequences represented by any of the above. The wound dressing according to claim 2 or 3, comprising a polypeptide (XM), wherein (XM) has at least one auxiliary amino acid sequence (Y) in one molecule of the polypeptide (XM). . The wound covering according to any one of claims 2 to 4, comprising a compound (AM) containing a functional group (A), wherein the weight average molecular weight (MW) of (AM) is 1,000 to 100,000. Wood.

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