JP4848224B2 - Nanofiber assembly - Google Patents

Description

  The present invention relates to a nanofiber assembly containing a functional material.

Until now, the technique which manufactures a nanometer-sized polymer fiber by the electrospinning method is known (for example, patent document 1). This method will be briefly described as follows. As shown in FIG. 1, a container 2 storing a polymer solution 1 as a raw material and a target electrode 3 are provided. A nozzle 4 capable of discharging the polymer solution 1 is provided at the tip of the container 2. Here, when the polymer solution 1 is released from the nozzle 4 with a high voltage applied between the nozzle 4 and the target electrode 3, the polymer solution 1 moves from the nozzle 4 to the target electrode 3. A fibrous fiber is formed along the lines of electric force, and the nanofiber 5 is produced on the target electrode 3.
This method is characterized in that a fiber of the order of 10 nm to several hundred nm and a sheet or mat (nanofiber aggregate) in which the fibers are integrated can be produced.

  Since the diameter of the nanofiber is thin and the sheet or mat that is an aggregate of the nanofiber has a large porosity, a wide range of applications for various uses is considered. Since the nanofiber aggregate has a fine internal structure and surface structure, it has a large specific surface area, and exhibits excellent properties for use as, for example, an adsorbent. However, the polymer nanofibers produced by the electrospinning method have a smooth surface, and it is not always easy to decorate the surface and add functions. For example, if an inorganic substance is mixed in a polymer for the purpose of utilizing the function of the inorganic substance, the inorganic substance is embedded in the polymer of the nanofiber, and the original function of the inorganic substance may not be exhibited. is there.

In addition, research and development for producing nanofibers from materials such as polylactic acid and collagen using the electrospinning method has been carried out. Since such materials are excellent in biodegradability and are derived from living organisms, application to automobiles and home appliances, and applications to medicines, cosmetics, foods and hygiene products are considered.
US Pat. No. 6,656,394

Furthermore, in order to improve the functionality of the nanofiber, there may be a concept of combining a functional material with the nanofiber manufactured from polylactic acid or collagen. However, sufficient studies have not been made as to whether such products can actually be manufactured.
The present invention has been made in view of the above-described circumstances, and an object thereof is to provide a nanofiber assembly containing a functional material and to provide a method for manufacturing such a nanofiber assembly. It is.

  As a result of intensive studies to solve the above-mentioned problems, the present inventors have applied the electrospinning method by combining a specific spinning base material and a functional substance, and thereby a functional material-containing nanofiber. The inventors have found that an assembly can be produced, and have basically completed the present invention.

That is, the present invention is as follows.
[1] From at least one spinning substrate selected from the group consisting of polylactic acid, egg white peptide, zein, gelatin, collagen, and chitosan, and catechin and catechin oligomers (hereinafter sometimes referred to collectively as “catechins”) A nanofiber assembly comprising at least one functional substance selected from the group consisting of:
[2] The nanofiber aggregate according to [1], wherein the functional substance is catechin.
[3] The catechin contains non-polymerized catechin (A) and polymerized catechin (B), and the content of non-polymerized catechin (A / (A + B)) is 0.5 to 1. [2] The nanofiber aggregate according to [2].

[4] The catechin includes non-polymerized catechin and polymerized catechin, and the non-polymerized catechin includes catechin gallate, epicatechin gallate, gallocatechin gallate, and epigallocatechin gallate (C). And a non-gallate body (D) containing catechin, epicatechin, gallocatechin and epigallocatechin, and having a gallate ratio (C / (C + D)) of 0.3 to 1 [ The nanofiber assembly according to [2] or [3].
[5] The catechin includes epicatechin, epigallocatechin, epicatechin gallate, and epigallocatechin gallate (E), and catechin, gallocatechin, catechin gallate, and gallocatechin gallate (E). The nanofiber assembly according to any one of [2] to [4], wherein an epi fraction (E / (E + F)) is 0.8 to 1.

[6] A nanofiber is produced by electrospinning at least one spinning substrate selected from the group consisting of polylactic acid, egg white peptide, zein, gelatin, collagen and chitosan, and the nanofiber is composed of catechin and catechin oligomer. A method for producing nanofibers, comprising at least one functional substance selected from the group.
[7] At least one spinning substrate selected from the group consisting of polylactic acid, egg white peptide, zein, gelatin, collagen and chitosan, and at least one functional substance selected from the group consisting of catechin and catechin oligomers A method for producing nanofibers, characterized in that nanofibers are produced from the contained material by electrospinning.

  According to the present invention, a nanofiber assembly containing a functional substance can be produced. This nanofiber assembly has both the functions of nanofibers and the functions of catechins, which are functional substances, and therefore has antioxidant, antibacterial, antiviral, deodorant, and harmful substance adsorption effects. is doing. Therefore, medical parts such as artificial skin, filters for air conditioners, automobiles, air cleaners, hygiene products such as masks, gauze, packs, sheets, medical care, NDS (Nutrition Delivery System: Minerals, Vitamins, etc.) New system aimed at supplying nutritional materials safely and efficiently into the living body) Food materials such as base materials, deoxygenated base materials, and fast-dissolving base materials, functional bases, bases for dirt removal It can be applied to cosmetics such as materials.

  Next, embodiments of the present invention will be described in detail. The technical scope of the present invention is not limited by the following embodiments, and various modifications can be made without changing the gist of the invention. Further, the technical scope of the present invention extends to an equivalent range.

The polylactic acid used in the present invention is a kind of biodegradable plastic in which lactic acid (CH ₃ CH (OH) COOH) is used as a unit and a plurality of lactic acids are linked to have a high molecular weight. Lactic acid as a material for producing polylactic acid can be produced from plants (for example, corn, bonito, sugar cane, beet, sweet potato, etc.). In order to produce polylactic acid, lactic acid is generally cyclized into lactide, which is subjected to ring-opening polymerization to obtain polylactic acid, but in the present invention, it is not dependent on the method for producing polylactic acid.

Two types of optical isomers, L-type and D-type, are known for lactic acid as a simple substance constituting polylactic acid. The present invention can also be used for polylactic acid produced using either L-type or D-type lactic acid as a unit (or for polylactic acid containing L-type and D-type in any ratio).
Egg white peptide means a peptide obtained by appropriately hydrolyzing egg white. Egg whites are those obtained by removing egg yolk from whole eggs (for example, chicken eggs), those obtained by splitting eggs with shells, those obtained by thawing frozen egg whites, those obtained by concentrating them, and egg white powder being returned with water. Any of them may be used, and is not particularly limited, but raw egg white liquid or frozen egg white liquid is preferable from the viewpoint of processability. In order to hydrolyze the egg white, for example, it can be treated with a suitable hydrolase. Such an enzyme is not particularly limited as long as it is a plant-derived, animal-derived, or bacterial-derived protease (proteolytic enzyme), but a protease extracted from cells of the genus Bacillus may be used. It is preferable to use a protease extracted from one or more cells selected from the group consisting of Bacillus licheniformis, Bacillus thermoproteolyticus Roko, and Bacillus subtils.

  The conditions for the protease treatment are not particularly limited, but the hydrolysis temperature is preferably 55 ° C. to 65 ° C. within a pH range of 7-10. The degree of degradation of the enzyme hydrolyzate is not particularly limited, but it is preferably degraded until the amino group amount is 7 to 20 times that of the raw protein, and the average amino acid chain length of the enzyme hydrolyzate is 2 or more and a free amino acid content of 20% or less is preferable, and the average amino acid chain length of the enzyme hydrolyzate is preferably 2-15. The amount of amino groups means the amount of amino groups of the end of the protein or protein hydrolyzate and the lysine residue. The amino group amount, average amino acid chain length, and free amino acid content can be measured by a formol titration method, a TNBS coloring method, a ninhydrin coloring method, or the like. Moreover, as an egg white peptide, Profix (trademark registration: Taiyo Kagaku Co., Ltd.) can be used.

Zein is also called zein. It is a major protein of corn seed and belongs to prolamin. Prolamin is a generic term for proteins belonging to simple proteins, soluble in 60% to 90% ethanol, and insoluble in 90% or more ethanol, water, and neutral salt solutions. In addition, wheat gliadin, barley hordein and so on. Zein is composed of various molecular species, and main components include those having a molecular weight of 22,000 and a molecular weight of 19000.
Gelatin is a general term for substances in which the three-dimensional structure of collagen contained in higher animals is broken. Gelatin is very soluble in water. In the present invention, any animal species and collagen type can be used.

Collagen means the main protein component that constitutes the connective tissue of animals. Mammals account for nearly 30% of the body's total protein. Collagen is composed of 10 or more protein super families such as type I and type II, and has a high content of glycine and proline. In the present invention, any type of collagen or mixture can be used.
Chitosan mainly means β-1,4-polyglucosamine and is a deacetylated product of chitin. Although a β-1,4-polyglucosamine structure is main, it is produced by melting chitin with concentrated alkali, so there may be some structural changes.

Catechin is a water-soluble polyhydric phenol that is abundant in plants such as tea. There are two types of catechin: non-polymerized catechin and polymerized catechin. The non-polymerized catechin includes a gallate body containing catechin gallate, epicatechin gallate, gallocatechin gallate and epigallocatechin gallate, and a non-gallate body containing catechin, epicatechin, gallocatechin and epigallocatechin. Furthermore, catechin has a classification method of epicatechin, epigallocatechin, epicatechin gallate, and epigallocatechin gallate, and a non-epi form including catechin, gallocatechin, catechin gallate, and gallocatechin gallate.
The catechin oligomer means a product in which 3 to 10 catechins are bonded. Catechin includes a plurality of types as described above. In the present invention, a catechin oligomer to which any of these molecules (including the same type or different types) is bound can be used.

The nanofiber of the present invention means a fiber having a single yarn diameter of about 1 nm to about 800 nm, and a collection of the fibers is called a nanofiber aggregate. The nanofiber aggregate can be formed into a one-dimensionally oriented long fiber or spun yarn shape, but when it is produced by electrospinning, it is obtained as a two-dimensional aggregate as a nonwoven fabric. The nanofiber aggregate obtained as a non-woven fabric can be used as an artificial skin, filter, mask, gauze, or other base material.
Although the nanofiber aggregate of the present invention can be produced by various methods for producing nanofibers, it is particularly preferable to use an electrospinning method (also referred to as an electrospinning method or an electrostatic spinning method). In this case, the catechins are a method of producing a nanofiber aggregate in the state of coexisting with a spinning substrate in advance, and a method of producing a nanofiber aggregate in advance and containing the catechins therein. is there.

In order to produce a nanofiber aggregate in advance and include catechins therein, first, a spinning substrate is made of an appropriate solvent (an inorganic solvent such as water, an organic solvent (protic polar solvent, aprotic polar solvent). And the like, and the electrospinning method is carried out with this solution. The electrospinning method can be influenced by factors such as the concentration of the spinning substrate, the type of solvent, the diameter of the needle, the injection distance, the rotation speed, the voltage, and the injection speed. In order to actually manufacture a nanofiber assembly, the above factors can be combined appropriately.
Catechin is contained in the nanofiber assembly thus manufactured. Examples of such a method include a method of immersing a nanofiber aggregate in a solution in which catechins are dissolved, or a method of dropping and spraying a solution in which catechins are dissolved in a nanofiber aggregate. .
Moreover, when producing a nanofiber aggregate by the above method, it is preferable to use a protein-based substance (for example, egg white peptide, zein, gelatin, etc.) as a spinning substrate. This is because catechins easily bind to nanofibers when producing a nanofiber assembly from such a substance.

  Further, in order to produce a nanofiber aggregate in the state of coexisting with the spinning base material in advance, the spinning base material and the functional material are mixed with an appropriate solvent (inorganic solvent such as water, organic solvent (protic polar solvent, non-polar solvent, etc.). In a protic polar solvent). At this time, (1) a method in which only the spinning substrate is dissolved in an appropriate solvent, while a functional substance is dissolved in another solvent, and both solutions are mixed later, or (2) the spinning substrate and functionality There is a method in which both substances are dissolved in the same solvent. The appropriate method (1) or (2) can be selected depending on the combination of the spinning substrate and the functional substance. Thus, the electrospinning method is performed on the material containing the spinning base material and the functional substance. The electrospinning method can be influenced by factors such as the concentration of the spinning substrate, the type of solvent, the diameter of the needle, the injection distance, the rotation speed, the voltage, and the injection speed. In order to actually manufacture a nanofiber assembly, the above factors can be combined appropriately.

EXAMPLES Next, although an Example and a test example demonstrate this invention in detail, this invention is not limited by these Examples and a test example.
<Example 1> Production of nanofiber assembly containing polylactic acid and catechin 90 mg of polylactic acid (weight average molecular weight (Mw) = 13,000, number average molecular weight (Mn) = 61,000, trade name Lacia ( Trademark registration) H-900 (manufactured by Mitsui Chemicals) and 819 mg of 1,1,1,3,3,3-hexafluoroisopropanol (HFIP) were added to sample bottle 1. Magnetic stirrer was added to sample bottle 1. The polylactic acid was completely dissolved using a stirrer at room temperature (20 ° C.-25 ° C.) with stirring overnight (about 10 hours or more).

In another sample bottle 2, 4.5 mg of epigallocatechin gallate (EGCg: Sunphenon EGCg manufactured by Taiyo Kagaku Co., Ltd.) and 86.5 mg of dimethylformamide (DMF) were added to the sample bottle 2. A magnetic stirrer was placed in the sample bottle 2, and EGCg was completely dissolved while stirring at room temperature for 1 hour to 2 hours using the stirrer.
909 mg of the polylactic acid solution in the sample bottle 1 and 91 mg of the EGCg solution in the sample bottle 2 were mixed in another sample bottle 3 and vibrated and stirred using a vortex mixer until they were mixed. After stirring, the solution became cloudy.

After injecting the sample solution in the sample bottle 3 into the syringe, the bubbles in the syringe were expelled. This syringe was set in a syringe pump of an electrospinning apparatus, and an electrospinning method was performed under the following conditions.
As the electrospinning apparatus, those of Imoto Seisakusho Co., Ltd. (model numbers: IMC-1631 type, IMC-163E type, and IMC-164E type) were used. A syringe needle of 25G (outer diameter 0.5 mm, inner diameter 0.32 mm: manufactured by Hoshiseido Medical Equipment Co., Ltd.) was used, and the injection distance was 10 cm, the rotation speed was 100 rpm, the voltage was 10 kv, and the injection speed was 3 mL / hr. .
The results are shown in FIGS. As shown in the figure, when the polylactic acid-catechin system was used, good nanofibers were produced by the electrospinning method. This nanofiber had an outer diameter of about 100 nm to 500 nm.

<Example 2> Production of nanofiber assembly containing zein 60 mg of zein (manufactured by Taiyo Chemical Co., Ltd.) and 3 mL of HFIP were added to a sample bottle. A magnetic stirrer was placed in the sample bottle, and the zein was completely dissolved using the stirrer while stirring at room temperature (20 ° C.-25 ° C.) for 1 hour-2 hours.
After injecting the sample solution in the sample bottle into the syringe, the bubbles in the syringe were expelled. This syringe was set in a syringe pump of an electrospinning apparatus, and an electrospinning method was performed under the following conditions.

As the electrospinning apparatus, those of Imoto Seisakusho Co., Ltd. (model numbers: IMC-1631 type, IMC-163E type, and IMC-164E type) were used. A 21G syringe needle (outer diameter 0.8 mm, inner diameter 0.57 mm: manufactured by Hoshiseido Medical Equipment Co., Ltd.) was used, and the injection distance was 10 cm, the rotation speed was 100 rpm, the voltage was 20 kv, and the injection speed was 1 mL / hr. .
The results are shown in FIGS. As shown in the figure, when zein was used, good nanofibers were produced by electrospinning. This nanofiber had an outer diameter of about 500 nm to 750 nm.
The nanofiber aggregate containing catechins can be produced by immersing the thus produced zein-containing nanofiber aggregate in a solution in which catechins are dissolved.

Example 3 Production of Nanofiber Assembly Containing Egg White Peptide and Gelatin 45 mg of gelatin and 15 mg of egg white peptide (Profix O: manufactured by Taiyo Kagaku Co., Ltd.) are placed in a sample bottle, and 1.8 mL is added here. Of tetrafluoroethylene (TFE) and 1.2 mL of purified water were added. A magnetic stirrer was placed in a sample bottle, and gelatin was completely dissolved using the stirrer while stirring at room temperature (20 ° C.-25 ° C.) for 1 hour-2 hours.
After injecting the sample solution in the sample bottle into the syringe, the bubbles in the syringe were expelled. This syringe was set in a syringe pump of an electrospinning apparatus, and an electrospinning method was performed under the following conditions.

As the electrospinning apparatus, those of Imoto Seisakusho Co., Ltd. (model numbers: IMC-1631 type, IMC-163E type, and IMC-164E type) were used. A 21G syringe needle (described above) was used, and the injection distance was 10 cm, the rotation speed was 100 rpm, the voltage was 25 kv, and the injection speed was 1 mL / hr.
The results are shown in FIG. As shown in the figure, when the egg white peptide-gelatin system was used, good nanofibers were produced by the electrospinning method. This nanofiber had an outer diameter of about 500 nm to 750 nm.
The nanofiber assembly containing catechins can be produced by immersing the thus produced egg white peptide and gelatin-containing nanofiber assembly in a solution in which catechins are dissolved.

  Thus, according to the present Example, the nanofiber assembly containing a functional substance was able to be manufactured. This nanofiber assembly has both the functions of nanofibers and the functions of catechins, which are functional substances, and therefore has antioxidant, antibacterial, antiviral, deodorant, and harmful substance adsorption effects. is doing. Therefore, medical parts such as artificial skin, filters for air conditioners, automobiles, air cleaners, hygiene products such as masks, gauze, packs, sheets, medical care, NDS (Nutrition Delivery System: Minerals, Vitamins, etc.) New system aimed at supplying nutritional materials safely and efficiently into the living body) Food materials such as base materials, deoxygenated base materials, and fast-dissolving base materials, functional bases, bases for dirt removal It can be applied to cosmetics such as materials.

It is a figure explaining the outline | summary of an electrospinning method. 2 is an electron micrograph showing the results of Example 1. FIG. The total width of 11 squares in the figure is 50 μm (the space between adjacent squares is 5 μm). 2 is an electron micrograph showing the results of Example 1. FIG. The total width of 11 squares in the figure is 10 μm (the space between adjacent squares is 1 μm). 6 is an electron micrograph showing the results of Example 2. FIG. The total width of 11 squares in the figure is 100 μm (the space between adjacent squares is 10 μm). 6 is an electron micrograph showing the results of Example 2. FIG. The total width of 11 squares in the figure is 10 μm (the space between adjacent squares is 1 μm). 6 is an electron micrograph showing the results of Example 3. FIG. The total width of 11 squares in the figure is 10 μm (the space between adjacent squares is 1 μm).

Claims (5)

A nanofiber assembly comprising at least one spinning substrate selected from the group consisting of polylactic acid, egg white peptide, and zein , and catechin . The catechin contains non-polymerized catechin (A) and polymerized catechin (B), and the content of non-polymerized catechin (A / (A + B)) is 0.5 to 1. The nanofiber assembly according to claim 1 . The catechin includes a non-polymerized catechin and a polymerized catechin, and the non-polymerized catechin includes a gallate body (C) containing catechin gallate, epicatechin gallate, gallocatechin gallate, and epigallocatechin gallate, and catechin. Or non-gallate body (D) containing epicatechin, gallocatechin and epigallocatechin, and the gallate rate (C / (C + D)) is 0.3 to 1. 2. The nanofiber assembly according to 2 . The catechin includes an epimer (E) containing epicatechin, epigallocatechin, epicatechin gallate and epigallocatechin gallate, and a non-epimer (F) containing catechin, gallocatechin, catechin gallate and gallocatechin gallate. The nanofiber aggregate according to any one of claims 1 to 3 , wherein the nanofiber aggregate is contained and has an epimer ratio (E / (E + F)) of 0.8 to 1 . A method for producing nanofibers, comprising producing nanofibers by electrospinning from a material containing at least one spinning substrate selected from the group consisting of polylactic acid, egg white peptide, and zein and catechin .